TMEM16A modulator for treating respiratory diseases

Abstract

General formula (I) The compound of TIFF2022535986000120.tif21170, and 2,3,4-trimethylamino- ... Compound defined as TIFF2022535986000121.tif23170.

Description

[Technical Field]

[0001] This invention relates to novel compounds having activity as positive modulators of calcium-activated chloride channels (CaCC), specifically TMEM16A. The invention also relates to methods for preparing these compounds and pharmaceutical compositions containing them, as well as the use of these compounds in treating diseases and conditions modulated by TMEM16A, particularly respiratory diseases and conditions. [Background technology]

[0002] Humans can inhale up to 12,000 liters of air daily, which means that airborne pathogens (such as bacteria, viruses, and fungal spores) can enter the respiratory tract. To protect against these airborne pathogens, the lungs have evolved natural defense mechanisms to minimize the possibility of respiratory tract infection and colonization. One such mechanism is the mucous clearance system, in which secreted mucus is propelled in and out of the respiratory tract by the coordinated pulsation of cilia along with coughing. This ongoing "washing" of the lungs constantly removes inhaled particles and microorganisms, thereby reducing the risk of infection.

[0003] In recent years, it has become clear that hydration of the mucous gel is important for enabling mucus clearance (Boucher 2007; Matsui et al., 1998). In a normal, healthy airway, the mucous gel is typically 97% water and 3% w / v solid, under which mucus is removed by mucociliary action. Hydration of the airway mucosa is regulated by the coordinated activity of several ion channels and transporters, including cystic fibrosis membrane conductance regulator (CFTR) and calcium-activated chloride conductance (CaCC; TMEM16A), and anion (Cl) mediated through these channels. - / HCO3 - ) secretion and epithelial Na + Na via channel (ENaC) +The balance with absorption determines the hydration state of the airway mucosa. When ions are transported across the epithelium, water must follow osmotically, and therefore the fluid is secreted or absorbed.

[0004] In respiratory diseases such as chronic bronchitis and cystic fibrosis, the solids percentage of the mucus gel increases as hydration decreases and mucus clearance decreases (Boucher, 2007). In cystic fibrosis, where loss-of-function mutations in CFTR reduce the airway's ability to secrete fluid, the solids percentage can increase to 15%, which is thought to contribute to small airway obstruction and mucus efflux. Strategies to increase airway mucus hydration include anion stimulation, resulting in fluid secretion or Na + This includes any inhibition of absorption. For this purpose, stimulating the activity of the TMEM16A channel increases anion secretion, and therefore increases fluid accumulation in the airway mucosa, hydrates mucus, and enhances the mucus clearance mechanism.

[0005] TMEM16A, also known as anoctamin-1 (Ano1), is the same molecule as the calcium-activated chloride channel (Caputo et al., 2008; Yang et al., 2008). The TMEM16A channel opens in response to an increase in intracellular calcium levels, allowing bidirectional flow of chloride, bicarbonate, and other anions across the cell membrane. Functional TMEM16A channels have been proposed to regulate transepithelial ion transport, gastrointestinal peristalsis, nociception, and cell migration / proliferation (Pedemonte & Galietta, 2014).

[0006] The TMEM16A channel is expressed by epithelial cells in various organs, including the lungs, liver, kidneys, pancreas, and salivary glands. In the airway epithelium, TMEM16A is highly expressed in mucus-producing goblet cells, ciliated cells, and submucosal glands. Physiologically, TMEM16A is activated by stimuli that recruit intracellular calcium, particularly purinergic agonists (ATP, UTP), released by the respiratory epithelium in response to periodic shear stress caused by respiration and other mechanical stimuli such as coughing. In addition to increased anion secretion leading to enhanced airway hydration, TMEM16A activation plays a crucial role in bicarbonate secretion. Bicarbonate secretion is an important regulator of mucin properties and has been reported to be important in controlling airway lumen pH and, therefore, the activity of natural antimicrobial agents such as defensins (Pezzulo et al., 2012).

[0007] Indirect regulation of TMEM16A via elevated intracellular calcium has been clinically investigated, for example, with denufosol (Kunzelmann & Mall, 2003). While promising initial outcomes were observed in small patient cohorts, this approach did not yield clinical benefits in larger cohorts (Accurso et al. 2011; Kellerman et al. 2008). This lack of clinical efficacy was thought to be due to the transient increase in anion secretion, the short half-life of denufosol on the epithelial surface and resulting receptor / pathway desensitization, as well as undesirable effects of elevated intracellular calcium, such as increased mucus release from goblet cells (Moss, 2013). Compounds that directly act on TMEM16A to enhance channel opening with low levels of elevated calcium are expected to sustainably enhance anion secretion and mucociliary clearance in patients, thereby improving congenital defense. Since TMEM16A activity is independent of CFTR function, a positive modulator of TMEM16A has the potential to provide clinical benefits to all CF patients and non-CF respiratory diseases characterized by mucocongestion, including chronic bronchitis and severe asthma.

[0008] TMEM16A modulation is involved as a treatment for salivary gland dysfunction in Sjögren's syndrome and xerostomia (dry mouth), dry eye, biliary stasis and gastrointestinal motility disorders resulting from radiotherapy.

Summary of the Invention

[0009] Our application, PCT / GB2019 / 050209, relates to compounds that are positive modulators of TMEM16A and are therefore used in the treatment of diseases and conditions in which the modulation of TMEM16A plays a role, particularly respiratory diseases and symptoms. We have developed further compounds that are positive modulators of TMEM16A.

[0010] In a first aspect of the invention, General formula (I), including all tautomers, all enantiomers and isotopic variants: TIFF0007862174000001.tif21170[where, A is TIFF0007862174000002.tif28170; X 1 、X 2 、X 3 One of them is S, and the other two of X 1 、X 2 and X 3 are CH; TIFF0007862174000003.tif6170 represents a single bond or a double bond such that ring A is aromatic; X 4 and X 5 each independently is CH or N; *1 indicates the point of attachment to Z 1 and *2 indicates the point of attachment to Z 2 ; Z 1 and Z 2 each independently is * -C(O)NH- and * -NHC(O)-, * indicates the point of attachment to ring A; R1 H, CN, C(O)OR 12 , C 1-3 Alkyl, C 2-3 Alkenyl or C 2-3 These are alkynyl groups, and any of these alkyl, alkenyl, or alkynyl groups are fluoro, OR 12 , N(R 12 )2, C(O)OR 12 , C(O)N(R 12 )2, C(O)R 12 and N(R 13 )C(O)R 12 One or more substituents selected from, which may be substituted with a single substituent if appropriate; R 12 and R 13 These are H and C, respectively, independently. 1-6 Alkyl and C 1-6 Selected from fluoroalkyl groups, R 2 is either H or OR 12 C arbitrarily replaced by 1-6 It is alkyl; and, R 3 teeth, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 These C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Any of the alkynyl groups are fluoro, CN, or R 14 Ure 14 , OR 15 , N(R 15 )2, C(O)OR 15 , C(O)N(R 15 )2, N(R 16 )C(O)R 15 , N(R 15 )S(O)2R 14 , N(R 15 )S(O)2R 16 and N(R 15 )C(O)OR 16 They may be substituted with one or more substituents selected from; or, R 3 teeth, A 3- to 7-membered carbocyclic or heterocyclic ring system, or a 6- to 10-membered aryl ring system or a 5- to 10-membered heteroaryl ring system, any of which may be substituted with one or more substituents selected from halo, CN, C 1-4 alkyl, C 1-4 haloalkyl, OR 17 and N(R 17 )2; R 14 is a 6- to 10-membered aryl ring system or a 5- to 10-membered heteroaryl ring system or a 3- to 7-membered carbocyclic or heterocyclic ring system, any of which aryl ring system, heteroaryl ring system, carbocyclic ring system or heterocyclic ring system may be substituted with one or more substituents selected from halo, C 1-4 alkyl, C 1-4 haloalkyl, OR 17 and N(R 17 )2; R 17 are each independently H, C 1-4 alkyl or C 1-4 haloalkyl; R 15 and R 16 are each independently H, C 1-6 alkyl or C 1-6 haloalkyl; or R 2 and R 3 [[ID=4I]]together with the carbon atom to which they are attached form a 3- to 10-membered carbocyclic or heterocyclic ring system which may be substituted with one or more substituents selected from halo, CN, OR 9 , N(R 9 )2, C(O)OR 9 , C(O)N(R 9 )2, C(O)R 9 , N(R 9 )C(O)R 9 , and halo, OR 9 or N(R 9 )2-substituted C 1-4 alkyl; or R 1 , R 2 and R 3together with the carbon atoms to which they are attached form a bridged 5- to 10-membered carbocyclic or heterocyclic ring system or phenyl, and any of these carbocyclic or heterocyclic ring systems or phenyl groups which may be substituted with halo, CN, OR 9 , N(R 9 )2, C(O)OR 9 , C(O)N(R 9 )2, C(O)R 9 , N(R 9 )C(O)R 9 , and may be substituted with one or more substituents selected from halo, OR 9 or N(R 9 )2-substituted C 1-4 alkyl; R 9 are each independently selected from H, C 1-6 alkyl or C 1-6 haloalkyl; Y is -CH2- or -CH(CH3)-; R 4 is a 6- to 14-membered aryl, 5- to 14-membered heteroaryl or 5- to 10-membered carbocyclic ring system, any of which may be substituted with one or more substituents selected from: halo, CN, nitro, R 19 , OR 19 , OR 6 , SR 6 , NR 6 R 7 , C(O)R 6 , C(O)R 19 , C(O)OR 6 , C(O)N(R 6 )(R 7 ), N(R 7 )C(O)R 6 ; any of which may be substituted with halo, CN, nitro, R 19 , OR 6 , SR 6 , NR 6 R 7 , C(O)R 6 C(O)OR 6 , C(O)N(R 6 )(R 7 ) and N(R 7)C(O)R 6 C may be substituted with one or more substituents selected from 1-6 Alkyl or O(C) 1-6 Alkyl); and R 4 If it is not completely aromatic, it is an oxo; R 19 These are 5- or 6-membered aryl ring systems or heteroaryl ring systems, or 3- to 7-membered carbocyclic or heterocyclic systems, and any of these aryl ring systems, heteroaryl ring systems, carbocyclic systems, or heterocyclic systems are halo, C 1-4 Alkyl, C 1-4 Haloalkyl, OH, O(C) 1-4 Alkyl), O(C 1-4 It may be substituted with one or more substituents selected from haloalkyl groups; R 6 H, C 1-6 Alkyl, C 1-6 They are haloalkyl, benzyl, 3- to 7-membered carbocyclyl, or 3- to 7-membered heterocyclyl; R 7 H, C 1-6 Alkyl or C 1-6 It is a haloalkyl; or R 6 and R 7 These, together with the nitrogen atoms to which they are bonded, optionally contain one or more further heteroatoms, C 1-4 It forms a 4- to 7-membered heterocycle which may be substituted with one or more substituents selected from alkyl, oxo, and halo; however, iA, TIFF0007862174000004.tif20170, Z 1 but, * When it is -C(O)NH-, Z 2 teeth, * -NHC(O)- not; and ii.A is, The filename is TIFF0007862174000005.tif20170. Z 1 but, *When -NHC(O)-, Z 2 teeth, * -C(O)NH- and R 1 and R 2 H is R 3 is a carbocyclic system, heterocyclic system, arylcyclic system, or heteroarylcyclic system which may be substituted as defined above; or R 1 H is R 2 and R 3 They together form a carbocyclic or heterocyclic system, which may be substituted as defined above; or R 1 , R 2 and R 3 Together with the carbon atoms to which they are bonded, they form a bridged 5-10 membered carbocyclic or heterocyclic system or phenyl, which may be substituted as defined above; and, R 4 Compounds of a 6- to 14-membered aryl group (which may be substituted as defined above), as well as salts and solvates thereof.

[0011] Summary of the Invention Throughout this specification and the following claims, unless contextually required, the word “comprise,” and variations such as “comprises” and “comprising,” will be understood to mean the inclusion of any integer, process, group of integers, or group of processes described, but not the exclusion of any other integer, process, group of integers, or group of processes.

[0012] All documents and patent references mentioned herein are incorporated by reference to the greatest extent possible.

[0013] In this specification, “medical use” refers to use for administration to humans or animals, particularly humans or mammals, such as livestock or domestic mammals, for the treatment or prevention of disease or symptoms. The term “medical composition” refers to a composition suitable for a medicinal use, and “pharmaceutically acceptable” refers to an agent suitable for use in a medicinal composition. Other similar terms should be interpreted accordingly.

[0014] In this specification, "C 1-6 The term "alkyl" refers to a straight-chain or branched fully saturated hydrocarbon group having 1 to 6 carbon atoms. This term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and t-butyl. Other alkyl groups, for example, C 1-10 Alkyl compounds are defined as described above, but they contain a different number of carbon atoms.

[0015] "C 2-6 The term "alkenyl" refers to a straight-chain or branched hydrocarbon group having 1 to 6 carbon atoms and at least one carbon-carbon double bond. This term includes ethenyl, propen-1-yl, propen-2-yl, buten-1-yl, and buten-2-yl. Other alkenyl groups, e.g., C 2-10 Alkenyls are defined as described above, but they contain a different number of carbon atoms.

[0016] "C 2-6 The term "alkynyl" refers to a linear or branched hydrocarbon group having 1 to 6 carbon atoms and at least one carbon-carbon triple bond. This term includes ethynyl, propyne-1-yl, propyne-2-yl, butyne-1-yl, and butyne-2-yl. Other alkynyl groups, e.g., C 2-10 Alkynnyls are defined as described above, but they contain a different number of carbon atoms.

[0017] The terms "carbocyclic" and "carbocykyl" refer to non-aromatic hydrocarbon ring systems containing 3 to 10 ring carbon atoms and optionally one or more double bonds, unless otherwise specified. A carbocyclic group may be a monocyclic or may contain two or three rings that are condensed or bridged, with carbon atoms in the bridges being included in the number of ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, and cyclohexenyl, as well as bridged systems such as bicyclo[1.1.1]pentyl, bicyclo-[2.2.1]heptyl, bicyclo-[2.2.2]octyl, and adamantyl.

[0018] In the context of this specification, the terms “heterocyclic” and “heterocyclyl” refer to non-aromatic ring systems containing 3 to 10 ring atoms, each containing at least one heteroatom selected from N, O, and S, unless otherwise specified. The heterocyclic group may be monocyclic or may contain two or three rings that are condensed or bridged, with the number of bridged atoms being included in the number of ring atoms. Examples include tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, and thiomorpholinyl, as well as condensed systems such as cyclopropyl condensed pyrrolidine.

[0019] In the context of this specification, the terms “aryl” and “aromatic” refer to aromatic ring systems having 5 to 14 ring carbon atoms and containing up to 3 rings, unless otherwise specified. If an aryl group contains two or more rings, not all rings are necessarily fully aromatic. Examples of aromatic moieties are benzene, naphthalene, fluorene, tetrahydronaphthalene, indane, and indene.

[0020] In the context of this specification, the terms “heteroaryl” and “heteroaromatic” refer to aromatic ring systems having 5 to 14 ring atoms, at least one of which is a heteroatom selected from N, O, and S, and containing up to 3 rings, unless otherwise specified. If a heteroaryl group contains two or more rings, not all rings are aromatic. Examples of heteroaryl groups include pyridine, pyrimidine, indole, indazole, thiophene, benzothiophene, benzoxazole, benzofuran, dihydrobenzofuran, tetrahydrobenzofuran, benzimidazole, benzimidazolin, quinoline, and indolene.

[0021] The term "oxo" refers to a C=O substituent, where the carbon atom is a ring atom of a carbocyryl or heterocyclyl group, or a ring of a non-aromatic aryl or heteroaryl group.

[0022] The term "halogen" refers to fluorine, chlorine, bromine, or iodine, while the term "halo" refers to the fluoro, chloro, bromo, or iodine group. Similarly, "halogenated compound" refers to fluoride, chloride, bromide, or iodide.

[0023] The term "C" used herein 1-6 The term "haloalkyl" refers to a C group defined above where one or more hydrogen atoms are replaced by a halo group. 1-6 This refers to alkyl groups. Any number of hydrogen atoms may be substituted up to perhalo substitution. Examples include trifluoromethyl, chloroethyl, and 1,1-difluoroethyl. Fluoroalkyl groups are haloalkyl groups where the halo is fluoro.

[0024] The term “isotope variant” refers to an isotope-labeled compound that is identical to those enumerated in formula (I), except that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those most commonly found in nature, or that the proportion of atoms having atomic masses or mass numbers less common in nature is increased (the latter concept is called “isotope enrichment”). Examples of isotopes that may be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine, and chlorine, e.g., 2H (deuterium), 3H, 11C, 13C, 14C, 18F, 123I, or 125I (e.g., 3H, 11C, 14C, 18F, 123I, or 125I), which may or may not be naturally occurring isotopes.

[0025] In some compounds of general formula (I), A is The filename is TIFF0007862174000006.tif25170.

[0026] In this case, the compound of general formula (I) is general formula (Ia), (Ib), or (Ic). TIFF0007862174000007.tif74170[where, R 1 , R 2 , R 3 , R 4 , Z 1 , Z 2 And Y may be a compound of the general formula (I) as defined.

[0027] In some cases, a compound of general formula (I) is a compound of general formula (Ia).

[0028] In some cases, a compound of general formula (I) is a compound of general formula (Ib).

[0029] Alternatively, A is The filename is TIFF0007862174000008.tif20170.

[0030] In this case, the compound of general formula (I) is (Id), (Ie), (If), or (Ig) TIFF0007862174000009.tif98170[where, R 1 , R 2 , R 3 , R 4 , Z 1 , Z 2 And Y may be a compound of the general formula (I) as defined.

[0031] In some cases, a compound of general formula (I) is a compound of general formula (Id).

[0032] In some cases, a compound of general formula (I) is a compound of general formula (Ie).

[0033] In some cases, a compound of general formula (I) is a compound of general formula (If).

[0034] In some cases, a compound of general formula (I) is a compound of general formula (Ig).

[0035] In some compounds of (Id), (Ie), (If), and (Ig), R 1 R 2 It's not H.

[0036] In some compounds of (Id), (Ie), (If), and (Ig), R 1 , R 2 and R 3 It does not bond to form a phenyl compound.

[0037] In some compounds of (Id), (Ie), (If), and (Ig), R 1 However, H is R 2 and R 3 However, when these combine with the carbon atoms to which they are bonded to form a 3- to 10-membered carbocyclic or heterocyclic system that is optionally substituted as described above, R 4 is a phenyl substituted with OH and halo. More preferably, R4 This is 2-hydroxy-5-halophenyl, for example, 2-hydroxy-5-chlorophenyl.

[0038] In some compounds of (Id), (Ie), (If), and (Ig), Z 1 but * -NHC(O)- and Z 2 but * If it is -C(O)NH-, then R 1 R 2 It's not H.

[0039] In some compounds of (Id), (Ie), (If), and (Ig), Z 1 but * -NHC(O)- and Z 2 but * If it is -C(O)NH-, then R 1 , R 2 and R 3 It does not bond to form a phenyl compound.

[0040] In some compounds of (Id), (Ie), (If), and (Ig), R 1 However, H is R 2 and R 3 However, when these combine with the carbon atoms to which they are bonded to form a 3- to 10-membered carbocyclic or heterocyclic system with arbitrary substitutions as described above, Z 1 teeth, * -NHC(O)- and Z 2 teeth, * -C(O)NH- and R 4 is a phenyl substituted with OH and halo. More preferably, R 4 This is 2-hydroxy-5-halophenyl, for example, 2-hydroxy-5-chlorophenyl.

[0041] In compounds of general formula (I), Y is appropriately -CH2-.

[0042] For a more appropriate compound of general formula (I), R 1 H, CN, C(O)OR 12 , C1-3 Alkyl, C 2-3 Alkenyl or C 2-3 It is an alkynyl, and any of these alkyl, alkenyl, or alkynyl groups is fluoro and OR 12 They may be substituted with one or more substituents selected from; In the formula, R 12 These are defined above, but more appropriately, H and C 1-3 Alkyl or C 1-3 Fluoroalkyls, such as H, methyl, or trifluoromethyl.

[0043] More preferably, R 1 If it is an alkyl, alkenyl, or alkynyl group, it is one or more halo substituents and / or a single OR 12 They may be substituted with substituents.

[0044] More precisely, R 1 is H, CN, or C 1-3 Alkyl, C 2-3 Alkenyl or C 2-3 These are alkynnyls, and all of them are either unsubstituted or substituted as defined above.

[0045] Furthermore, in more suitable compounds, R 1 This is H, CN, or C, which are arbitrarily substituted as described above. 1-3 It is an alkyl group, more specifically CN, unsubstituted C 1-3 Alkyl, for example, methyl or C 1-3 Haloalkyls, such as trifluoromethyl, are examples of this type of element.

[0046] In some suitable compounds, R 2 is OR 12 It is either H arbitrarily substituted with C 1-3 It is alkyl, R 12 These are defined above, but more appropriately, H and C 1-3 Alkyl or C 1-3 Fluoroalkyls, especially C 1-3 Alkyl or C 1-3These are fluoroalkyl groups, such as methyl or trifluoromethyl.

[0047] More precisely, R 2 is either H or unsubstituted C 1-3 Alkyl, especially unsubstituted C such as methyl, is a type of alkyl group. 1-3 It is alkyl.

[0048] In some suitable compounds of the present invention, R 3 Fluoro, CN, R 14 Ure 14 , OR 15 , N(R 15 )2, C(O)OR 15 , C(O)N(R 15 )2, N(R 16 )C(O)R 15 , N(R 15 )S(O)2R 14 , N(R 15 )S(O)2R 16 and N(R 15 )C(O)OR 16 One or more substituents selected from, which may appropriately be substituted with one substituent, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 It is alkinyl;

[0049] R 15 and R 16 Each of these is independent and as defined above, but in particular, H and C 1-3 Alkyl or C 1-3 The haloalkyl group is, for example, H, methyl, or trifluoromethyl.

[0050] More precisely, R 3 C 1-3 Alkyl, C 2-3 Alkenyl or C 2-3 These are alkynyl compounds, and any of these may be substituted as described above. In particularly suitable compounds of the present invention, R 3 This is C, which is arbitrarily substituted as explained above. 1-3 Alkyl, especially unsubstituted C such as methyl, is a type of alkyl group.1-3 It is alkyl.

[0051] Alternatively, R 2 and R 3 These, together with the carbon atoms to which they are bonded, can form 3- to 10 membered carbocyclic or heterocyclic systems that are either unsubstituted or substituted as defined above. More suitable substituents for such cyclic systems include halo, CN, OR 9 , N(R 9 )2, and, halo, OR 9 or N(R 9 )C arbitrarily substituted in 2 1-4 Examples include those having one or more substituents selected from alkyl groups; R 9 As defined above, but more appropriately, H and C 1-3 Alkyl or C 1-3 Haloalkyls, particularly H, methyl, or trifluoromethyl.

[0052] R 2 and R 3 Particularly preferred substituents for the ring system formed by include halo, OH, methoxy, trifluoromethoxy, methyl, and trifluoromethyl.

[0053] R 2 and R 3 Suitable heterocyclic rings formed by these include: tetrahydropyran-yl, e.g., tetrahydropyran-4-yl; tetrahydrofuranyl, e.g., tetrahydrofuran-3-yl; oxetanyl, e.g., oxetan-3-yl; piperidinyl, e.g., piperidinyl-2-yl and piperidinyl-4-yl; morpholinyl, piperazinyl and cyclopropyl condensed pyrrolidine.

[0054] More preferably, R 2 and R 3The ring formed by this is a carbocyclic ring. Examples of such carbocyclic rings include: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; and crosslinking systems such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, and adamantyl (however, R 2 and R 3 (The atoms to which it is bonded are not bridging atoms.)

[0055] R 2 and R 3 More suitable carbocyclic compounds formed by this process include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0056] In yet another suitable compound of the present invention, R 1 , R 2 and R 3 These, together with the carbon atoms to which they are bonded, form bridging carbocyclic or heterocyclic systems, particularly carbocyclic systems such as bicyclo[1.1.1]pentanyl, bicyclo[2.1.1]hexanyl, bicyclo-[2.2.1]heptanyl, bicyclo-[2.2.2]octanyl, or adamantyl.

[0057] In some cases, the carbocyclic system can be selected from 3-bicyclo[1.1.1]pentanyl, bicyclo-[2.2.1]heptanyl, bicyclo-[2.2.2]octanyl, and 1-adamantyl.

[0058] In other cases, the carbocyclic system may be selected from bicyclo[1.1.1]pentanyl, bicyclo[2.1.1]hexanyl, and adamantyl, in particular from 3-bicyclo[1.1.1]pentanyl, 1-bicyclo[2.1.1]hexanyl, and 1-adamantyl.

[0059] Ideally, the ring system is either unsubstituted or OR 9 and C(O)OR 9 It is substituted with a single substituent selected from R 9As defined above, but more preferably, H, methyl, or ethyl, in particular H or methyl, more preferably methyl. More preferably, the ring system is unsubstituted.

[0060] In some particularly suitable compounds of the present invention, R 1 , R 2 and R 3 Each of these is either unsubstituted or C is substituted as described above. 1-3 It is alkyl. More specifically, R 1 , R 2 and R 3 Each of these is a non-substituted C 1-3 Alkyl, for example, methyl.

[0061] In other particularly suitable compounds of the present invention, R 1 It is cyano, and R 2 and R 3 Each of these is either unsubstituted or C is substituted as described above. 1-3 It is alkyl. More specifically, R 2 and R 3 Each of these is a non-substituted C 1-3 Alkyl, for example, methyl.

[0062] In other particularly suitable compounds of the present invention, R 1 is either H or CF3, and R 2 and R 3 These, together with the carbon atoms to which they are bonded, form a 3- to 10-membered carbocyclic or heterocyclic ring system that is either unsubstituted or substituted as defined above. More specifically, R 1 is either H or CF3, and R 2 and R 3 These, together with the carbon atoms to which they are bonded, form a 3- to 10-membered carbocyclic system, particularly a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl ring.

[0063] In the compound of the present invention, R 4R is an unsubstituted or substituted 6-14 member aryl, 5-14 member heteroaryl, or 5-10 member carbocyclic system as defined above. More preferably, 4 These are a 6-10 member aryl, a 5-10 member heteroaryl, or a 5-10 member carbocyclic or heterocyclic system, which may be substituted with one or more substituents as described above.

[0064] In some more appropriate compounds of general formula (I), R 4 This is a 6- to 11-membered aryl group, which is either unsubstituted or substituted as described above, selected from, for example, phenyl, naphthyl, indanyl, 1,2,3,4-tetrahydronaphthyl, and benzocycloheptanyl.

[0065] In other, more suitable compounds of general formula (I), R 4 This is a 5-10 membered heteroaryl group, which is either unsubstituted or substituted as described above, selected from, for example, pyridyl, quinolinyl, quinoxalinyl, indazolyl, indolyl, benzoxazolyl, dihydrobenzofuranyl, furyl, and thienyl.

[0066] In other, more suitable compounds of general formula (I), R 4 This group is either unsubstituted or substituted as described above, and is selected from carbocyclyl groups, such as cyclohexyl and adamantyl.

[0067] In some suitable compounds of the present invention, R 4 This is a phenyl compound optionally substituted with one or more of the above substituents.

[0068] Alternatively, R 4 is a 5-10 member heteroaryl substituted with one or more of the above substituents. More preferably, in this case, R 4 These are pyridyl, pyrrolyl, thienyl, furyl, benzoxazolyl, imidazolyl, indolyl, or indazolyl.

[0069] In some suitable compounds, R 4 It is replaced by one or more substituents selected from the following: Hello, CN; Ure 6 , NR 6 R 7 , C(O)OR 6 , C(O)N(R 6 )(R 7 ); Hello, CN, OR 6 , NR 6 R 7 , C(O)OR 6 and C(O)N(R 6 )(R 7 C optionally substituted with one or more substituents selected from ) 1-6 alkyl; Here, R 6 H, C 1-6 Alkyl, C 1-6 They are haloalkyl, 3- to 7-membered carbocyclyl, or 3- to 7-membered heterocyclyl; R 7 H, C 1-6 Alkyl or C 1-6 It is a haloalkyl; or R 6 and R 7 These may, together with the nitrogen atom to which they are bonded, form a 5- or 6-membered heterocycle, which optionally contains one or more further heteroatoms and may be substituted with one or more oxo substituents.

[0070] R 4 In other suitable compounds where is an aryl group, it is preferable that they are unsubstituted or substituted with one or more substituents selected from the following: Hello, CN, R 19 , OR 19 ; Ure 6 , C(O)OR 6 ; Hello, CN, R 19 , OR 19 , OR 6 and NR 6 R 7C optionally substituted with one or more substituents selected from 1-4 alkyl or O(C 1-4 alkyl); where R 6 , R 7 and R 19 are as defined above.

[0071] However, more preferably, R 6 is H, C 1-4 alkyl or C 1-4 haloalkyl, or for the moiety NR 6 R 7 where R 6 and R 7 are bonded to the nitrogen atom to which they are attached to form a 5- or 6-membered heterocyclic ring, optionally containing one or more additional heteroatoms and optionally substituted with one or more halo substituents.

[0072] R 19 is more preferably either a 3- to 6-membered carbocyclic group optionally substituted with one or more substituents selected from halo, methyl and methoxy or phenyl.

[0073] In particularly suitable compounds, R 4 is phenyl, optionally substituted at either the 2- or 3-position with an OH group and optionally with one or more additional substituents as defined for general formula (I), more preferably one or two additional substituents selected from, for example, those defined immediately above. Suitably, when additional substituents are present, one of the additional substituents is halo, for example chloro or fluoro. The halo substituent is suitably disposed para to the OH group. When a third substituent is present, this is most preferably unsubstituted or substituted as defined above, most preferably unsubstituted or substituted with an OH group, and is C 1-4 alkyl).

[0074] R 4When it is a bicyclic aryl group, it is more preferably unsubstituted or substituted with one or two substituents selected from OH and halo.

[0075] R 4 When it is a heteroaryl group, it is more preferably unsubstituted or substituted with one or more substituents selected from OH and halo.

[0076] R 4 When it is a carbocyclic group, it is more preferably unsubstituted or substituted with one or more substituents selected from OH and halo. Even more preferably, it is unsubstituted.

[0077] In a particularly suitable compound of the present invention, Y is -CH2-, and R 4 is phenyl, and the compound is of the general formula (Ii) TIFF0007862174000010.tif32170[wherein, R 1 R 2 R 3 A, Z 1 and Z 2 are as defined for the general formula (I); R 10 is H, OH, halo, C 1-6 alkyl, -O(C 1-6 alkyl); R 11 are each independently H, halo, OH, CN, C 1-6 alkyl, C 1-6 haloalkyl, -O(C 1-6 alkyl) or C(O)O-(C 1-6 alkyl); and n is 1 or 2] compound.

[0078] Even more preferably, the compound is of the general formula (Iii) or (Iiii) TIFF0007862174000011.tif87170[wherein, R 1 R 2 R 3A, Z 1 and Z 2 This is defined for general formula (I); R 11a H, Halo, C 1-4 Alkyl, C 1-4 Haloalkyl or C(O)O(C) 1-4 Alkyl) is; R 11b H, Halo, C 1-4 Alkyl or C 1-4 It is a haloalkyl; and R 11c H, Haro, CN, C 1-4 Alkyl or C 1-4 It is a compound that is a haloalkyl group.

[0079] A compound of general formula (Ii), wherein A is It is TIFF0007862174000012.tif27170, Compounds of general formula (Ii) are also known as (Iai), (Ibi), or (Ici). It may be a compound of TIFF0007862174000013.tif99170.

[0080] A If it is TIFF0007862174000014.tif20170, Compounds of general formula (Ii) are also known as (Idi), (Iei), (Ifi), or (Igi). It may be a compound of TIFF0007862174000015.tif152170.

[0081] In particularly suitable compounds of general formulas (Ii), (Iai), (Ibi), (Ici), (Idi), (Iei), (Ifi), and (Igi), R 11a and R 11b One or both of them are H.

[0082] In some such compounds, R 11a H, Halo, C 1-4 Alkyl or C(O)O(C 1-4is (alkyl), and R 11b is H.

[0083] More preferably, R 11a is H, chloro, C 1-4 alkyl or C(O)OCH3, and R 11b is H.

[0084] In some compounds of general formulas (Ii), (Iai), (Ibi), (Ici), (Idi), (Iei), (Ifi) and (Igi), R 11a is chloro and R 11b is H.

[0085] In other such compounds of general formulas (Ii), (Iai), (Ibi), (Ici), (Idi), (Iei), (Ifi) and (Igi), R 11a is H and R 11b is H, halo or C 1-6 haloalkyl.

[0086] More preferably, R 11a is H and R 11b is H, chloro, bromo or trifluoromethyl.

[0087] Alternatively, in other particularly suitable compounds of general formulas (Ii), (Iai), (Ibi), (Ici), (Idi), (Iei), (Ifi) and (Igi), both R 11a and R 11b are halo, particularly chloro or bromo.

[0088] For a compound of general formula (Iii) where A is TIFF0007862174000016.tif25170, the compound of general formula (Iii) may be a compound of general formula (Iaii), (Ibii) or (Icii) TIFF0007862174000017.tif127170.

[0089] When A is If the file is TIFF0007862174000018.tif19170, Compounds of general formula (III) are also known as compounds of general formula (Idii), (Ieii), (Ifii), or (Igii). It may be a compound of TIFF0007862174000019.tif183170.

[0090] In some suitable compounds of general formulas (Iii), (Iaii), (Ibii), (Icii), (Idii), (Ieii), (Ifii), and (Igii), R 11a H is R 11b C 1-4 Alkyl, OH, or C 1-4 C substituted with haloalkyl 1-4 It is alkyl, R 11c The elements are H, halo, methyl, or ethyl.

[0091] For more appropriate compounds of general formulas (Iii), (Iaii), (Ibii), (Icii), (Idii), (Ieii), (Ifii), and (Igii), R 11a H is R 11b C 1-4 Alkyl, OH, or C 1-4 C substituted with haloalkyl 1-4 Alkyl, for example, t-butyl, or 2-hydroxy-1,1-dimethylethyl, R 11c It is chloro or fluoro.

[0092] Specific examples of compounds with general formula (I) include: N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]thiophene-2-carboxamide (compound 1); N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]thiophene-3-carboxamide (compound 1.2); N-tert-butyl-4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]thiophene-2-carboxamide (compound 1.3); N-[(5-chloro-2-hydroxyphenyl)methyl]-3-(2,2-dimethylpropanoylamide)benzamino(compound 2); N-[(5-chloro-2-hydroxyphenyl)methyl]-4-(2,2-dimethylpropanoylamino)pyridine-2-carboxamide (compound 3); N2-tert-butyl-N4-[(2-hydroxyphenyl)methyl]pyridine-2,4-dicarboxamide (compound 4); N1-tert-butyl-N3-[(2-hydroxyphenyl)methyl]benzene-1,3-dicarboxamide (compound 5); N-[3-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminophenyl]phenyl]-2,2-dimethylpropanamide (compound 6); N-[4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]-2-pyridyl]-2,2-dimethylpropanamide (compound 7); N-[4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]-2-pyridyl]cyclohexanecarboxamide (compound 7.1); 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]amino]-N-(1-cyano-1-methyl-ethyl)thiophene-2-carboxamide (compound 8); 5-[[2-[2-fluoro-5-hydroxy-4-(2-hydroxy-1,1-dimethyl-ethyl)phenyl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide (compound 9); and The above-mentioned salts and solvates are examples.

[0093] Z 1 but * -C(O)NH- and Z 2 but * -NHC(O)- and A Compounds of general formula (I) less than or equal to TIFF0007862174000020.tif25170 are: General formula (II): TIFF0007862174000021.tif29170[where, X 1 , X 2 , X 3 , Y and R 4 The compound of the general formula (I) is defined as follows: General formula (III): TIFF0007862174000022.tif21170[where, R 1 , R 2 and R 3 It can be prepared by reacting it with a compound of the general formula (I) as defined by the formula (I).

[0094] Ideally, the reaction is carried out in the presence of a coupling reagent and under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA) and in an organic solvent such as DMF.

[0095] Suitable coupling reagents include known peptide coupling agents, such as O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU). Examples include tramethyluronium tetrafluoroborate (TATU), (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazole-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP)carbodiimide, such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), and triazoles, such as 1-hydroxy-7-azabenzotriazole (HOAt) or hydroxybenzotriazole (HOBt). Preferably, the reaction is carried out under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA) and in an organic solvent such as DMF.

[0096] Alternatively, the coupling reagent may be propylphosphonic anhydride (T3P®). When using T3P as the coupling reagent, the reaction should be carried out under basic conditions in an organic solvent such as dioxane, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA).

[0097] Compounds of general formula (III) are readily available or can be synthesized by known methods. Compounds of general formula (II) are of general formula (IV): TIFF0007862174000023.tif29170[where, X 1 , X 2 , X 3 , Y and R 4This is defined for general formula (I), and R 26 is C 1-6 It can be prepared by hydrolysis of a compound that is alkyl or benzyl.

[0098] Ideally, hydrolysis is carried out under basic conditions, for example, by using a metal hydroxide such as lithium hydroxide, sodium hydroxide, or potassium hydroxide in an aqueous organic solvent.

[0099] Compounds of general formula (IV) are of general formula (V): TIFF0007862174000024.tif26170[where, X 1 , X 2 and X 3 This is defined for general formula (I), and R 26 The compound of the general formula (IV) is defined as follows: General formula (VI): TIFF0007862174000025.tif20170[In the formula, Y and R 4 [is defined for general formula (I)] carboxylic acid; or General formula (VII): TIFF0007862174000026.tif21170[In the formula, Y and R 4 This is defined for general formula (I), and R 27 It can be prepared by reacting it with an acid halide, such as a halide, for example, a chloride or bromide.

[0100] When reacting a compound of general formula (V) with a carboxylic acid of general formula (VI), the reaction is preferably carried out under basic conditions, in the presence of a coupling agent, such as an amine such as diisopropylethylamine (DIPEA), and in an organic solvent such as DMF. Suitable coupling agents are as described above.

[0101] When a compound of general formula (V) is reacted with an acid halide of general formula (VII), the reaction is preferably carried out under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA), and in an organic solvent such as dichloromethane.

[0102] Compounds of general formulas (V) and (VI) are readily available or can be prepared by known methods. Compounds of general formula (VII) can be prepared from compounds of general formula (VI) by known methods. For example, acid chlorides of general formula (VII) can be obtained by treating the corresponding acid of general formula (VI) with thionyl chloride, preferably at high temperatures, for example, about 60–80°C, typically about 70°C.

[0103] Z 1 but * -NHC(O)- and Z 2 but * Compounds with the general formula (I) -C(O)NH- include those with the general formula (X): TIFF0007862174000027.tif31170[where, R 1 , R 2 , R 3 And A is as defined for general formula (I)], General formula (XI): H2N-YR 4 (XI) [In the formula, Y and R 4 It can be prepared by reacting it with a compound of the general formula (I) as defined below.

[0104] Ideally, the reaction is carried out under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as dioxane, using a coupling agent such as T3P®.

[0105] Compounds with general formula (XI) where Y is -CH2- are general formula (XII): N≡CR 4 (XII) [In the formula, Y and R 4 It can be prepared by reducing a nitrile of the general formula (I) as defined.

[0106] Reduction can be carried out using a reducing agent such as a hydride, for example, lithium aluminum hydride, in an organic solvent such as tetrahydrofuran.

[0107] Compounds of general formula (XII) are known and readily available, or can be prepared by methods known to those skilled in the art.

[0108] The carboxyl group of general formula (X) is general formula (XIII): TIFF0007862174000028.tif30170[where, R 1 , R 2 , R 3 And A is defined for general formula (I), and R 28 C 1-6 It can be prepared by hydrolysis of an ester of alkyl or benzyl.

[0109] Ideally, hydrolysis is carried out under basic conditions, for example, by using a metal hydroxide such as lithium hydroxide, sodium hydroxide, or potassium hydroxide in an aqueous organic solvent.

[0110] Compounds of general formula (XIII) are of general formula (XIV): TIFF0007862174000029.tif22170[In the formula, A is as defined for general formula (I), R 28 The compound of the general formula (XIII) is defined as follows: General formula (XV): TIFF0007862174000030.tif31170[where, R 1 , R 2 and R 3 This is defined for general formula (I), and R 29It can be prepared by reacting it with a compound of halo, for example, chloro or bromo.

[0111] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as triethylamine or diisopropylethylamine, in an organic solvent such as dichloromethane.

[0112] Compounds of general formulas (XIV) and (XV) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0113] Z 1 but * -NHC(O)- and Z 2 but * For compounds of general formula (I) which are -C(O)NH-, alternatively, for compounds of general formula (XV) as defined above, general formula (XVI): TIFF0007862174000031.tif25170[In the formula, A, Y and R 4 It can be prepared by reacting it with a compound of the general formula (I) as defined below.

[0114] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as triethylamine or diisopropylethylamine, in an organic solvent such as dichloromethane.

[0115] Compounds with general formula (XVI) are those with general formula (XVII): The compound of TIFF0007862174000032.tif21170 [wherein A is as defined for general formula (I)] It can be prepared by reacting it with a compound of the general formula (XI) defined above.

[0116] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as N,N'-dimethylformamide, using a coupling agent such as HATU.

[0117] Compounds of general formula (XVII) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0118] Z 1 but * -C(O)NH- and Z 2 but * Compounds with the general formula (I) -C(O)NH- are general formula (XX): TIFF0007862174000033.tif22170[In the formula, A, Y and R 4 Compounds of the general formula (I) can be prepared by reacting them with compounds of the general formula (III) as defined above.

[0119] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as N,N'-dimethylformamide, using a coupling agent such as TBTU.

[0120] Carboxylic acids of general formula (XX) are those of general formula (XXI): TIFF0007862174000034.tif22170[In the formula, A, Y and R 4 This is defined for general formula (I), and R 30 C 1-6 It can be prepared by hydrolysis of a compound that is alkyl or benzyl.

[0121] Ideally, hydrolysis is carried out under basic conditions, for example, by using a metal hydroxide such as lithium hydroxide, sodium hydroxide, or potassium hydroxide in an aqueous organic solvent.

[0122] Compounds with general formula (XXI) are those with general formula (XXII): TIFF0007862174000035.tif20170[In the formula, A and Y are as defined for general formula (I), R30 Compounds of the general formula (XXI) can be prepared by reacting a compound of the general formula (XI) as defined above with a compound of the general formula (XI).

[0123] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as N,N'-dimethylformamide, using a coupling agent such as HATU.

[0124] Compounds of general formula (XXII) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0125] Or, Z 1 but * -C(O)NH- and Z 2 but * Compounds with the general formula (I) -C(O)NH- are general formula (XXV): TIFF0007862174000036.tif24170[where, R 1 , R 2 , R 3 Compounds of formula (I) and A can be prepared by reacting a compound of formula (XI) as defined above with a compound of formula (XI).

[0126] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as N,N'-dimethylformamide, using a coupling agent such as HATU.

[0127] Carboxylic acids of general formula (XXV) are those of general formula (XXVI): TIFF0007862174000037.tif25170[where, R 1 , R 2 , R 3 And A is defined for general formula (I), and R 31 C1-6 It can be prepared by hydrolysis of an ester of alkyl or benzyl.

[0128] Ideally, hydrolysis is carried out under basic conditions, for example, by using a metal hydroxide such as lithium hydroxide, sodium hydroxide, or potassium hydroxide in an aqueous organic solvent.

[0129] Compounds with general formula (XXVI) are those with general formula (XXVII): TIFF0007862174000038.tif20170[In the formula, A is as defined for general formula (I), R 31 The compound of the general formula (XXVI) is defined as follows: It can be prepared by reacting it with a compound of the general formula (III) defined above.

[0130] Ideally, the reaction is carried out under basic conditions, in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as N,N'-dimethylformamide, using a coupling agent such as HATU.

[0131] Compounds of general formula (XXVII) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0132] Z 1 but * -NHC(O)- and Z 2 but * Compounds of general formula (I) that are -NHC(O)- include those of general formula (XXX): TIFF0007862174000039.tif22170[In the formula, A, Y and R 4 This is defined for general formula (I), and R 32 The compound is a halo, for example, chloro or bromo. General formula (XXXI): TIFF0007862174000040.tif28170[where, R 1, R 2 and R 3 The compound is defined as follows for general formula (I): It can be prepared by reacting in the presence of a phosphorus ligand such as XantPhos, a palladium catalyst, and a base such as potassium carbonate.

[0133] Compounds of general formula (XXXI) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0134] Compounds of general formula (XXX) are of general formula (XXXII): TIFF0007862174000041.tif11170[In the formula, A is as defined for general formula (I), R 32 The compound of the general formula (XXX) is defined as follows: It can be prepared by reacting it with a compound of general formula (VI) or (VII) as defined above.

[0135] When a compound of general formula (XXXII) is reacted with a carboxylic acid of general formula (VI), the reaction is preferably carried out under basic conditions, in the presence of a coupling agent, such as an amine (DIPEA), and in an organic solvent such as DMF. Suitable coupling agents are as described above.

[0136] When a compound of general formula (XXXII) is reacted with an acid halide of general formula (VII), the reaction is preferably carried out under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA), and in an organic solvent such as dichloromethane.

[0137] Compounds of general formula (XXXII) are known and readily available, or can be prepared by methods well known to those skilled in the art.

[0138] Z 1 but * -NHC(O)- and Z 2 but* It is -NHC(O)-, Y is -CH2-, and R 4 The 2nd position is replaced by OH and the 5th position is R 11a Compounds of general formula (I) that are phenyl substituted with are general formula (XXXV): TIFF0007862174000042.tif28170[where, R 1 , R 2 , R 3 And A is as defined for general formula (I)], General formula (XXXIV): TIFF0007862174000043.tif25170[where, R 11a This refers to H, halo, C1-4 alkyl, C1-4 haloalkyl or C(O)O(C 1-4 It can be prepared by reacting it with a 5-substituted 3H-benzofuran-2-one of an alkyl group, but especially a halo, especially a fluoro, or chloro group.

[0139] Ideally, the reaction is carried out in an organic solvent such as toluene at a high temperature, for example, about 100-140°C, typically about 120°C.

[0140] 5-Chloro-3H-benzofuran-2-one can be prepared by the method described in Journal of Fluorine Chemistry 99(1999)189-195.

[0141] Compounds of general formula (XXXV) are also of general formula (XXXIX): TIFF0007862174000044.tif48170[where, R 11c H, Haro, CN, C 1-4 Alkyl or C 1-4 Reacting with a haloalkyl compound, especially a halo, such as fluoro or chloro, Next, it is reduced with a hydride reducing agent such as lithium borohydride, R 4 The 3rd position is replaced by OH, the 4th position is replaced by -C(CH3)2-CH2OH, and the 6th position is R 11Compounds of general formula (I), which are phenyls substituted with , can be obtained.

[0142] The reaction between compounds of general formula (XXXV) and (XXXIX) can be carried out appropriately under basic conditions in the presence of a coupling reagent such as T3P®, for example, in the presence of DIPEA.

[0143] Compounds with general formula (XXXV) are those with general formula (XXXVI): TIFF0007862174000045.tif29170[where, R 1 , R 2 and R 3 The compound of the general formula (I) is defined as follows: General formula (XXXVII): H2N-A-NH2(XXXVII) [In the formula, A is as defined for general formula (I)] and the compound, It can be prepared by reacting a base, typically an amine such as DIPEA.

[0144] The reaction may be carried out at a low temperature, for example, around 0°C, before being heated to room temperature (approximately 15-25°C). Suitable reaction solvents include organic solvents such as dichloromethane.

[0145] Compounds of general formulas (XXXVI) and (XXXVII) are known and readily available, or can be prepared by methods well known to those skilled in the art. Benzene-1,3-diamine is an example of a compound of general formula (XXXVII).

[0146] Compounds with general formula (XXXIX) are those with general formula (XL): TIFF0007862174000046.tif43170[where, R 11c As defined above for the general formula (XXXIX), R 35 C 1-6 Alkyl or benzyl, R 36 C 1-6Compounds that are alkyl or benzyl, It can be prepared by reacting it with boron tribromide, as described in Example 9 below.

[0147] Compounds with general formula (XL) are those with general formula (XLI): TIFF0007862174000047.tif43170[where, R 11c As defined above for the general formula (XXXIX), R 35 and R 36 As defined above for the general formula (XL), R 37 [The compound is a halo, for example, chloro or bromo, especially bromo] It can be prepared by reacting with (1-methoxy-2-methyl-propa-1-enoxy)-trimethyl-silane.

[0148] The reaction is carried out in the presence of ZnF2, preferably under inert conditions, for example, under a nitrogen atmosphere, in a degassed organic solvent such as N,N-dimethylformamide, at a high temperature, for example, 60-100°C, and in the presence of a palladium catalyst, for example, Pd(PtBu3)2).

[0149] Compounds with general formula (XLI) are those with general formula (XLII): TIFF0007862174000048.tif43170[where, R 11c As defined above for the general formula (XXXIX), R 35 As defined above for the general formula (XL), R 37 The compound of the general formula (XLI) is defined above. General formula (XLIII): R 36 -R 38 (XLIII) [In the formula, R 36 As defined above for the general formula (XL), R 38 It can be prepared by reacting it with a compound of halo, particularly bromo.

[0150] Compounds of general formula (XLIII) are readily available or can be prepared by methods familiar to those skilled in the art.

[0151] Compounds with general formula (XLII) are those with general formula (XLIV): TIFF0007862174000049.tif42170[where, R 11c As defined above for the general formula (XXXIX), R 35 Compounds of the general formula (XL) as defined above can be prepared by halogenation.

[0152] Suitable halogenating agents are well known to those skilled in the art, for example, R 37 If the halogenating agent is bromo, the halogenating agent may be bromine, for example, a solution of bromine in acetonitrile as described in Example 9 below.

[0153] An alternative method for preparing a compound of general formula (I), Z 1 but * -C(O)NH- and Z 2 but * -NHC(O)- and A TIFF0007862174000050.tif25170 is: R 4 The 3rd position is substituted with OH, the 4th position is substituted with -C(CH3)2-CH2OH, and the 6th position is R 11 The method for phenyl substituted with is the general formula (XLV): TIFF0007862174000051.tif27170[where, R 1 , R 2 , R 3 , X 1 , X 2 and X 3 The compound of the general formula (I) is defined as follows: This method involves reacting the compound with the general formula (XXXIX) defined above, and then reducing it using a hydride reducing agent, such as lithium borohydride.

[0154] The process is appropriately carried out under basic conditions in the presence of a coupling reagent such as T3P®, for example, in the presence of DIPEA.

[0155] Compounds with general formula (XLV) are those with general formula (XLVI): TIFF0007862174000052.tif27170[where, R 1 , R 2 , R 3 , X 1 , X 2 and X 3 It can be prepared by reducing a compound of the general formula (I) as defined.

[0156] Ideally, reduction is carried out by catalytic hydrogenation, for example, using a palladium-carbon catalyst.

[0157] Compounds with general formula (XLVI) are those with general formula (XLVII): TIFF0007862174000053.tif27170[In the formula, X 1 , X 2 and X 3 The compound of the general formula (I) is defined as follows: It can be prepared by reacting it with a compound of general formula (III) as defined above. Preferably, the reaction is carried out in the presence of a coupling reagent under the same conditions as described above for the reaction of a compound of general formula (III) with a compound of general formula (II).

[0158] Compounds of general formulas (II), (X), (XVI), (XX), (XXV), (XXX), or (XXXV) form further embodiments of the present invention.

[0159] Compounds of general formula (I) can also be prepared from other compounds of general formula (I). For example, any of R 3 ;R 1 , R 2 and R 3 together; or R 4However, if the compound contains phenyl groups substituted with alkoxy groups, they can be converted to hydroxyl-substituted phenyl groups by treatment with boron tribromide.

[0160] Other interconversions of various substituents can be carried out by methods familiar to those skilled in the art.

[0161] Since the compounds of general formula (I) are modulators of TMEM16A, further aspects of the present invention provide compounds of general formula (I) as defined above for use in medicine, particularly in the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A.

[0162] The use of compounds of general formula (I) in the manufacture of pharmaceuticals for the treatment or prevention of diseases and symptoms affected by the modification of TMEM16A is also provided.

[0163] Methods for treating or preventing diseases and conditions affected by the modification of TMEM16A are also provided, comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment.

[0164] Diseases and symptoms affected by the regulation of TMEM16A include respiratory diseases and symptoms, dry mouth (xerostomia), hypermotility of the intestines, cholestasis, and ocular symptoms.

[0165] The following are also provided: • Compounds of general formula (I) for use in the treatment or prevention of respiratory diseases and symptoms. A compound of general formula (I) for use in the treatment or prevention of dry mouth (xerostomia). • Compounds of general formula (I) for use in the treatment or prevention of excessive bowel motility. • Compounds of general formula (I) for use in the treatment or prevention of cholestasis. • Compounds of general formula (I) for use in the treatment or prevention of ocular symptoms.

[0166] This invention also provides the following: • Use of compounds of general formula (I) in the manufacture of medicinal products for the treatment or prevention of respiratory diseases and symptoms. • Use of compounds of general formula (I) in the manufacture of pharmaceuticals for the treatment or prevention of dry mouth (xerostomia). • Use of compounds of general formula (I) in the manufacture of pharmaceuticals for the treatment or prevention of excessive bowel motility. • Use of compounds of general formula (I) in the manufacture of pharmaceuticals for the treatment or prevention of cholestasis. • Use of compounds of general formula (I) in the manufacture of pharmaceuticals for the treatment or prevention of eye conditions.

[0167] Furthermore, the following are provided: A method for treating or preventing respiratory diseases and symptoms, comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment. A method for treating or preventing dry mouth (xerostomia), comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment. A method for treating or preventing excessive bowel motility, comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment. A method for treating or preventing cholestasis, comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment. A method for treating or preventing an eye condition, comprising administering an effective amount of a compound of general formula (I) to a patient in need of such treatment.

[0168] Respiratory diseases and conditions that can be treated or prevented by compounds of general formula (I) include cystic fibrosis, chronic obstructive pulmonary disease (COPD), chronic bronchitis, emphysema, bronchiectasis (including non-cystic fibrosis and bronchiectasis), asthma, and primary ciliary dyskinesia.

[0169] Dry mouth (xerostomia) that can be treated or prevented by compounds of general formula (I) may be caused by Sjögren's syndrome, radiotherapy, and xerogenic drugs.

[0170] Excessive bowel motility, which can be treated or prevented by compounds of general formula (I), may be associated with gastric indigestion, gastroparesis, chronic constipation, and irritable bowel syndrome.

[0171] Ocular conditions that can be treated or prevented by compounds of general formula (I) include dry eye disease.

[0172] The compounds of the present invention are generally administered as part of a pharmaceutical composition, and therefore, the present invention further provides pharmaceutical compositions comprising a compound of general formula (I) together with pharmaceutically acceptable excipients.

[0173] The compounds of general formula (I) that are more suitable for use in pharmaceutical compositions are as described above.

[0174] Pharmaceutical compositions may be formulated for oral, rectal, nasal, bronchial (inhalation), topical (including skin, transdermal, eye drops, buccal, and sublingual), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, and intradermal) administration, and may be prepared by any method well known in the field of pharmacy.

[0175] The composition may be prepared by associating the active agent defined above with an excipient. Generally, formulations are prepared by homogeneously and closely associating the active agent with a liquid carrier, a micronized solid carrier, or both, and then, if necessary, shaping the product. The present invention relates to a method for preparing pharmaceutical compositions comprising combining or associating a compound of general formula (I) with a pharmaceutically acceptable carrier or vehicle.

[0176] The preparations for oral administration in the present invention are provided as individual units such as capsules, sachets, or tablets, each containing a predetermined amount of the active agent; as powder or granules; as a solution or suspension of the active agent in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil liquid emulsion; or as a bolus, etc.

[0177] In the case of compositions for oral administration (e.g., tablets and capsules), the term "acceptable carrier" includes common excipients, such as binders, vehicles such as syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sucrose, and starch; fillers and carriers, such as corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, and alginic acid; and lubricants, such as magnesium stearate, sodium stearate, and other metal stearates, glycerol stearate, stearic acid, silicone fluids, talc wax, oils, and colloidal silica. Flavoring agents such as peppermint, wintergreen oil, and cherry flavoring agents may also be used. It may be desirable to add colorants to make the dosage form easily identifiable. Tablets may also be coated by methods well known in the art.

[0178] Tablets may be prepared by compression or molding, along with one or more auxiliary components as desired. Compressed tablets may be prepared by compressing an active agent in a free-flowing form, such as a powder or granules, mixed optionally with a binder, lubricant, inert diluent, preservative, surfactant, or dispersant, in a suitable machine. Molded tablets may be prepared by molding a mixture of a moistened powdered compound and an inert liquid diluent in a suitable machine. Tablets may optionally be coated or grooved, and may be formulated to provide a slowed or controlled release of the active agent.

[0179] Other formulations suitable for oral administration include flavored bases, medicinal candies usually containing the active agent in sucrose and gum arabic or tragacanth, scented tablets containing the active agent in an inert base such as gelatin and glycerin or sucrose and gum arabic, and mouthwashes containing the active agent in a suitable liquid carrier.

[0180] For topical application to the skin, compounds of general formula (I) can be formulated as creams, ointments, jellies, solutions, or suspensions. Cream or ointment formulations that can be used as drugs are conventional formulations well known in the art, such as those described in standard pharmaceutical textbooks, such as the British Pharmacopoeia.

[0181] Local administration to the lungs can be achieved by the use of aerosol formulations. Aerosol formulations typically contain an active ingredient suspended or dissolved in a suitable aerosol propellant such as a chlorofluorocarbon (CFC) or hydrofluorocarbon (HFC). Suitable CFC propellants include trichloromonofluoromethane (propellant 11), dichlorotetrafluoromethane (propellant 114), and dichlorodifluoromethane (propellant 12). Suitable HFC propellants include tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227). The propellant typically constitutes 40% to 99.5% by weight, for example, 40% to 90% by weight, of the total inhalation composition. The formulation may contain excipients including a cosolvent (e.g., ethanol) and a surfactant (e.g., lecithin, sorbitan trioleate, etc.). Other possible excipients include polyethylene glycol, polyvinylpyrrolidone, and glycerin. The aerosol formulation is packaged in a canister, and the appropriate dose is delivered by a dispensing valve (e.g., supplied by Bespak, Valois, or 3M, or by Aptar, Coster, or Vari).

[0182] Local administration to the lungs can also be achieved by using non-pressurized formulations such as aqueous solutions or suspensions. These can be administered by nebulizers, such as handheld and portable nebulizers, or by home or hospital (i.e., non-portable) nebulizers. The formulations may contain water, buffers, isotonic modifiers, pH adjusters, surfactants, and excipients such as cosolvents. Suspensions and aerosol formulations (whether pressurized or non-pressurized) typically contain, for example, 0.5–10 μm, or for example, about 1–5 μm of D 50 The compound of the present invention is contained in a finely powdered form having the particle size distribution, D 10 , D 50 and D 90 It can be expressed using a value. D of particle size distribution 50 The median is defined as the particle size in microns that divides the distribution in half. Measurements derived from laser diffraction are more accurately described as volume distributions, and therefore, the D obtained using this procedure 50 The value is Dv 50 The value (median of the volume distribution) is more meaningful. As used herein, the Dv value refers to the particle size distribution measured using laser diffraction. Similarly, D used in the context of laser diffraction 10 and D 90 The value is Dv 10 and Dv 90 It is interpreted as meaning a value, referring to particle size, and thus 10% of the distribution is D 10 The values ​​are below the threshold, and 90% of the distribution is D 90 It falls below the value.

[0183] Local administration to the lungs can also be achieved by using a dry powder formulation. Dry powder formulations typically have a mass-average diameter (MMAD) of 1–10 μm or 0.5–10 μm, for example, about 1–5 μm. 50The formulation contains the compound of the present disclosure in a micronized form having the following characteristics. The powder of the compound of the present invention in a micronized form can be prepared by a micronization process or a similar size reduction process. Micronization may be performed using a jet mill such as that manufactured by Hosokawa Alpine. The resulting particle size distribution can be measured using laser diffraction (e.g., with a Malvern Mastersizer 2000S instrument). The formulation typically has a relatively large particle size, e.g., 50 μm or larger, e.g., a mass-average diameter (MMAD) of 100 μm or larger, or a D of 40-150 μm. 50 It contains a topically acceptable diluent such as lactose, glucose, or mannitol (preferably lactose). As used herein, the term “lactose” refers to lactose-containing components, including α-lactose monohydrate, β-lactose monohydrate, α-lactose anhydrous, β-lactose anhydrous, and amorphous lactose. Lactose components can be processed by micronization, sieving, grinding, compression, agglomeration, or spray drying. This also includes commercially available forms of lactose in various forms, such as Lactohale® (inhalation-grade lactose; DFE Pharma), InhaLac® 70 (sieved lactose for dry powder inhalation; Meggle), Pharmatose® (DFE Pharma), and Respitose® (sieved inhalation-grade lactose; DFE Pharma) products. In one embodiment, the lactose component is selected from the group consisting of α-lactose monohydrate, α-lactose anhydrous, and amorphous lactose. Preferably, the lactose is α-lactose monohydrate.

[0184] Dry powder formulations may also contain other excipients. Therefore, in one embodiment, the dry powder formulation according to this disclosure comprises magnesium stearate or calcium stearate. Such formulations may have excellent chemical and / or physical stability, particularly if they also contain lactose.

[0185] Dry powder formulations are typically delivered using dry powder inhaler (DPI) devices. Examples of dry powder delivery systems include SPINHALER®, DISKHALER®, TURBOHALER®, DISKUS®, SKYEHALER®, ACCUHALER®, and CLICKHALER®. Further examples of dry powder delivery systems include ECLIPSE, NEXT, ROTAHALER, HANDIHALER, AEROLISER, CYCLOHALER, BREEZHALER / NEOHALER, MONODOSE, FLOWCAPS, TWINCAPS, X-CAPS, TURBOSPIN, ELPENHALER, MIATHALER, TWISTHALER, NOVOLIZER, PRESSAIR, ELLIPTA, ORIEL dry powder inhaler, MICRODOSE, PULVINAL, EASYHALER, ULTRAHALER, TAIFUN, PULMOJET, OMNIHALER, GYROHALER, TAPER, CONIX, XCELOVAIR, and PROHALER.

[0186] In one embodiment, the compound of general formula (I) is provided as a micronized dried powder formulation containing, for example, a suitable grade of lactose.

[0187] Accordingly, in one aspect of the present invention, a pharmaceutical composition is provided which comprises a compound of general formula (I) in granular form combined with granular lactose, and optionally contains magnesium stearate.

[0188] In one embodiment, the compound of general formula (I) is provided as a micronized dry powder formulation containing appropriate grade lactose and magnesium stearate, which is filled into a device such as a DISKUS. Preferably, such a device is a multi-dose device, and for example, the formulation is filled into blisters for use in a multi-unit dosing device such as a DISKUS.

[0189] In another embodiment, the compound of general formula (I) is provided as a finely atomized dried powder formulation containing a suitable grade of lactose, filled into hard-shell capsules for use in single-dose devices such as AEROLISER.

[0190] In another embodiment, the compound of general formula (I) is provided as a finely atomized dried powder formulation containing appropriate grade lactose and magnesium stearate, filled into hard-shell capsules for use in single-dose devices such as AEROLISER.

[0191] In another embodiment, the compound of general formula (I) is provided as a finely pulverized powder for use in an inhalation dosage form, the powder being produced by a size reduction process other than jet milling, e.g., spray drying, spray freezing, microfluidization, high-pressure homogenization, supercritical fluid crystallization, ultrasonic crystallization, or a combination thereof, or by other suitable particle formation methods known in the art used to produce fine particles with an aerodynamic particle size of 0.5 to 10 μm, for example, about 1 to 5 μm. 50 These are fine particles having [a certain characteristic]. The resulting particle size distribution can be measured using laser diffraction (for example, with a Malvern Mastersizer 2000S instrument). The particles may contain the compound alone or in combination with other suitable excipients that can assist in processing. The resulting fine particles may form a final formulation for delivery to humans, or optionally, be further formulated with other suitable excipients to facilitate delivery in an acceptable dosage form.

[0192] The compounds of the present invention can also be administered rectally in the form of suppositories or enemas, including, for example, aqueous or oily solutions, as well as suspensions, emulsions, and foams. Such compositions are prepared according to standard procedures well known to those skilled in the art. For example, suppositories can be prepared by mixing the active ingredient with a conventional suppository base such as cocoa butter or other glycerides. In this case, the drug is mixed with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycol.

[0193] Generally, for compositions intended for topical administration to the eye in the form of eye drops or ointments, the total amount of the compound of general formula (I) is less than approximately 0.0001 to 4.0% (w / w).

[0194] Preferably, for topical ocular administration, the composition administered according to general formula (I) is formulated as a solution, suspension, emulsion, or other dosage form. Aqueous solutions are generally preferred because they are easy to formulate and allow the patient to easily administer such a composition by dropping one or two drops of the solution onto the affected eye. However, the composition may also be a suspension, a viscous or semi-viscous gel, or other types of solid or semi-solid compositions. Suspensions may be preferred for compounds that are poorly soluble in water.

[0195] An alternative method of administration to the eye is intravitreal injection of a solution or suspension of the compound of general formula (I). Furthermore, the compound of general formula (I) can also be introduced via an ocular implant or insert.

[0196] Compositions administered according to general formula (I) may also contain, but are not limited to, a variety of other components, including isotonic agents, buffers, surfactants, stabilizing polymers, preservatives, cosolvents, and viscosity enhancers. A suitable pharmaceutical composition of general formula (I) comprises the compound of the present invention formulated together with an isotonic agent and a buffer. The pharmaceutical composition of general formula (I) may optionally further contain a surfactant and / or a mitigating agent and / or a stabilizing polymer.

[0197] The isotonicity of a composition can be adjusted to the isotonicity of natural tears, preferably for ophthalmic compositions, using various isotonic agents. For example, physiological tonicity can be approximated by adding simple sugars such as sodium chloride, potassium chloride, magnesium chloride, calcium chloride, dextrose, fructose, and galactose, and / or sugar alcohols such as mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, and hydrolyzed starch to a composition. The amount of such isotonic agent varies depending on the specific agent added. However, generally, a composition contains enough isotonic agent to have an ophthalmologically acceptable osmolality by weight (generally about 150-450 mOsm, preferably 250-350 mOsm, most preferably about 290 mOsm) in the final composition. Generally, the isotonic agents of the present invention are present in the range of 2-4% (w / w). Preferred isotonic agents of the present invention include simple sugars or sugar alcohols, such as D-mannitol.

[0198] A suitable buffer system (e.g., sodium phosphate, sodium acetate, sodium citrate, sodium borate, or boric acid) can be added to the composition to prevent pH drift under storage conditions. The specific concentration will vary depending on the agent used. However, preferably, the buffer is selected to maintain the target pH within the range of pH 5 to 8, more preferably at a target pH of pH 5 to 7.

[0199] A surfactant may be used to deliver a higher concentration of the compound of general formula (I). The surfactant functions to solubilize the compound and stabilize colloidal dispersions such as micellar solutions, microemulsions, emulsions, and suspensions. Examples of surfactants that can be optionally used include polysorbate, poloxamer, polyosyl 40 stearate, polyoxyl castor oil, tyroxapole, Triton, and sorbitan monolaurate. Preferred surfactants used in the present invention have a hydrophilic / lipophilic / balance "HLB" in the range of 12.4 to 13.2 and are acceptable for ophthalmic use, such as Triton X114 and tyroxapole.

[0200] Further agents that can be added to ophthalmic compositions of compounds of general formula (I) are mitigating agents that function as stabilizing polymers. The stabilizing polymer should be an ionic / charged example preferred for topical ophthalmic use, and more specifically, a polymer having a negative charge on its surface that can exhibit a zeta potential of (-)10 to 50 mV for physical stability and can produce dispersions in water (i.e., water-soluble). Preferred stabilizing polymers of the present invention are 0.1 to 0.5% w / w of polyelectrolytes from the family of cross-linked polyacrylates such as carbomers and pemlen(R), particularly carbomer 974p (polyacrylic acid), or multiple polyelectrolytes in the case of multiple polymers.

[0201] To increase the viscosity of the carrier, other compounds may be added to the ophthalmic composition of the compound of general formula (I). Examples of thickeners include, but are not limited to, hyaluronic acid and its salts, chondroitin sulfate and its salts, dextran, various polymers of the cellulose family, vinyl polymers, and polysaccharides such as acrylic acid polymers.

[0202] Topical ophthalmic products are typically packaged in multi-dose forms. Therefore, preservatives are necessary to prevent microbial contamination during use. Suitable preservatives include benzalkonium chloride, chlorobutanol, benzododecinium bromide, methylparaben, propylparaben, phenylethyl alcohol, disodium edentate, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art. Such preservatives are typically used at levels of 0.001–1.0% w / v. Unit-dose compositions of general formula (I) are sterile but typically not preserved. Therefore, such compositions generally do not contain preservatives.

[0203] Parenteral formulations are generally sterile.

[0204] A physician or other person skilled in the art can determine the appropriate dosage of the compound of general formula (I), and therefore the amount of the compound of the present invention to be included in any particular pharmaceutical preparation (whether in unit dosage form or not).

[0205] Compounds of general formula (I) can be used in combination with one or more other active agents useful for the treatment or prevention of respiratory diseases and symptoms.

[0206] Further active agents of this type may be included in the above-described pharmaceutical composition, or they may be administered simultaneously with, or earlier than or later than, the compound of general formula (I).

[0207] Accordingly, in a further aspect of the present invention, a product is provided comprising a compound of general formula (I) and an additional agent useful for the treatment or prevention of respiratory symptoms, as a combination preparation for simultaneous, sequential, or separate use in the treatment of a disease or condition affected by the modification of TMEM16A, particularly respiratory diseases or conditions, for example.

[0208] Compounds of general formula (I) are also provided in combination with additional agents useful for the treatment or prevention of respiratory symptoms, for example, as combination preparations for simultaneous, sequential, or separate use in the treatment of one of the above-mentioned diseases and symptoms, particularly respiratory diseases or symptoms, which are affected by the modification of TMEM16A.

[0209] Suitable additional active agents that may be included in a pharmaceutical composition or preparation in combination with a compound of general formula (I) include: β2 adrenergic receptor agonists, such as metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol, formoterol, salmeterol, indacaterol, terbutaline, orciprenaline, bitolterol mesylate, pirbuterol, orodaterol, vilanterol, and abedeterol; Antihistamines, such as histamine H1 receptor antagonists or H4 receptor antagonists, including loratadine, cetirizine, desloratadine, levocetirizine, fexofenadine, astemizole, azelastine, and chlorpheniramine; Dornase α; Corticosteroids such as prednisone, prednisolone, flunisolide, triamcinolone acetonide, beclomethasone dipropionate, budesonide, fluticasone propionate, mometasone furoate, and fluticasone furoate; Leukotriene antagonists such as montelukast and zafirlukast; Anticholinergic compounds, particularly muscarinic antagonists such as ipratropium, tiotropium, glycopyrrolate, acridinium, and umeclidinium; CFTR repair therapies such as Ivacaftor, QBW251, Bamacaftor (VX659), Elexacaftor (VX445), VX561 / CPT-656, VX152, VX440, GLP2737, GLP2222, GLP2451, PTI438, PTI801, PTI808, FDL-169 and FDL-176 (e.g., CFTR enhancers, collectors or amplifiers), as well as CFTR collectors such as Lumacaftor and Tezacaftor, or combinations thereof (e.g., combinations of Ivacaftor, Tezacaftor and Elexacaftor); ENaC modulators, especially ENaC inhibitors; antibiotics; Antiviral agents, such as ribavirin and neuraminidase inhibitors, such as zanamivir; Antifungal agents such as PUR1900; Hypertonic saline and respiratory tract wettable powders such as mannitol (Bronchitol®) (osmoloytes); and Mucolytic agents, such as N-acetylcysteine.

[0210] If the additional activator is an ENaC modulator, it may be an ENaC inhibitor such as amiloride, VX-371, AZD5634, QBW276, SPX-101, BI443651, BI1265162, and ETD001. Other suitable ENaC blockers are disclosed in our application's International Publication Brochures 2017 / 221008, 2018 / 096325, 2019 / 077340, and 2019 / 220147, and any of the exemplary compounds in these applications may be used in combination with compounds of general formula (I), (Ix), (IA), (IB), (IC), (ID), or (IE). Compounds particularly suitable for use in combination with compounds of general formula (I), (Ix), (IA), (IB), (IC), (ID), or (IE) include compounds having cations selected from the following: 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)ethyl]-6-(4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}piperidine-1-carbonyl)-1,3-diethyl-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-6-{[2-(4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}piperidine-1-yl)ethyl]carbamoyl}-1,3-diethyl-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-5-[4-({bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}methyl)piperidine-1-carbonyl]-1,3-diethyl-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-6-[(3R)-3-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}pyrroridine-1-carbonyl]-1,3-diethyl-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-6-[(3S)-3-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}pyrroridine-1-carbonyl]-1,3-diethyl-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-1,3-diethyl-6-{[(1r,4r)-4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}cyclohexyl]carbamoyl}-1H-1,3-benzodiazole-3-ium; 2-[({3-amino-5H-pyrrolo[2,3-b]pyrazine-2-yl}formamide)methyl]-1,3-diethyl-6-{[(1s,4s)-4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}cyclohexyl]carbamoyl}-1H-1,3-benzodiazole-3-ium; And suitable anions, such as halides, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, citric acid, tartaric acid, oxalic acid, succinic acid, mandelic acid, methanesulfonic acid, or p-toluenesulfonic acid.

[0211] The present invention is illustrated by the following non-limiting embodiments and drawings. [Brief explanation of the drawing]

[0212] [Figure 1] This is an exemplary trace from a whole-cell patch-clamp (Qpatch) TMEM16A potentiometer assay, as used in Biological Example 95, illustrating the methodology used in this assay. [Modes for carrying out the invention] [Examples]

[0213] General conditions: Mass spectra were performed using an LC-MS system with electrospray ionization. These were performed using either a Waters Acquity uPLC system with Waters PDA and ELS detectors or a Shimadzu LCMS-2010 EV system. [M+H]+ refers to the monoisotope molecular weight.

[0214] NMR spectra were recorded using a Bruker Avance III HD 500 MHz with a 5 mm Broad Band Inverse probe, or a Bruker Avance III HD 250 MHz or 400 MHz Avance III HD Nanobay with a 5 mm Broad Band Observed SmartProbe, with the solvent used as an internal deuterium lock. Unless otherwise specified, spectra were recorded at room temperature and referenced using solvent peaks.

[0215] With reference to the following examples, compounds of preferred embodiments were synthesized using the methods described herein or other methods known in the art.

[0216] Various starting materials, intermediates, and compounds of preferred embodiments can be isolated and purified using conventional techniques such as precipitation, filtration, crystallization, evaporation, distillation, and chromatography, as needed. Unless otherwise specified, all starting materials are obtained from commercial suppliers and used without further purification. Salts can be prepared from compounds by known salt-forming procedures.

[0217] The compounds were purified by flash column chromatography on normal-phase silica using a suitable SNAP cartridge and gradient in a Biotage® Isolera system. Alternatively, the compounds were purified on reverse-phase silica using a Biotage® Isolera system equipped with a suitable SNAP C18 cartridge and reverse-phase eluent, or by preparative HPLC (as otherwise stated). Preparative HPLC using acidic pH, early elution method

[0218] Purification was performed on a Gilson LC system using a Waters Sunfire C18 column (30 mm x 100 mm, 10 μM; temperature: RT) with a gradient from 10 to 95% B (A = 0.1% formic acid in water; B = 0.1% formic acid in acetonitrile) over 14.44 minutes, followed by a gradient to 95% B over 2.11 minutes, at an injection volume of 1500 μL and a flow rate of 40 mL / min. The UV spectrum was recorded at 215 nm using a Gilson detector.

[0219] Preparative HPLC using acidic pH, standard elution method Purification by preparative HPLC (acidic pH, standard elution method) was performed on a Gilson LC system using a Waters Sunfire C18 column (30 mm x 100 mm, 10 μM; temperature: RT) with a gradient of 30–95% B (A = 0.1% formic acid in water; B = 0.1% formic acid in acetonitrile) over 11 minutes, followed by 95% B over 2.11 minutes, at an injection volume of 1500 μL and flow rate of 40 mL / min. The UV spectrum was recorded at 215 nm using a Gilson detector.

[0220] Preparative HPLC using basic pH, early elution method Purification by preparative HPLC (basic pH, early elution method) was performed on a Gilson LC system using a Waters Xbridge C18 column (30 mm x 100 mm, 10 μM; temperature: RT) with a gradient of 10–95% (A = 0.2% ammonium hydroxide in water; B = 0.2% ammonium hydroxide in acetonitrile) over 14.44 minutes, followed by 95% B over 2.11 minutes, at an injection volume of 1500 μL and flow rate of 40 mL / min. UV spectra were recorded at 215 nm using a Gilson detector.

[0221] Preparative HPLC using basic pH, standard elution method Purification by preparative HPLC (basic pH, standard elution method) was performed on a Gilson LC system using a Waters Xbridge C18 column (30 mm x 100 mm, 10 μM; temperature: RT) with a gradient of 30–95% (A = 0.2% ammonium hydroxide in water; B = 0.2% ammonium hydroxide in acetonitrile) over 11 minutes, followed by 95% B over 2.11 minutes, at an injection volume of 1500 μL and flow rate of 40 mL / min. UV spectra were recorded at 215 nm using a Gilson detector.

[0222] Unless otherwise specified, the analytical HPLC conditions are as follows: Method A Column: Phenomenex Kinetix-XB C18 2.1×100mm, 1.7μm Column temperature: 40℃ Eluents: A: H2O + 0.1% formic acid, B: Acetonitrile + 0.1% formic acid Flow rate: 0.6mL / min Gradient: 0-5.3 min 5-100% B, 5.3-5.8 min 100% B, 5.8-5.82 min 100-5% B, 5.82-7.00 min 5% B Method B Column: Waters UPLC (registered trademark) BEH (trademark) C18 2.1 × 100 mm, 1.7 μm Column temperature: 40℃ Eluent: A: 2 mM ammonium bicarbonate buffered to pH 10, B: acetonitrile Flow rate: 0.6mL / min Gradient: 0-5.3 min 5-100% B, 5.3-5.8 min 100% B, 5.8-5.82 min 100-5% B, 5.82-7.00 min 5% B Method C Column: Kinetex Core-Shell C18 2.1x50mm, 5μm Column temperature: 40℃ Eluents: A: H2O + 0.1% formic acid, B: Acetonitrile + 0.1% formic acid Flow rate: 1.2mL / min Gradient: 0-1.20 min 5-100% B, 1.20-1.30 min 100% B, 1.30-1.31 min 100-5% B, 1.31-1.7 min 5% B Method G Column: Waters UPLC (registered trademark) BEH (trademark) C18, 2.1 mm x 50 mm, 1.7 μm Column temperature: 40℃ Eluents: A: H2O + 0.1% formic acid, B: Acetonitrile + 0.1% formic acid Flow rate: 0.9mL / min Gradient: 0-1.10 min 5-100% B, 1.10-1.35 min 100% B, 1.35-1.40 min 100-1% B Method H Column: Kinetex Core-Shell C18 2.1x50mm, 5μm Column temperature: 40℃ Eluents: A: H2O + 0.1% formic acid, B: Acetonitrile + 0.1% formic acid Flow rate: 1.2mL / min Gradient: 0-1.83 min 5-100% B, 1.83-2.25 min 100% B, 2.25-2.26 min

[0223] The following examples are intended to illustrate the present invention and should not be construed as limitations thereon. Temperatures are given in Celsius. Unless otherwise specified, all evaporation is carried out in vacuum, preferably at about 15 mmHg to 100 mmHg (= 20 to 133 mbar). The structures of the final product, intermediates, and starting materials are confirmed by standard analytical methods, e.g., trace analysis and spectroscopic properties, e.g., MS, IR, and NMR. Abbreviations used are conventional in the art. Unless otherwise defined, terms have their generally accepted meanings.

[0224] Abbreviation aq.Aqueous solution br broad d doublet dd doublet doublet DCM Dichloromethane DIPEA Diisopropylethylamine DMF (N,N-dimethylformamide) HCl ethyl acetate HATU 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HPLC (High-Pressure Liquid Chromatography) MeCN acetonitrile MS mass spectrometry m multiplet min mL (milliliter) m / z mass-to-charge ratio NMR nuclear magnetic resonance Rt retention time s singlet t triplet TBTU N,N,N',N'-Tetramethyl-O-(benzotriazol-1-yl)uroniumtetrafluoroborate TEA (Triethylamine) TFA (Trifluoroacetic Acid) THF (Tetrahydrofuran)

[0225] Preparation example Example 1 N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophene-2-carboxamide TIFF0007862174000054.tif32170

[0226] Step 1: Methyl 5-[[2-(5-chloro-2-methoxyphenyl)acetyl]aminothiophene-2-carboxylate TIFF0007862174000055.tif381702-(5-chloro-2-methoxyphenyl)acetic acid (351 mg, 1.75 mmol) and DIPEA (306 μL, 1.75 mmol) were added to a solution of DMF (8 mL) to which HATU (665 mg, 1.75 mmol) was added, followed by the addition of methyl 5-aminothiophene-2-carboxylate (250 mg, 1.59 mmol). The mixture was stirred overnight at room temperature. The resulting mixture was diluted with HCl (30 mL) and washed with a 1 M HCl (30 mL) solution and brine (30 mL). The organic portion was separated, dried over Na₂SO₄, and concentrated under vacuum. The crude residue was absorbed onto silica, and the marked compound was obtained as a colorless waxy solid by chromatography on silica eluted with 0-50% HCl in heptane. LC-MS (Method C): Rt 1.15 min; MS m / z 340.0 / 341.9=[M+H]+(81% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.79(s,1H), 7.60(d,J=4.2Hz,1H), 7.33-7.26(m, 2H), 7.02-6.98(m, 1H), 6.72(d,J=4.2Hz,1H), 3.76(s,3H), 3.75(s,3H), 3.72(s,2H).

[0227] Step 2: 5-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophene-2-carboxylic acid TIFF0007862174000056.tif36170 In a solution of methyl 5-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophene-2-carboxylate (Step 1) (95%, 268 mg, 0.75 mmol) in siRNA (1.33 mL), THF (1.33 mL), and water (1.33 mL), LiOH.H2O (63 mg, 1.5 mmol) was added, and the mixture was stirred at room temperature for 4 hours. Further LiOH.H2O (95 mg, 2.25 mmol) was added, and the mixture was stirred at room temperature for 2 days, after which it was heated to 50°C for 2 hours. After cooling to room temperature, the mixture was acidified to pH 2 with 1 M HCl (4 mL). The resulting precipitate was filtered, washed with water, and dried in a vacuum oven to obtain the labeled compound. LC-MS (Method C): Rt 1.06 min; MS m / z 323.9 / 325.8=[MH] (96% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.70(s,1H), 7.51(d,J=4.1Hz,1H), 7.33-7.28(m, 2H), 7.01(d,J=9.4Hz,1H), 6.69(d,J=4.1Hz,1H), 3.75(s,3H), 3.71(s,2H).

[0228] Step 3: N-tert-butyl-5-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophene-2-carboxamide To a solution of TIFF0007862174000057.tif371705-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophene-2-carboxylic acid (Step 2) (130 mg, 0.40 mmol) and DIPEA (84 μL, 0.48 mmol) in DMF (2 mL), HATU (182 mg, 0.48 mmol), followed by 2-methylpropan-2-amine (50 μL, 0.48 mmol), the mixture was stirred overnight at room temperature. The resulting mixture was diluted with SiO2 (10 mL), washed with 1 M HCl (10 mL) and brine (10 mL), dried over Na2SO4, and concentrated under vacuum. The crude residue was absorbed onto silica and purified by chromatography on silica eluted with 0-50% SiO2 in heptane to obtain the marked compound as a colorless powder. LC-MS (Method C): Rt 1.17 min; MS m / z 381.0 / 383.0=[M+H]+(100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.44(s,1H), 7.54(d,J=4.1Hz,1H), 7.49(s,1H), 7.32-7.29(m, 2H), 7.01(d,J=8.4Hz,1H), 6.61(d,J=4.1Hz,1H), 3.75(s,3H), 3.68(s,2H), 1.34(s,9H).

[0229] Step 4: N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophene-2-carboxamide To a solution of N-tert-butyl-5-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophenyl-2-carboxamide (Step 3) (100%, 130 mg, 0.34 mmol) in 1.5 mL of DCM at 0°C, 1 M BBr3 (0.51 mL, 0.51 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction product was quenched by adding saturated NaHCO3 solution (10 mL), and the resulting mixture was extracted with CHCl3 / IPA (3:1, 2 × 10 mL). The combined organic extract was washed with brine (10 mL), dried over Na2SO4, and concentrated under vacuum. The crude residue was absorbed onto silica, and the labeled compound was obtained as a colorless powder by chromatography on silica eluted with 0-50% ethyl acetate in heptane. LC-MS (Method A): Rt 2.97 min; MS m / z 367.1 / 369.1=[M+H]+(98% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.41(s(br), J=14.4Hz, 1H), 9.83(s(br), 1H), 7.55(d,J=4.1Hz,1H), 7.50(s,1H), 7.21(d ,J=2.7Hz,1H), 7.13(dd,J=8.6,2.7Hz,1H), 6.81(d,J=8.6Hz,1H), 6.61(d,J=4.1Hz,1H), 3.64(s,2H), 1.35(s,9H).

[0230] Example 1.1 N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophene-3-carboxamide The title compound was prepared in the same manner as in Example 1 by replacing methyl 5-aminothiophene-2-carboxylate (Step 1) with methyl 5-aminothiophene-3-carboxylate. LC-MS (Method A): Rt 2.97 min; MS m / z 367.2 / 369.1=[M+H]+(97% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.29(s(br), 1H), 9.84(s(br), 1H), 7.53(d,J=1.7Hz,1H), 7.46(s,1H), 7.20(d,J=2. 7Hz,1H), 7.12(dd,J=8.6,2.7Hz,1H), 6.95(d,J=1.7Hz,1H), 6.80(d,J=8.6Hz,1H), 3.62(s,2H), 1.34(s,9H).

[0231] Example 1.2 N-tert-butyl-4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophene-2-carboxamide TIFF0007862174000059.tif27170

[0232] Step 1: Methyl 4-[[2-(5-chloro-2-methoxyphenyl)acetyl]aminothiophene-2-carboxylate TIFF0007862174000060.tif321702-(5-chloro-2-methoxyphenyl)acetic acid (0.16 g, 0.82 mmol) was dissolved in thionyl chloride (0.73 mL, 8.24 mmol), and the mixture was heated at 70°C for 1 hour. The resulting mixture was concentrated under vacuum, and the residue was azeotropically mixed with toluene (2 × 2 mL). The crude acid chloride was dissolved in DCM (1 mL) and added to a chilled (0°C) solution of methyl 4-aminothiophene-2-carboxylate (125 mg, 0.80 mmol) and DIPEA (0.28 mL, 1.59 mmol) in DCM (2 mL). The resulting mixture was stirred and warmed to room temperature. After 1 hour, the mixture was diluted with DCM (4 mL) and water (5 mL), the phases were separated, and the aqueous portion was extracted with DCM (5 mL). The combined organic extract was concentrated under vacuum to obtain a light brown solid. The product was purified by chromatography on silica eluted with 0-80% dimethyl ammonium in heptane, yielding the marked compound as a white, waxy solid. LC-MS (Method C): Rt 1.16 min; MS m / z 339.9 / 341.9=[M+H]+(100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ10.57(s,1H), 7.79(d,J=1.6Hz,1H), 7.78(d,J=1.6Hz,1 H), 7.31-7.27(m, 2H), 7.02-6.98(m, 1H), 3.81(s,3H), 3.76(s,3H), 3.62(s,2H).

[0233] Steps 2-4: N-tert-butyl-4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophen-2-carboxamide The title compound was prepared from methyl 4-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]thiophene-2-carboxylate (step 1) in the same manner as steps 2 to 4 of Example 1. LC-MS (Method A): Rt 3.09 min; MS m / z 367.2 / 369.1=[M+H]+(97% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ10.44(s,1H), 9.78(s,1H), 7.85(s,1H), 7.80(d,J=1.4Hz,1H), 7.49(d,J=1.4Hz, 1H), 7.19(d,J=2.6Hz,1H), 7.10(dd,J=8.6,2.7Hz,1H), 6.79(d,J=8.6Hz,1H), 3.56(s,2H), 1.35(s,9H).

[0234] Example 2 N-[(5-chloro-2-hydroxyphenyl)methyl]-3-(2,2-dimethylpropanoylamide)benzamide TIFF0007862174000061.tif27170

[0235] Step 1: Methyl 3-(2,2-dimethylpropanoylamino)benzoate TIFF0007862174000062.tif26170 A chilled (0°C) mixture of methyl 3-aminobenzoate (500 mg, 3.31 mmol) and TEA (1.73 mL, 9.92 mmol) in DCM (5.5 mL) was mixed with 2,2-dimethylpropanoyl chloride (0.61 mL, 4.96 mmol), and the resulting mixture was stirred for 2 hours. The reaction mixture was diluted with DCM (5 mL), the organic portion was washed with saturated sodium bicarbonate aqueous solution (5 mL) and brine (5 mL), dried over Na2SO4, and concentrated under vacuum to obtain the marked compound as a light brown solid. LC-MS (Method C): Rt 1.08 min; MS m / z 236.1=[M+H]+ (99% at 215 nm) 1H NMR (250MHz, DMSO-d6) δ9.43(s,1H), 8.30(t,J=1.9Hz,1H), 7.95(ddd,J=8.1,2.2,1.1 Hz,1H), 7.63(dt,J=7.7,1.2Hz,1H), 7.43(t,J=7.9Hz,1H), 3.85(s,3H), 1.23(s,9H).

[0236] Step 2: 3-(2,2-dimethylpropanoylamino)benzoic acid TIFF0007862174000063.tif26170 A 2:1 solution of methyl 3-(2,2-dimethylpropanoylamino)benzoate (Step 1) (99%, 706 mg, 2.97 mmol) was added to 1 M LiOH (14.85 mL, 14.85 mmol) in THF / water (11 mL), and the mixture was stirred for 1 hour. The resulting mixture was diluted with water (20 mL) and acidified to pH 1 by adding 6 M HCl dropwise. The mixture was extracted with SiO2 (3 × 15 mL), the combined organic extract was washed with brine (30 mL), dried with Na2SO4, and concentrated in vacuum to obtain the title compound as a light brown solid. LC-MS (Method C): Rt 0.97 min; MS m / z 222.0=[M+H]+(100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.38(s,1H), 8.25(t,J=1.8Hz,1H), 7.91(ddd,J=8.1,2 .2,1.0Hz,1H), 7.61(dt,J=7.7,1.3Hz,1H), 7.40(t,J=7.9Hz,1H), 1.23(s,9H).

[0237] Step 3: (5-chloro-2-methoxyphenyl)methaneamine TIFF0007862174000064.tif2717055555555555555655566555556 LC-MS (Method B): Rt 2.43 min; MS m / z 171.7 / 173.7=[M+H]+(89% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ7.38(d,J=2.7Hz,1H), 7.21(dd,J=8.7,2.7Hz,1H), 6.94(d,J=8.7Hz,1H), 3.77(s,3H), 3.64(s,2H), 1.89(s(br), 2H).

[0238] Step 4: N-[(5-chloro-2-methoxyphenyl)methyl]-3-(2,2-dimethylpropanoylamide)benzamide TIFF0007862174000065.tif351701,4-Dioxane (3 mL) was used to a solution of 3-(2,2-dimethylpropanoylamino)benzoic acid (Step 2) (100 mg, 0.45 mmol) and (5-chloro-2-methoxyphenyl)methaneamine (Step 3) (89%, 95.9 mg, 0.50 mmol). TEA (0.20 mL, 1.13 mmol) and a 50% T3P® solution in ethyl acetate (0.27 mL, 0.45 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (3 × 5 mL). The combined organic extract was washed with saturated sodium bicarbonate aqueous solution (10 mL) and brine (10 mL), dried over Na₂SO₄, and concentrated under vacuum to obtain the title compound as a yellow oil. LC-MS (Method C): Rt 1.18 min; MS m / z 375.1 / 377.0=[M+H]+(95% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.37(s,1H), 8.86(t,J=5.9Hz,1H), 8.11(t,J=1.8Hz,1H), 7.88-7.85(m, 1H), 7.60-7.56(m, 1H), 7.40(t,J=7 .9Hz,1H), 7.28(dd,J=8.7,2.7Hz,1H), 7.13(d,J=2.7Hz,1H), 7.03(d,J=8.8Hz,1H), 4.41(d,J=5.9Hz,2H), 3.83(s,3H), 1.23(s,9H).

[0239] Step 5: N-[(5-chloro-2-hydroxyphenyl)methyl]-3-(2,2-dimethylpropanoylamide)benzamide In 3.23 mL of cooled (0°C) DCM, a solution of N-[(5-chloro-2-methoxyphenyl)methyl]-3-(2,2-dimethylpropanoylamide)benzamide (step 4) (95%, 150 mg, 0.38 mmol) was added dropwise to 1.14 mL of DCM (1.14 mmol). The mixture was heated to room temperature with stirring for 100 minutes. The reaction product was quenched with 5 mL of saturated sodium bicarbonate aqueous solution, and the mixture was extracted in RINKAN (3 × 5 mL). The combined organic extract was dried over Na₂SO₄ and concentrated under vacuum. The crude residue was purified by preparative HPLC (acidic pH, early elution method) to obtain the title compound as a white solid. LC-MS (Method A): Rt 3.20 min; MS m / z 361.1 / 363.2=[M+H]+(98% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.91(s,1H), 9.37(s,1H), 8.90(t,J=5.9Hz,1H), 8.11(t,J=1.8Hz,1H), 7.88-7.84(m, 1H), 7.57(d,J=7.9Hz,1H) ), 7.39(t,J=7.9Hz,1H), 7.11(dd,J=8.5,2.7Hz,1H), 7.08(d,J=2.6Hz,1H), 6.83(d,J=8.5Hz,1H), 4.38(d,J=5.9Hz,2H), 1.23(s,9H).

[0240] Example 3 N-[(5-chloro-2-hydroxyphenyl)methyl]-4-(2,2-dimethylpropanoylamide)pyridine-2-carboxamide TIFF0007862174000066.tif26170

[0241] Step 1: 4-amino-N-[(5-chloro-2-methoxyphenyl)methyl]pyridine-2-carboxamide A mixture of (5-chloro-2-methoxyphenyl)methaneamine (0.34 mL, 2.72 mmol) and 4-aminopyridine-2-carboxylic acid (250 mg, 1.81 mmol) in DMF (5 mL) was treated with DIPEA (0.63 mL, 3.62 mmol), followed by HATU (1.03 g, 2.72 mmol), and stirred at room temperature for 16 hours. The resulting mixture was diluted with HCl (25 mL) and water (25 mL), and the phases were separated. The organic portion was washed with water (2 × 25 mL) and brine (25 mL), dried over Na₂SO₄, and concentrated under vacuum. The crude residue was purified by chromatography on silica eluted with 0-100% HCl in heptane (Biotage® SNAP KP-NH), yielding the marked compound as a yellow oil. LC-MS (Method C): Rt 0.89 min; MS m / z 292.0 / 294.0=[M+H]+(89% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ8.95(t,J=6.4Hz,1H), 8.04(d,J=5.5Hz,1H), 7.27(dd,J=8.7,2.7Hz,1H), 7.22(d,J=2.3Hz,1H), 7 .07(d,J=2.7Hz,1H), 7.02(d,J=8.8Hz,1H), 6.61(dd,J=5.6,2.4Hz,1H), 6.33(s,2H), 4.39(d,J=6.4Hz,2H), 3.83(s,3H).

[0242] Step 2: N-[(5-chloro-2-methoxyphenyl)methyl]-4-(2,2-dimethylpropanoylamide)pyridine-2-carboxamide A chilled (0°C) mixture of 4-amino-N-[(5-chloro-2-methoxyphenyl)methyl]pyridine-2-carboxamide (Step 1) (89%, 58 mg, 0.18 mmol) and DIPEA (47 μL, 0.27 mmol) in DCM (4 mL) was treated with a solution of pivaloyl chloride (26 μL, 0.21 mmol) in DCM (1 mL). The mixture was warmed to room temperature and stirred for 24 hours. After adding an additional 47 μL, 0.27 mmol of DIPEA, a solution of pivaloyl chloride (26 μL, 0.21 mmol) in DCM (1 mL) was added. The resulting mixture was stirred for a further 6 hours and then allowed to stand at room temperature for 2 days. The mixture was concentrated under vacuum, and the crude residue was purified by chromatography on silica eluted with 0-80% dimethyl heptane, yielding the indicated compound as a yellow film. LC-MS (Method C): Rt 1.23 min; MS m / z 376.1,378.1=[M+H]+(95% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.80(s,1H), 9.14(t,J=6.4Hz,1H), 8.51(d,J=5.5Hz,1H), 8.34(d,J=2.1Hz,1H), 8.01(dd,J=5.6,2.2Hz ,1H), 7.28(dd,J=8.7,2.7Hz,1H), 7.09(d,J=2.7Hz,1H), 7.03(d,J=8.8Hz,1H), 4.44(d,J=6.3Hz,2H), 3.84(s,3H), 1.24(s,9H).

[0243] Step 3: N-[(5-chloro-2-hydroxyphenyl)methyl]-4-(2,2-dimethylpropanoylamide)pyridine-2-carboxamide The title compound was prepared in the same manner as in step 4 of Example 1, from N-[(5-chloro-2-methoxyphenyl)methyl]-4-(2,2-dimethylpropanoylamino)pyridine-2-carboxamide (step 2) and 1M BBr3 in DCM. LC-MS (Method A): Rt 3.28 min; MS m / z 362.2,364.2=[M+H]+(100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.99(s,1H), 9.81(s,1H), 9.14(t,J=6.4Hz,1H), 8.50(d,J=5.6Hz,1H), 8.34(d,J=2.1Hz,1H), 8.01(dd, J=5.6,2.2Hz,1H), 7.11(dd,J=8.5,2.7Hz,1H), 7.07(d,J=2.7Hz,1H), 6.83(d,J=8.5Hz,1H), 4.42(d,J=6.3Hz,2H), 1.24(s,9H).

[0244] Example 4 N2-tert-butyl-N4-[(2-hydroxyphenyl)methyl]pyridine-2,4-dicarboxamide TIFF0007862174000069.tif28170

[0245] Step 1: Methyl 4-[(2-methoxyphenyl)methylcarbamoyl]pyridine-2-carboxylate The title compound was prepared from 2-methoxycarbonylpyridine-4-carboxylic acid and (2-methoxyphenyl)methaneamine in the same manner as in step 1 of Example 3. LC-MS (Method C): Rt 1.01 min; MS m / z 301.1=[M+H]+ (97% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.34(t,J=5.7Hz,1H), 8.87(dd,J=4.9,0.7Hz,1H), 8.48(dd,J=1.7,0.8Hz,1H), 8.06(dd,J=5.0,1.7Hz,1H), 7.26(td,J=8.2 ,1.7Hz,1H), 7.21(dd,J=7.5,1.5Hz,1H), 7.01(dd,J=8.2,0.7Hz,1H), 6.9 1(td,J=7.4,1.0Hz,1H), 4.48(d,J=5.8Hz,2H), 3.92(s,3H), 3.83(s,3H).

[0246] Step 2: 4-[(2-methoxyphenyl)methylcarbamoyl]pyridine-2-carboxylic acid LiOH (0.85 mL, 0.85 mmol) of TIFF0007862174000071.tif321701M was added to a mixture of methyl 4-[(2-methoxyphenyl)methylcarbamoyl]pyridine-2-carboxylate (Step 1) (97%, 220 mg, 0.71 mmol) in THF (2 mL) and water (1 mL). After stirring at room temperature for 1 hour, the mixture was acidified to pH 1 with 1 M HCl and diluted with SiO (15 ml) and water (15 ml). The phases were separated, the organic layer was washed with brine (15 ml), dried over Na2SO4, and concentrated under vacuum to obtain the title compound as a beige foamy solid. LC-MS (Method C): Rt 0.89 min; MS m / z 287.0=[M+H]+ (98% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.32(t,J=5.8Hz,1H), 8.85(d,J=4.9Hz,1H), 8.50-8.45(m, 1H), 8.02(dd,J=5.0,1.7Hz,1H), 7.2 8-7.23(m, 1H), 7.22-7.19(m, 1H), 7.01(d,J=7.8Hz,1H), 6.91(td,J=7.4,0.9Hz,1H), 4.47(d,J=5.8Hz,2H), 3.83(s,3H).

[0247] Step 3: N2-tert-butyl-N4-[(2-methoxyphenyl)methyl]pyridine-2,4-dicarboxamide TIFF0007862174000072.tif34170TEA (0.18 mL, 1.03 mmol) was added to a mixture of 4-[(2-methoxyphenyl)methylcarbamoyl]pyridine-2-carboxylic acid (step 2) (98%, 100 mg, 0.34 mmol) and TBTU (131.9 mg, 0.41 mmol) in DMF (2 mL). The resulting mixture was stirred for 5 minutes, then treated with 2-methylpropan-2-amine (0.04 mL, 0.41 mmol), and stirred at room temperature for 1 hour. The resulting mixture was diluted with SiO2 (20 ml) and water (20 ml), and the phases were separated. The organic portion was washed with water (2 × 20 ml) and brine (20 ml), dried over Na2SO4, and concentrated under vacuum to obtain a yellow oily substance. When the crude product was purified by chromatography on silica eluted at 0-60% methoxy in heptane, the labeled compound was obtained as a concentrated yellow rubber. LC-MS (Method C): Rt 1.18 min; MS m / z 342.1=[M+H]+ (100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.33(t,J=5.7Hz,1H), 8.76(d,J=4.9Hz,1H), 8.49-8.44(m, 1H), 8.08(s,1H), 7.98(dd,J=5.0,1.7Hz,1H), 7.26(dd,J=7 .6,1.6Hz,1H), 7.20(dd,J=7.5,1.3Hz,1H), 7.01(d,J=7.8Hz,1H), 6.91(dd,J=7.4,0.7Hz,1H), 4.47(d,J=5.8Hz,2H), 3.83(s,3H), 1.43(s,9H).

[0248] Step 4: N2-tert-butyl-N4-[(2-hydroxyphenyl)methyl]pyridine-2,4-dicarboxamide The title compound was prepared in the same manner as in step 4 of Example 1, from N2-tert-butyl-N4-[(2-methoxyphenyl)methyl]pyridine-2,4-dicarboxamide (step 3) and 1M BBr3 in DCM. LC-MS (Method A): Rt 2.94 min; MS m / z 328.1=[M+H]+ (100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ9.56(s,1H), 9.35(t,J=5.8Hz,1H), 8.76(dd,J=5.0, 0.7Hz,1H), 8.46(dd,J=1.7,0.7Hz,1H), 8.08(s,1H), 7.98(dd,J=5.0,1.8Hz, 1H), 7.14(dd,J=7.5,1.4Hz,1H), 7.08(td,J=7.9,1.7Hz,1H), 6.82(dd,J=8.0 ,1.0Hz,1H), 6.76(td,J=7.4,1.1Hz,1H), 4.44(d,J=5.8Hz,2H), 1.42(s,9H).

[0249] Example 5 N1-tert-butyl-N3-[(2-hydroxyphenyl)methyl]benzene-1,3-dicarboxamide TIFF0007862174000073.tif26170

[0250] Step 1: Methyl 3-(tert-butylcarbamoyl)benzoate The title compound was prepared from 3-methoxycarbonylbenzoic acid and 2-methylpropan-2-amine in the same manner as in step 3 of Example 1. LC-MS (Method C): Rt 1.09 min; MS m / z 236.1=[M+H]+ (64% at 215 nm) ¹H NMR (500 MHz, chloroform-d): δ 8.30 (t, J=1.7 Hz, 1H), 8.13 (dt, J=7.8, 1.3 Hz, 1H), 7.99 (dt, J=7.8, 1.4 Hz, 1H), 7.51 (t, J=7.8 Hz, 1H), 6.01 (s, 1H), 3.94 (s, 3H), 1.48 (s, 9H).

[0251] Step 2: 3-(tert-butylcarbamoyl)benzoic acid TIFF0007862174000075.tif241701,4-Dioxane (3 mL) was used to a solution of methyl 3-(tert-butylcarbamoyl)benzoate (Step 1) (219 mg, 0.93 mmol). 2 M LiOH (0.93 mL, 1.86 mmol) was added, and the mixture was shaken at room temperature for 80 minutes. The solvent was removed under vacuum, and the residue was acidified to pH 1 with 1 M HCl (2 mL). The mixture was extracted with DCM (2 × 5 mL), and the organic extract was passed through a hydrophobic frit. The filtrate was concentrated under vacuum to obtain the title compound as a white solid. LC-MS (Method C): Rt 0.93 min; MS m / z 222.2=[M+H]+ (97% at 215 nm) 1H NMR (500MHz, methylene chloride-d2) δ 8.33 (t, J = 1.6 Hz, 1 H), 8.06-7.96 (m, 3 H), 7.55 (t, J = 7.7 Hz, 1 H), 1.39 (s, 9 H).

[0252] Step 3: N1-tert-butyl-N3-[(2-hydroxyphenyl)methyl]benzene-1,3-dicarboxamide In a solution of 3-(tert-butylcarbamoyl)benzoic acid (step 2) (70 mg, 0.32 mmol) and 2-(aminomethyl)phenol (0.13 mL, 0.35 mmol) in DCM (5 mL), DIPEA (0.06 mL, 0.32 mmol), followed by HATU (120.3 mg, 0.32 mmol), was added, and the mixture was shaken overnight at room temperature. The resulting mixture was washed with water (3 mL) and passed through hydrophobic frit. The filtrate was concentrated under vacuum to obtain the title compound as a grayish-white solid. LC-MS (Method A): Rt 2.79 min; MS m / z 327.2=[M+H]+ (95% at 215 nm) 1H NMR (500MHz, chloroform-d) δ8.10(d,J=1.6Hz,1H), 7.88(d,J=7.8Hz,1H), 7.83(d,J=7.8Hz,1H), 7.46(t,J=7.8Hz,1H), 7.23(td,J=8.1,1. 7Hz,1H), 7.20-7.13(m, 2H), 6.97(dd,J=8.1,1.0Hz,1H), 6.85(td,J=7.4,1.1Hz,1H), 6.06(s,1H), 4.56(d,J=6.5Hz,2H), 1.47(s,9H).

[0253] Example 6 N-[3-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminophenyl]phenyl]-2,2-dimethyl-propanamide TIFF0007862174000076.tif31170

[0254] Step 1: N-(3-aminophenyl)-2,2-dimethylpropanamide A chilled (0°C) solution of benzene-1,3-diamine (1.0 g, 9.25 mmol) in DCM (5 mL) was treated with DIPEA (4.85 mL, 27.74 mmol) and 2,2-dimethylpropanoyl-2,2-dimethylpropanoate (2.06 mL, 10.17 mmol), and stirred at 0°C for 2 hours. The resulting mixture was warmed to room temperature and stirred for 20 hours. The mixture was diluted with water (12.5 mL), saturated NaHCO3 (25 mL), and DCM (30 mL) and passed through hydrophobic frit. The organic portion was further washed with saturated brine (25 mL) and recovered using hydrophobic frit. The crude product was purified by chromatography on silica eluted with 25-100% siRNA in heptane, yielding the marked compound as a pale reddish-brown solid. LC-MS (Method C): Rt 0.69 min; MS m / z 193=[M+H]+(100% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ8.84(s,1H), 6.93(t,J=2.1Hz,1H), 6.88(t,J=7.9Hz,1H), 6.71( ddd,J=8.0,1.9,0.9Hz,1H), 6.24(ddd,J=7.9,2.2,0.9Hz,1H), 4.97(s,2H), 1.19(s,9H).

[0255] Step 2: N-[3-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]phenyl]-2,2-dimethyl-propanamide A solution of N-(3-aminophenyl)-2,2-dimethyl-propanamide (Step 1) (48 mg, 0.25 mmol) and 5-chloro-3H-benzofuran-2-one (prepared according to the literature procedure in Journal of Fluorine Chemistry 99 (1999) 189-195) (40 mg, 0.24 mmol) was stirred in toluene (2 mL) at 120 °C for 2 hours. The resulting mixture was concentrated under vacuum, and the residue was purified by preparative HPLC (acidic pH, early elution method) to obtain the indicated compound as a grayish-white powder. LC-MS (Method A): Rt 3.24 min; MS m / z 361 / 363=[M+H]+ (98% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ10.04(s,1H), 9.80(s,1H), 9.18(s,1H), 7.97(t,J=1.9Hz,1H), 7.30-7.24(m, 2H), 7.21-7.14(m, 2H), 7.10(dd,J=8.6,2.7Hz,1H), 6.80(d,J=8.6Hz,1H), 3.59(s,2H), 1.21(s,9H).

[0256] Example 7 N-[4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]-2-pyridyl]-2,2-dimethylpropanamide TIFF0007862174000078.tif26170

[0257] Step 1: 2-(5-chloro-2-methoxyphenyl)-N-(2-chloro-4-pyridyl)acetamide The title compound was prepared from 2-(5-chloro-2-methoxyphenyl)acetic acid and 2-chloropyridine-4-amine in the same manner as in Example 1.2, Step 1. LC-MS (Method C): Rt 1.12 min; MS m / z 311.1=[M+H]+ (100% at 215 nm) ¹H NMR (500 MHz, chloroform-d): δ 8.23 ​​(d, J=5.6 Hz, ¹H), 7.91 (s, ¹H), 7.49 (d, J=1.7 Hz, ¹H), 7.32-7.27 (m, ³H), 6.91 (d, J=8.7 Hz, ¹H), 3.95 (s, ³H), 3.69 (s, ²H).

[0258] Step 2: N-[4-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]-2-pyridyl]-2,2-dimethyl-propanamide TIFF0007862174000080.tif341701,4-Dioxane (1 mL) and DMF (0.2 mL) were used to purify a suspension of 2-(5-chloro-2-methoxyphenyl)-N-(2-chloro-4-pyridyl)acetamide (Step 1) (100%, 50 mg, 0.16 mmol), 2,2-dimethylpropanamide (33 mg, 0.32 mmol), and potassium carbonate (44 mg, 0.32 mmol). The suspension was degassed with N2 for 5 minutes. XantPhos Pd-G3 (15 mg, 0.02 mmol) was added, and the sealed tube was heated to 120°C for 45 minutes using microwave radiation, followed by heating to 130°C for 30 minutes. The resulting mixture was concentrated under vacuum, and the crude residue was purified by chromatography on silica eluted with phenyl in heptane, yielding the marked compound as colorless glass. LC-MS (Method C): Rt 1.06 min; MS m / z 376.0=[M+H]+(54% at 215 nm) ¹H NMR (500 MHz, chloroform-d): δ 8.16 (d, J=6.1 Hz, 1H), 7.90 (t, J=6.4 Hz, 2H), 7.29-7.27 (m, 1H), 7.24 (d, J=2.5 Hz, 1H), 6.89 (d, J=8.8 Hz, 1H), 3.95 (s, 3H), 3.81 (s, 2H), 1.39 (s, 9H).

[0259] Step 3: N-[4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]-2-pyridyl]-2,2-dimethylpropanamide The title compound was prepared in the same manner as in step 4 of Example 1, from N-[4-[[2-(5-chloro-2-methoxyphenyl)acetyl]amino]-2-pyridyl]-(2,2-dimethylpropanamide (step 2) and 1M BBr3 in DCM. LC-MS (Method A): Rt 2.17 min; MS m / z 362.1=[M+H]+(100% at 215 nm) 1H NMR (500MHz, methanol-d4) δ8.25(d,J=1.8Hz,1H), 8.15(d,J=6.3Hz,1H), 7.50(dd,J=6.3,2.0Hz,1H), 7.18(d,J=2.6Hz,1H), 7.09(dd,J=8.6,2.6Hz,1H), 6.78(d,J=8.6Hz,1H), 3.73(s,2H), 1.32(s,9H).

[0260] Example 7.1 N-[4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]-2-pyridyl]cyclohexanecarboxamide The title compound was prepared from 2-(5-chloro-2-methoxyphenyl)-N-(2-chloro-4-pyridyl)acetamide (step 1 of Example 7) and cyclohexanecarboxamide, in the same manner as steps 2 and 3 of Example 7. LC-MS (Method A): Rt 2.51 min; MS m / z 388.3=[M+H]+ (99% at 215 nm) 1H NMR (500MHz, methanol-d4) δ 8.14 (d, J=6.7Hz, 1H), 8.11 (d, J=2.0Hz, 1H), 7.43 (dd, J=6.7, 2.0Hz, 1H), 7.18 (d, J=2.6Hz, 1H), 7.10 (dd, J=8.6, 2.6Hz, 1H), 6.78 (d, J=8.6Hz, 1H), 3.75(s,2H), 2.47(tt,J=11.6,3.5Hz,1H), 1.97-1.89(m, 2H), 1.85(dt,J=12.5, 3.1Hz,2H), 1.73(d,J=12.5Hz,1H), 1.53(qd,J=12.5,3.1Hz,2H), 1.43-1.22(m, 3H).

[0261] Example 8 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]amino]-N-(1-cyano-1-methyl-ethyl)thiophene-2-carboxamide TIFF0007862174000082.tif35170

[0262] Step 1: 2-(4-tert-butyl-2-fluoro-5-methoxyphenyl)acetic acid A solution of 2-(2-fluoro-5-methoxyphenyl)acetic acid (5.0 g, 27.15 mmol) in DCE (181.01 mL) was treated with tert-butanol (31.16 mL, 325.8 mmol) and concentrated sulfuric acid (17.37 mL, 325.8 mmol). After stirring for 1 hour, further tert-butanol (10.0 mL, 105 mmol) and concentrated sulfuric acid (5.8 mL, 109 mmol) were added, and the mixture was stirred overnight. The resulting mixture was diluted with water (150 mL), and the phases were separated. The aqueous layer was extracted with DCM (3 × 150 mL). The combined organic extracts were washed with brine, dried over Na₂SO₄, and concentrated under vacuum. The residue was diluted with MeOH (100 mL), treated with 2 M HCl (100 mL, freshly prepared from thionyl chloride) in MeOH, and the mixture was heated under reflux for 40 minutes. The resulting mixture was concentrated under vacuum, the residue was dissolved in DCM (150 mL), and washed with saturated sodium bicarbonate aqueous solution (150 mL). The aqueous washing solution was further extracted with DCM (3 × 100 mL), the combined organic extract was dried over Na2SO4, and concentrated under vacuum. Purification by chromatography on silica eluted with 0-10% HCl in heptane yielded a methyl ester intermediate. The material was dissolved in 1 M LiOH (80 mL) and THF (80 mL) and stirred for 1 hour. Volatile substances were removed under vacuum, and the aqueous solution was acidified with HCl to obtain a solid precipitate. The solid was washed with excess water and dried to obtain the title compound as a pale yellow solid. LC-MS (Method C): Rt 1.16 min; no ionization (98% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ6.93(d,J=6.1Hz,1H), 6.92(s,1H), 3.77(s,3H), 3.54(s,2H), 1.31(s,9H).

[0263] Step 2: 2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetic acid A solution of 2-(4-tert-butyl-2-fluoro-5-methoxyphenyl)acetic acid (Step 1) (600 mg, 2.5 mmol) in cooled (0°C) DCM (12 mL) was treated with 1 M BBr3 (7.49 mL, 7.49 mmol) in DCM, and the mixture was warmed to room temperature and stirred for 3 days. The resulting mixture was diluted with water (20 mL), stirred for 5 minutes, and then diluted with DCM (30 mL) and water (20 mL). The phases were separated, and the aqueous phase was further extracted with DCM (30 mL). The combined organic extracts were dried over Na2SO4 and concentrated in vacuum to obtain a brown oil. The oil was dissolved in THF (5 mL) and treated with 2 M LiOH aqueous solution (2 mL) while stirring at room temperature for 20 minutes. The resulting mixture was acidified to pH 1-2 by the dropwise addition of 1 M HCl, and then diluted with HCl (40 mL) and water (30 mL). The phases were separated, the organic portion was washed with brine (30 mL), dried over Na₂SO₄, and concentrated under vacuum to obtain the title compound as a brown oily substance. LC-MS (Method C): Rt 1.07 min; no ionization (97% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ12.30(s(br), 1H), 9.27(s,1H), 6.85(d,J=11.9Hz,1H), 6.67(d,J=7.0Hz,1H), 3.45(s,2H), 1.32(s,9H).

[0264] Step 3: Methyl 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]aminothiophene-2-carboxylate In TIFF0007862174000085.tif34170DMF (4 mL), a solution of methyl 5-aminothiophene-2-carboxylate (219 mg, 1.39 mmol), 2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetic acid (Step 2) (75%, 350 mg, 1.16 mmol), and DIPEA (0.26 mL, 1.51 mmol) was added, to which HATU (485 mg, 1.28 mmol) was added, and the mixture was stirred at room temperature for 4 hours. The resulting mixture was diluted with SiO2 (30 mL) and water (30 mL), and the phases were separated. The organic portion was washed with water (30 mL) and brine (30 mL), dried over Na2SO4, and 真空中で The compound was concentrated. Purification of the crude substance by chromatography using silica eluted at 0-100% methoxy in heptane yielded the indicated compound as an orange solid. LC-MS (Method C): Rt 1.21 min; MS m / z 366.1=[M+H]+ (84% at 215 nm) 1H NMR (400MHz, DMSO-d6) δ11.87(s,1H), 9.33(s,1H), 7.60(d,J=4.2Hz,1H), 6.8 8(d,J=11.9Hz,1H), 6.74-6.69(m, 2H), 3.76(s,3H), 3.65(s,2H), 1.32(s,9H).

[0265] Step 4: 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]aminothiophene-2-carboxylic acid TIFF0007862174000086.tif341702M LiOH (0.72 mL, 1.44 mmol) was added to a solution of methyl 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]aminothiophene-2-carboxylate (step 3) (84%, 314 mg, 0.72 mmol) in THF (2 mL) and water (2 mL), and the mixture was heated overnight at 50°C. The resulting mixture was concentrated under vacuum to remove most of the THF, and then acidified to pH 1-2 by adding 1 M HCl dropwise. The mixture was partitioned into SiO2 (15 mL) and water (15 mL) to separate the phases. The organic portion was washed with brine (15 mL), dried over Na2SO4, and concentrated under vacuum to obtain the title compound as an orange oil. LC-MS (Method C): Rt 1.11 min; MS m / z 352.0=[M+H]+ (98% at 215 nm) 1H NMR (400MHz, DMSO-d6) δ11.78(s,1H), 9.33(s,1H), 7.52(d,J=4.1Hz,1H), 6.87(d,J= 11.9Hz,1H), 6.73(d,J=7.0Hz,1H), 6.70(d,J=4.2Hz,1H), 3.64(s,2H), 1.32(s,9H).

[0266] Step 5: 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]amino]-N-(1-cyano-1-methyl-ethyl)thiophene-2-carboxamide HATU (133 mg, 0.35 mmol) was added to a solution of 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]aminothiophene-2-carboxylic acid (step 4) (70%, 175 mg, 0.35 mmol), 2-amino-2-methyl-propanenitrile hydrochloride (63 mg, 0.52 mmol), and DIPEA (0.15 mL, 0.87 mmol) in DMF (3 mL), and the mixture was stirred overnight at room temperature. The resulting mixture was partitioned into SiO2 (20 mL) and water (20 mL) to separate the phases. The organic portion was washed with brine (15 mL), dried over Na2SO4, and concentrated under vacuum. The crude substance was purified by preparative HPLC (acidic pH, standard elution method), yielding the marked compound as a grayish-white solid. LC-MS (Method A): Rt 3.30 min; MS m / z 418.2=[M+H]+ (99% at 215 nm) 1H NMR (500MHz, DMSO-d6) δ11.69(s,1H), 9.33(s,1H), 8.49(s,1H), 7.65(d,J=4.2Hz,1H), 6.87(d,J= 11.9Hz,1H), 6.74(d,J=6.9Hz,1H), 6.68(d,J=4.2Hz,1H), 3.64(s,2H), 1.66(s,6H), 1.32(s,9H).

[0267] Example 9 5-[[2-[2-fluoro-5-hydroxy-4-(2-hydroxy-1,1-dimethyl-ethyl)phenyl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide TIFF0007862174000087.tif36170 Intermediate A: 2-(5-fluoro-3,3-dimethyl-2-oxo-benzofuran-6-yl)acetic acid

[0268] Process A1: 2-(4-bromo-2-fluoro-5-methoxyphenyl)acetic acid TIFF0007862174000088.tif41170 To a solution of 2-(2-fluoro-5-methoxyphenyl)acetic acid (45 g, 244.4 mmol) in cooled (0°C) MeCN (1.2 L), a solution of bromine (12.63 mL, 219.9 mmol) in MeCN (100 mL) was added dropwise over 10 minutes. The resulting mixture was gradually warmed to room temperature without removing the ice bath (approximately 1.5 hours). Further bromine (4.21 mL, 73.3 mmol) in MeCN (50 mL) was added dropwise at 0°C, and the mixture was stirred at room temperature for a further 3.5 hours. Further bromine (4.21 mL, 73.3 mmol) in MeCN (50 mL) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The reaction was carefully quenched with saturated sodium sulfite aqueous solution (approximately 700 mL) until the bright orange color disappeared. The colorless solution was diluted with brine (200 mL) and SiO (200 mL) and stirred vigorously for 10 minutes. The organic layer was separated, and the aqueous layer was further extracted with SiO (200 mL). The organic layers were combined, dried over Na₂SO₄, and concentrated in vacuum to obtain the crude product as a white solid. The crude product was recrystallized by dissolving the solid in AcOH (700 mL) and then treated with water (4 L). When the mixture was stirred and the solvent was mixed, crystals gradually appeared. The mixture was maintained at room temperature for 1 hour, then at 0°C for 3 hours. After filtration and subsequent vacuum drying at 40°C, the marked compound was obtained as a fluffy white solid. LC-MS (Method C): Rt 1.07 min; (99% at 215 nm) 1 H NMR (500MHz, DMSO-d6) δ12.55(br s, 1H), 7.50(d,J=8.9Hz,1H), 7.13(d,J=6.6Hz,1H), 3.81(s,3H), 3.61(d,J=1.3Hz,2H).

[0269] Process A2: Benzyl 2-(4-bromo-2-fluoro-5-methoxyphenyl) acetate In 140 mL of DMF, a mixture of 2-(4-bromo-2-fluoro-5-methoxyphenyl)acetic acid (step A1) (15 g, 57.02 mmol) and K2CO3 (15.76 g, 114.0 mmol) was treated with benzyl bromide (7.45 mL, 62.7 mmol) and stirred at room temperature for 18 hours. The resulting mixture was filtered and concentrated under vacuum. The residue was dissolved in siRNA (300 mL) and sequentially washed with brine (200 mL) and saturated aqueous sodium bicarbonate (2 × 200 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. Purification by chromatography on silica eluted with 0-20% siRNA in heptane yielded the marked compound as a colorless solid. LC-MS (Method G): Rt 1.13 mins; (95% at 215 nm) 1 H NMR (500MHz, DMSO-d6) δ7.53 (d, J = 8.9 Hz, 1H), 7.40-7.31 (m, 5H), 7.15 (d, J = 6.6Hz, 1H), 5.14 (s, 2H), 3.81-3.78 (m, 5H).

[0270] Process A3: Methyl 2-[4-(2-benzyloxy-2-oxoethyl)-5-fluoro-2-methoxyphenyl]-2-methyl-propanoate TIFF0007862174000090.tif41170 Benzyl 2-(4-bromo-2-fluoro-5-methoxyphenyl) acetate (step A2) (5.0 g, 14.16 mmol), ZnF2 (1.1 g, 10.62 mmol), and Pd(PtBu3)2 (0.36 g, 0.71 mmol) were added to the reaction vessel and placed under a nitrogen atmosphere. A solution of (1-methoxy-2-methyl-propa-1-enoxy)-trimethyl-silane (5.75 mL, 28.31 mmol) was added to degassed DMF (50 mL), and the reaction mixture was heated at 80°C for 18 hours. The resulting mixture was filtered and concentrated under vacuum. The residue was dissolved in siRNA (100 mL), washed with brine (2 × 50 mL), and the organic layer was dried over Na2SO4 and concentrated under vacuum. The title compound was purified by chromatography on silica eluted with 0-30% toluene in heptane, yielding a pale yellow oily substance. LC-MS (Method A): Rt 3.99 min; MS m / z 375.3=[M+H]+ (92% at 215 nm) 1 H NMR (400MHz, DMSO-d6) δ7.40-7.30(m, 5H), 7.09(d,J=11.0Hz,1H), 6.98(d,J= 6.5Hz,1H), 5.14(s,2H), 3.76(s,2H), 3.66(s,3H), 3.53(s,3H), 1.40(s,6H).

[0271] Process A4: 2-(5-fluoro-3,3-dimethyl-2-oxobenzofuran-6-yl)acetic acid TIFF0007862174000091.tif40170 In DCM (34.13 mL, 34.1 mmol), 1 M BBr3 was added to a chilled (0°C) mixture of methyl 2-[4-(2-benzyloxy-2-oxo-ethyl)-5-fluoro-2-methoxyphenyl]-2-methyl-propanoate (Step A3) (92%, 2777 mg, 6.83 mmol) in dry DCM (60 mL). The resulting mixture was warmed to room temperature and stirred for 4.5 hours. The reaction mixture was cooled again to 0°C and water (50 mL) was added. Stirring was continued while gradually warming to room temperature over 30 minutes. The resulting mixture was diluted with DCM (80 mL) and water (80 mL) and the phases were separated. The aqueous portion was extracted with dimethylammonium phosphate (80 mL), and the combined organic extract was then dried with Na2SO4 and concentrated in vacuum to obtain the crude product as a brown oily substance. The crude product was purified by C18 reversed-phase chromatography using elution with 10-100% MeCN / water (+0.1% formic acid), and the labeled compound was obtained as a pale yellow solid. LC-MS (Method H): Rt 1.19 min; (98% at 215 nm) 1 H NMR (500MHz, DMSO-d6) δ12.56(br.s,1H), 7.42(d,J=8.9Hz,1H), 7.25(d,J=5.8Hz,1H), 3.64(d,J=1.5Hz,2H), 1.44(s,6H).

[0272] Intermediate B: 5-amino-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide Process B1: 5-Nitro-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide In TIFF0007862174000092.tif26170DMF (10 mL), a mixture of 5-nitrothiophene-2-carboxylic acid (400 mg, 2.31 mmol) and HATU (1054 mg, 2.77 mmol) was mixed with 1-(trifluoromethyl)cyclopropanamine hydrochloride (411 mg, 2.54 mmol), followed by DIPEA (0.97 mL, 5.54 mmol). The reaction mixture was stirred at room temperature for 1 hour. The resulting mixture was diluted with siRNA (40 mL) and water (40 mL), and the phases were separated. The organic portion was washed with water (40 mL) and brine (40 mL), and then dried over Na2SO4. 真空中で The compound was concentrated. Purification of the crude substance by chromatography using silica eluted at 0-100% dimethyl in heptane yielded the indicated compound as a grayish-white solid. LC-MS (Method C): Rt 1.12 min; MS m / z 280.9=[M+H]+(100% at 215 nm) 1H NMR (400MHz, DMSO-d6) δ9.66(s,1H), 8.14(d,J=4.4Hz,1H), 7.86(d,J=4.4Hz,1H), 1.38-1.29(m, 2H), 1.23-1.15(m, 2H).

[0273] Process B2: 5-amino-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide A mixture of 5-nitro-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide (step B1) (596 mg, 2.13 mmol) in EtOH (20 mL) under a nitrogen atmosphere was treated with carbon-supported Pd (10%, 225 mg, 0.21 mmol) and placed under a hydrogen atmosphere. After stirring at room temperature for 2 hours, the resulting mixture was filtered through Celite® (filter material) and washed with EtOH. The filtrate was concentrated under vacuum, and the crude product was purified by chromatography on KP-NH silica eluted with siRNA in heptane, yielding the marked compound as a yellow oil. LC-MS (Method G): Rt 0.67 min; MS m / z 251.1=[M+H]+ (97% at 215 nm) 1H NMR (400MHz, DMSO-d6) δ8.55(s,1H), 7.38(d,J=4.1Hz,1H), 6.34(s,2H), 5.81(d,J=4.1Hz,1H), 1.27-1.23(m, 2H), 1.10-1.05(m, 2H).

[0274] Final step: Intermediate A + Intermediate B: 5-[[2-[2-fluoro-5-hydroxy-4-(2-hydroxy-1,1-dimethyl-ethyl)phenyl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide In 3 mL of DMF, a solution of 5-amino-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide (intermediate B) (80%, 134 mg, 0.43 mmol) and 2-(5-fluoro-3,3-dimethyl-2-oxobenzofuran-6-yl)acetic acid (intermediate A) (98%, 114 mg, 0.47 mmol) was added. DIPEA (149 μL, 0.85 mmol), followed by a 50% T3P® solution in SiO (0.38 mL, 0.64 mmol), and the reaction mixture was stirred at room temperature for 30 minutes. The resulting mixture was diluted with SiO (15 mL) and water (15 mL), and the phases were separated. The organic portion was washed with water (15 mL) and brine (15 mL), dried over Na₂SO₄, and concentrated under vacuum to obtain the crude intermediate as a yellow / orange oily substance. The oil was dissolved in THF (3 mL) and cooled to -78°C. 4 M LiBH4 (107 μL, 0.43 mmol) was added dropwise to the THF, and the resulting mixture was heated to room temperature with stirring for 1 hour. The mixture was cooled to 0°C, and the reaction was quenched by adding 1 M HCl (5 mL) dropwise. The resulting mixture was diluted with HCl (20 mL) and water (15 mL), and the phases were separated. The aqueous portion was further extracted with HCl (20 mL), and the combined organic extract was dried over Na2SO4 and concentrated under vacuum. The crude substance was purified by preparative HPLC (acidic pH, early elution method) to obtain the title compound as a white solid. LC-MS (Method A): Rt 2.76 min; MS m / z 475.3=[M+H]+(100% at 215 nm) 1H NMR (400MHz, DMSO-d6) δ11.65(s,1H), 9.38(s,1H), 8.95(s,1H), 7.61(d,J=4.2Hz,1H), 6.89(d,J=12.1Hz,1H), 6.71( d,J=7.0Hz,1H), 6.65(d,J=4.2Hz,1H), 4.77(s,1H), 3.63(s,2H), 3.60(s,2H), 1.31-1.22(m, 8H), 1.15-1.08(m, 2H).

[0275] Biological examples Automated whole-cell patch-clamp assay for detecting TMEM16A activity in recombinant cells Cell culture and preparation Fisher rat thyroid (FRT) cells stably expressing human TMEM16A (TMEM16Aabc variant; Dr. Luis Galietta, Insituto Giannina, Italy) were cultured in T-75 flasks in Hams F-12 medium containing Coon Modified (Sigma) supplemented with 10% (v / v) fetal bovine serum, penicillin-streptomycin (10,000 U / mL / 10,000 μg / mL), G-418 (750 μg / mL), L-glutamine (2 mM), and sodium bicarbonate solution (7.5% v / v). Cells were harvested for experimentation by detachment using a 2:1 (v / v) mixture of Detachin (BMS Biotechnology) and 0.25% (w / v) trypsin-EDTA at approximately 90% confluence. Cells were cultured in a medium consisting of CHO-S-SFM II (Sigma), 25 mM HEPES (Sigma), and a soy trypsin inhibitor (Sigma) in a volume of 3.5–4.5 × 10⁴ cells. 6 The cells were diluted to a density of cells / mL.

[0276] Whole cell patch clamp records Whole-cell patch clamping of FRT-TMEM16A cells was performed using an automated planar patch clamp system (Qpatch, Sophion). Briefly, once a high-resistance (GOhm) seal was established between the cell and the planar recording array, the patch was ruptured using an aspiration pulse to establish the whole-cell recording configuration of the patch clamp technique. This assay used the following solutions (all reagents are Sigma): Intracellular solution (mM): N-methyl-D-glucamine 130, CaCl2 18.2, MgCl2 1, HEPES 10, EGTA 10, BAPTA 20, Mg-ATP 2, pH 7.25, sucrose 325 mOsm. Extracellular solution (mM): N-methyl-D-glucamine 130, CaCl22, MgCl21, HEPES 10, pH 7.3, sucrose 320 mOsm.

[0277] Intracellular solution has a maximum TMEM16A mediated current (EC for calcium ions). 20 Intracellular calcium was buffered to the level necessary to provide approximately 20% activation of the compound. Cells were voltage-clamped at a holding potential of -70mV, and a synthesis voltage step (up to +70mV) / ramp (-90mV to +90mV) was applied at 0.05Hz. After a period of current stabilization testing, the compound was solubilized in 100% (v / v) DMSO, followed by application of the diluted compound in extracellular solution to create a cumulative concentration response curve. After incubating each concentration of the test compound for 5 minutes, the next concentration was added. After testing the final concentration, the upper and lower limits of the assay were defined by adding either a known hypermaximal concentration of an active positive modulator or the TMEM16A inhibitor CaCCinhA01 (Del La Fuente et al., 2008).

[0278] Compound activity was quantified by measuring the increase in current upon compound addition and expressing this as the percentage increase in the baseline TMEM16A current level. The percentage increase in current was determined for each concentration, and the data was plotted as a function of concentration using either Qpatch software or Graphpad Prism v6.05, with the maximum effect at 50% (EC) 50) and concentrations that provide maximum efficacy (rate of increase from baseline) were obtained.

[0279] The method for calculating the results is shown in Figure 1, which illustrates the trace from the Qpatch TMEM16A assay. In Figure 1, I BL This is equal to the baseline current, I [#1] This is equal to the peak current during the incubation period of the test compound at a concentration of 1, and so on. The peak TMEM16A current at +70mV was plotted as a function of time over the assay period. Baseline current (I BL The current was measured after the stabilization period. The increase in current with each compound addition was determined by obtaining the peak current during the incubation period, subtracting the current from the previous recording period, and then expressing this as a percentage of the baseline current. For test compound concentration 1 in Figure 1: (I [#1] -I BL / I BL ) x 100 For each additional concentration tested, the increase in current was determined by subtracting the current from the previous incubation period and normalizing the baseline value - for test concentration 2 in Figure 1: (I [#2] -I [#1} / I BL ) x 100 The values ​​for each test concentration were plotted as a cumulative function of concentration. For example, for test concentration 2, this represents the sum of the peak changes measured between concentration 1 and concentration 2.

[0280] The results obtained for the compounds in the examples are shown in Table 1. From this table, it can be seen that the compounds of the present invention can significantly increase the TMEM16A current level.

[0281] Table 1 - Percentage enhancement and calculated EC values ​​shown by a 3.33 μM solution of the test compound. 50 value TIFF0007862174000094.tif92170

[0282] References Accurso FJ,Moss RB,Wilmott RW,Anbar RD,Schaberg AE,Durham TA,Ramsay BW;TIGER-1 Investigator Study Group(2011)Denufosol tetrasodium in patients with cystic fibrosis and normal to mildly impaired lung function.Am J Respir Crit Care Med,183(5):627-634. Boucher RC(2007)Evidence for airway surface dehydration as the initiating event in CF airway disease.J Intern Med.,261(1):5-16. Caputo A,Caci E,Ferrera L,Pedemonte N,Barsanti C,Sondo E,Pfeffer U,Ravazzolo R,Zegarra-Moran O&Galietta LJ(2008)TMEM16A,a membrane protein associated with calcium-dependent chloride channel activity.Science,322(5901):590-594. Del La Fuente R,Namkung W,Mills A&Verkman AS(2008)Small molecule screen identifies inhibitors of a human intestinal calcium-activated chloride channel.Mol Pharmacol,73(3):758-768. Kellerman D,Rossi Mospan A,Engels J,Schaberg A,Gorden J&Smiley L(2008)Denufosol:a review of studies with inhaled P2Y(2)agonists that led to Phase 2.Pulm Pharmacol Ther,21(4):600-607. Kunzelmann K&Mall M(2003)Pharmacotherapy of the ion transport defect in cystic fibrosis:role of purinergic receptor agonists and other potential therapeutics.Am J Respir Med,2(4):299-309. Matsui H,Grubb BR,Tarran R,Randell SH,Gatzy JT,Davis CW and Boucher RC(1998)Evidence for periciliary liquid layer depletion,not abnormal ion composition,in the pathogenesis of cystic fibrosis airways disease.Cell,95(7):1005-15. Moss RB(2013)Pitfalls of drug development:lessons learned from trials of denufosol in cystic fibrosis.J Pediatr,162(4):676-680. Pedemonte N&Galietta LJ(2014)Structure and function of TMEM16 proteins(anoctamins).Physiol Rev,94(2):419-459. Pezzulo AA,Tang XX,Hoegger MJ,Abou Alaiwa MH,Ramachandran S,Moninger TO,Karp PH,Wohlford-Lenan CL,Haagsman HP,van Eijk M,Banfi B,Horswill AR,Stoltz DA,McCray PB Jr,Welsh MJ&Zabner J(2012)redcued airway surface pH impairs bacterial killing in the porcine cystic fibrosis lung.Nature,487(7405):109-113. Yang YD,Cho H,Koo JY,Tak MH,Cho Y,Shim WS,Park SP,Lee J,Lee B,Kim BM,Raouf R,Shin YK&Oh U(2008)TMEM16 confers receptor-activated calcium-dependent chloride conductance.Nature,455(7217):1210-1215.

Claims

1. The salts of the general formulas (II), (III), or (III) [In the formula, A is And, X １ , X ２ , X ３ One of them is S, and X １ , X ２ and X ３ The other two of them are CH; This represents a single or double bond, such that ring A is aromatic; ＊１ indicates the connection point to Z １ and ＊２ indicates the connection point to Z ２ ; Z １ and Z ２ Each of them is independent, ＊ -C(O)NH- and ＊ Selected from -NHC(O)- ＊ indicates the bonding point to ring A; R １ , CN, C １－３ Alkyl, C ２－３ Alkenyl or C ２－３ They are alkynyl groups, and any of these alkyl, alkenyl, or alkynyl groups may be substituted with fluoropolymer one or more times; R ２ C １－６ It is alkyl; and, R ３ teeth, C １－６ Alkyl, C ２－６ Alkenyl or C ２－６ These C １－６ Alkyl, C ２－６ Alkenyl or C ２－６ Any of the alkynyl groups may be substituted with fluoropolymer one or more times; R 10 is H, OH, halo, C １－６ Alkyl, -O(C) １－６ It is alkyl; R １１ Each of these is independently H, Halogen, OH, CN, and C. １－６ Alkyl, C １－６ Haloalkyl, -O(C) １－６ Alkyl) or C(O)O-(C １－６ It is alkyl; n is either 1 or 2; R １１ａ H, Halo, C １－４ Alkyl, C １－４ Haloalkyl or C(O)O(C) １－４ It is alkyl; R １１ｂ H, Halo, C １－４ Alkyl or C １－４ It is a haloalkyl; and R １１ｃ H, Halo, CN, C １－４ Alkyl or C １－４ [It is a haloalkyl] A compound of [this].

2. General formula (Iai), (Ibi), or (Ici) Compounds of the general formula (Iaii), (Ibii), or (Icii) It is a compound of which, in the formula, R １１ａ , R １１ｂ and R １１ｃ The compound according to claim 1, wherein the compound is as defined in claim 1.

3. The aforementioned compound is of the general formula (Iai), (Ibi), or (Ici) It is a compound of, R １１ａ However, H, Haro, C １－４ Alkyl or C(O)O(C １－４ Alkyl) and R １１ｂ Is H; or R １１ａ and R １１ｂ Are both of them halo? It is either one of the following; or The aforementioned compound is of the general formula (Iaii), (Ibii), or (Icii) It is a compound of, R １１ａ is H; and R １１ｂ C １－４ C substituted with alkyl and OH groups １－４ Alkyl, or C １－４ It is a haloalkyl; and R １１ｃ is H, halo, methyl, or ethyl. The compound according to claim 2.

4. N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]aminothiophene-2-carboxamide (compound 1); N-tert-butyl-5-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]thiophene-3-carboxamide (compound 1.2); N-tert-butyl-4-[[2-(5-chloro-2-hydroxyphenyl)acetyl]amino]thiophene-2-carboxamide (compound 1.3); 5-[[2-(4-tert-butyl-2-fluoro-5-hydroxyphenyl)acetyl]amino]-N-(1-cyano-1-methyl-ethyl)thiophene-2-carboxamide (compound 8); 5-[[2-[2-fluoro-5-hydroxy-4-(2-hydroxy-1,1-dimethyl-ethyl)phenyl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]thiophene-2-carboxamide (compound 9); and its salts and solvates A compound selected from the following.

5. A compound according to any one of claims 1 to 4 for use in pharmaceuticals.

6. A compound according to any one of claims 1 to 4, for use in the treatment or prevention of diseases and symptoms affected by the modification of TMEM16A.

7. Use of the compound according to any one of claims 1 to 4 in the manufacture of a pharmaceutical product for the treatment or prevention of diseases and symptoms affected by the regulation of TMEM16A.

8. A pharmaceutical product for the treatment or prevention of diseases and symptoms affected by the regulation of TMEM16A, comprising an effective amount of the compound described in any one of claims 1 to 4.

9. The use according to claim 7, wherein the diseases and symptoms affected by the regulation of TMEM16A are selected from respiratory diseases and symptoms, dry mouth (xerostomia), hypermotility, cholestasis, and ocular symptoms.

10. The pharmaceutical product according to claim 8, wherein the diseases and symptoms affected by the regulation of TMEM16A are selected from respiratory diseases and symptoms, dry mouth (xerostomia), hypermotility of the intestines, cholestasis, and ocular symptoms.

11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4 and a pharmaceutically acceptable excipient.

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