Pyridine derivatives as calcium-activated chloride channel modulators

Abstract

compound Compounds of general formula (I): TIFF2022541638000121.tif48170 (in the formula, R 1 , R 2 , R 3 , R 4 , R 5 , X 1 , X 2 and X 3 (wherein TMEM16A is as defined herein) are useful in the treatment of respiratory diseases and other diseases and conditions modulated by TMEM16A.

Description

Technical Field

[0001] The present invention relates to novel compounds having activity as positive modulators of calcium-activated chloride channels (CaCC), TMEM16A. The present invention also relates to methods for preparing these compounds and pharmaceutical compositions containing them and to their use in the treatment of diseases and conditions in which TMEM16A plays a role, particularly respiratory diseases and conditions.

Background Art

[0002] Humans can inhale up to 12,000 L of air per day, and along with that, airborne pathogens (such as bacteria, viruses and fungal spores) may enter the airways. To protect against these airborne pathogens, the lungs have evolved innate defense mechanisms to minimize the potential for infection and colonization of the airways. One such mechanism is the mucociliary clearance system, by which secreted mucus is advanced through the airways and expelled from the airways by ciliary synchronous movement together with cough clearance. This continuous "washing" of the lungs removes inhaled particles and microorganisms, thereby reducing the risk of infection.

[0003] In recent years, it has become clear that water supply to the mucus gel is important for enabling mucociliary clearance (Boucher 2007; Matsui et al., 1998). In normal and healthy airways, the mucus gel typically consists of 97% water and 3 w / v% solids, and under these conditions mucus is removed by mucociliary action. Water supply to the airway mucosa is controlled by the coordinated activity of several ion channels and transporters. Anion (Cl - / HCO 3 - ) secretion mediated by cystic fibrosis transmembrane conductance regulator (CFTR) and calcium-activated chloride conductance (CaCC; TMEM16A), and Na + through epithelial Na +The balance with absorption determines the water supply state of the airway mucosa. Since ions are transported across the epithelium, water has to follow the osmotic pressure, and thus, body fluids are secreted or absorbed.

[0004] In respiratory diseases such as chronic bronchitis and cystic fibrosis, the solid content ratio of the mucus gel increases as water supply decreases, and mucus clearance decreases (Boucher, 2007). In cystic fibrosis, where loss-of-function mutations in CFTR attenuate the ability of the airway to secrete body fluids, the solid content ratio can increase up to 15%, which is thought to cause peripheral airway obstruction and failure of mucus clearance. Strategies to increase water supply to airway mucus include either inducing body fluid secretion by anion stimulation or inhibiting Na + absorption. To achieve this goal, stimulating the activity of the TMEM16A channel will increase anion secretion, thereby increasing body fluid accumulation in the airway mucosa, supplying water to the mucus, and improving the mucus clearance mechanism.

[0005] TMEM16A, also known as anoctamin-1 (Ano1), is the molecular identity of 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 influx of chloride ions, bicarbonate ions, and other anions across the cell membrane. Functionally, the TMEM16A channel has 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 of different organs, including the lung, liver, kidney, pancreas, and salivary gland. 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 mobilize intracellular calcium, particularly purinergic agonists (ATP, UTP), which are released by the respiratory epithelium in response to repetitive shear stress caused by other mechanical stimuli such as breathing and coughing. In addition to increasing anion secretion, which improves water supply to the airway, activation of TMEM16A plays an important role in bicarbonate secretion. Bicarbonate secretion has been reported to be an important regulator of mucus properties in the control of airway lumen pH and thus the activation of natural antibacterial substances such as defensins (Pezzulo et al., 2012).

[0007] For example, the indirect regulation of TMEM16A via the elevation of intracellular calcium by denufosol has been clinically investigated (Kunzelmann & Mall, 2003). Promising initial results were observed in small patient cohorts, but this approach did not result in clinical benefit in larger patient cohorts (Accurso et al., 2011; Kellerman et al., 2008). The cause of this lack of clinical effect has been attributed to only a transient increase in anion secretion, as a result of the short half-life of denufosol on the epithelial surface and receptor / pathway desensitization, as well as the unwanted effects of the elevation of intracellular calcium such as increased release of mucus from goblet cells. Compounds that act directly on TMEM16A to improve channel opening with low-level calcium elevation are expected to permanently improve anion secretion and mucociliary clearance in patients and improve innate defense. Since TMEM16A activity is independent of CFTR function, TMEM16A-positive modulators may have clinical benefit in all CF patients and non-CF respiratory diseases characterized by mucus plugging, including chronic bronchitis and severe asthma.

[0008] TMEM16A modulation is implicated in the therapy of dry mouth (xerostomia) resulting from salivary gland dysfunction in Sjögren's syndrome and radiotherapy, dry eye, biliary stasis as well as gastrointestinal motility disorders.

[0009] Our application, International Publication No. WO 2019 / 145726, relates to positive modulators of TMEM16A and thus to compounds for use in the treatment of diseases and conditions in which TMEM16A plays a role, in particular respiratory diseases and conditions.

[0010] The inventors have further developed compounds having excellent activity as positive modulators of TMEM16A. In addition, at least some of these compounds have one or more advantages compared to the compounds exemplified in International Publication No. WO 2019 / 145726. Some of the compounds of the present invention have increased metabolic stability, as represented by their low clearance rates from human microsomes and hepatocytes. This makes them particularly suitable for oral administration as it results in improved oral bioavailability and lower doses for certain pharmacological effects due to lower first-pass metabolism. Other compounds of the present invention have decreased metabolic stability to such an extent that they are particularly suitable for administration by inhalation compared to the compounds exemplified in International Publication No. WO 2019 / 145726. The decreased in vivo half-life in the blood ensures that the compound administered directly to the lung is less likely to interact with TMEM16A at other sites in the body and thus minimizes potential side effects. The compounds of the present invention also have appropriate water solubility as represented by their log D values, which also results in moderate lipophilicity that is advantageous when preparing pharmaceutical formulations. SUMMARY OF THE INVENTION

[0011] In a first aspect of the invention, there is provided a compound of general formula (I), including all its tautomeric forms, all its enantiomers, isotope variants, as well as salts and solvates: TIFF0007682850000001.tif48170(wherein, R1 is selected from H, ethynyl, CN, methyl, fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl, dichlorofluoromethyl and hydroxymethyl, R 2 is methyl and CH 2 OH, R 3 is selected from H and methyl, or R 2 and R 3 together with the carbon atom to which they are attached form a 3- to 10-membered carbocyclic ring system or an oxygen-containing heterocyclic ring system, and any of these ring systems may be substituted with one or more substituents selected from C 1 alkyl, C 1~4 haloalkyl, halo and OH in addition to the R 1~4 group, or R 1 R 2 and R 3 combine together with the carbon atom to which they are attached to form a 5- to 8-membered bridged carbocyclic or heterocyclic ring system which may be substituted with one or more substituents selected from OH, halo, C 1~4 alkyl and C 1~4 haloalkyl, R 4 is H or halo, R 5 is selected from H, halo, CN and C 1~4 alkyl which may be substituted with one or more substituents selected from halo and OH, X 1 is CR 6 or N, R 6 is selected from H, halo, CN and C 1~4 alkyl which may be substituted with one or more substituents selected from halo and OH, X 2 is CR 7 or N, R 7is a 3- to 7-membered carbocyclic or heterocyclic ring system which may be substituted with one or more substituents selected from H, halo, CN; halo and OH; or is substituted with one or more substituents selected from halo, OH and one or more substituents selected from halo and OH and may be substituted with a C 1~4 alkyl, X 3 is CR 8 or N, R 8 is H, halo, CN and may be substituted with one or more substituents selected from halo and OH and C 1~4 alkyl selected).

[0012] Since the compounds of general formula (I) are positive modulators of TMEM16A, they are useful for the treatment of diseases and conditions in which the regulation of TMEM16A plays a role, particularly respiratory diseases and conditions.

BEST MODE FOR CARRYING OUT THE INVENTION

[0013] As used herein, unless the language or context requires a different interpretation to express the necessary meaning, the term "comprise", or variations such as "comprises" or "comprising", is used in an inclusive sense, i.e., it is used not only to specify the presence of the recited features, but also to not exclude the presence or addition of further features in various embodiments of the present invention.

[0014] All documents and patent documents referred to herein are incorporated by reference to the maximum extent possible.

[0015] As used herein, reference to "pharmaceutical use" refers to use for administration to a human or animal, particularly a human or mammal, such as a domesticated mammal or livestock animal, for the treatment or prevention of a disease or medical condition. The term "pharmaceutical composition" refers to a composition suitable for pharmaceutical use, and "pharmaceutically acceptable" refers to an agent suitable for use in a pharmaceutical composition. Other similar terms shall be construed according to this definition.

[0016] Salts and solvates (such as hydrates) of the compounds of general formula (I) are preferably pharmaceutically acceptable. Suitable pharmaceutically acceptable salts are well known to those skilled in the art and are described, for example, in Gupta et al. (2018). Some particularly suitable salts of the compounds of general formula (I) include base addition salts such as sodium, potassium, calcium, aluminum, zinc, magnesium and other metal salts, as well as choline, diethanolamine, ethanolamine, ethyldiamine and meglumine salts. Alternatively, acid addition salts can form, for example, hydrochloride, mesylate, hydrobromide, sulfate, and fumarate salts. Salts of synthetic intermediates need not be pharmaceutically acceptable.

[0017] As used herein, the term "C 1~4 " alkyl refers to a straight-chain or branched-chain fully saturated hydrocarbon group having 1 to 4 carbon atoms. This term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl and t-butyl. Other alkyl groups, such as C 1~6 alkyl are as defined above but contain the stated number of carbon atoms.

[0018] The term "3- to 10-membered carbocyclic" refers to a non-aromatic hydrocarbon ring system containing 3 to 10 ring carbon atoms. The carbocyclic ring system may be monocyclic, or may contain two rings that are fused, or in a spiro configuration, or may be bridged, and the carbon atoms within the bridge are included in the count of the ring carbon atoms. Examples include cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, as well as spiro systems and bridged systems such as bicyclo[1.1.1]pentyl. The carbocyclic ring system may contain other numbers of ring atoms as specified, for example, 5 to 8 ring atoms or 3 to 7 ring atoms.

[0019] In the context of this specification, the term "cycloalkyl" refers to a fully saturated carbocyclic ring system as defined above. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, as well as bridged cycloalkyl systems such as bicyclo[1.1.1]pentyl.

[0020] In the context of this specification, the terms "3- to 10-membered heterocyclic" and "3- to 10-membered heterosilyl" refer to non-aromatic ring systems containing 3 to 10 ring atoms including at least one heteroatom selected from N, O, and S. The heterocyclic ring system may contain one or more carbon-carbon double bonds, but is preferably fully saturated. The heterocyclic ring system may be monocyclic, or may contain two or three rings that are fused, or in a spiro configuration, or may be bridged, and the bridging atoms are included in the count of the ring atoms. Oxygen-containing heterocyclic ring systems contain at least one oxygen as a ring atom and optionally one or two additional heteroatoms selected from O, N, and S. Examples of 3- to 10-membered heterocyclic ring systems include oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, and 2-oxaspiro[3.3]heptan-6-yl. The heterocyclic ring system may contain other numbers of ring atoms as specified, for example, 5 to 8 ring atoms or 3 to 7 ring atoms.

[0021] The term "halogen" refers to fluorine, chlorine, bromine or iodine, and the term "halo" refers to fluoro, chloro, bromo or iodo groups. Similarly, "halide" refers to fluoride, chloride, bromide or iodide.

[0022] As used herein, the term "C 1~4 haloalkyl" refers to a C 1~4 alkyl group as defined above, wherein one or more of the hydrogen atoms are replaced by halo groups. Any number of hydrogen atoms may be replaced up to perhalo substitution. Examples include trifluoromethyl, chloroethyl and 1,1-difluoroethyl. A fluoroalkyl group is a haloalkyl group in which the halo is fluoro. Other haloalkyl groups, e.g., C 1~3 haloalkyl are as defined above but contain the recited number of carbon atoms.

[0023] The term "isotope variant" refers to a compound that is identical to those listed in formula (I), but in which in fact one or more atoms have been replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature, or in which the proportion of atoms having an atomic mass or mass number not commonly found in nature has increased (the latter concept is referred to as "isotope enrichment"). Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine and chlorine such as 2H (deuterium), 3H, 11C, 13C, 14C, 18F, 123I or 125I (e.g., 3H, 11C, 14C, 18F, 123I or 125I), which may be either naturally occurring isotopes or non-naturally occurring isotopes.

[0024] In particularly preferred isotope variants of the compounds of general formula (I), some or all of the methyl groups are replaced by CD 3 . For example, R 1 , R 2 and R 3One, two, or all of them may be CDs 3 may also be.

[0025] In some preferred compounds of the present invention, R 1 is H, methyl, difluoromethyl, trifluoromethyl, ethynyl or CN, and in particular, methyl, difluoromethyl, trifluoromethyl, ethynyl or CN.

[0026] In some compounds of the present invention, R 2 is methyl. In other compounds of the present invention, R 2 is CH 2 OH. Preferably, R 3 is methyl.

[0027] In some preferred compounds of the present invention, R 2 is methyl and R 3 is methyl.

[0028] In some such compounds, R 1 is not H and is preferably ethynyl or methyl optionally substituted with one or more substituents selected from CN, fluoro and OH.

[0029] More preferably, in these compounds, R 1 is methyl, difluoromethyl, trifluoromethyl, ethynyl or CN.

[0030] In other preferred compounds of the present invention, R 2 and R 3 together with the carbon atom to which they are attached form an optionally substituted 3- to 10-membered carbocyclic ring system or oxygen-containing heterocyclic ring as described above. Preferred substituents for such rings (not including the R 1 group) include fluoro, chloro, methyl, ethyl, trifluoromethyl and OH. Even more preferably, R 2 and R 3and the ring systems formed by the carbon atoms to which they are attached are unsubstituted (except for R 1 groups) or substituted with one or more, for example one or two, substituents selected from methyl and fluoro in addition to the R 1 groups.

[0031] R 2 and R 3 and the ring systems formed by the carbon atoms to which they are attached include C 3 ~ 6 cycloalkyl rings, especially cyclopropyl, cyclobutyl and cyclopentyl, any of which may be unsubstituted or substituted as described above.

[0032] R 2 and R 3 and other rings formed by the carbon atoms to which they are attached include 3- to 8-membered oxygen-containing heterocyclic ring systems, which may be monocyclic ring systems such as tetrahydropyranyl, tetrahydrofuranyl and oxetanyl, or alternatively may be fused ring systems or spiro-bonded ring systems such as oxaspiro[3.3]heptan-6-yl. These ring systems may be unsubstituted or substituted as described above.

[0033] In still other compounds of general formula (I), R 1 , R 2 and R 3 combine together with the carbon atoms to which they are attached to form a 5- to 8-membered bridged carbocyclic or heterocyclic ring system. In this case, the carbon atoms to which R 1 , R 2 and R 3 are attached are bridgehead atoms. Examples of such ring systems include bicyclo[1.1.1]pentyl.

[0034] R 1 , R 2 and R 3The ring system formed by them and the carbon atoms to which they are attached may be unsubstituted or substituted as described above, but is more preferably unsubstituted.

[0035] In some preferred compounds of general formula (I), R 4 is H.

[0036] In some preferred compounds of general formula (I), R 5 is H or halo, especially H, fluoro or chloro. More preferably, R 5 is H.

[0037] In some preferred compounds of general formula (I), X 1 is N. However, more preferably, X 1 is CR 6 R 6 is preferably H or halo, for example, H, fluoro or chloro. More preferably, R 6 is H.

[0038] In the compounds of general formula (I), X 2 can be CR 7 or N. More preferably, X 2 is CR 7 and in this case, R 7 is preferably H; C 1~3 alkyl optionally substituted with OH; C 1~3 haloalkyl optionally substituted with OH; methyl substituted with a 3- to 6-membered carbocyclic or heterocyclic ring system; or a 3- to 6-membered carbocyclic or heterocyclic ring system.

[0039] In some cases, R 7 is H, C 1~3 alkyl, C 1~3 haloalkyl, C 1~3 alkyl substituted with OH, C 1~3 haloalkyl substituted with OH, a 3- to 6-membered carbocyclic or heterocyclic ring system or CH 2 -R 11and in the formula, R 11 is a nitrogen-containing 3- to 6-membered heterocyclic ring bonded to the CH 2 moiety through a ring nitrogen atom.

[0040] When R 7 is C substituted with OH 1~3 alkyl or C substituted with OH 1~3 haloalkyl, it can take the form of -C(CH 3 )(OH)-R 12 , where R 12 is C 1~2 alkyl or C 1~2 haloalkyl.

[0041] More preferably, R 7 is H, methyl, ethyl, isopropyl, trifluoromethyl, morpholinylmethyl, tetrahydrofuryl or 1-hydroxy-1-trifluoromethyl(ethyl), and in particular, H, methyl, ethyl, isopropyl and trifluoromethyl.

[0042] In particularly preferred compounds, R 7 is H.

[0043] In the compounds of the present invention, X 3 is CR 8 or N.

[0044] In some preferred compounds of general formula (I), X 3 is CR 8 . In these compounds, R 8 is preferably H, methyl or trifluoromethyl, in particular H or trifluoromethyl, and more specifically H.

[0045] However, in other preferred compounds, R 8 is N.

[0046] In some preferred compounds of the present invention, X 1 is CR 6 , and X2 is CR 7 and thereby the compound is of general formula (Ia): TIFF0007682850000002.tif48170(wherein X 3 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are as defined for general formula (I)).

[0047] In some preferred compounds of the present invention, X 1 is CR 6 and X 2 is CR 7 and X 3 is N, and thereby the compound is of general formula (Ib): TIFF0007682850000003.tif45170(wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are as defined for general formula (I)).

[0048] In some preferred compounds of the present invention, X 1 is CR 6 and X 2 is CR 7 and X 3 is CR 8 and thereby the compound is of general formula (Ic): TIFF0007682850000004.tif57170(wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are as defined for general formula (I)).

[0049] Preferably, in the compounds of general formulas (Ia), (Ib) and (Ic), R 5 , R 6 and R 7 at least one of which is H.

[0050] Preferably, in the compounds of general formulas (Ia), (Ib) and (Ic), when R 5 is halo, R 6 is H, and when R 6 is halo, R 5 is H.

[0051] In some compounds of general formulas (Ia), (Ib) and (Ic), when R 5 is halo, both R 6 and R 7 are H.

[0052] In some compounds of general formulas (Ia), (Ib) and (Ic), when R 6 is halo, both R 5 and R 7 are H.

[0053] In some compounds of general formulas (Ia), (Ib) and (Ic), when R 7 is other than halo, both R 5 and R 6 are H.

[0054] Specific examples of the compounds of general formula (I) include the following: N-(1,1-Dimethylprop-2-ynyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 1); 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 1.1) N-tert-Butyl-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 2); N-(1-cyano-1-methyl-ethyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 2.1); N-(1,1-dimethylprop-2-ynyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 3); N-(1-ethynylcyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 4); 4-[[2-(1H-indazol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide (Compound 4.1); 4-[[2-(1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 4.2); N-(3,3-difluoro-1-methyl-cyclobutyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 4.3); N-(2,2-difluoro-1,1-dimethyl-ethyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5); 4-[[2-(1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclobutyl]pyridine-2-carboxamide (Compound 5.1); N-(3-fluoro-3-methyl-cyclobutyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.2); N-(2,2-difluorocyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.3); N-(4-cyanotetrahydropyran-4-yl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.4); N-tert-butyl-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 6); 4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 6.1); N-(2,2-difluorocyclopentyl)-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 6.2); 4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 6.3); 4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]-N-(1-ethynylcyclopentyl)pyridine-2-carboxamide (Compound 6.4); N-tert-butyl-4-[[2-(4-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7); N-tert-butyl-4-[[2-(5-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7.1); N-tert-butyl-4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7.2); N-tert-butyl-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 8); N-[1-(trifluoromethyl)cyclopropyl]-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Compound 8.1); N-tert-butyl-4-[[2-(3-isopropyl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 9); 4-[[2-(3-Tetrahydrofuran-2-yl-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 9.1); 4-[[2-[3-(Morpholinomethyl)-1H-indazol-6-yl]acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide (Compound 10); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(3-methyloxetan-3-yl)pyridine-2-carboxamide (Compound 11); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(2-oxaspiro[3.3]heptan-6-yl)pyridine-2-carboxamide (Compound 11.1); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[4-(trifluoromethyl)tetrahydropyran-4-yl]pyridine-2-carboxamide (Compound 11.2); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[3-(trifluoromethyl)oxetan-3-yl]pyridine-2-carboxamide (Compound 11.3); N-[1-(Difluoromethyl)cyclopropyl]-4-[[2-(1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Compound 11.4); N-(3-Fluoro-1-bicyclo[1.1.1]pentanyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 11.5); 4-[[2-[3-(2,2,2-Trifluoro-1-hydroxy-1-methyl-ethyl)-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 12); 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide; (Compound 13); N-(3,3-Difluoro-1-methyl-cyclobutyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 13.1); N-(4-Cyanotetrahydropyran-4-yl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 13.2); And salts and solvates of the above.

[0055] The compounds of the present invention are novel and can be prepared by the methods described below which form further aspects of the present invention.

[0056] The compound of general formula (I) is a compound of general formula (II): TIFF0007682850000005.tif37170(wherein R 1 , R 2 , R 3 and R 4 are as defined for general formula (I)) is reacted with a compound of general formula (III): TIFF0007682850000006.tif50170(wherein R 5 , X 1 , X 2 and X 3 are as defined for general formula (I)) and can be prepared by reacting.

[0057] Preferably, the reaction is carried out in the presence of a coupling reagent, under basic conditions, for example, in the presence of an amine such as diisopropylethylamine (DIPEA), and in an organic solvent such as DMF.

[0058] Suitable coupling reagents include known peptide coupling reagents 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), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TATU), (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), carbodiimides such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and triazoles such as 1-hydroxy-7-azabenzotriazole (HOAt) or hydroxybenzotriazole (HOBt). Preferably, when these coupling agents are used, the reaction is carried out in an organic solvent such as DMF in the presence of a base, for example, an amine such as diisopropylethylamine (DIPEA).

[0059] Alternatively, the coupling reagent may be propylphosphonic anhydride (T3P®). When T3P is used as the coupling reagent, the reaction can be carried out in an organic solvent such as 1,4-dioxane in the presence of a base, for example, an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA).

[0060] The compounds of general formula (III) are known, commercially available, or may be obtained by known methods.

[0061] The compound of general formula (II) is a compound of general formula (IV): TIFF0007682850000007.tif27170(wherein R 1, R 2 , and R 3 is as defined for general formula (I)). with a compound of general formula (V): TIFF0007682850000008.tif38170(wherein R 4 is as defined for general formula (I)). It can be prepared by reacting.

[0062] This reaction is also preferably carried out in the presence of a coupling reagent such as TBTU as described above and in the presence of a base such as triethylamine.

[0063] Compounds of general formula (IV) and (V) are known, already commercially available, or may be prepared by known methods.

[0064] Alternatively, a compound of general formula (I) is a protected compound of general formula (Iz): TIFF0007682850000009.tif54170(wherein R 1 , R 2 , R 3 , R 4 , R 5 , X 1 , X 2 and X 3 are as defined for general formula (I), and R 15 is an amine protecting group, for example, a cyclic ether such as tetrahydropyran-2-yl or a substituted benzyl group such as benzyl or 4-methoxybenzyl)), It can be prepared by deprotecting.

[0065] Deprotection can be achieved by treatment with an acid, for example, trifluoroacetic acid.

[0066] The protected compound of general formula (Iz) is obtained by reacting a compound of general formula (II) with a protected compound of general formula (IIIz): TIFF0007682850000010.tif50170(wherein, R5 , X 1 , X 2 and X 3 are as defined for general formula (I), and R 15 is as defined for general formula (Iz)). It can be prepared by reacting with

[0067] The reaction can be carried out in the presence of a coupling agent as described above for the reaction between a compound of general formula (II) and a compound of general formula (III). The crude product may be used in subsequent deprotection steps without further purification.

[0068] Some compounds of general formula (IIIz) can be prepared from compounds of general formula (III) by known methods depending on the nature of the protecting group.

[0069] For example, when the group R 15 is a tetrahydropyran-2-yl group, the compound of general formula (III) can be reacted with 3,4-dihydro-2H-pyran in the presence of pyridinium p-toluenesulfonate. When the group R 15 is benzyl or a benzyl derivative, the compound of general formula (III) can be reacted with its substituted derivatives such as chloromethylbenzene or 1-chloromethyl-4-methoxybenzene. This reaction usually results in a mixture of the necessary protected compounds of general formula (IIIz) as a mixture with the ester of the compound of formula (IIIz). This ester can be converted to the compound of general formula (IIIz) by hydrolysis using an aqueous base such as lithium hydroxide or sodium hydroxide. X 2 and / or X 3 is N, a mixture of isomers may be formed, but since this will result in the product that requires final removal of the protecting group, there is no need to separate them.

[0070] Methods for preparing other compounds of general formula (IIIz) are described below (see the method for preparing the compound of formula (IIIaz)).

[0071] An alternative method for preparing a compound of general formula (Iz) is to react a compound of general formula (IV) as defined above with a protected compound of general formula (XIIz): TIFF0007682850000011.tif50170(wherein R 4 , R 5 , X 1 , X 2 and X 3 are as defined for general formula (I), and R 15 is as defined for general formula (Iz)) by reacting them together.

[0072] Preferably, this reaction is carried out in the presence of a coupling agent under the same conditions as described above for the reaction of a compound of general formula (II) with a compound of general formula (III).

[0073] The compound of general formula (XIIz) is a compound of general formula (XIIIz): TIFF0007682850000012.tif51170(wherein R 4 , R 5 , X 1 , X 2 and X 3 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 16 is C 1~6 alkyl or benzyl, for example methyl) and can be prepared by hydrolysis thereof.

[0074] Preferably, the hydrolysis is carried out under basic conditions, for example, in the presence of lithium hydroxide, sodium hydroxide or potassium hydroxide and in a mixed solvent containing an alcohol such as methanol, water and an organic solvent such as tetrahydrofuran. The reaction may be carried out at a temperature of about 15 to 25 ° C, typically at room temperature.

[0075] The compound of general formula (XIIIz) is obtained by reacting the compound of general formula (IIIz) defined above with a compound of general formula (XIV): TIFF0007682850000013.tif38170(wherein R 4 is as defined for general formula (I), and R 16 is as defined for general formula (XIIIz)) and can be prepared by reacting.

[0076] Preferably, the reaction is carried out in the presence of a coupling reagent as described above.

[0077] The compound of general formula (XIV) is known, commercially available, or may be prepared by known methods, for example, by esterification of the carboxylic acid of general formula (V) described above.

[0078] X 1 is CR 6 and X 2 is CR 7 The compound of general formula (XIIz) in which is is prepared from a compound of general formula (XIIaz): TIFF0007682850000014.tif50170(wherein X 3 , R 4 , R 5 , R 6 , and R 7 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 16 is as defined for general formula (XIIIz)) is shown as.

[0079] The compound of general formula (XIIaZ) is R 5 and R 6 wherein one of them is H, and R 5 and R 6 wherein the other is halo, is particularly useful for preparing the compound of general formula (I) and the compound wherein R 7 is other than H.

[0080] The compound of general formula (XIIaZ) is a compound of general formula (IIIz) wherein X 1 is CR 6 and X 2 is CR 7 (i.e., the compound of general formula (IIIaz)): TIFF0007682850000015.tif51170(wherein X 3 , R 5 , R 6 and R 7 are as defined for general formula (I), and R 15 is as defined for general formula (Iz)) can be prepared by reacting with the compound of general formula (XIV) defined above.

[0081] The compound of general formula (IIIaz) is a compound of general formula (XXI): TIFF0007682850000016.tif53170(wherein X 3 , R 5 , R 6 , and R 7 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 20 is halo, for example, bromo) can be prepared by carbonylation of

[0082] The carbonylation is carried out with [Pd(allyl)Cl)] 2The reaction of an alkali metal salt of a monoester of malonic acid, such as potassium methyl malonate, in the presence of a Pd catalyst such as, a phosphine ligand such as 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), and an organic base such as DMAP, followed by hydrolysis of the resulting ester with an aqueous base, such as an alkali metal hydroxide such as lithium hydroxide, can be achieved.

[0083] Preferably, this reaction is carried out under an inert atmosphere such as nitrogen and at a high temperature, typically 120 °C to 160 °C.

[0084] An alternative method for preparing a compound of general formula (IIIaz) is to react a compound of general formula (XXI) as defined above with an alkali metal salt of cyanoacetic acid, such as potassium 2-cyanoacetate, in the presence of a palladium catalyst such as a phosphine ligand such as SPhos and dichloride [Pd(allyl)Cl] 2 followed by hydrolysis of the resulting nitrile with an aqueous base, such as an alkali metal hydroxide such as sodium hydroxide. Preferably, this reaction is carried out under an inert atmosphere such as nitrogen and at a high temperature, typically 120 °C to 160 °C.

[0085] The compound of general formula (XXI) can be prepared by protecting a compound of general formula (XXII): TIFF0007682850000017.tif48170(wherein X 3 , R 5 , R 6 and R 7 are as defined for general formula (I), and R 20 is as defined for general formula (XXI)). The protection can be achieved by using the method described above for the preparation of a compound of general formula (IIIz) from a compound of general formula (III).

[0086] The protection can be achieved by using the method described above for the preparation of a compound of general formula (IIIz) from a compound of general formula (III).

[0087] The compounds of general formula (XXII) are known, commercially available, or may be prepared by known methods.

[0088] R 7 is CH 2 -R 11 and, where R 11 is a nitrogen-containing 3- to 6-membered heterocyclic ring linked to the CH 2 moiety via a ring nitrogen atom), an alternative method for the preparation of compounds of general formula (XXI) which are particularly suitable for compounds: TIFF0007682850000018.tif54170(wherein X 3 , R 5 and R 6 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 20 is as defined for general formula (XXI)) with a compound of general formula (XXXI): R 11 -H (XXXI) (wherein R 11 is a nitrogen-containing 3- to 6-membered heterocyclic ring linked to an H atom via a ring nitrogen atom) by reaction therewith.

[0089] This reaction can be carried out under reducing conditions, for example, in an organic solvent such as tetrahydrofuran, in the presence of a hydride such as sodium triacetoxyborohydride, at a temperature of about 15 - 25 °C, typically at room temperature and in the presence of an acid such as acetic acid.

[0090] The compounds of general formula (XXXI) are known, commercially available, or may be prepared by known methods.

[0091] The compounds of general formula (XXX) are prepared by the method described above for the preparation of compounds of general formula (IIIz) from a compound of general formula (XXXII): TIFF0007682850000019.tif45170(wherein X3 , R 5 , and R 6 is as defined for general formula (I), and R 20 is as defined for general formula (XXI)). can be prepared by protecting

[0092] Compounds of general formula (XXXII) are known, commercially available, or may be prepared by known methods.

[0093] R 7 being -C(CH 3 )(OH)-R 12 wherein R 12 is CF 3 or CHF 2 An alternative method for preparing general formula (XXI), which is particularly suitable for compounds wherein TIFF0007682850000020.tif51170 (wherein X 3 , R 5 and R 6 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 20 is as defined for general formula (XXI)). with a compound of general formula (XXXVI): (R 25 ) 3 Si-R 12 (XXXVI) (wherein each R 25 is independently C 1~6 alkyl, and R 12 is C 1~2 alkyl or C 1~2 haloalkyl). by reaction with

[0094] This reaction can be carried out in the presence of fluoride ions such as tetrabutylammonium fluoride (TBAF).

[0095] R 7 is -C(CH 3 )(OH)-R 12 and here R 12 is CF 3 or CHF 3 compounds of general formula (XXI) other than can be synthesized by reacting a compound of general formula (XXXV) with a suitable Grignard reagent under suitable reaction conditions.

[0096] Compounds of general formula (XXXVI) are known, commercially available, or may be prepared by known methods.

[0097] Compounds of general formula (XXXV) are compounds of general formula (XXXVII): TIFF0007682850000021.tif49170(wherein X 3 , R 5 and R 6 are as defined for general formula (I), R 15 is as defined for general formula (Iz), R 20 is as defined for general formula (XXI)) can be prepared by oxidation of.

[0098] Suitable oxidizing agents include Dess-Martin periodinane. In this case, the oxidation is preferably carried out in an organic solvent such as dichloromethane.

[0099] Compounds of general formula (XXXVII) can be prepared by reacting a compound of general formula (XXX) as defined above with a methyl Grignard reagent.

[0100] R 7 is -C(CH 3 )(OH)-R 12 -C(CH 3 )(OH)-R 12The OH group of the compound of general formula (XXI) will generally be protected, for example, as a benzyl ether before the carbonylation to obtain the compound of general formula (IIIaz). The protecting group is retained in the compounds of general formulas (XIIaz) and (Iaz) and can be removed simultaneously with the conversion of the compound of general formula (Iaz) to the compound of general formula (Ia), where R 15 can be removed simultaneously.

[0101] The compound of general formula (IIIaz) is a protected compound of general formula (III) where X 1 is CR 6 and X 2 is CR 7 and thus this is reacted with the compound of general formula (II) described above to obtain the compound of general formula (Iaz): TIFF0007682850000022.tif54170(wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are as defined for general formula (I) and R 15 is as defined for general formula (Iz)) can be obtained; this can be deprotected to obtain the compound of general formula (Ia).

[0102] A further alternative method for preparing the compound of general formula (I) is by reacting the compound of general formula (IV) defined above with the compound of general formula (XII): TIFF0007682850000023.tif44170(wherein R 4 , R 5 , X 1 , X 2 and X 3 are as defined for general formula (I)). by reacting them.

[0103] Preferably, this reaction is carried out in the presence of a coupling agent under the same conditions as described above for the reaction of a compound of general formula (II) with a compound of general formula (III).

[0104] The compound of general formula (XII) can be prepared by deprotecting the above-described compound of general formula (XIIz) by reacting it with an acid such as trifluoroacetic acid.

[0105] R 7 For a compound of general formula (Ia) in which R is alkyl or heterocyclyl, a compound of general formula (XXVaz): TIFF0007682850000024.tif54170(wherein X 3 , R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 21 is halo, for example, bromo or iodo) from under basic conditions, for example, in the presence of K 2 PO 4 , in the presence of a palladium catalyst such as palladium acetate, reacting with a suitable dioxaborolane and then performing catalytic hydrogenation on the palladium catalyst. When the protecting group R 15 is a derivative such as benzyl or paramethoxybenzyl, it will be removed by the hydrogenation process. In another situation, an additional deprotection step, for example, treatment with an acid such as TFA, may be required.

[0106] R 7 For a compound of general formula (Ib) in which R is isopropyl, the borolane used in the first step may be 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, while R 7For a compound where it is tetrahydrofuran-2-yl, 2-(2,3-dihydrofuran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane may be used. For the introduction of an alternative R 7 group, other diborolanes may be selected.

[0107] A compound of general formula (XXVaz) is obtained by reacting a compound of general formula (II) as defined above with a compound of general formula (XXVI): TIFF0007682850000025.tif53170(wherein X 3 , R 5 and R 6 are as defined for general formula (I), R 15 is as defined for general formula (Iz), and R 21 is as defined for general formula (XXVaz)) and can be prepared by reacting them.

[0108] This reaction may be carried out in the presence of a coupling reagent under the conditions described above for the reaction of a compound of general formula (II) with a compound of general formula (III).

[0109] A compound of general formula (XXVI) can be prepared from a compound of general formula (XXVII): TIFF0007682850000026.tif49170(wherein X 3 , R 5 , and R 6 are as defined for general formula (I), and R 21 is as defined for general formula (XXVaz)) by a method similar to that described above for preparing compounds of general formula (IIIz) and (IIIaz).

[0110] A compound of general formula (XXVII) can be prepared from a compound of general formula (XXVIII) according to the procedure of Tang et al., J. Org. Chem. 2018, 83(2), 930 - 938: TIFF0007682850000027.tif45170(wherein X 3 , R 5 , and R 6 are as defined for general formula (I)) can be prepared by halogenating

[0111] Compounds of general formula (XXVIII) are known, commercially available, or may be prepared by known methods.

[0112] Compounds of general formula (I) are positive modulators of TMEM16A, and thus, in a further aspect of the invention, there is provided a compound of general formula (I) as defined above for use in medicine, in particular for the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A.

[0113] Also provided is the use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A.

[0114] Also provided is a method for the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A, the method comprising administering to a patient in need of such treatment an effective amount of a compound of general formula (I).

[0115] Diseases and conditions affected by the modulation of TMEM16A include respiratory diseases and conditions, dry mouth (xerostomia), intestinal hypermotility, cholestasis, and ocular symptoms.

[0116] Also provided are: · A compound of general formula (I) for use in the treatment or prevention of respiratory diseases and conditions. · A compound of general formula (I) for use in the treatment or prevention of dry mouth (xerostomia). · A compound of general formula (I) for use in the treatment or prevention of intestinal hypermotility. · Compounds of general formula (I) for use in the treatment or prevention of biliary stasis. · Compounds of general formula (I) for use in the treatment or prevention of eye symptoms.

[0117] The present invention also provides the following: · Use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of respiratory diseases and conditions. · Use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of dry mouth (xerostomia). · Use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of intestinal hypermotility. · Use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of biliary stasis. · Use of a compound of general formula (I) in the manufacture of a medicament for the treatment or prevention of eye symptoms.

[0118] Furthermore, the following are provided: · A method for the treatment or prevention of respiratory diseases and conditions, the method comprising administering to a patient in need thereof an effective amount of a compound of general formula (I). · A method for the treatment or prevention of dry mouth (xerostomia), the method comprising administering to a patient in need thereof an effective amount of a compound of general formula (I). · A method for the treatment or prevention of intestinal hypermotility, the method comprising administering to a patient in need thereof an effective amount of a compound of general formula (I). · A method for the treatment or prevention of biliary stasis, the method comprising administering to a patient in need thereof an effective amount of a compound of general formula (I). · A method for the treatment or prevention of eye symptoms, the method comprising administering to a patient in need thereof an effective amount of a compound of general formula (I).

[0119] Respiratory diseases and conditions that can be treated or prevented by the compounds of general formula (I) include cystic fibrosis, chronic obstructive pulmonary disease (COPD), chronic bronchitis, emphysema, bronchiectasis including non-cystic fibrosis bronchiectasis, asthma and primary ciliary dyskinesia.

[0120] Dry mouth (xerostomia) that can be treated or prevented by the compounds of general formula (I) can be caused by Sjogren's syndrome, radiation therapy and drugs that cause dry mouth (xerogenic drug).

[0121] Intestinal hypermotility that can be treated or prevented by the compounds of general formula (I) can be associated with dyspepsia, gastroparesis, chronic constipation and irritable bowel syndrome.

[0122] Eye symptoms that can be treated or prevented by the compounds of general formula (I) include dry eye disease.

[0123] The compounds of the present invention will generally be administered as part of a pharmaceutical composition and accordingly the present invention further provides a pharmaceutical composition comprising a compound of general formula (I) together with pharmaceutically acceptable additives.

[0124] The pharmaceutical composition can be formulated for oral, rectal, nasal, bronchial (inhalation), topical (including skin, transdermal, ophthalmic, oral, sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, and intradermal) administration and can be prepared by any method well known in the art of pharmacy.

[0125] The composition can be prepared by combining the active agent as defined above with additives. Generally, the formulation is prepared by uniformly and intimately bringing the active agent into association with a liquid carrier or a finely divided solid carrier or both and then, if necessary, shaping the product. The present invention extends to a method for preparing a pharmaceutical composition comprising combining or associating a compound of general formula (I) with a pharmaceutically acceptable carrier or vehicle.

[0126] The pharmaceutical preparations for oral administration in the present invention can be presented as individual units such as capsules, sachets or tablets, each containing a predetermined amount of the active agent, as powders or granules, as solutions or suspensions of the active agent in aqueous or non-aqueous liquids, or as water-in-oil or oil-in-water liquid emulsions, or as a bolus, etc.

[0127] In the case of pharmaceutical preparations for oral administration (for example, tablets and capsules), the term "acceptable carrier" includes vehicles such as common additives, for example, binders 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 fluid, talc, wax, oil and colloidal silica. For example, flavoring agents such as peppermint, wintergreen oil, cherry flavoring, etc. can also be used. It may be desirable to add coloring agents to make the dosage form easily distinguishable. Tablets can also be coated by methods well known in the art.

[0128] Tablets can be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets are prepared by compressing in a suitable machine the active ingredient in free-flowing form such as a powder or granules, optionally mixed with a binder, a lubricant, an inert diluent, a preservative, a surfactant, or a dispersing agent. Molded tablets can be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert diluent. Tablets can optionally be coated or scored and formulated to provide for slow or controlled release of the active ingredient.

[0129] Formulations suitable for oral administration include flavored bases, usually lozenges containing the active ingredient in sucrose and acacia or tragacanth, pastilles containing the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia, and mouthwashes containing the active ingredient in a suitable liquid carrier.

[0130] For topical administration to the skin, the compounds of general formula (I) may be formulated in, for example, creams, ointments, jellies, solutions, or suspensions. Cream or ointment formulations which can be used for pharmaceuticals are conventional formulations well known in the art and are described, for example, in standard texts on pharmaceuticals such as the British Pharmacopoeia.

[0131] Local administration to the lungs can be achieved by using an aerosol formulation. An aerosol formulation typically contains an active ingredient suspended or dissolved in a suitable aerosol propellant such as 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 additives such as a co-solvent (e.g., ethanol) and a surfactant (e.g., lecithin, sorbitan trioleate, etc.). Other possible additives include polyethylene glycol, polyvinylpyrrolidone, glycerin, etc. The aerosol formulation is packaged in a canister, and a suitable dose is delivered by a metering valve (e.g., supplied by Bespak, Valois or 3M, or supplied by Aptar, Coster or Vari).

[0132] Local administration to the lungs can be achieved by using a non-pressurized formulation such as an aqueous solution or suspension. These may be administered by a nebulizer, for example, a portable one or one for use at home or in a hospital (i.e., a non-portable one). The formulation may contain additives such as water, a buffer, an isotonic agent, a pH adjuster, a surfactant and a co-solvent. Suspensions and aerosol formulations (whether pressurized or non-pressurized) will typically contain the compound of the invention in the form of fine powder having, for example, a D 50 of, for example, 0.5 to 10 μm, for example, about 1 to 5 μm. The particle size distribution can be represented using the D 10 , D 50 , and D 90 values. The D 50The average value is defined as the particle size in microns when the distribution is divided by 2. Since the measured values derived from laser diffraction are more accurately described as volume distributions, the D 50 value obtained using this procedure indicates the Dv 50 value (median of the volume distribution) in a more meaningful way. As used herein, the Dv value refers to the particle size distribution measured using laser diffraction. Similarly, the D 10 and D 90 values are taken to mean the Dv10 and Dv 90 values, respectively, where 10% of the distribution is below the D 10 value and 90% of the distribution is below the D 90 value, referring to the particle size.

[0133] Local administration to the lungs can be achieved by using dry powder formulations. The dry powder formulations will contain the compounds of the present disclosure in the form of fine powders typically having a mass median aerodynamic diameter (MMAD) of 1 to 10 μm or a D 50 of 0.5 to 10 μm, for example, about 1 to 5 μm. The powders of the compounds of the invention in fine powder form may be prepared by a micronization process or a similar size reduction process. Micronization may be carried out using a jet mill such as those manufactured by Hosokawa Alpine. The resulting particle size distribution can be measured using laser diffraction (e.g., by a Malvern Mastersizer 2000S instrument). The formulations typically usually have a relatively large particle size, for example, a mass median aerodynamic diameter (MMAD) of 50 μm or more, for example, 100 μm or more, or a D 50It will contain a locally 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, anhydrous α-lactose, anhydrous β-lactose and amorphous lactose. The lactose component may be treated by micronization, sieving, milling, compression, agglomeration or spray drying. Commercially available forms of various forms of lactose are also included, for example, Lactohale® (inhalation grade lactose; DFE Pharma), InhaLac® 70 (sieved lactose for dry powder inhalers; 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, anhydrous α-lactose and amorphous lactose. Preferably, the lactose is α-lactose monohydrate.

[0134] The dry powder formulation may also contain other additives. Thus, in one embodiment, the dry powder formulation according to the present disclosure contains magnesium stearate or calcium stearate. Such formulations may have excellent chemical and / or physical stability, especially when such formulations contain lactose.

[0135] Dry powder formulations are typically delivered using a dry powder inhaler (DPI) device. Exemplary 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.

[0136] In one embodiment, the compound of general formula (I) is provided as a micronized dry powder formulation comprising, for example, a suitable grade of lactose.

[0137] Accordingly, as an aspect of the present invention, there is provided a pharmaceutical composition comprising the compound of general formula (I) in particulate form in combination with particulate lactose, which composition optionally comprises magnesium stearate.

[0138] In one embodiment, the compound of general formula (I) is provided as a micronized dry powder formulation comprising a suitable grade of lactose and magnesium stearate filled into a device such as DISKUS. Suitably, such a device is a multi-dose device, for example, in which the formulation is filled into blisters for use in a multi-unit dose device such as DISKUS.

[0139] In another embodiment, the compound of general formula (I) is provided as a micronized dry powder formulation containing a suitable grade of lactose, filled in a hard shell capsule for use in a single-dose device such as an AEROLISER.

[0140] In another embodiment, the compound of general formula (I) is provided as a micronized dry powder formulation containing a suitable grade of lactose and magnesium stearate, filled in a hard shell capsule for use in a single-dose device such as an AEROLISER.

[0141] In another embodiment, the compound of general formula (I) is provided as a fine powder for use in an inhaled dosage form, the powder being produced by a size reduction process other than jet milling micronization, such as spray drying, spray freezing, microfluidization, high-pressure homogenization, supercritical fluid crystallization, ultrasonic crystallization or a combination of these methods, or by other suitable particle formation methods known in the art for producing fine particles having an aerodynamic particle size of 0.5 to 10 μm, for example about 1 to 5 μm D 50 which are fine particles having. The resulting particle size distribution can be measured using laser diffraction (e.g., by a Malvern Mastersizer 2000S instrument). The particles can be included either alone or in combination with other suitable additives that can assist in processing. The resulting fine particles may form the final formulation for delivery to humans or, optionally, may be further formulated with other suitable additives to facilitate delivery in an acceptable dosage form.

[0142] The compounds of the present invention may also be administered rectally, for example, in the form of suppositories or enemas, which include 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 conventional suppository bases such as cocoa butter or other glycerides. In this case, the drug is solid at room temperature but liquid at rectal temperature and is thus mixed with a suitable non-irritating additive that dissolves in the rectum and releases the drug. Such materials are cocoa butter and polyethylene glycol.

[0143] Generally, in compositions intended for topical administration to the eye in the form of eye drops or eye ointments, the total amount of the compound of general formula (I) will be less than about 0.0001 - 4.0 (w / w)%.

[0144] Preferably, for topical ocular administration, the compositions administered according to general formula (I) will be formulated as solutions, suspensions, emulsions and other dosage forms. Aqueous solutions are generally preferred based on ease of formulation and the patient's ability to easily administer such compositions by instilling 1 - 2 drops of the solution into 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 only slightly soluble in water.

[0145] An alternative for administration to the eye is intravitreal injection of a solution or suspension of the compound of general formula (I). In addition, the compound of general formula (I) may also be introduced using an intraocular implant or insert. I

[0146] The compositions administered according to general formula (I) may also contain various other components including, but not limited to, tonicity agents, buffers, surfactants, stabilizing polymers, preservatives, co-solvents, and viscosity builders. Suitable pharmaceutical compositions of general formula (I) contain the compounds of the invention formulated with a tonicity agent and a buffer. The pharmaceutical compositions of general formula (I) may optionally further contain a surfactant and / or a soothing agent and / or a stabilizing polymer.

[0147] Various tonicity agents can be used to adjust the tonicity of the composition, preferably to adjust the tonicity of natural tears of an ophthalmic composition. For example, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, monosaccharides such as dextrose, fructose, galactose, and / or polyols such as mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, and hydrolyzed hydrogenated starch can be added to the composition to approach physiological tonicity. Such amounts of the tonicity agent will vary depending on the particular agent being added. However, generally, the composition will have a tonicity agent in an amount sufficient to make the final composition have an ophthalmically acceptable osmotic pressure (generally about 150 - 450 mOsm, preferably 250 - 350 mOsm and most preferably approximately 290 mOsm). Generally, the tonicity agent of the present invention will be present in the range of 2 - 4 w / w%. Preferred tonicity agents of the present invention include monosaccharides or sugar alcohols, for example, D-mannitol.

[0148] Suitable buffer systems (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 being used. However, preferably, the buffer will be selected to maintain the target pH within the range of pH 5 - 8, and more preferably to maintain a target pH of pH 5 - 7.

[0149] The surfactant can optionally 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, polyoxyl 40 stearate, polyoxyl castor oil, tyloxapol, Triton, and sorbitan monolaurate. Preferred surfactants to be used in the present invention have a hydrophilic-lipophilic balance "HLB" in the range of 12.4 to 13.2, such as Triton X114 and tyloxapol, and are acceptable for ophthalmic use.

[0150] An additional agent that can be added to the ophthalmic composition of the compound of general formula (I) is a viscous agent that functions as a stabilizing polymer. The stabilizing polymer should be an ionic / charged example that is preferred for topical ocular use, and more specifically, it can exhibit a zeta potential of (-)10 to 50 mV for physical stability and can form a dispersion in water (i.e., be water-soluble), and can be a polymer carrying a negative charge on its surface. Preferred stabilizing polymers of the present invention will be polyelectrolytes at 0.1 to 0.5 w / w%, or in the case of two or more, polyelectrolytes from the family of cross-linked polyacrylates such as carbomer and Pemulen® (registered trademark), specifically carbomer 974p (polyacrylic acid).

[0151] Other compounds may also be added to the ophthalmic composition of the compound of general formula (I) to increase the viscosity of the carrier. Examples of viscosity enhancing agents include, but are not limited to, hyaluronic acid and its salts, chondroitin sulfate and its salts, polysaccharides such as dextran, various polymers of the cellulose family, vinyl polymers, and acrylic acid polymers.

[0152] Ophthalmic products for local use are typically packaged in multiple-dose forms. Therefore, preservatives are required to prevent microbial contamination during use. Suitable preservatives include benzalkonium chloride, chlorobutanol, benzododecinium bromide, methylparaben, propylparaben, phenylethyl alcohol, disodium edetate, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art. Such preservatives are typically used at levels of 0.001 to 1.0 w / v%. The unit-dose compositions of general formula (I) will be sterile but will typically not be preserved. Such compositions will generally not contain preservatives.

[0153] Parenteral formulations will generally be sterile.

[0154] A physician, or other person skilled in the art, can determine a suitable dosage for the compounds of general formula (I) and thus can determine the amount of the compounds of the present invention that should be included in any particular pharmaceutical formulation, whether in unit-dose form or otherwise.

[0155] The compounds of general formula (I) may be used in combination with one or more other active agents useful in the treatment or prevention of respiratory diseases and conditions.

[0156] This type of additional active agent may be included in the pharmaceutical compositions described above, or it may be administered separately, either simultaneously with, or at an earlier or later time than, the compounds of general formula (I).

[0157] Accordingly, in a further aspect of the invention, there is provided a product comprising the compounds of general formula (I) and an additional agent useful in the treatment or prevention of respiratory conditions, as a combined preparation for simultaneous, sequential or separate use in the treatment of a disease affected by the modulation of TMEM16A, particularly a respiratory disease or condition, for example one of the diseases and conditions described above.

[0158] Also, a compound of general formula (I) in combination with an additional agent useful for the treatment or prevention of respiratory conditions is provided as a combination preparation for simultaneous, sequential or separate use in the treatment of particularly respiratory diseases or conditions affected by the modulation of TMEM16A, for example, one of the diseases and conditions described above.

[0159] Suitable additional active agents that may be included in pharmaceutical compositions or combination preparations with compounds of general formula (I), (Ix), (IA), (IB), (IC), (ID) or (IE) include the following: β2 - adrenergic receptor agonists such as metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol, formoterol, salmeterol, indacaterol, terbutaline, orciprenaline, bitolterol mesylate, pirbuterol, olodaterol, vilanterol and abediterol; Antihistamines, for example, histamine H 1 receptor antagonists such as loratadine, cetirizine, desloratadine, levocetirizine, fexofenadine, astemizole, azelastine, and chlorpheniramine, or H 4 receptor antagonists; Dornase alfa; 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, aclidinium and umeclidinium; CFTR repair therapies (e.g., CFTR potentiators, correctors or amplifiers), such as ivacaftor, QBW251, vafacaftor (VX659), elexacaftor (VX445), VX561 / CPT-656, VX152, VX440, GLP2737, GLP2222, GLP2451, PTI438, PTI801, PTI808, FDL-169 and FDL-176, and CFTR correctors such as lumacaftor and tezacaftor, or combinations thereof (e.g., a combination of ivacaftor, tezacaftor and elexacaftor); ENaC modulators, particularly ENaC inhibitors; Antibiotics; Antiviral agents such as ribavirin and neuraminidase inhibitors such as zanamivir; Antifungal agents such as PUR1900; Airway water supply agents (osmolytes) such as hypertonic saline and mannitol (Bronchitol®); and Mucolytics such as N-acetylcysteine.

[0160] If the additional active agent is an ENaC modulator, it can be an ENaC inhibitor such as amiloride, VX-371, AZD5634, QBW276, SPX-101, BI443651, BI1265162 and ETD001. Other suitable ENaC blockers are described in our applications, International Publication No. WO2017 / 221008, International Publication No. WO2018 / 096325, International Publication No. WO2019 / 077340 and International Publication No. WO2019 / 220147, and exemplary compounds of those applications may all be used in combination with the compounds of general formula (I). Particularly suitable compounds for use in combination with the compounds of general formula (I) include 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)ethyl]-6-(4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}piperidine-1-carbonyl)-1,3-diethyl-1H-1,3-benzodiazol-3-ium; 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)methyl]-6-{[2-(4-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}piperidin-1-yl)ethyl]carbamoyl}-1,3-diethyl-1H-1,3-benzodiazol-3-ium; 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)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-benzodiazol-3-ium; 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)methyl]-6-[(3R)-3-{bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino}pyrrolidine-1-carbonyl]-1,3-diethyl-1H-1,3-benzodiazol-3-ium; 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)methyl]-6-[(3S)-3-{bis[(2S,3R,4R,5R )-2,3,4,5,6-pentahydroxyhexyl]amino}pyrrolidine-1-carbonyl]-1,3-diethyl-1H-1,3-benzodiazol-3-ium; 2-[({3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl}formamido)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-benzodiazol-3-ium; 2-[(3-Amino-5H-pyrrolo[2,3-b]pyrazin-2-yl)formamidomethyl]-1,3-diethyl-6-{[(1s,4s)-4-bis[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]amino]cyclohexyl}carbamoyl}-1H-1,3-benzodiazol-3-ium A cation selected from the following; And suitable anions such as halides, sulfates, nitrates, phosphates, formates, acetates, trifluoroacetates, fumarates, citrates, tartrates, oxalates, succinates, mandelates, methanesulfonates or p-toluenesulfonates Compounds having are included.

[0161] The present invention is illustrated by the following non-limiting examples and drawings.

Brief Description of the Drawings

[0162]

Figure 1

[0163] The present invention is illustrated by the following examples.

Examples

[0164] General conditions: Mass spectra were run on an LC-MS system using electrospray ionization. These were run on either a Waters Acquity uPLC system equipped with Waters PDA and ELS detectors or a Shimadzu LCMS-2010EV system. [M+H]+ refers to the monoisotopic molecular weight.

[0165] NMR spectra were recorded on a Bruker Avance III HD 500 MHz equipped with a 5 mm Broad Band Inverse probe, a Bruker Avance III HD 250 MHz, or a 400 MHz Avance III HD Nanobay equipped with a 5 mm Broad Band Observed SmartProbe using the solvent as an internal deuterium lock. Unless otherwise specified, spectra were recorded at room temperature and referenced using the solvent peak.

[0166] The compounds of the preferred embodiments were synthesized using the methods described herein or other methods known in the art with reference to the following examples.

[0167] The various starting materials, intermediates, and compounds of the 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 were obtained from suppliers and used without further purification. Salts may be prepared from the compounds by known salt-forming procedures.

[0168] Compounds were purified by flash column chromatography on normal-phase silica of a Biotage® Isolera system using an appropriate SNAP cartridge and gradient. Alternatively, compounds were purified on reverse-phase silica using a Biotage® Isolera system with an appropriate SNAP C18 cartridge and reverse-phase eluent, or by preparative HPLC (where otherwise specified).

[0169] Preparative HPLC using acidic pH, early elution method Purification by preparative HPLC was carried out on a Gilson LC system using a Waters Sunfire C18 column (30 mm × 100 mm, 10 μM; temperature: room temperature), and a gradient of 10 - 95% B over 14.44 minutes (A = 0.1% formic acid in water; B = 0.1% formic acid in MeCN), followed by 95% B for 2.11 minutes, with 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.

[0170] Preparative HPLC using acidic pH, standard elution method Purification by preparative HPLC (acidic pH, standard elution method) was carried out on a Gilson LC system using a Waters Sunfire C18 column (30 mm × 100 mm, 10 μM; temperature: room temperature), and a gradient of 30 - 95% B over 11 minutes (A = 0.1% formic acid in water; B = 0.1% formic acid in MeCN), followed by 95% B for 2.11 minutes, with 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.

[0171] Preparative HPLC using basic pH, early elution method Purification by preparative HPLC (basic pH, early elution method) was carried out on a Gilson LC system using a Waters Xbridge C18 column (30 mm × 100 mm, 10 μM; temperature: room temperature), and a gradient of 10 - 95% B over 14.44 minutes (A = 0.2% ammonium hydroxide in water; B = 0.2% ammonium hydroxide in MeCN), followed by 95% B for 2.11 minutes, with 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.

[0172] Preparative HPLC using basic pH, standard elution method Purification by preparative HPLC (basic pH, standard dissolution method) was carried out on a Waters Xbridge C18 column (30 mm × 100 mm, 10 μM; temperature: room temperature) and a Gilson LC system using a gradient of 30 - 95% B (A = 0.2% ammonium hydroxide in water; B = 0.2% ammonium hydroxide in MeCN) over 11 minutes, followed by a gradient of 95% B for 2.11 minutes, with 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.

[0173] Unless otherwise specified, the HPLC analysis conditions are as follows:

[0174] Method A Column: Phenomenex Kinetix-XB C18 2.1×100 mm, 1.7 μm Column temperature 40 °C Eluent: A: H 2 0.1% formic acid, B: MeCN, 0.1% formic acid Flow rate: 0.6 mL / 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

[0175] Method B Column: Waters UPLC® CSH™ C18 2.1×100 mm 1.7 μm Column temperature 40 °C Eluent: A: 2 mM ammonium bicarbonate buffered to pH 10, B: MeCN Flow rate: 0.6 mL / 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

[0176] Method C Column: Waters UPLC® BEH™ C18 2.1×100 mm 1.7 μm Column temperature 40 °C Eluent: A: 2 mM ammonium bicarbonate buffered to pH 10, B: MeCN Flow rate: 0.6 mL / 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

[0177] Method D Column: Waters Atlantis dC18 2.1×100 mm 3 μm Column temperature 40 °C Eluent: A: H 2 O + 0.1% formic acid, B: MeCN + 0.1% formic acid Flow rate: 0.6 mL / min Gradient: 0 - 5 min 5 - 100% B, 5 - 5.4 min 100% B, 5.4 - 5.42 min 100 - 5% B, 5.42 - 7.00 min 5% B

[0178] Method E Column: Kinetex Core - Shell C18 2.1×50 mm 5 μm Column temperature 40 °C Eluent: A: H 2 O + 0.1% formic acid, B: MeCN + 0.1% formic acid Flow rate: 1.2 mL / min Gradient: 0 - 1.20 min 5 - 100% B, 1.20 - 1.30 min 100% B, 1.30 - 1.31 min 100 - 5% B

[0179] Method F Column: Phenomenex Gemini - NX C18 2×50 mm 3 μm Column temperature 40 °C Eluent: A: 2 mM ammonium bicarbonate buffered to pH 10, B: MeCN Flow rate: 1 mL / min Gradient: 0 - 1.80 min 1 - 100% B, 1.80 - 2.10 min 100% B, 2.10 - 2.30 min 100 - 1% B

[0180] The following examples are intended to illustrate the present invention and should not be construed as limiting it thereto. Temperatures are given in degrees Celsius. Unless otherwise specified, all evaporations are carried out in vacuo, preferably at about 15 mmHg to 100 mmHg (= 20 to 133 mbar). The structures of the final products, intermediates, and starting materials are confirmed by standard analytical methods, such as microanalysis and spectroscopic properties, such as MS, IR, and NMR. The abbreviations used are conventional in the art. Where not defined, terms have their generally accepted meanings.

[0181] Abbreviations aq. Aqueous Br Broad d Doublet dd Double doublet DCM Dichloromethane DIPEA Diisopropylethylamine DMF N,N-Dimethylformamide EDCl 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide EtOAc Ethyl acetate HOAt 1-Hydroxy-7-azabenzotriazole HATU 2-(7-Aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HPLC High-performance liquid chromatography MeCN Acetonitrile MeOH Methanol MS Mass spectrometry m Multiplet min Minute(s) mL Milliliter(s) m / z Mass-to-charge ratio NCS N-Chlorosuccinimide NMR Nuclear magnetic resonance PTFE Polytetrafluoroethylene Rt Retention time s Singlet sPhos 2-Dicyclohexylphosphino-2’,6’-dimethoxybiphenyl t Triplet TBME Methyl tert-butyl ether TBTU N,N,N’,N’-Tetramethyl-O-(benzotriazol-1-yl)uronium tetrafluoroborate TEA Triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran T3P® Propylphosphonic anhydride

[0182] Example 1 N-(1,1-Dimethylprop-2-ynyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000028.tif301701,4-Dioxane (2 mL) was added to a stirred solution of 2-(1H-indol-6-yl)acetic acid (70 mg, 0.40 mmol), 4-amino-N-(1,1-dimethylprop-2-ynyl)pyridine-2-carboxamide (Intermediate A) (70 mg, 0.36 mmol) and TEA (95 μL, 0.54 mmol), and a 50% solution of T3P® in EtOAc (0.32 mL, 0.54 mmol) was added. The mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with EtOAc (5 mL) and washed successively with 1 M HCl (5 mL), saturated NaHCO 3 solution (5 mL) and brine. The organic layer was separated, dried over Na 2 SO 4 and concentrated in vacuo. The crude residue was purified by chromatography on silica (10 g, KP-Sil) eluting with 0–100% EtOAc in heptane to afford the title compound as a pale pink powder. LC-MS (Method A): Rt 3.01 min; MS m / z 361.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 11.03 (s, 1H), 10.76 (s, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.31 (s, 1H), 8.22 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.48 (d, J = 8.1 Hz, 1H), 7.37 (s, 1H), 7.30 (t, J = 2.7 Hz, 1H), 6.98 (dd, J = 8.1, 1.3 Hz, 1H), 6.40 - 6.37 (m, 1H), 3.76 (s, 2H), 3.21 (s, 1H), 1.65 (s, 6H).

[0183] Example 1.1 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide The title compound was prepared from 4-amino-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Intermediate AD) and 2-(1H-indol-6-yl)acetic acid in the same manner as in Example 1, using TIFF0007682850000029.tif34170 LC-MS (Method A): Rt 3.08 min; MS m / z 403.2 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 10.76 (s, 1H), 9.38 (s, 1H), 8.48 (d, J = 5.5 Hz, 1H), 8.22 (d, J = 2.0 Hz, 1H), 7.86 (dd, J = 5.5, 2.2 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 7.37 (s, 1H), 7.30 (t, 1H), 6.97 (dd, J = 8.1, 1.4 Hz, 1H), 6.39 - 6.36 (m, 1H), 3.75 (s, 2H), 1.31 - 1.27 (m, 2H), 1.20 - 1.16 (m, 2H).

[0184] Example 2 N-tert-Butyl-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000030.tif331701, A mixture of 2-(1-tetrahydropyran-2-ylindazol-6-yl)acetic acid (Intermediate B) (154 mg, 0.59 mmol) and 4-amino-N-tert-butyl-pyridine-2-carboxamide (Intermediate AB) (95 mg, 0.49 mmol) in 1,4-dioxane (4.9 mL) was treated with TEA (0.17 mL, 0.98 mmol) and a 50% solution of T3P® (1.17 mL, 0.98 mmol) in EtOAc and stirred at room temperature for 2 h. The resulting mixture was concentrated in vacuo and the residue was dissolved in EtOAc (10 mL). The organic matter was washed with saturated aqueous sodium bicarbonate (2 × 10 mL) and the combined aqueous washes were re-extracted with EtOAc (2 × 10 mL). The combined organic matter was dried over Na 2 SO 4 and concentrated in vacuo. The resulting residue was dissolved in DCM (4.9 mL), cooled (0 °C), and then treated with TFA (0.94 mL, 12.29 mmol). The mixture was warmed to room temperature and stirred for 3 h. The resulting mixture was concentrated in vacuo and the crude residue was dissolved in EtOAc (10 mL). The solution was washed with saturated aqueous sodium bicarbonate (10 mL) and dried over Na 2 SO 4 and concentrated in vacuo. Purification by preparative HPLC (acidic pH, early elution method) afforded the title compound as a pale yellow solid. LC-MS (Method A): Rt 2.71 min; MS m / z 352.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 12.99 (s, 1H), 10.79 (s, 1H), 8.44 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 8.03 - 8.01 (m, 2H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 3.84 (s, 2H), 1.39 (s, 9H).

[0185] Example 2.1 N-(1-Cyano-1-methyl-ethyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000031.tif36170 The title compound was prepared from 4-amino-N-(1-cyano-1-methyl-ethyl)pyridine-2-carboxamide (Intermediate AC) and 2-(1-tetrahydropyran-2-ylindazol-6-yl)acetic acid (Intermediate B) in the same manner as in Example 2. LC-MS (Method A): Rt 2.32 min; MS m / z 363.2 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6 ) δ 12.99 (s, 1H), 10.84 (s, 1H), 8.83 (s, 1H), 8.51 (d, J = 5.5 Hz, 1H), 8.24 (d, J = 2.1 Hz, 1H), 8.02 (s, 1H), 7.87 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.50 (s, 1H), 7.09 (dd, J = 8.3, 1.2 Hz, 1H), 3.85 (s, 2H), 1.72 (s, 6H).

[0186] Example 3 N-(1,1-Dimethylprop-2-ynyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000032.tif38170

[0187] Process 1: N-(1,1-dimethylprop-2-ynyl)-4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000033.tif50170 The title compound was prepared from 4-amino-N-(1,1-dimethylprop-2-ynyl)pyridine-2-carboxamide (Intermediate A) and 2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetic acid (Intermediate B) in the same manner as in Example 1. LC-MS (Method E): Rt 1.15 min; MS m / z 446.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.84 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 8.31 (s, 1H), 8.21 (d, J = 1.5 Hz, 1H), 8.07 (s, 1H), 7.84 (dd, J = 5.5, 1.7 Hz, 1H), 7.72 (d, J = 8.3 Hz, 1H), 7.66 (s, 1H), 7.16 (d, J = 8.3 Hz, 1H), 5.81 (d, J = 9.6 Hz, 1H), 3.92 - 3.86 (m, 3H), 3.78 - 3.68 (m, 1H), 3.20 (s, 1H), 2.46 - 2.37 (m, 1H), 2.09 - 1.93 (m, 2H), 1.82 - 1.69 (m, 1H), 1.64 (s, 6H), 1.60 - 1.55 (m, 2H).

[0188] Process 2: N-(1,1-dimethylprop-2-ynyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide A cooled (0 °C) solution of N-(1,1-dimethylprop-2-ynyl)-4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide in DCM (12 mL) (step 1) (83%, 950 mg, 1.77 mmol) was treated with TFA (1.74 mL, 22.79 mmol). The mixture was warmed to room temperature and stirred for 3 h. Additional TFA (0.87 mL, 11.40 mmol) was added and the reaction was continued for 3 h. The resulting mixture was concentrated in vacuo. The crude residue was dissolved in EtOAc (30 mL) and washed with saturated NaHCO 3 solution (20 mL), brine (20 mL), dried over Na 2 SO 4 and concentrated in vacuo. Purification by C18 reverse-phase chromatography eluting with 10 - 100% MeCN (0.1% formic acid) and water (0.1% formic acid) afforded the title compound as an off-white solid. LC-MS (method A): Rt 2.52 min; MS m / z 362.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.99 (s, 1H), 10.82 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 8.31 (s, 1H), 8.21 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.84 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.09 (dd, J = 8.3, 1.2 Hz, 1H), 3.84 (s, 2H), 3.21 (s, 1H), 1.64 (s, 6H).

[0189] Example 4 N-(1-Ethynylcyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000034.tif37170

[0190] Project 1: Mixture of methyl 4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylate and methyl 4-[[2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylate TIFF0007682850000035.tif103170The title compound was prepared from methyl 4-aminopyridine-2-carboxylate and 2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetic acid (Intermediate B1) in the same manner as in Example 1. LC-MS (Method E): Rt 0.97, 0.99 min; MS m / z 395.2 = [M+H]+: two peaks for positional isomers

[0191] Project 2: Mixture of 4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid TIFF0007682850000036.tif50170To a solution of a mixture of methyl 4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylate and methyl 4-[[2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylate (Project 1) (80%, 1.76 g, 3.57 mmol) in THF (6 mL), MeOH (6 mL) and water (6 mL) was added LiOH.H 2 O (180 mg, 4.28 mmol), and the mixture was stirred at room temperature for 2 h. Additional LiOH.H 2O (45 mg, 1.07 mmol) was added and the mixture was stirred for an additional 1 h at room temperature. The resulting mixture was acidified to pH 5 using 1 M HCl and then diluted with brine (15 mL) and EtOAc (15 mL), whereupon a precipitate formed, which was collected by vacuum filtration. The phases of the filtrate were separated, the aqueous phase was extracted with EtOAc (15 mL), and then the combined organics were concentrated in vacuo. The resulting residue was triturated in ether (10 mL), filtered, and the solid was combined with that from the previous filtration. The aqueous phase was cooled to 2 - 8 °C and allowed to stand for 3 days. The resulting suspension was filtered and the solid was combined with that from the previous filtration. The combined solids were dried in a vacuum oven to afford the title compound as a colorless solid. LC-MS (Method E): Rt 0.86 min; MS m / z 381.1 = [M+H]+

[0192] Step 3: N-(1-Ethynylcyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide To a solution of a mixture of 4-[[2-(1-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 2) (99%, 150 mg, 0.39 mmol) and 1-ethynylcyclopentanamine hydrochloride (68 mg, 0.47 mmol) in DMF (2 mL), DIPEA (136 μL, 0.78 mmol) was added, followed by HATU (163 mg, 0.43 mmol), and the mixture was stirred at room temperature for 3 h. The resulting mixture was diluted with EtOAc (10 mL) and washed with saturated NaHCO 3 solution (10 ml), brine (10 mL), and then dried over Na 2 SO 4 and concentrated in vacuo. The crude material was dissolved in DCM (2 mL), treated with TFA (1.0 mL, 13.07 mmol), and stirred at room temperature for 2 h. The reaction mixture was concentrated in vacuo and the residue was dissolved in EtOAc. The organic mixture was washed with saturated NaHCO 3 solution and Na2 SO 4 It was dried above and concentrated in vacuo. Purification of the crude residue by preparative HPLC (acidic pH, early elution method) gave the title compound as a colorless powder. LC-MS (Method A): Rt 2.83 min; MS m / z 388.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.00 (s, 1H), 10.82 (s, 1H), 8.52 - 8.44 (m, 2H), 8.21 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.71 (d, J = 8.3 Hz, 1H), 7.50 (s, 1H), 7.09 (dd, J = 8.3, 1.2 Hz, 1H), 3.85 (s, 2H), 3.17 (s, 1H), 2.30 - 2.21 (m, 2H), 2.13 - 2.06 (m, 2H), 1.77 - 1.65 (m, 4H).

[0193] The compounds of the following table of Examples (Table 1) were prepared in the same manner as in Step 3 of Example 4 from a mixture of 4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 2 of Example 4), and appropriate commercially available amines. TIFF0007682850000037.tif253170TIFF0007682850000038.tif252170

[0194] Example 5 N-(2,2-Difluoro-1,1-dimethyl-ethyl)-4-[[2-( 1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000039.tif341701, A mixture of 4-[[2-(1-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (step 2 of Example 4) (60 mg, 0.16 mmol) in 1,4-dioxane (1.2 mL), 1,1-difluoro-2-methyl-propan-2-amine hydrochloride (28 mg, 0.19 mmol) and a suspension of DIPEA (69 μL, 0.39 mmol) were treated with a 50% solution of T3P® in EtOAc (235 μL, 0.39 mmol) and stirred at room temperature for 2 hours. The resulting mixture was partitioned between DCM (5 mL) and water (5 mL), and the organic portion was separated by filtering through a hydrophobic PTFE frit tube. The filtrate was concentrated in vacuo and the residue was dissolved in TFA (50% in DCM) (2.0 mL, 0.16 mmol). The resulting mixture was stirred for 4 hours and then diluted with DCM (10 mL). Saturated NaHCO 3 An aqueous solution (10 mL) was added slowly and the free base mixture was separated by filtering through a hydrophobic PTFE frit tube. The organic portion was concentrated in vacuo and the residue was purified by chromatography on basic silica eluting with 0 - 100% EtOAc in heptane followed by 0 - 100% MeOH in EtOAc. The resulting material was further purified by C18 reverse phase chromatography eluting with 10 - 100% MeCN in water with 0.1% formic acid modifier. The fractions containing the product were combined, concentrated in vacuo to remove the volatile solvents and then treated with saturated NaHCO 3 An aqueous solution (10 mL) and DCM (10 mL). The organic portion was separated by filtering through a hydrophobic PTFE frit tube and concentrated in vacuo to give the title compound as a white crystalline solid. LC-MS (method A): Rt 2.83 min; MS m / z 388.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 13.00 (s, 1H), 10.83 (s, 1H), 8.48 (d, J = 5.5 Hz, 1H), 8.29 (s, 1H), 8.23 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 7.83 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 6.46 (t, J = 57.0 Hz, 1H), 3.84 (s, 2H), 1.44 (s, 6H).

[0195] The compounds of the following examples (Table 2) were prepared in the same manner as in Example 5 from a mixture of 4-[[2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 2 of Example 4), and appropriate commercially available amines. TIFF0007682850000040.tif255170TIFF0007682850000041.tif255170

[0196] Example 6 N-tert-butyl-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000042.tif33170

[0197] Step 1: A mixture of 6-bromo-5-fluoro-1-[(4-methoxyphenyl)methyl]indazole and 6-bromo-5-fluoro-2-[(4-methoxyphenyl)methyl]indazole TIFF0007682850000043.tif39170 6-Bromo-5-fluoro-1H-indazole (1.0 g, 4.65 mmol), K in acetone (100 mL) 2 CO 3(964 mg, 6.98 mmol) and potassium iodide (849 mg, 5.12 mmol) were treated with 1-(chloromethyl)-4-methoxy-benzene (694 μL, 5.12 mmol), and the mixture was heated at 45 °C for 4 h and then stirred at room temperature for 2 days. The resulting mixture was concentrated in vacuo and the residue was dissolved in EtOAc (50 mL). The organic mixture was washed with water (2 × 50 mL), brine (2 × 50 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with a gradient of 0 - 100% EtOAc in heptane to afford the title positional isomer mixture as a yellow solid. LC-MS (Method E): Rt 1.22, 1.26 min; MS m / z 335.0, 337.0 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.47 (d, J = 0.7 Hz, 0.4H), 8.25 (d, J = 5.6 Hz, 0.6H), 8.10 (d, J = 0.9 Hz, 0.6H), 8.04 (d, J = 6.2 Hz, 0.4H), 7.74 (d, J = 8.8 Hz, 0.6H), 7.66 (d, J = 9.3 Hz, 0.4H), 7.34 - 7.29 (m, 0.8H), 7.25 - 7.19 (m, 1.2H), 6.93 - 6.89 (m, 0.8H), 6.89 - 6.84 (m, 1.2H), 5.58 (s, 1.2H), 5.55 (s, 0.8H), 3.72 (s, 1.2H), 3.70 (s, 1.8H). (Ratio of positional isomers 0.4:0.6)

[0198] Step 2: Mixture of 2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid and 2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid A solution of a mixture of 6-bromo-5-fluoro-1-[(4-methoxyphenyl)methyl]indazole and 6-bromo-5-fluoro-2-[(4-methoxyphenyl)methyl]indazole (1200 mg, 3.58 mmol) and potassium 3-ethoxy-3-oxopropanoate (914 mg, 5.37 mmol) in mesitylene (30 mL) was added with DMAP (44 mg, 0.36 mmol). Nitrogen was injected into the resulting mixture for 5 minutes and the mixture was stirred at room temperature. Diallyldichloropalladium (26 mg, 0.07 mmol) and BINAP (134 mg, 0.21 mmol) were added and nitrogen was further injected into the mixture for 5 minutes. The resulting mixture was stirred at 140 °C for 16 hours. After cooling to room temperature, the mixture was diluted with MeOH (50 mL), treated with 2 M LiOH (25 mL) and stirred at room temperature for 2 hours. The resulting mixture was concentrated in vacuo and the crude material was partitioned between water and EtOAc. The layers were separated and the organic portion was extracted with water. The combined aqueous portions were acidified to pH 4 using 2 M HCl and then re-extracted with EtOAc (2 × 50 mL). The organic extracts were dried over Na 2 SO 4 and concentrated in vacuo to afford the title mixture of positional isomers as a yellow solid. LC-MS (Method E): Rt 1.02, 1.04 min; MS m / z 315.1 = [M+H]+; two positional isomers

[0199] Step 3: A mixture of methyl 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylate and methyl 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylate TIFF0007682850000045.tifA solution of a mixture of 2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid and 2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid (975 mg, 3.1 mmol) in 1,4-dioxane (25 mL) (step 2) was added with methyl 4-aminopyridine-2-carboxylate (472 mg, 3.1 mmol), DIPEA (1.08 mL, 6.2 mmol), and then a 50% solution of T3P® in EtOAc (1.02 mL, 3.41 mmol), and the reaction mixture was stirred at room temperature for 16 h under an inert atmosphere. The resulting mixture was diluted with EtOAc (100 mL) and washed with water (2 × 50 mL). The organic extracts were combined, dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with a gradient of 0 - 100% EtOAc in heptane to afford the title positional isomer mixture as an off-white solid. LC-MS (method E): Rt 1.04, 1.06 min; MS m / z 449.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 10.85 (s, 0.75H), 10.80 (s, 0.25H), 8.57 - 8.54 (m, 1H), 8.38 (s, 0.25H), 8.30 (d, J = 2.0 Hz, 1H), 8.06 - 8.05 (m, 0.75H), 7.78 (dd, J = 5.5, 2.1 Hz, 1H), 7.74 (d, J = 5.9 Hz, 0.75H), 7.62 (d, J = 6.6 Hz, 0.25H), 7.54 (d, J = 9.9 Hz, 0.75H), 7.42 (d, J = 10.5 Hz, 0.25H), 7.31 - 7.27 (m, 0.5H), 7.21 - 7.17 (m, 1.5H), 6.92 - 6.88 (m, 0.5H), 6.86 - 6.81 (m, 1.5H), 5.55 (s, 1.5H), 5.54 (s, 0.5H), 3.91 (s, 2H), 3.86 (s, 2.25H), 3.86 (s, 0.75H), 3.72 (s, 0.75H), 3.68 (s, 2.25H). Ratio of positional isomers 0.75:0.25

[0200] Step 4: A mixture of 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid A solution of a mixture of methyl 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and methyl 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (830 mg, 1.85 mmol) in THF (20 mL) was added 1 M LiOH (4.63 mL, 4.63 mmol), and the reaction mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated in vacuo, and the residue was redissolved in water (20 mL). The aqueous portion was acidified to pH 4 using 6 M HCl. The resulting white precipitate was filtered, washed with water (20 mL), and dried under vacuum at 40 °C to give the title positional isomer mixture as a white solid. LC-MS (Method E): Rt 0.95 min; MS m / z 435.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.84 (s, 0.7H), 10.79 (s, 0.3H), 8.54 - 8.49 (m, 1H), 8.38 - 8.36 (m, 0.3H), 8.25 - 8.23 (m, 1H), 8.04 (d, J = 0.8 Hz, 0.7H), 7.80 - 7.77 (m, 1H), 7.74 (d, J = 5.9 Hz, 0.7H), 7.61 (d, J = 6.5 Hz, 0.3H), 7.53 (d, J = 9.9 Hz, 0.7H), 7.41 (d, J = 10.5 Hz, 0.3H), 7.30 - 7.26 (m, 0.6H), 7.21 - 7.14 (m, 1.4H), 6.91 - 6.88 (m, 0.6H), 6.85 - 6.81 (m, 1.4H), 5.54 (s, 1.4H), 5.54 (s, 0.6H), 3.90 (s, 1.4H), 3.86 (s, 0.6H), 3.71 (s, 0.9H), 3.67 (s, 2.1H). Ratio of positional isomers 0.3:0.7.

[0201] Step 5: N-tert-Butyl-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide To a solution of a mixture of 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Step 4) (200 mg, 0.46 mmol) in DMF (5 mL) was added HATU (193 mg, 0.51 mmol), followed by DIPEA (161 μL, 0.92 mmol). After stirring for 5 minutes, 2-methylpropan-2-amine (37 mg, 0.51 mmol) was added and the mixture was stirred at room temperature for 2 hours under an inert atmosphere. The resulting mixture was diluted with EtOAc (20 mL) and washed with water (2 × 20 mL), brine (20 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude material was dissolved in DCE (5 mL), TFA (1.76 mL, 23.02 mmol) was added, and the mixture was stirred at 75 °C for 16 hours. The resulting mixture was concentrated in vacuo and azeotroped with toluene (2 mL). The crude material was purified by C18 reverse-phase chromatography eluting with 10 - 100% MeCN in water with 0.1% formic acid modifier to afford the title compound as a pale yellow solid. LC-MS (Method A): Rt 2.76 min; MS m / z 370.2 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6 ) δ 13.12 (br. s, 1H), 10.84 (s, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 8.06 - 8.02 (m, 2H), 7.82 (dd, J = 5.6, 2.2 Hz, 1H), 7.57 (d, J = 6.1 Hz, 1H), 7.52 (d, J = 10.1 Hz, 1H), 3.93 (s, 2H), 1.40 (s, 9H).

[0202] Example 6.1 4-[[2-(5-Fluoro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000047.tif33170 The title compound was prepared from a mixture of 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Example 6, Step 4) and 1-(trifluoromethyl)cyclopropanamine hydrochloride in the same manner as in Step 5 of Example 6. LC-MS (Method A): Rt 2.68 min; MS m / z 422.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.12 (s, 1H), 10.85 (s, 1H), 9.38 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.57 (d, J = 6.0 Hz, 1H), 7.52 (d, J = 10.1 Hz, 1H), 3.93 (s, 2H), 1.33 - 1.27 (m, 2H), 1.21-1.17 (m, 2H).

[0203] Example 6.2 N-(2,2-Difluorocyclopentyl)-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000048.tif The title compound was prepared from a mixture of 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Example 6, step 4) and 2,2-difluorocyclopentanamine hydrochloride in the same manner as in step 5 of Example 6. LC-MS (Method A): Rt 2.73 min; MS m / z 418.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.12 (s, 1H), 10.85 (s, 1H), 8.61 (d, J = 9.3 Hz, 1H), 8.52 (d, J = 5.5 Hz, 1H), 8.25 (d, J = 2.1 Hz, 1H), 8.04 (s, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.58 (d, J = 6.0 Hz, 1H), 7.52 (d, J = 10.1 Hz, 1H), 4.67 - 4.52 (m, 1H), 3.94 (s, 2H), 2.24 - 2.05 (m, 3H), 1.91 - 1.65 (m, 3H).

[0204] Example 6.3 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000049.tif36170

[0205] Steps 1 - 4: 4-[[2-[4-Chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid TIFF0007682850000050.tif The title compound was prepared in the same manner as the mixture of 4-[[2-[5-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-[5-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Steps 1-4 of Example 6) by replacing 6-bromo-5-fluoro-1H-indazole (Step 1) with 6-bromo-4-chloro-1H-indazole. LC-MS (Method E): Rt 1.00 min; MS m / z 451.1, 453.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.98 (s, 1H), 8.55 (d, J = 5.6 Hz, 1H), 8.30 (d, J = 2.0 Hz, 1H), 8.12 (d, J = 0.8 Hz, 1H), 7.84 (dd, J = 5.6, 2.1 Hz, 1H), 7.69 (s, 1H), 7.22 (d, J = 0.8 Hz, 1H), 7.22 - 7.18 (m, 2H), 6.85 - 6.81 (m, 2H), 5.57 (s, 2H), 3.89 (s, 2H), 3.67 (s, 3H).

[0206] Step 5: 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide The title compound was prepared from 4-[[2-[4-chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Steps 1-4) and 1-(trifluoromethyl)cyclopropanamine hydrochloride in the same manner as in Step 5 of Example 6. LC-MS (Method A): Rt 3.03 min; MS m / z 438.1, 440.1 = [M+H]+ 11H NMR (500 MHz, DMSO-d 6 ) δ 13.38 (s, 1H), 10.83 (s, 1H), 9.39 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.20 (d, J = 2.0 Hz, 1H), 8.09 (s, 1H), 7.86 (dd, J = 5.5, 2.2 Hz, 1H), 7.48 (s, 1H), 7.19 (d, J = 0.9 Hz, 1H), 3.87 (s, 2H), 1.32 - 1.27 (m, 2H), 1.21 - 1.15 (m, 2H).

[0207] Example 6.4 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]-N-(1-ethynylcyclopentyl)pyridine-2-carboxamide TIFF0007682850000051.tif34170

[0208] Step 1: 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid TIFF0007682850000052.tif35170TFA (0.5 mL, 6.53 mmol) was added to a solution of 4-[[2-[4-chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxylic acid (Steps 1 - 4 of Example 6.3) (80 mg, 0.18 mmol) in DCE (1 mL), and the mixture was heated at 75 °C for 40 h. The resulting mixture was diluted with DCM (2 mL), concentrated in vacuo, and azeotroped with DCM (3 × 5 mL). The crude residue was suspended in MeCN (1 mL), filtered, washed with MeCN (3 × 1 mL), and dried to afford the title compound as a light brown solid. LC-MS (Method E): Rt 0.86 min; MS m / z 331.0, 333.0 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6) δ 13.40 (s, 1H), 10.96 (s, 1H), 8.55 (d, J = 5.7 Hz, 1H), 8.30 (d, J = 2.0 Hz, 1H), 8.09 (s, 1H), 7.86 (dd, J = 5.7, 2.1 Hz, 1H), 7.48 (s, 1H), 7.19 (s, 1H), 1H), 3.89 (s, 2H).

[0209] Step 2: 4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]-N-(1-ethynylcyclopentyl)pyridine-2-carboxamide The title compound was prepared from 4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 1) and 1-ethynylcyclopropanamine hydrochloride in the same manner as in Step 5 of Example 6. LC-MS (Method A): Rt 3.22 min; MS m / z 422.2, 424.2 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6 ) δ 13.38 (s, 1H), 10.82 (s, 1H), 8.50 - 8.44 (m, 2H), 8.19 (d, J = 2.0 Hz, 1H), 8.09 (s, 1H), 7.84 (dd, J = 5.5, 2.2 Hz, 1H), 7.48 (s, 1H), 7.19 (s, 1H), 3.87 (s, 2H), 3.16 (s, 1H), 2.28 - 2.20 (m, 2H), 2.13 - 2.05 (m, 2H), 1.77 - 1.63 (m, 4H).

[0210] Example 7 N-tert-butyl-4-[[2-(4-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000053.tif33170

[0211] Project 1: Mixture of 6-bromo-4-fluoro-1-[(4-methoxyphenyl)methyl]indazole and 6-bromo-4-fluoro-2-[(4-methoxyphenyl)methyl]indazole TIFF0007682850000054.tif41170 The title compound was prepared from 6-bromo-4-fluoro-1H-indazole and 1-(chloromethyl)-4-methoxy-benzene in the same manner as in Step 1 of Example 6. LC-MS (Method E): Rt 1.31, 1.35 min; MS m / z 335.0, 337.0 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.73 (d, J = 0.7 Hz, 0.4H), 8.24 (d, J = 0.8 Hz, 0.6H), 8.00 - 7.97 (m, 0.6H), 7.75 - 7.71 (m, 0.4H), 7.36 - 7.31 (m, 0.8H), 7.25 - 7.21 (m, 1.2H), 7.19 (dd, J = 9.6, 1.2 Hz, 0.6H), 7.03 (dd, J = 9.9, 1.2 Hz, 0.4H), 6.93 - 6.89 (m, 0.8H), 6.89 - 6.85 (m, 1.2H), 5.60 (s, 1.2H), 5.56 (s, 0.8H), 3.72 (s, 1.2H), 3.70 (s, 1.8H). Ratio of positional isomers 0.4:0.6.

[0212] Project 2: Mixture of 2-[4-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid and 2-[4-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid. TIFF0007682850000055.tif48170 The title compound was prepared from a mixture of 6-bromo-4-fluoro-1-[(4-methoxyphenyl)methyl]indazole and 6-bromo-4-fluoro-2-[(4-methoxyphenyl)methyl]indazole (Step 1), and potassium 3-ethoxy-3-oxo-propanoate in the same manner as in Step 2 of Example 6. LC-MS (Method E): Rt 1.07, 1.10 min; MS m / z 315.0 = [M+H]+; two positional isomers

[0213] Step 3: N-tert-butyl-4-[[2-(4-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide To a solution of the mixture of 2-[4-fluoro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid and 2-[4-fluoro-2-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid from Step 2 (87%, 114 mg, 0.32 mmol), 4-amino-N-tert-butyl-pyridine-2-carboxamide (Intermediate AB) (61 mg, 0.32 mmol) and DIPEA (110 μL, 0.63 mmol) in 1,4-dioxane (5 mL) was added a 50% solution of T3P® in EtOAc (103 μL, 0.35 mmol), and the reaction mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated in vacuo and the residue was dissolved in EtOAc (20 mL). The organic mixture was washed with water (20 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude material was dissolved in DCE (2 mL), then TFA (1.21 mL, 15.78 mmol) was added and the mixture was heated at 75 °C for 48 h. The resulting mixture was concentrated in vacuo and the crude residue was purified by C18 reverse phase chromatography eluting with 10–100% MeCN in water containing 0.1% formic acid modifier to afford the title compound as an off-white solid. LC-MS (Method A): Rt 2.91 min; MS m / z 370.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 13.35 (s, 1H), 10.81 (s, 1H), 8.45 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.1 Hz, 1H), 8.13 (s, 1H), 8.02 (s, 1H), 7.82 (dd, J = 5.5, 2.1 Hz, 1H), 7.34 (s, 1H), 6.88 (d, J = 11.2 Hz, 1H), 3.86 (s, 2H), 1.39 (s, 9H).

[0214] Example 7.1 N-tert-butyl-4-[[2-(5-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000056.tif36170 The title compound was prepared in the same manner as N-tert-butyl-4-[[2-(4-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide [Example 7 (Steps 1 to 3)] by replacing 6-bromo-5-fluoro-1H-indazole (Step 1) with 6-bromo-5-chloro-1H-indazole. LC-MS (Method A): Rt 2.96 min; MS m / z 386.1, 388.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.22 (s, 1H), 10.86 (s, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.20 (d, J = 2.0 Hz, 1H), 8.04 (d, J = 7.3 Hz, 2H), 7.88 (s, 1H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.64 (s, 1H), 4.03 (s, 2H), 1.40 (s, 9H).

[0215] Example 7.2 N-tert-butyl-4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000057.tif34170

[0216] Step 1: 6-Bromo-4-chloro-1-[(4-methoxyphenyl)methyl]indazole TIFF0007682850000058.tif34170 A mixture of 6-bromo-4-chloro-1H-indazole (500 mg, 2.16 mmol) and Cs 2 CO 3 (1.06 mg, 3.24 mmol) in DMF (10 mL) was treated with 1-(chloromethyl)-4-methoxy-benzene (0.35 mL, 2.59 mmol), and the resulting mixture was stirred at room temperature for 16 h. The resulting mixture was diluted with EtOAc (50 mL) and water (50 mL), and the phases were separated. The organic portion was washed with brine (50 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with 0 - 80% EtOAc in heptane to afford the title compound as an orange solid. LC-MS (Method E): Rt 1.40 min; MS m / z 350.9, 352.9, 354.9 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.18 (d, J = 0.9 Hz, 1H), 8.13 (t, J = 1.1 Hz, 1H), 7.43 (d, J = 1.3 Hz, 1H), 7.24 - 7.20 (m, 2H), 6.89 - 6.85 (m, 2H), 5.60 (s, 2H), 3.70 (s, 3H).

[0217] Step 2: 2-[4-Chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid TIFF0007682850000059.tif36170 The title compound was prepared from 6-bromo-4-chloro-1-[(4-methoxyphenyl)methyl]indazole (Step 1) and potassium 3-ethoxy-3-oxo-propanoate in the same manner as in Step 2 of Example 6. LC-MS (Method E): Rt 1.16 min; MS m / z 331.0, 333.0 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.44 (s (br), 1H), 8.10 (d, J = 0.8 Hz, 1H), 7.63 (s, 1H), 7.22 - 7.19 (m, 2H), 7.15 (d, J = 0.9 Hz, 1H), 6.88 - 6.84 (m, 2H), 5.56 (s, 2H), 3.71 (s, 2H), 3.70 (s, 3H).

[0218] Step 3: N-tert-butyl-4-[[2-[4-chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide TIFF0007682850000060.tif441704-Amino-N-tert-butyl-pyridine-2-carboxamide (Intermediate AB) (28 mg, 0.14 mmol), a mixture of 2-[4-chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid (Step 2) (99%, 43 mg, 0.13 mmol) and DIPEA (0.045 mL, 0.26 mmol) in 1,4-dioxane (1 mL) was treated with a 50% solution of T3P® in EtOAc (50%, 0.093 mL, 0.16 mmol), and the mixture was stirred at room temperature for 30 minutes. The resulting mixture was diluted with EtOAc (15 mL) and 1:1 water / brine (15 mL), and the phases were separated. The organic portion was dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with 0 - 100% EtOAc in heptane to afford the title compound as a pale yellow waxy solid. LC-MS (Method E): Rt 1.34 min; MS m / z 506.2, 508.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6) δ 10.80 (s, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.18 (dd, J = 1.8, 0.4 Hz, 1H), 8.12 (d, J = 0.9 Hz, 1H), 8.03 (s, 1H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.68 (s, 1H), 7.23 - 7.18 (m, 3H), 6.86 - 6.81 (m, 2H), 5.58 (s, 2H), 3.87 (s, 2H), 3.67 (s, 3H), 1.40 (s, 9H).

[0219] Step 4: N-tert-Butyl-4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide A solution of N-tert-butyl-4-[[2-[4-chloro-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Step 3) (100%, 40 mg, 0.08 mmol) in DCE (1 mL) was treated with TFA (0.3 mL, 3.91 mmol), and the mixture was heated at 75 °C overnight. Additional TFA (0.3 mL, 3.91 mmol) was added, and stirring was continued at 75 °C for an additional 24 h. The resulting mixture was concentrated in vacuo, azeotroped with DCM (2 × 5 mL), and purified by C18 reverse-phase chromatography eluting with 10 - 100% MeCN in water with 0.1% formic acid modifier to afford the title compound as a pale yellow solid. LC-MS (Method A): Rt 3.12 min; MS m / z 386.2, 388.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 13.38 (s, 1H), 10.80 (s, 1H), 8.45 (d, J = 5.5 Hz, 1H), 8.18 (d, J = 2.0 Hz, 1H), 8.09 (s, 1H), 8.03 (s, 1H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.48 (s, 1H), 7.19 (d, J = 0.7 Hz, 1H), 3.87 (s, 2H), 1.39 (s, 9H).

[0220] Example 8 N-tert-Butyl-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide TIFF0007682850000061.tif36170

[0221] Step 1: 6-Bromo-1-[(4-methoxyphenyl)methyl]-3-(trifluoromethyl)indazole TIFF0007682850000062.tif631706-bromo-3-(trifluoromethyl)-1H-indazole (1.0 g, 3.77 mmol), K 2 CO 3 (782 mg, 5.66 mmol) and potassium iodide (689 mg, 4.15 mmol) in a mixture of acetone (100 mL), followed by 1-(chloromethyl)-4-methoxy-benzene (0.56 mL, 4.15 mmol) was added, and the mixture was heated at 40 °C for 2 hours. The resulting mixture was concentrated in vacuo. The dry crude residue was dissolved in EtOAc (30 mL), washed with brine (30 mL), and dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica (50 g of KP-Sil) eluting with 0 - 6% EtOAc in heptane to give the title compound as an off-white solid. LC-MS (Method E): Rt 1.44 min; no MS m / z ions observed 1 H NMR (400 MHz, DMSO- d6 ) δ 8.35 (dd, J = 1.6, 0.6 Hz, 1H), 7.76 (dt, J = 8.7, 0.8, 1H), 7.49 (dd, J = 8.7, 1.6 Hz, 1H), 7.31 - 7.25 (m, 2H), 6.93 - 6.88 (m, 2H), 5.70 (s, 2H), 3.71 (s, 3H).

[0222] Step 2: 2-[1-[(4-Methoxyphenyl)methyl]-3-(trifluoromethyl)-1H-indazol-6-yl]acetic acid A mixture of 6-bromo-1-[(4-methoxyphenyl)methyl]-3-(trifluoromethyl)-1H-indazole (Step 1) (250 mg, 0.65 mmol), potassium 2-cyanoacetate (120 mg, 0.97 mmol), diallyldipalladium dichloride (10 mg, 0.03 mmol) and SPhos (16 mg, 0.04 mmol) in toluene (5 mL) was degassed with nitrogen for 5 minutes. The resulting mixture was heated at 140 °C for 1.5 h using microwave irradiation. After cooling to room temperature, the mixture was concentrated in vacuo and the residue was dissolved in 1,4-dioxane (5 mL) and 1 M aqueous NaOH (5 mL). The resulting mixture was heated to reflux (130 °C) for 2 h, then the volatile organic solvents were removed in vacuo. The remaining basic aqueous solution was extracted with EtOAc (2 × 10 mL) and the organics were discarded. The aqueous phase was acidified to pH 2 using 1 M HCl and then extracted with EtOAc (3 × 15 mL). The combined organic extracts were dried over Na 2 SO 4 and concentrated in vacuo to afford the title compound as an off-white solid. LC-MS (Method E): Rt 1.17 min; MS m / z 365.0 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 7.83 (s, 1H), 7.74 (d, J = 8.4 Hz, 1H), 7.29 - 7.23 (m, 3H), 6.92 - 6.87 (m, 2H), 5.66 (s, 2H), 3.75 (s, 2H), 3.70 (s, 3H).

[0223] Step 3: N-tert-Butyl-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide The title compound was prepared from 2-[1-[(4-methoxyphenyl)methyl]-3-(trifluoromethyl)indazol-6-yl]acetic acid (Step 2) and (4-amino-N-tert-butyl-pyridine-2-carboxamide (Intermediate AB) in the same manner as in Step 3 of Example 7. LC-MS (Method A): Rt 3.44 min; MS m / z 420.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.96 (s, 1H), 10.86 (s, 1H), 9.39 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 1.6 Hz, 1H), 7.85 (dd, J = 5.4, 1.9 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.66 (s, 1H), 7.31 (d, J = 8.4 Hz, 1H), 3.91 (s, 2H), 1.32 - 1.27 (m, 2H), 1.21 - 1.16 (m, 2H).

[0224] Example 8.1 N-[1-(Trifluoromethyl)cyclopropyl]-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide TIFF0007682850000064.tif The title compound was prepared from 2-[1-[(4-methoxyphenyl)methyl]-3-(trifluoromethyl)-1H-indazol-6-yl]acetic acid (Example 8, Step 2) and 4-amino-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Intermediate AD) in the same manner as in Step 3 of Example 7. LC-MS (Method A): Rt 3.44 min; MS m / z 472.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.96 (s, 1H), 10.86 (s, 1H), 9.39 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 1.6 Hz, 1H), 7.85 (dd, J = 5.4, 1.9 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.66 (s, 1H), 7.31 (d, J = 8.4 Hz, 1H), 3.91 (s, 2H), 1.30 (t, J = 6.8 Hz, 2H), 1.18 (s, 2H).

[0225] Example 9 N-tert-Butyl-4-[[2-(3-isopropyl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000065.tif34170

[0226] Step 1: 2-[3-Iodo-1-[(4-methoxyphenyl)methyl]-1H-indazol-6-yl]acetic acid A mixture of 2-(3-iodo-1H-indazol-6-yl)acetic acid (Intermediate C) (728 mg, 2.41 mmol), potassium iodide (880 mg, 5.3 mmol) and potassium carbonate (999 mg, 7.23 mmol) in acetone (12.0 mL) was treated with 1-(chloromethyl)-4-methoxy-benzene (830 mg, 5.3 mmol), heated at 45 °C for 24 h, then stirred at room temperature for 2 days. The resulting mixture was filtered and washed with EtOAc (30 mL). The filtrate was diluted with water (30 mL) and the phases were separated. The organic layer was washed with brine (30 mL) and then concentrated in vacuo. The residue was treated with 1 M LiOH (7.23 mL, 7.23 mmol) and THF (8 mL). After stirring at room temperature for 2 h, the volatile solvents were removed in vacuo and the pH was adjusted to about 5 / 6 with 1 M HCl to precipitate. The solid was filtered, washed with water and dried to give the title compound as an off-white solid. LC-MS (Method E): Rt 1.16 min; MS m / z 422.9 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.58 (s, 1H), 7.29 (d, J = 8.3 Hz, 1H), 7.23 - 7.16 (m, 2H), 7.12 (dd, J = 8.4, 1.2 Hz, 1H), 6.91 - 6.83 (m, 2H), 5.52 (s, 2H), 3.69 (s, 3H), 3.56 (s, 2H).

[0227] Step 2: N-tert-butyl-4-[[2-[3-iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide TIFF0007682850000067.tif661701, 2-[3-Iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid (Step 1) (84%, 428 mg, 0.85 mmol) and 4-amino-N-tert-butyl-pyridine-2-carboxamide (Intermediate AB) (181.09 mg, 0.94 mmol) in dioxane (8.5 mL) were treated with TEA (0.30 mL, 1.7 mmol), followed by a 50% solution of T3P® in EtOAc (1.01 mL, 1.7 mmol), and the reaction mixture was stirred at room temperature for 18 h. The volatile solvents were removed in vacuo, and the residue was dissolved in EtOAc (20 mL). The organic solution was washed with saturated aqueous sodium bicarbonate (2 × 15 mL), and the combined aqueous washes were extracted with EtOAc (3 × 10 mL). The combined organic extracts were dried over Na 2 SO 4 and concentrated in vacuo. Purification by chromatography on silica eluting with 0 - 100% EtOAc in heptane afforded the title compound as a colorless solid. LC-MS (Method E): Rt 1.33 min; MS m / z 598.0 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.82 (s, 1H), 8.45 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.71 (s, 1H), 7.38 (d, J = 8.3 Hz, 1H), 7.24 - 7.17 (m, 3H), 6.87 - 6.82 (m, 2H), 5.56 (s, 2H), 3.87 (s, 2H), 3.68 (s, 3H), 1.39 (s, 9H).

[0228] Step 3: N-tert-Butyl-4-[[2-[3-isopropenyl-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide TIFF0007682850000068.tif66170N-tert-butyl-4-[[2-[3-iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Step 2) (158 mg, 0.26 mmol), Pd(OAc) 2 (12 mg, 0.05 mmol), P(Cy) 3 (30 mg, 0.11 mmol) and tripotassium phosphate (225 mg, 1.06 mmol) in a mixture of nitrogen atmosphere, toluene (2.7 mL), followed by 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.10 mL, 0.53 mmol) was added, and the reaction mixture was heated at 100 °C for 4 hours. After cooling to room temperature, the resulting mixture was purified by chromatography on silica eluting with 0 - 100% EtOAc in heptane to give the title compound as a pale yellow solid. LC-MS (Method E): Rt 1.35 min; MS m / z 512.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.79 (s, 1H), 8.45 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.1 Hz, 1H), 8.03 (s, 1H), 7.93 (d, J = 8.5 Hz, 1H), 7.82 (dd, J = 5.5, 2.2 Hz, 1H), 7.64 (s, 1H), 7.23 - 7.18 (m, 2H), 7.16 (dd, J = 8.5, 1.2 Hz, 1H), 6.86 - 6.77 (m, 2H), 5.76 (s, 1H), 5.54 (s, 2H), 5.37 - 5.30 (m, 1H), 3.84 (s, 2H), 3.67 (s, 3H), 2.23 (s, 3H), 1.40 (s, 9H).

[0229] Step 4: N-tert-butyl-4-[[2-(3-isopropyl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide N-tert-Butyl-4-[[2-[3-isopropenyl-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Step 3) (98%, 113 mg, 0.22 mmol) and 10% Pd-C (23 mg, 0.02 mmol) were dissolved in EtOH (2.2 mL), placed under hydrogen, and stirred for 4 h. The resulting mixture was filtered through diatomaceous earth and concentrated in vacuo. The residue was treated with DCE (2.16 mL), followed by TFA (0.017 mL, 0.22 mmol), and the mixture was heated at 70 °C overnight. Additional TFA (1.2 mL) was added and stirring was continued at 70 °C for a further 24 h. The resulting mixture was concentrated in vacuo and the crude material was purified by preparative HPLC (acidic pH, early elution method) to give the title compound as a colorless solid. LC-MS (Method A): Rt 3.27 min; MS m / z 394.3 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 12.51 (s, 1H), 10.78 (s, 1H), 8.44 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.1 Hz, 1H), 8.02 (s, 1H), 7.81 (dd, J = 5.5, 2.2 Hz, 1H), 7.71 (d, J = 8.3 Hz, 1H), 7.40 (s, 1H), 7.03 (dd, J = 8.3, 1.2 Hz, 1H), 3.81 (s, 2H), 1.39 (s, 9H), 1.35 (d, J = 6.9 Hz, 6H).

[0230] Example 9.1 4-[[2-(3-Tetrahydrofuran-2-yl-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000069.tif36170

[0231] Project 1: 4-[[2-(3-Iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000070.tif65170 The title compound was prepared from 2-[3-iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid (Example 9, Step 1) and 4-amino-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Intermediate AD) in the same manner as in Step 2 of Example 9. LC-MS (Method E): Rt 1.30 min; MS m / z 650.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.83 (s, 1H), 9.39 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.20 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.71 (s, 1H), 7.38 (d, J = 8.3 Hz, 1H), 7.24 - 7.16 (m, 3H), 6.86 - 6.81 (m, 2H), 5.56 (s, 2H), 3.87 (s, 2H), 3.67 (s, 3H), 1.32-1.27 (m, 2H), 1.21-1.16 (m, 2H).

[0232] Project 2: 4-[[2-[3-(2,3-Dihydrofuran-4-yl)-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000071.tif60170 The title compound was prepared from 4-[[2-[3-iodo-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Step 1) and 2-(2,3-dihydrofuran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane in the same manner as in Step 3 of Example 9. LC-MS (Method E): Rt 1.25 min; MS m / z 592.3 = [M+H]+

[0233] Step 3: 4-[[2-(3-Tetrahydrofuran-2-yl-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide The title compound was prepared from 4-[[2-[3-(2,3-dihydrofuran-5-yl)-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Step 2) and 10% Pd / C in the same manner as in Step 4 of Example 9. LC-MS (Method A): Rt 2.83 min; MS m / z 474.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 12.78 (s, 1H), 10.83 (s, 1H), 9.38 (s, 1H), 8.49 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 5.5, 2.1 Hz, 1H), 7.72 (d, J = 8.3 Hz, 1H), 7.44 (s, 1H), 7.06 (dd, J = 8.4, 1.1 Hz, 1H), 5.17 (t, J = 7.2 Hz, 1H), 3.94 (q, J = 7.2 Hz, 1H), 3.84 - 3.81 (m, 3H), 2.29 - 2.15 (m, 2H), 2.10 - 1.93 (m, 2H), 1.32 - 1.26 (m, 2H), 1.21 - 1.14 (m, 2H).

[0234] Example 10 4-[[2-[3-(Morpholinomethyl)-1H-indazol-6-yl]acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide TIFF0007682850000072.tif44170

[0235] Step 1: 6-Bromo-1-[(4-methoxyphenyl)methyl]indazole-3-carbaldehyde TIFF0007682850000073.tif341701-(Chloromethyl)-4-methoxy-benzene (0.49 mL, 3.67 mmol) was added to a mixture of 6-bromo-1H-indazole-3-carbaldehyde (750 mg, 3.33 mmol) and Cs 2 CO 3 (1629 mg, 5.0 mmol) in DMF (12 mL), and the reaction mixture was stirred at room temperature for 2 hours and 30 minutes. The resulting mixture was diluted with EtOAc (60 mL) and brine (60 mL). The phases were separated and the aqueous portion was extracted with EtOAc (60 mL). The combined organic extracts were washed with Na 2 SO 4It was dried above and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with 0 - 80% EtOAc in heptane to afford the title compound as an orange solid. LC-MS (Method E): Rt 1.30 min; MS m / z 345.0, 347.0 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.14 (s, 1H), 8.31 (d, J = 1.2 Hz, 1H), 8.06 (d, J = 8.6 Hz, 1H), 7.53 (dd, J = 8.6, 1.6 Hz, 1H), 7.33 - 7.30 (m, 2H), 6.92 - 6.88 (m, 2H), 5.74 (s, 2H), 3.71 (s, 3H).

[0236] Step 2: 4-[[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]methyl]morpholine To a solution of 6-bromo-1-[(4-methoxyphenyl)methyl]indazole-3-carbaldehyde (Step 1) (600 mg, 1.74 mmol) and AcOH (0.15 mL, 2.61 mmol) in THF (10 mL) were added morpholine (0.30 mL, 3.48 mmol) and sodium triacetoxyborohydride (442 mg, 2.09 mmol), and the reaction mixture was stirred at room temperature for 16 h. The resulting mixture was diluted with EtOAc (80 mL) and saturated sodium bicarbonate solution (80 mL), and the phases were separated. The organic portion was washed with brine (80 mL) and dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by chromatography on silica eluting with 0 - 100% EtOAc in heptane, followed by 0 - 5% MeOH in EtOAc to afford the title compound as a pale orange oil. LC-MS (Method E): Rt 0.98 min; MS m / z 416.0, 418.0 = [M+H]+ 11H NMR (500 MHz, DMSO-d 6 ) δ 8.00 (d, J = 1.2 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.25 (dd, J = 8.6, 1.6 Hz, 1H), 7.21 - 7.16 (m, 2H), 6.88 - 6.84 (m, 2H), 5.52 (s, 2H), 3.80 (s, 2H), 3.69 (s, 3H), 3.57 - 3.52 (m, 4H), 2.42 - 2.37 (m, 4H).

[0237] Step 3: 2-[1-[(4-Methoxyphenyl)methyl]-3-(morpholin-4-ium-4-ylmethyl)indazol-6-yl]acetate TIFF0007682850000075.tif42170 The title compound was prepared from 6-[[6-bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]methyl]morpholine (Step 2) and potassium 3-ethoxy-3-oxopropanoate in the same manner as in Step 2 of Example 6. LC-MS (Method E): Rt 0.81 min; MS m / z 396.1 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6 ) δ 7.65 (d, J = 8.3 Hz, 1H), 7.32 (s, 1H), 7.17 - 7.13 (m, 2H), 7.03 (dd, J = 8.4, 1.0 Hz, 1H), 6.87 - 6.82 (m, 2H), 5.45 (s, 2H), 3.77 (s, 2H), 3.70 (s, 3H), 3.58 - 3.53 (m, 4H), 3.26 (s, 2H), 2.43 - 2.38 (m, 4H).

[0238] Step 4: Methyl 4-[[2-[1-[(4-methoxyphenyl)methyl]-3-(morpholinomethyl)indazol-6-yl]acetyl]amino]pyridine-2-carboxylate The title compound was prepared from 2-[1-[(4-methoxyphenyl)methyl]-3-(morpholin-4-ium-4-ylmethyl)-1H-indazol-6-yl]acetate (Step 3) and methyl 4-aminopyridine-2-carboxylate in the same manner as in Step 3 of Example 6. LC-MS (Method E): Rt 0.88 min; MS m / z 530.2 = [M+H]+ 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.81 (s, 1H), 8.55 (d, J = 5.7 Hz, 1H), 8.29 (d, J = 1.9 Hz, 1H), 7.82 - 7.77 (m, 2H), 7.56 (s, 1H), 7.18 - 7.13 (m, 2H), 7.09 (dd, J = 8.4, 1.2 Hz, 1H), 6.84 - 6.78 (m, 2H), 5.49 (s, 2H), 3.86 (s, 3H), 3.82 (s, 2H), 3.79 (s, 2H), 3.66 (s, 3H), 3.55 - 3.52 (m, 4H), 2.41 - 2.38 (m, 4H).

[0239] Step 5: 4-[[2-[1-[(4-methoxyphenyl)methyl]-3-(morpholin-4-ium-4-ylmethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxylate The title compound was prepared from methyl 4-[[2-[1-[(4-methoxyphenyl)methyl]-3-(morpholinomethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxylate (Step 4) and 1 M LiOH in the same manner as in Step 4 of Example 6. LC-MS (Method E): Rt 0.80 min; MS m / z 516.3 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 10.79 (s, 1H), 8.49 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 1.7 Hz, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.78 (dd, J = 5.6, 2.2 Hz, 1H), 7.58 (s, 1H), 7.19 - 7.13 (m, 2H), 7.09 (dd, J = 8.4, 1.1 Hz, 1H), 6.83 - 6.80 (m, 2H), 5.49 (s, 2H), 3.82 (s, 2H), 3.79 (s, 2H), 3.66 (s, 3H), 3.55 - 3.53 (m, 4H), 2.41 - 2.38 (m, 4H).

[0240] Step 6: 4-[[2-[3-(Morpholinomethyl)-1H-indazol-6-yl]acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide The title compound was prepared from 4-[[2-[1-[(4-methoxyphenyl)methyl]-3-(morpholin-4-ium-4-ylmethyl)indazol-6-yl]acetyl]amino]pyridine-2-carboxylate (Step 5) and 1,1,1-trifluoro-2-methyl-propan-2-amine hydrochloride in the same manner as in Step 5 of Example 6. LC-MS (Method A): Rt 1.91 min; MS m / z 505.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 12.78 (s, 1H), 10.84 (s, 1H), 8.49 (d, J = 5.6 Hz, 1H), 8.33 (s, 1H), 8.23 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 5.6, 2.2 Hz, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.43 (s, 1H), 7.07 (dd, J = 8.3, 1.2 Hz, 1H), 3.83 (s, 2H), 3.79 (s, 2H), 3.56 - 3.52 (m, 4H), 2.42 - 2.37 (m, 4H), 1.65 (s, 6H).

[0241] Example 11 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(3-methyloxetan-3-yl)pyridine-2-carboxamide TIFF0007682850000078.tif36170

[0242] Step 1: 4-[[2-(1H-Indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid A mixture of 4-[[2-(1-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid and 4-[[2-(2-tetrahydropyran-2-ylindazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 4, Step 2) (50%, 1.9 g, 2.5 mmol) in DCM (100 mL) was treated with TFA (1.91 mL, 24.97 mmol), and the reaction mixture was stirred at room temperature overnight. The resulting mixture was concentrated in vacuo, and the crude residue was suspended in DCM (50 mL). The suspension was sonicated for 10 minutes, filtered, washed with DCM (20 mL), and dried to obtain the title compound as an off-white solid. LC-MS (Method E): Rt 0.78 min; MS m / z 297.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 13.10 (s, 1H), 11.96 (s, 1H), 8.58 (d, J = 6.0 Hz, 1H), 8.50 (s, 1H), 8.08 (d, J = 6.3 Hz, 1H), 8.02 (d, J = 0.8 Hz, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.56 (s, 1H), 7.13 (dd, J = 8.3, 1.0 Hz, 1H), 3.96 (s, 2H).

[0243] Step 2: 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(3-methyloxetan-3-yl)pyridine-2-carboxamide HATU (83 mg, 0.22 mmol) was added to a mixture of 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 1) (40%, 125 mg, 0.17 mmol), 3-methyloxetan-3-amine hydrochloride (42 mg, 0.34 mmol) and DIPEA (88 μL, 0.51 mmol) in DMF (1 mL), and the reaction mixture was stirred at room temperature overnight. The resulting mixture was diluted with EtOAc (10 mL) and water (10 mL), and the phases were separated. The aqueous portion was extracted with EtOAc (10 mL), and the combined organic extracts were dried over Na 2 SO 4 and concentrated in vacuo. The crude material was purified by C18 reverse-phase chromatography eluting with 10 - 100% MeCN (+0.1% formic acid) and H 2 O (+0.1% formic acid) to afford the title compound as an off-white solid. LC-MS (Method A): Rt 1.90 min; MS m / z 366.2 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6) δ 9.21 (s, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.17 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.83 (dd, J = 5.5, 2.1 Hz, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.1 Hz, 1H), 4.72 (d, J = 6.5 Hz, 2H), 4.35 (d, J = 6.5 Hz, 2H), 3.83 (s, 2H), 1.58 (s, 3H).

[0244] Example 11.1 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(2-oxaspiro[3.3]heptan-6-yl)pyridine-2-carboxamide The title compound was prepared from 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 11 Step 1) and 2-oxaspiro[3.3]heptan-6-amine hydrochloride in the same manner as in Step 2 of Example 11. LC-MS (Method A): Rt 2.01 min; MS m / z 392.3 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.88 (d, J = 8.3 Hz, 1H), 8.46 (d, J = 5.5 Hz, 1H), 8.18 (d, J = 1.9 Hz, 1H), 8.02 (s, 1H), 7.82 (dd, J = 5.5, 2.1 Hz, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.1 Hz, 1H), 4.62 (s, 2H), 4.48 (s, 2H), 4.25 (1H, J = 8.3 Hz, 1H), 3.83 (s, 2H), 2.57 - 2.50 (m, 2H), 2.38 - 2.30 (m, 2H).

[0245] Example 11.2 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[4-(trifluoromethyl)tetrahydropyran-4-yl]pyridine-2-carboxamide TIFF0007682850000081.tif34170 The title compound was prepared from 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 11 Step 1) and 4-(trifluoromethyl)tetrahydropyran-4-amine hydrochloride in the same manner as in Step 2 of Example 11. LC-MS (Method A): Rt 2.72 min; MS m / z 448.3 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.03 (s, 1H), 10.89 (s, 1H), 8.52 (d, J = 5.5 Hz, 1H), 8.44 (s, 1H), 8.25 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 7.86 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 3.88 - 3.81 (m, 4H), 3.45 - 3.39 (m, 2H), 2.71 (d, J = 13.1 Hz, 2H), 1.79 (td, J = 12.9, 4.5 Hz, 2H).

[0246] Example 11.3 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[3-(trifluoromethyl)oxetan-3-yl]pyridine-2-carboxamide TIFF0007682850000082.tif34170 The title compound was prepared from 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 11 Step 1) and 3-(trifluoromethyl)oxetan-3-amine hydrochloride in the same manner as in Step 2 of Example 11. LC-MS (Method A): Rt 2.38 min; MS m / z 420.2 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 13.00 (s, 1H), 10.84 (s, 1H), 9.88 (s, 1H), 8.53 (d, J = 5.5 Hz, 1H), 8.22 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.88 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 4.92 (d, J = 8.0 Hz, 2H), 4.72 (d, J = 8.4 Hz, 2H), 3.84 (s, 2H).

[0247] Example 11.4 N-[1-(Difluoromethyl)cyclopropyl]-4-[[2-(1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide The title compound TIFF0007682850000083.tif33170 was prepared from 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 11 Step 1) and 1-(difluoromethyl)cyclopropanamine hydrochloride in the same manner as in Step 2 of Example 11. LC-MS (Method A): Rt 2.41 min; MS m / z 386.2 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6) δ 13.00 (s, 1H), 10.82 (s, 1H), 9.09 (s, 1H), 8.48 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 6.07 (t, J = 57.1 Hz, 1H), 3.84 (s, 2H), 1.11 - 1.04 (m, 2H), 1.06 - 0.97 (m, 2H).

[0248] Example 11.5 N-(3-Fluoro-1-bicyclo[1.1.1]pentanyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide TIFF0007682850000084.tif39170 The title compound was prepared from 4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 11 Step 1) and 3-fluorobicyclo[1.1.1]pentan-1-amine hydrochloride in the same manner as in Step 2 of Example 11. LC-MS (Method A): Rt 2.56 min; MS m / z 380.2 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 13.00 (s, 1H), 10.83 (s, 1H), 9.37 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 8.16 (d, J = 2.0 Hz, 1H), 8.02 (s, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.49 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 3.84 (s, 2H), 2.41 (d, J = 2.2 Hz, 6H).

[0249] Example 12 4-[[2-[3-(2,2,2-Trifluoro-1-hydroxy-1-methyl-ethyl)-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000085.tif45170

[0250] Step 1: 6-Bromo-1-[(4-methoxyphenyl)methyl]indazole-3-carboxaldehyde TIFF0007682850000086.tif371704-Methoxybenzyl chloride (1.45 mL, 10.75 mmol) was added to a mixture of 6-bromo-1H-indazole-3-carboxaldehyde (2.2 g, 9.78 mmol) and Cs 2 CO 3 (4.78 g, 14.66 mmol) in DMF (44.4 mL), and the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with EtOAc (100 mL) and water (100 mL), and the phases were separated. The organic layer was washed with water (100 mL), brine (100 mL), dried over Na 2 SO 4 and concentrated in vacuo. Purification by chromatography on silica eluting with 0 - 100% EtOAc in heptane afforded the title compound as a red solid. LC-MS (Method E): Rt 1.31 min; MS m / z 344.9, 347.0 = [M+H]+ 1H NMR (400 MHz, DMSO-d 6 ) δ 10.15 (s, 1H), 8.32 (d, J = 1.0 Hz, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.53 (dd, J = 8.6, 1.6 Hz, 1H), 7.36 - 7.29 (m, 2H), 6.94 - 6.88 (m, 2H), 5.75 (s, 2H), 3.71 (s, 3H).

[0251] Step 2: 1-[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]ethanol 6-Bromo-1-[(4-methoxyphenyl)methyl]indazole-3-carbaldehyde (Step 1) (73%, 3.06 g, 6.48 mmol) in THF (64.76 mL) was cooled to -78 °C and treated dropwise with 3 M bromo(methyl)magnesium in diethyl ether (2.37 mL, 7.12 mmol). The mixture was warmed to room temperature and stirred for 4 h. The reaction was quenched with saturated ammonium chloride (50 mL) and the volatile solvents were removed in vacuo. The resulting suspension was diluted with EtOAc (50 mL) and the phases were separated. The aqueous phase was extracted with EtOAc (3 × 50 mL) and the combined organic extracts were dried over Na 2 SO 4 and filtered, then concentrated in vacuo. Purification by chromatography on silica eluting with 0 - 100% EtOAc in heptane afforded the title compound as an orange oil. LC-MS (Method E): Rt 1.24 min; MS m / z 360.9, 363.0 = [M+H]+ 1H NMR (400 MHz, DMSO-d 6 ) δ 8.00 - 7.96 (m, 1H), 7.88 - 7.84 (m, 1H), 7.24 - 7.18 (m, 3H), 6.89 - 6.84 (m, 2H), 5.52 (d, J = 15.5 Hz, 1H), 5.47 (d, J = 15.5 Hz, 1H), 5.41 (d, J = 4.8 Hz, 1H), 5.08 (qd, J = 6.5, 4.9 Hz, 1H), 3.70 (s, 3H), 1.51 (d, J = 6.5 Hz, 3H).

[0252] Step 3: 1-[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]ethanone TIFF0007682850000088.tif A mixture of 1-[6-bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]ethanol (step 2) (87%, 2.45 g, 5.9 mmol) in DCM (28 mL) at 341700 °C was treated with Dess-Martin periodinane (3.0 g, 7.08 mmol). The mixture was warmed to room temperature and stirred for 18 h. Aqueous saturated sodium bisulfite solution (20 mL) was added and the phases were separated. The organic layer was washed with aqueous saturated sodium bicarbonate solution (2 × 20 mL) and the combined aqueous washes were re-extracted with DCM (2 × 30 mL). The combined organic extracts were dried over Na 2 SO 4 and filtered, then concentrated in vacuo. Purification by chromatography on silica eluting with 0 - 100% EtOAc in heptane afforded the title compound as an orange solid. LC-MS (method E): Rt 1.36 min; MS m / z 358.8, 360.8 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 8.23 (d, J = 1.3 Hz, 1H), 8.09 (d, J = 8.6 Hz, 1H), 7.47 (dd, J = 8.6, 1.6 Hz, 1H), 7.32 - 7.26 (m, 2H), 6.92 - 6.87 (m, 2H), 5.71 (s, 2H), 3.70 (s, 3H), 2.63 (s, 3H).

[0253] Step 4: 2-[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]-1,1,1-trifluoro-propan-2-ol TIFF0007682850000089.tif1-[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]ethenone (Step 3) (500 mg, 1.39 mmol) in THF (10 mL) at -78 °C was treated with trimethyl(trifluoromethyl)silane (411 μL, 2.78 mmol), followed by treatment with 1 M TBAF in THF (14 μL, 0.014 mmol), and stirred for 30 minutes. The mixture was warmed to room temperature and stirred for 2 hours. 1 M TBAF in THF (2.78 mL, 2.78 mmol) was added and stirring was continued for an additional 2 hours. The resulting mixture was concentrated in vacuo, and the residue was dissolved in EtOAc (10 mL) and washed with saturated aqueous sodium bicarbonate (2 x 10 mL). The combined aqueous washes were extracted with EtOAc (2 x 10 mL), and the combined organic extracts were dried over 2 SO 4 Na and concentrated in vacuo to afford the title compound as a brown oil. LC-MS (Method E): Rt 1.37 min; MS m / z 429.0, 431.0 = [M+H]+ 1H NMR (400 MHz, DMSO-d 6 ) δ 8.04 (d, J = 1.2 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.28 (dd, J = 8.7, 1.6 Hz, 1H), 7.22 - 7.17 (m, 2H), 6.98 - 6.89 (m, 1H), 6.89 - 6.85 (m, 2H), 5.60 (d, J = 15.6 Hz, 1H), 5.56 (d, J = 15.6 Hz, 1H), 3.70 (s, 3H), 1.83 (s, 3H).

[0254] Step 5: 3-(1-Benzyloxy-2,2,2-trifluoro-1-methyl-ethyl)-6-bromo-1-[(4-methoxyphenyl)methyl]indazole TIFF0007682850000090.tif 2-[6-Bromo-1-[(4-methoxyphenyl)methyl]indazol-3-yl]-1,1,1-trifluoro-propan-2-ol (658 mg, 1.53 mmol) in 46170 DMF (15.3 mL) in step 4 was treated with a 60% dispersion of NaH in mineral oil (123 mg, 3.07 mmol) and bromomethylbenzene (0.36 mL, 3.07 mmol), and stirred for 18 h. The reaction was quenched with brine (20 mL), and the mixture was extracted with EtOAc (3 × 20 mL). The combined organic extracts were dried over Na 2 SO 4 and concentrated in vacuo. Purification by chromatography on silica eluting with 0 - 35% EtOAc in heptane gave the title compound as a colorless oil. LC-MS (method E): Rt 1.64 min; MS m / z 519.1, 521.1 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 8.12 (d, J = 1.4 Hz, 1H), 7.73 (d, J = 8.7 Hz, 1H), 7.36 - 7.27 (m, 6H), 7.24 - 7.20 (m, 2H), 6.91 - 6.86 (m, 2H), 5.67 (d, J = 15.6 Hz, 1H), 5.63 (d, J = 15.6 Hz, 1H), 4.58 (d, J = 11.2 Hz, 1H), 4.14 (d, J = 11.2 Hz, 1H), 3.70 (s, 3H), 2.01 (s, 3H).

[0255] Step 6: 2-[3-(1-Benzyloxy-2,2,2-trifluoro-1-methyl-ethyl)-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid TIFF0007682850000091.tif431703 - (1 - Benzyloxy - 2,2,2 - trifluoro - 1 - methyl - ethyl) - 6 - bromo - 1 - [(4 - methoxyphenyl)methyl]indazole (Step 5) (632 mg, 1.22 mmol), potassium 3 - ethoxy - 3 - oxo - propanoate (414 mg, 2.43 mmol), DMAP (15 mg, 0.12 mmol), BINAP (76 mg, 0.12 mmol) and diallyldipalladium dichloride (22 mg, 0.06 mmol) were added to a sealed tube and placed under a nitrogen atmosphere. Toluene (6.1 mL) was added, nitrogen was injected into the mixture for 10 minutes, and then the mixture was stirred at 140 °C for 20 hours. The resulting mixture was purified by chromatography on silica eluting with 0 - 100% EtOAc in heptane to obtain an ester intermediate. The intermediate was dissolved in a 1:1 mixture of MeOH and 2 M LiOH (12 mL) and stirred for 18 hours. The volatile solvents were removed in vacuo, and the aqueous mixture was acidified to pH 1 with 3 M HCl. The aqueous suspension was extracted with EtOAc (3 × 50 mL), and the combined organic extracts were dried over Na 2 SO 4 and concentrated in vacuo to obtain the title compound as a pale yellow gum. LC - MS (Method E): Rt 1.40 min; MS m / z 499.2 = [M + H]+ 1H NMR (400 MHz, DMSO - d 6 ) δ 7.75 (d, J = 8.4 Hz, 1H), 7.39 - 7.26 (m, 6H), 7.24 - 7.15 (m, 2H), 7.07 (dd, J = 8.5, 1.2 Hz, 1H), 6.91 - 6.83 (m, 2H), 5.61 (s, 2H), 4.56 (d, J = 11.3 Hz, 1H), 4.17 (d, J = 11.3 Hz, 1H), 3.71 - 3.65 (m, 5H), 2.01 (s, 3H).

[0256] Step 7: 4-[[2-[3-(2,2,2-Trifluoro-1-hydroxy-1-methyl-ethyl)-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide A mixture of 2-[3-(1-benzyloxy-2,2,2-trifluoro-1-methyl-ethyl)-1-[(4-methoxyphenyl)methyl]indazol-6-yl]acetic acid (Step 6) (85%, 312 mg, 0.53 mmol) and 4-amino-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Intermediate AD) (143 mg, 0.58 mmol) in 1,4-dioxane (5.3 mL) was treated with DIPEA (0.19 mL, 1.06 mmol) and a 50% solution of T3P® in EtOAc (1.27 mL, 1.06 mmol), and the reaction mixture was stirred for 2 h. The resulting mixture was diluted with EtOAc (15 mL) and washed with saturated aqueous sodium bicarbonate (15 mL). The organic layer was dried over Na 2 SO 4 and concentrated in vacuo. The residue was dissolved in DCE (5.3 mL), treated with TFA (2.5 mL, 32.67 mmol), and the reaction mixture was heated at 75 °C for 18 h. The resulting mixture was concentrated in vacuo, the residue was dissolved in EtOAc (20 mL), and washed with saturated aqueous sodium bicarbonate (20 mL). The aqueous phase was extracted with EtOAc (2 × 20 mL), and the combined organic extracts were dried over Na 2 SO 4 and filtered through a plug of silica and concentrated in vacuo. Purification by preparative HPLC (acidic pH, early elution method) afforded the title compound as a colorless solid. LC-MS (Method A): Rt 2.97 min; MS m / z 516.3 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6) δ 13.04 (s, 1H), 10.81 (s, 1H), 9.38 (s, 1H), 8.49 (d, J = 5.5 Hz, 1H), 8.21 (d, J = 2.0 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.85 (dd, J = 5.5, 2.2 Hz, 1H), 7.46 (s, 1H), 7.09 (dd, J = 8.5, 1.3 Hz, 1H), 6.74 (s, 1H), 3.83 (s, 2H), 1.82 (s, 3H), 1.32 - 1.27 (m, 2H), 1.21 - 1.15 (m, 2H).

[0257] Example 13 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide TIFF0007682850000092.tif37170

[0258] Step 1: 4-[[2-(1H-Indol-6-yl)acetyl]amino]pyridine-2-carboxylic acid TIFF0007682850000093.tif34170A solution of methyl 4-aminopyridine-2-carboxylate (751 mg, 4.93 mmol) and 2-(1H-indol-6-yl)acetic acid (786 mg, 4.48 mmol) in 1,4-dioxane (5 mL) was treated with DIPEA (1.57 mL, 8.97 mmol) and a 50% solution of T3P® in EtOAc (5.33 mL, 8.97 mmol), and the reaction mixture was stirred at room temperature for 90 minutes. The resulting mixture was concentrated in vacuo, and the residue obtained was dissolved in EtOAc (50 mL) and washed with saturated aqueous sodium bicarbonate (2 × 50 mL). The organic layer was dried over Na 2 SO 4It was dried above and concentrated in vacuo. The residue was dissolved in 1,4-dioxane (10 mL) and 1 M LiOH (10 mL) and stirred for 2 h. The volatile solvents were removed in vacuo and the resulting aqueous solution was acidified to pH ca. 3 with 3 M HCl. The resulting precipitate was collected by filtration to give the title compound as an orange solid. LC-MS (Method E): Rt 0.86 min; MS m / z 296.1 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 10.74 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 8.23 (s, 1H), 7.80 (dd, J = 5.6, 2.1 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 7.36 (s, 1H), 7.32 - 7.29 (m, 1H), 6.97 (dd, J = 8.1, 1.4 Hz, 1H), 6.40 - 6.37 (m, 1H), 3.75 (s, 2H). The signal of COOH was not observed.

[0259] Step 2: 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide 4-[[2-(1H-Indol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Step 1) (79%, 100 mg, 0.27 mmol) and 1,1,1-trifluoro-2-methyl-propan-2-amine hydrochloride (48 mg, 0.29 mmol) in DMF (3 mL) were treated with DIPEA (0.094 mL, 0.54 mmol) and HATU (112 mg, 0.29 mmol) and the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with EtOAc (10 mL) and washed with saturated aqueous sodium hydrogen carbonate (2×10 mL). The organic layer was dried over Na 2 SO 4 and concentrated in vacuo. Purification by preparative HPLC (acidic pH, early elution method) gave the title compound as an off-white solid. LC-MS (Method A): Rt 3.46 min; MS m / z 405.4 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 10.79 (s, 1H), 8.48 (d, J = 5.5 Hz, 1H), 8.33 (s, 1H), 8.24 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 5.6, 2.2 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 7.36 (s, 1H), 7.32 - 7.27 (m, 1H), 6.97 (dd, J = 8.1, 1.4 Hz, 1H), 6.40 - 6.35 (m, 1H), 3.75 (s, 2H), 1.64 (s, 6H).

[0260] Example 13.1 N-(3,3-Difluoro-1-methyl-cyclobutyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide The title compound was prepared from 4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 13, Step 1) and 3,3-difluoro-1-methyl-cyclobutaneamine hydrochloride in the same manner as in Step 2 of Example 13. LC-MS (Method A): Rt 3.12 min; MS m / z 399.4 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6) δ 11.03 (s, 1H), 10.74 (s, 1H), 9.02 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.84 (dd, J = 5.5, 2.2 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 7.36 (s, 1H), 7.33 - 7.28 (m, 1H), 6.97 (dd, J = 8.1, 1.4 Hz, 1H), 6.40 - 6.35 (m, 1H), 3.75 (s, 2H), 3.11 - 2.98 (m, 2H), 2.73 - 2.62 (m, 2H), 1.51 (s, 3H).

[0261] Example 13.2 N-(4-Cyanotetrahydropyran-4-yl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide The title compound was prepared from 4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxylic acid (Example 13, Step 1) and 4-aminotetrahydropyran-4-carbonitrile hydrochloride in the same manner as in Step 2 of Example 13. LC-MS (Method A): Rt 2.61 min; MS m / z 404.4 = [M+H]+ 1H NMR (500 MHz, DMSO-d 6) δ 11.04 (s, 1H), 10.79 (s, 1H), 9.01 (s, 1H), 8.52 (d, J = 5.5 Hz, 1H), 8.25 (d, J = 2.0 Hz, 1H), 7.88 (dd, J = 5.6, 2.2 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 7.37 (s, 1H), 7.32 - 7.27 (m, 1H), 6.97 (dd, J = 8.1, 1.4 Hz, 1H), 6.41 - 6.35 (m, 1H), 3.86 (dt, J = 12.3, 3.8 Hz, 2H), 3.76 (s, 2H), 3.62 - 3.54 (m, 2H), 2.40 - 2.31 (m, 2H), 2.06 (ddd, J = 13.8, 12.0, 3.9 Hz, 2H).

[0262] Preparation of Intermediate Intermediate A 4-Amino-N-(1,1-dimethylprop-2-ynyl)pyridine-2-carboxamide To a mixture of 4-aminopyridine-2-carboxylic acid (2 g, 14.48 mmol), TBTU (5.58 g, 17.38 mmol) and TEA (2.42 mL, 17.38 mmol) in DMF (36 mL), 2-methylbut-3-yn-2-amine (22.82 mL, 17.38 mmol) was added and the mixture was stirred at room temperature for 3 days. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The crude residue was dissolved in EtOAc (40 mL) and washed with saturated NaHCO 3 solution (40 mL). The aqueous phase was further extracted with EtOAc (40 mL), and the combined organic portions were washed with brine (2 × 40 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude material was triturated with a minimum amount of ether at 0 °C to afford the title compound as an off-white crystalline solid. LC-MS (Method F): Rt 1.28 min; MS m / z 204.3 = [M+H]+ 11H NMR (500 MHz, DMSO-d 6 ) δ 8.23 (s, 1H), 7.99 (d, J = 5.6 Hz, 1H), 7.19 (d, J = 2.3 Hz, 1H), 6.59 (dd, J = 5.6, 2.4 Hz, 1H), 6.36 (s, 2H), 3.19 (s, 1H), 1.62 (s, 6H).

[0263] Intermediate AB 4-Amino-N-tert-butyl-pyridine-2-carboxamide The title compound TIFF0007682850000097.tif27170 was prepared from 4-aminopyridine-2-carboxylic acid and 2-methylpropan-2-amine in the same manner as Intermediate A. LC-MS (Method E): Rt 0.48 min; MS m / z 194.0 = [M+H]+ 1 1H NMR (500 MHz, DMSO-d 6 ) δ 7.99 (s, 1H), 7.97 (d, J = 5.6 Hz, 1H), 7.18 (d, J = 2.2 Hz, 1H), 6.56 (dd, J = 5.6, 2.4 Hz, 1H), 6.32 (s, 2H), 1.37 (s, 9H).

[0264] Intermediate AC 4-Amino-N-(1-cyano-1-methyl-ethyl)pyridine-2-carboxamide To a mixture of 4-aminopyridine-2-carboxylic acid (15 g, 108.6 mmol), TBTU (41.84 g, 130.32 mmol) and triethylamine (37.84 mL, 271.5 mmol) in DMF (271.52 mL) was added 2-amino-2-methyl-propanenitrile hydrochloride (14.4 g, 119.46 mmol), and the mixture was stirred at room temperature for 3 days. The reaction mixture was filtered and the solid was washed with DMF (2 × 30 mL). The combined filtrate was concentrated in vacuo and the crude residue was dissolved in EtOAc (300 mL) and saturated NaHCO3 It was washed with solution (2 × 300 mL). The aqueous portion was re-extracted with EtOAc (30 mL), the organic layers were combined, washed with brine (160 mL × 2), and dried over anhydrous Na 2 SO 4 and concentrated in vacuo. The solid obtained was purified by chromatography on silica eluting with 0 - 100% EtOAc in heptane, and it was triturated and mixed with ice-cold TBME:heptane (3:1 mixture) to give the title compound as a colorless crystalline solid. 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.60 (s, 1H), 8.03 (d, J = 5.6 Hz, 1H), 7.21 (d, J = 2.3 Hz, 1H), 6.62 (dd, J = 5.6, 2.4 Hz, 1H), 6.40 (s, 2H), 1.70 (s, 6H). LCMS (Method E) Rt 0.35 min; MS m / z 205.0 = [M+H]+

[0265] Intermediate AD 4-Amino-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide TIFF0007682850000099.tif251704-Aminopyridine-2-carboxylic acid (1 g, 7.24 mmol) in DMF (36.2 mL) was treated with TEA (3.68 mL, 26.43 mmol), 1-(trifluoromethyl)cyclopropanamine hydrochloride (1.29 g, 7.96 mmol), followed by TBTU (3.14 g, 9.77 mmol) and stirred at room temperature for 4 days. The resulting mixture was filtered and the solid was washed with DMF (2 × 30 mL). The filtrate was concentrated in vacuo, the crude residue was dissolved in EtOAc (300 mL), and washed with saturated NaHCO 3 solution (2 × 300 mL). The aqueous portion was re-extracted with EtOAc (30 mL), the combined organic extracts were washed with brine (2 × 160 mL), and dried over Na 2 SO 4It was dried above and concentrated in vacuo. It was purified by C18 reverse-phase column chromatography eluting with 10 - 100% MeCN in water to give the title compound as an off-white solid. LC-MS (Method F): Rt 1.31 min; MS m / z 246.1 = [M+H]+ 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.13 (s, 1H), 8.02 (d, J = 5.6 Hz, 1H), 7.20 (d, J = 2.3 Hz, 1H), 6.61 (dd, J = 5.6, 2.4 Hz, 1H), 6.36 (s, 2H), 1.33 - 1.20 (m, 2H), 1.19 - 1.06 (m, 2H).

[0266] Intermediate B 2-(1-Tetrahydropyran-2-yl-1H-indazol-6-yl)acetic acid To a solution of 2-(1H-indazol-6-yl)acetic acid (750 mg, 4.26 mmol) and 3,4-dihydro-2H-pyran (1.17 mL, 12.77 mmol) in DCE (14.2 mL) was added pyridinium p-toluenesulfonate (1.18 g, 4.68 mmol), and the mixture was stirred at room temperature for 16 h. Additional 3,4-dihydro-2H-pyran (1.17 mL, 12.77 mmol) and pyridinium p-toluenesulfonate (1.18 g, 4.68 mmol) were added and stirring was continued for a further 8 h. The resulting mixture was diluted with DCM (10 mL) and washed with water and brine. The organic portion was separated and dried over Na 2 SO 4 and concentrated in vacuo. The crude mixture was dissolved in THF (10 mL) and treated with 1 M LiOH (10 mL). After stirring at room temperature for 1 h, the pH was adjusted to pH 5 with 1 M HCl (2.5 mL). The mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL). The organic extract was dried over Na 2 SO 4 and concentrated in vacuo to give the title compound as a yellow viscous oil. LC-MS (Method E): Rt 0.94 min; MS m / z 261.1 = [M+H]+ 1 1H NMR (400 MHz, DMF-d 7 ) δ 12.28 (s, 1H), 8.06 (s, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.59 (s, 1H), 7.08 (dd, J = 8.3, 1.2 Hz, 1H), 5.80 (dd, J = 9.7, 2.5 Hz, 1H), 3.95 - 3.84 (m, 1H), 3.78 - 3.68 (m, 3H), 2.47 - 2.36 (m, 1H), 2.09 - 2.00 (m, 1H), 1.98 - 1.91 (m, 1H), 1.80 - 1.69 (m, 1H), 1.62 - 1.53 (m, 2H).

[0267] Intermediate B1 Mixture of 2-(1-tetrahydropyran-2-yl-1H-indazol-6-yl)acetic acid and 2-(2-tetrahydropyran-2-yl-1H-indazol-6-yl)acetic acid To a solution of 2-(1H-indazol-6-yl)acetic acid (1.2 g, 6.81 mmol) and 3,4-dihydro-2H-pyran (1864 uL, 20.43 mmol) in DCE (35 mL), pyridinium p-toluenesulfonate (1.88 g, 7.49 mmol) was added and the mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with DCM (80 mL) and washed with water (80 mL) and brine (80 mL). The organic layer was dried over Na 2 SO 4 and concentrated in vacuo. The crude mixture was dissolved in THF (35 mL), 2M LiOH (3.4 mL) was added and the mixture was stirred at room temperature for 1 h. The resulting mixture was acidified to pH 5 using 1M HCl, then diluted with water (100 mL) and extracted with EtOAc (100 mL). The organic layer was separated, dried over Na 2 SO 4 and concentrated in vacuo to afford the title compound as a yellow viscous oil. LC-MS (Method E): Rt 0.94, 0.97 min; MS m / z 261.00 = [M+H]+

[0268] Intermediate C 2-(3-Iodo-1H-indazol-6-yl)acetic acid TIFF0007682850000102.tif28170 This compound was prepared according to the procedure of J. Org Chem. 2018, 83, 2, 930-938. LC-MS (Method E): Rt 0.94 min; MS m / z 302.9 = [M+H]+ 1H NMR (400 MHz, DMSO-d 6 ) δ 13.44 (s, 1H), 12.39 (s, 1H), 7.44 (s, 1H), 7.35 (d, J = 8.3 Hz, 1H), 7.09 (d, J = 8.4 Hz, 1H), 3.72 (s, 2H).

[0269] Biological Examples In the following examples, the compounds of the present invention are compared with Examples 79.2 and 79.3 of International Publication No. WO 2019 / 145726 of our previous application (designated as Compound A and Compound B, respectively). These compounds are as follows: Compound A (Example 79.2 of International Publication No. WO 2019 / 145726): TIFF0007682850000103.tif34170 Compound B (Example 79.3 of International Publication No. WO 2019 / 145726) TIFF0007682850000104.tif33170

[0270] Example 14 - Automated Whole-Cell Patch-Clamp Assay for Detecting TMEM16A Activity in Recombinant Cells Cell Cultures 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 Coon's modified Hams F-12 medium (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 experiments by separating them at approximately 90% confluence using a 2:1 (v / v) mixture of detaching solution (BMS Biotechnology) and 0.25% (w / v) trypsin-EDTA. The cells were diluted to a density of 3.5 - 4.5×10 6 cells / mL in a medium consisting of CHO-S-SFM II (Sigma), 25 mM HEPES (Sigma), and soybean trypsin inhibitor (Sigma).

[0271] Whole-cell patch-clamp recording FRT-TMEM16A cells were whole-cell patch-clamped 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 a suction impulse to establish the whole-cell recording configuration of the patch-clamp technique. The assay used the following solutions (all reagents from Sigma): Intracellular solution (mM): IN-methyl-D-glucamine 130, CaCl 2 18.2, MgCl 2 1, HEPES 10, EGTA 10, BAPTA 20, Mg-ATP 2, pH 7.25, 325 mOsm using sucrose. Extracellular solution (mM): N-methyl-D-glucamine 130, CaCl 2 2, MgCl 2 1, HEPES 10, pH 7.3, 320 mOsm using sucrose.

[0272] The intracellular solution buffers intracellular calcium at a level necessary to activate the maximal TMEM16A-mediated current (for calcium ions, EC 20 ) by approximately 20%. Cells were voltage-clamped at a holding potential of -70 mV, and a composite voltage step (+70 mV) / ramp (-90 mV to +90 mV) was applied at 0.05 Hz. After a period of current stabilization, test compounds dissolved in 100% (v / v) DMSO and then diluted in the extracellular solution were applied to generate a cumulative concentration-response curve. Each concentration of the test compound was incubated for 5 minutes before adding the next concentration. After testing the final concentration, either a known active positive modulator or a TMEM16A inhibitor at the maximum concentration, the maximum concentration of CaCCinhA01 (Del La Fuente et al., 2008) was added to define the upper and lower limits of the assay.

[0273] The activity of the compound was quantified by measuring the increase in current upon addition of the compound and expressing this as the rate of increase in the baseline TMEM16A current level. The rate of increase in current was determined for each concentration, and the concentration that gives 50% of its maximum effect (EC 50 ) and the maximum efficiency (rate of increase from baseline) were used with either Qpatch software or Graphpad Prism v6.05 to plot the data as a function of concentration.

[0274] The method of calculating the results is illustrated in Figure 1, which shows an exemplary trace from the Qpatch TMEM16A assay. In Figure 1, I BL is equal to the baseline current, I [#1] is equal to the peak current during the incubation period of concentration 1 of the test compound, etc.

[0275] The peak TMEM16A current at +70 mV was plotted as a function of time over the assay period. The baseline current (I BL) was measured after stabilization for a certain period. The increase in current with the addition of each compound was determined by taking the peak current during the incubation period, subtracting the current from the previous recording period, and then expressing this as a ratio (enhancement rate) to the baseline current. In Figure 1, for test compound concentration 1, this is as follows: (I [#1] -I BL / I BL )×100

[0276] For each additional concentration tested, the increase in current was determined by subtracting the current from the previous incubation period and normalizing to the baseline value. In Figure 1, for test concentration 2, this is as follows: (I [#2] -I [#1] / I BL )×100

[0277] The values for each test concentration were plotted as a cumulative function of concentration. For example, for test concentration 2, this is the sum of the peak changes measured during the period of concentration 1 + concentration 2.

[0278] The results obtained for the compounds of the examples are shown in Table 3, from which it can be seen that the compounds of the present invention can significantly increase the current level of TMEM16A. TIFF0007682850000105.tif255170

[0279] The results presented in Table 3 demonstrate that the compounds of the present invention have good excellent EC 50 values, similar to those of compound A and better than those of compound B.

[0280] Compared to compound B (a more potent comparator compound), many of the compounds of the present invention have a significantly improved maximum enhancement effect on TMEM16A, as shown, for example, at a concentration of 3.3 mM.

[0281] Compounds 1, 1.1, 13, 13.1 and 13.2 are indoles and can thus be compared to Compound B. Compounds 1, 2.2, 13.1 and 13.2 have lower EC50 values than Compound B, and it can be seen that Compounds 1.1, 13.1 and 13.2 have an improved maximum enhancement effect on TMEM16A.

[0282] The compounds of the remaining examples are indazoles and can thus be compared to Compound A. All of the indazole compounds of the examples have a significantly lower EC50 than Compound A and also have a similar or improved maximum enhancement effect on TMEM16A, especially at a concentration of 370 mM.

[0283] Example 15 - Physicochemical Assays LogD (pH 7.4 Flask Shaking Method) All compounds were tested in a 'cassette' containing a mixture of four test compounds each initially dissolved at 5 mmol in DMSO. Phosphate buffer (1 M) was diluted to 20 mM with deionized water and adjusted to pH 7.4 using phosphoric acid or sodium hydroxide. 1-Octanol and phosphate buffer (20 mM) were saturated by tumbling overnight. The two phases were separated using a separating funnel. A 5 μL cassette of 5 mM compound was added to 495 μL of octanol-saturated buffer and 495 μL of buffer-saturated octanol in a 96-well plate (maximum concentration 50 μM). The plate was shaken for 1 hour and centrifuged at 25 °C for 5 minutes. 200 μL of each phase was transferred to another plate. To avoid cross-contamination, the octanol layer was sampled first. 5 μL of the solution was transferred to 495 μL of quenching solution * (maximum concentration 0.5 μM). In addition, 40 μL of buffer was added to 360 μL of quenching solution (maximum concentration 5 μM). The samples were analyzed by LC-MS / MS. Evaluation was performed with respect to the calibration curve for each compound and with reference to the control compounds sulpiride, diclofenac, chlorpromazine and tamoxifen. The quenching solution was a 1:3:1 (v / v / v) mixture of acetonitrile:acetonitrile:water containing 0.1% formic acid and imipramine / labetalol, 200 nM.

[0284] The results of the LogD assay are shown in Table 4. Compounds having an mLogD value at pH 7.4 of 5 or less are generally sufficiently soluble in pharmaceutical formulations, and preferably have an mLogD value of 4.2 or less. All of the compounds of the examples fall within this range. TIFF0007682850000106.tif255170TIFF0007682850000107.tif63170

[0285] Example 16 - Microsomal Stability Microsomes (human) were obtained from Bioreclamation.

[0286] All test compounds and reference control compounds (raloxifene, diclofenac, terfenadine, propranolol, dextromethorphan and metoprolol) were dissolved to create a 100 μM stock (final concentration; 91.5% acetonitrile:85% DMSO). The final test compound concentration in the incubation was 1 μM (<0.1% DMSO).

[0287] The assay buffer was prepared from Potassium Phosphate Solution 1 and 2 by mixing to form a pH 7.42 solution at 37°C. Solution 1: 17.4 g of dipotassium phosphate anhydrous (K 2 HPO 4 , 0.1 M) dissolved in 1 L of deionized water. Solution 2: 13.6 g of potassium dihydrogen phosphate anhydrous (KH 2 PO 4 , 0.1 M) dissolved in 1 L of deionized water. Adjusted to pH 7.4 with 2 mM magnesium chloride. NADPH (10 mM) was prepared in deionized water.

[0288] Microsomes (from all species) were removed at -80 °C and thawed at 37 °C. The microsomes were diluted with assay buffer to achieve a final protein concentration of 0.5 mg / mL and 1 mM NADPH.

[0289] The following procedure was completed on a Perkin Elmer Janus robotic platform in 96-well format: The microsome incubation plate was transferred to a heater shaker at 300 rpm and the solution was heated to 37 °C for 10 minutes pre-warming. There were no cofactor controls at 0 and 45 minutes and one set of each test compound was included per assay. The microsomes were incubated at 37 °C on a shaker set at 300 rpm throughout the assay. At each time point (0, 5, 15, 30, 45 minutes), 50 μL of the sample was removed from the 96-well plate and added to 200 μL of quenching solution (acetonitrile containing 0.1% formic acid and imipramine / labetalol, 200 nM). The samples were diluted 1:1 with water using the Janus robot and analyzed by LC-MS / MS. The results obtained were quantified by a standard calibration curve generated for each test sample and the results were controlled by analysis of a reference control compound.

[0290] The microsome clearance data are shown in Table 5. TIFF0007682850000108.tif255170TIFF0007682850000109.tif72170

[0291] Although not essential, compounds intended for oral administration preferably have a low microsome clearance rate. The target value for Cl int for such compounds in the microsome stability assay is less than 30 μL / min / mg, preferably less than 20 μL / min / mg and particularly preferably less than 10 μL / min / mg.

[0292] Compounds having a clearance rate within this range are particularly suitable for oral administration because they tend to have a lower propensity for increased oral bioavailability leading to metabolism contributing to first-pass clearance resulting in higher systemic exposure and lower doses for a given pharmacological effect.

[0293] Although not essential, compounds administered by inhalation preferably have a high clearance rate. The target value of Cl for such compounds in a microsomal stability assay is greater than 35 μL / min / mg, preferably greater than 40 μL / min / mg and particularly preferably greater than 50 μL / min / mg. int is greater than 35 μL / min / mg, preferably greater than 40 μL / min / mg and particularly preferably greater than 50 μL / min / mg.

[0294] A high microsomal clearance rate corresponding to a low in vivo half-life after absorption into the systemic circulation ensures that a compound administered directly to the lung is less likely to interact with TMEM16A at other sites in the body and thus potential side effects are minimized.

[0295] 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. Gupta D, Bhatia D, Dave V, Sutariya V & Gupta SV (2018) Salts of Therapeutic Agents: Chemical, Physicochemical, and Biological Considerations. Molecules, 23, 1719. 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) reduced airway surface pH impairs bacterial killing in the porcine cystic fibrosis lung. Nature, 487(7405):109 - 113. Tang R-J, Milcent T, Crousse B (2018) Regioselective Halogenation of Arenes and Heterocycles in Hexafluoroisopropanol.J. Org. Chem. 2018, 83(2), 930-938. 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. A compound of general formula (I), including all its tautomeric forms, all enantiomers, isotope variants, as well as salts and solvates: (wherein, R １ is selected from H, ethynyl, CN, methyl, fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl, dichlorofluoromethyl and hydroxymethyl, R ２ is selected from methyl and CH ２ OH, R ３ is selected from H and methyl, or R ２ and R ３ together with the carbon atom to which they are attached, in addition to the R １ group, form a 3- to 10-membered carbocyclic ring or an oxygen-containing heterocyclic ring which may be substituted with one or more substituents selected from C １～４ alkyl, C １～４ haloalkyl, halo and OH, or R １ , R ２ and R ３ together with the carbon atoms to which they are attached combine to form a 5- to 8-membered bridged carbocyclic or bridged heterocyclic ring optionally substituted with one or more substituents selected from OH, halo, C １～４ alkyl and C １～４ haloalkyl, R ４ is H or halo, R ５ is selected from C １～４ alkyl, which may be substituted with one or more substituents selected from H, halo, CN and halo and OH X １ is either CR ６ or N, and R ６ is selected from C alkyl optionally substituted with one or more substituents selected from H, halo, CN and halo and OH, １～４ and X ２ is either CR ７ or N, and R ７ is a 3- to 7-membered carbocyclic or heterocyclic ring optionally substituted with one or more substituents selected from H, halo, CN; halo and OH; or a C optionally substituted with one or more substituents selected from halo, OH and one or more substituents selected from halo and OH, which are optionally substituted with one or more substituents selected from a 3- to 7-membered carbocyclic or heterocyclic ring １～４ is alkyl, X ３ is either CR ８ or N, and R ８ is C optionally substituted with one or more substituents selected from H, halo, CN and halo and OH １～４ alkyl).

2. R １ The compound according to claim 1, wherein R is H, methyl, difluoromethyl, trifluoromethyl, ethynyl or CN.

3. R ２ is methyl and R ３ is methyl, the compound according to claim 1.

4. R １ The compound according to claim 3, wherein R is methyl, difluoromethyl, trifluoromethyl, ethynyl or CN.

5. R ２ and R ３ together with the carbon atom to which they are attached form an optionally substituted 3- to 10-membered carbocyclic or oxygen-containing heterocyclic ring as defined in claim 1, a compound according to claim 1 or claim 2.

6. R ２ and R ３ are, together with the carbon atom to which they are attached, unsubstituted (except for the R １ group) or substituted with one or two substituents selected from methyl and fluoro in addition to the R １ group, forming a 3- to 10-membered carbocyclic ring or an oxygen-containing heterocyclic ring, the compound according to claim 5.

7. R １ 、 R ２ and R ３ which, together with the carbon atoms to which they are attached, combine to form a 5- to 8-membered bridged carbocyclic or heterocyclic ring, the compound according to claim 1.

8. R ４ The compound according to any one of claims 1 to 7, wherein R is H.

9. R ５ is H, fluoro or chloro; and / or X １ is CR ６ where R ６ is H or halo; and / or X ２ is CR ７ where R ７ is H; C alkyl optionally substituted with OH; C １～３ haloalkyl; methyl substituted with a 3- to 6-membered carbocyclic or heterocyclic ring; or a 3- to 6-membered carbocyclic or heterocyclic ring; and / or １～３ ​ X ３ is N, A compound according to any one of claims 1 to 8.

10. A compound of general formula (Ia): (wherein X ３ , R １ , R ２ , R ３ , R ４ , R ５ , R ６ and R ７ are as defined in claim 1); or A compound of general formula (Ib): (wherein, R １ , R ２ , R ３ , R ４ , R ５ , R ６ and R ７ are as defined in claim 1); or A compound of general formula (Ic): (wherein, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ and R ８ are as defined in claim 1) A compound according to any one of claims 1 to 9, which is as defined above.

11. R ５ , R ６ and R ７ wherein at least one of the R ５ , R ６ and R ７ is H, the compound according to claim 10.

12. R ５ is a halo and R ６ is H; or R ６ is a halo and R ５ is H; or R ５ is a halo, and R ６ and R ７ are both H; or R ６ is a halo and R ５ and R ７ are both H; or R ７ is other than H, and R ５ and R ６ are both H, A compound according to claim 11.

13. N-(1,1-Dimethylprop-2-ynyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 1); 4-[[2-(1H-Indol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 1.1) N-tert-Butyl-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 2); N-(1-Cyano-1-methyl-ethyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 2.1); N-(1,1-Dimethylprop-2-ynyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 3); N-(1-Ethynylcyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 4); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide (Compound 4.1); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 4.2); N-(3,3-Difluoro-1-methyl-cyclobutyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 4.3); N-(2,2-Difluoro-1,1-dimethyl-ethyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclobutyl]pyridine-2-carboxamide (Compound 5.1); N-(3-Fluoro-3-methyl-cyclobutyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.2); N-(2,2-Difluorocyclopentyl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.3); N-(4-Cyanotetrahydropyran-4-yl)-4-[[2-(1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 5.4); N-tert-Butyl-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 6); 4-[[2-(5-Fluoro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 6.1); N-(2,2-Difluorocyclopentyl)-4-[[2-(5-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 6.2); 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 6.3); 4-[[2-(4-Chloro-1H-indazol-6-yl)acetyl]amino]-N-(1-ethynylcyclopentyl)pyridine-2-carboxamide (Compound 6.4); N-tert-Butyl-4-[[2-(4-fluoro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7); N-tert-Butyl-4-[[2-(5-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7.1); N-tert-Butyl-4-[[2-(4-chloro-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 7.2); N-tert-Butyl-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Compound 8); N-[1-(Trifluoromethyl)cyclopropyl]-4-[[2-[3-(trifluoromethyl)-1H-indazol-6-yl]acetyl]amino]pyridine-2-carboxamide (Compound 8.1); N-tert-Butyl-4-[[2-(3-isopropyl-1H-indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 9); 4-[[2-(3-Tetrahydrofuran-2-yl-1H-indazol-6-yl)acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 9.1); 4-[[2-[3-(Morpholinomethyl)-1H-indazol-6-yl]acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide (Compound 10); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(3-methyloxetan-3-yl)pyridine-2-carboxamide (Compound 11); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-(2-oxaspiro[3.3]heptan-6-yl)pyridine-2-carboxamide (Compound 11.1); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[4-(trifluoromethyl)tetrahydropyran-4-yl]pyridine-2-carboxamide (Compound 11.2); 4-[[2-(1H-Indazol-6-yl)acetyl]amino]-N-[3-(trifluoromethyl)oxetan-3-yl]pyridine-2-carboxamide (Compound 11.3); N-[1-(Difluoromethyl)cyclopropyl]-4-[[2-(1H-Indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 11.4); N-(3-Fluoro-1-bicyclo[1.1.1]pentanyl)-4-[[2-(1H-Indazol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 11.5); 4-[[2-[3-(2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl)-1H-indazol-6-yl]acetyl]amino]-N-[1-(trifluoromethyl)cyclopropyl]pyridine-2-carboxamide (Compound 12); 4-[[2-(1H-indol-6-yl)acetyl]amino]-N-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyridine-2-carboxamide; (Compound 13); N-(3,3-difluoro-1-methyl-cyclobutyl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 13.1); N-(4-cyanotetrahydropyran-4-yl)-4-[[2-(1H-indol-6-yl)acetyl]amino]pyridine-2-carboxamide (Compound 13.2) A compound according to claim 1, selected from the above, and salts and solvates of the above.

14. A compound according to any one of claims 1 to 13 for use in medicine.

15. A compound according to any one of claims 1 to 13 for use in the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A.

16. Use of a compound according to any one of claims 1 to 13 in the manufacture of a medicament for the treatment or prevention of diseases and conditions affected by the modulation of TMEM16A.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 and a pharmaceutically acceptable additive.

18. A method for the preparation of a compound according to any one of claims 1 to 13, comprising: A. Reacting a compound of general formula (II): (wherein, R １ , R ２ , R ３ and R ４ are as defined for general formula (I)) with a compound of general formula (III): (wherein R ５ , X １ , X ２ and X ３ are as defined in claim 1) or B. Deprotecting a protected compound of general formula (Iz): (wherein, R １ , R ２ , R ３ , R ４ , R ５ , X １ , X ２ and X ３ are as defined in claim 1, and R １５ is an amine protecting group) or C. Reacting a compound of general formula (IV): (wherein R １ , R ２ , and R ３ are as defined in claim 1) with a compound of general formula (XII): (wherein R ４ , R ５ , X １ , X ２ and X ３ are as defined in claim 1) or D.R ７ In the case of a compound of general formula (Ib) where D.R is alkyl or heterocyclyl: Reacting a compound of general formula (XXVaz): (wherein R １ , R ２ , R ３ , R ４ , R ５ , and R ６ are as defined in claim 1, R １５ is an amine protecting group, and R ２１ is halo) with a suitable alkyldioxaborolane or heterocyclyldioxaborolane in the presence of a palladium catalyst under basic conditions, followed by catalytic hydrogenation on a palladium catalyst.

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