Disubstituted pyrazole compounds as hexoketokinase inhibitors

Abstract

The present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof, and the use of the compound of formula I for treating metabolic disorders such as type 2 diabetes, heart failure, diabetic nephropathy and non-alcoholic steatohepatitis.

Description

[0001] The present invention relates to novel ketohexokinase (KHK) inhibitor compounds, pharmaceutical compositions comprising the compounds, and the use of the compounds for treating certain conditions, such as type 2 diabetes mellitus (T2DM), heart failure, diabetic nephropathy and non-alcoholic steatohepatitis (NASH).

[0002] KHK, also known as fructokinase, is a rate-limiting enzyme involved in fructose metabolism. It catalyzes the phosphorylation of fructose to fructose-1-phosphate (F1P), resulting in the concomitant consumption of cellular ATP levels. In contrast to glucose, fructose metabolism lacks feedback inhibition, and it triggers the accumulation of downstream intermediates involved in, for example, lipogenesis, gluconeogenesis, and oxidative phosphorylation (Hannou, SA, et al.; J. Clin. Invest., 128 (2), 544-555, 2018). This has negative metabolic consequences associated with many serious metabolic disorders.

[0003] KHK exists in two alternatively spliced ​​isoforms, including KHK-C and KHK-A, which are different in exon 3. KHK-C is mainly expressed in the liver, kidney and intestine, while KHK-A is more commonly present. Mice lacking both isoforms completely avoid fructose-induced metabolic syndrome. However, this adverse metabolic effect is exacerbated in mice lacking only KHK-A (Ishimoto T, et al.; Proc. Natl. Acad. Sci. USA, 109 (11), 4320-4325, 2012).

[0004] Several epidemiological and experimental studies have reported that increased fructose consumption, or more precisely, increased fructose metabolism, may play an important role in the development of several diseases, including metabolic syndrome, particularly T2DM (Softic et al.; J. Clin. Invest., 127(11), 4059-4074, 2017), heart failure (Mirtschink, P., et al.; Eur. Heart J., 39, 2497-2505, 2018), diabetic nephropathy (Cirillo, P., et al.; J. Am. Soc. Nephrol., 20, 545-553, 2009), and NAFLD / NASH (Vos, MB, et al.; Hepatology, 57, 2525-2531, 2013). Targeted inhibition of KHK is expected to limit fructose metabolism and provide an effective treatment option for many metabolic disorders.

[0005] US 2017 / 0183328 A1 discloses substituted 3-azabicyclo[3.1.0]hexanes as KHK inhibitors. Recently published data show that the hexokinases inhibitor PF-06835919 can reduce total liver fat after 6 weeks of administration in individuals with non-alcoholic fatty liver disease, as measured by magnetic resonance imaging proton density fat fraction (J. Hepatology. EASL International Liver Congress Abstracts, Supplement N°1S Vol. 70, April 2019).

[0006] Compounds containing carboxyl functional groups have the risk of forming acyl glucuronide metabolites (Vleet Van et al., Toxicology Letters, 272 (2017) 1-7). Acyl glucuronide metabolites are generally unstable and may undergo chemical reactions, resulting in covalent bonding to macromolecules and toxicity.

[0007] Alternative therapies for metabolic syndrome and related indications (including T2DM, heart failure, diabetic nephropathy and NASH) are needed. In particular, compounds of effective KHK inhibitors are needed. KHK inhibitor compounds with favorable properties are needed, for example, good oral bioavailability to support once-daily administration. In addition, KHK inhibitor compounds are also needed, which do not have a carboxylic acid moiety and lack the ability to form acyl glucuronides.

[0008] Therefore, in one embodiment, the present invention provides a compound of formula I:

[0009]

[0010] in

[0011] X is N or C substituted by CN;

[0012] R 1 Selected from: H.

[0013]

[0014] R 2 and R 3 Both are H, or one is H and the other is OH;

[0015] R 4 , R 5 , R 6 , R 7 and R 9 are independently H or CH3;

[0016] R 8is H, CH3, CH2CH2OH, C(=O)CH2NH2 or C(=O)CH3; and

[0017] R 10 It is OH or NH2;

[0018] or a pharmaceutically acceptable salt thereof.

[0019] In a specific embodiment, R 1 Selected from: H.

[0020]

[0021] or a pharmaceutically acceptable salt thereof.

[0022] In a specific embodiment, there is provided a compound of formula I, wherein X is N or C substituted with CN;

[0023] R 1 Is selected from:

[0024]

[0025] And among them

[0026] R 2 is H and R 3 Yes OH,

[0027] or R 2 is OH and R 3 It is H.

[0028] or R 2 and R 3 All are H;

[0029] or a pharmaceutically acceptable salt thereof.

[0030] In a specific embodiment, the compound is of Formula Ia:

[0031]

[0032] or a pharmaceutically acceptable salt thereof.

[0033] In a specific embodiment, the compound is of Formula Ic:

[0034]

[0035] or a pharmaceutically acceptable salt thereof.

[0036] In a specific embodiment, the compound is of Formula Ie:

[0037]

[0038] or a pharmaceutically acceptable salt thereof.

[0039] In one embodiment, X is N.

[0040] In one embodiment, X is C substituted with CN.

[0041] In one embodiment, R 1 yes

[0042] In one embodiment, the compound of formula I is:

[0043]

[0044] or a pharmaceutically acceptable salt thereof.

[0045] In a specific embodiment, it provides a succinate salt of the formula:

[0046]

[0047] In a preferred embodiment, the succinate salt is sesquisuccinate.

[0048] In one embodiment, the compound of formula I is:

[0049]

[0050] or a pharmaceutically acceptable salt thereof.

[0051] In one embodiment, the compound of formula I is:

[0052]

[0053] or a pharmaceutically acceptable salt thereof.

[0054] In one embodiment, the compound of formula I is:

[0055]

[0056] or a pharmaceutically acceptable salt thereof.

[0057] In one embodiment, the compound of formula I is:

[0058]

[0059] or a pharmaceutically acceptable salt thereof.

[0060] In one embodiment, the compound of formula I is:

[0061]

[0062] or a pharmaceutically acceptable salt thereof.

[0063] In one embodiment, the compound of formula I is:

[0064]

[0065] or a pharmaceutically acceptable salt thereof.

[0066] In a specific embodiment, the compound is selected from:

[0067] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone;

[0068] [(2R)-1-[4-[1-(4-piperidinyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;

[0069] [(2R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;

[0070] (2S,3R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol;

[0071] 2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-N-(2-hydroxyethyl)acetamide;

[0072] 2-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0073] 2-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0074] 2-[(2S)-2-methylazetidin-1-yl]-6-[1-(1-methyl-4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0075] 6-[1-[1-(2-hydroxyethyl)-4-piperidinyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0076] 6-[1-[2-(Dimethylamino)ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0077] 6-[1-(2-hydroxyethyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0078] 2-[(2S)-2-methylazetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0079] 2-[(2S)-2-methylazetidin-1-yl]-4-[1-(4-piperidinyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine;

[0080] (2S,3R)-2-methyl-1-[4-[1-[(3R)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol;

[0081] (2S,3R)-2-methyl-1-[4-[1-[(3S)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol;

[0082] [(2R)-1-[4-[1-[(3R)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;

[0083] 6-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0084] 6-[1-[1-(2-aminoacetyl)-4-piperidinyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0085] 6-[1-(1-acetyl-4-piperidinyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0086] 2-[(2S)-2-methylazetidin-1-yl]-6-[1-[2-(4-methylpiperazin-1-yl)-2-oxo-ethyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0087] 2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-N,N-bis(2-hydroxyethyl)acetamide;

[0088] 6-[1-[2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0089] 6-[1-[2-[(3S,4S)-3,4-dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0090] 6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0091] 2-[(2S)-2-Methylazetidin-1-yl]-6-(1H-pyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0092] 2-[(2S)-2-Methylazetidin-1-yl]-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0093] 6-[1-(3-hydroxy-4-piperidinyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;

[0094] N-(2-aminoethyl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetamide;

[0095] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(3S)-3-methylpiperazin-1-yl]ethanone;

[0096] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(2S)-2-methylpiperazin-1-yl]ethanone;

[0097] 1-(3,3-dimethylpiperazin-1-yl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]ethanone;

[0098] 1-[(2S,5R)-2,5-dimethylpiperazin-1-yl]-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]ethanone;

[0099] 4-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidine;

[0100] 2-[(2S)-2-methylazetidin-1-yl]-4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine;

[0101] (2S,3R)-2-methyl-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol; or

[0102] [(2R)-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;

[0103] or a pharmaceutically acceptable salt thereof.

[0104] In a specific embodiment, the compound is 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone sesquisuccinate, which is also known as ethanone, 2-[4-[2-[(2S)-2-methyl-1-azetidinyl]-6-(trifluoromethyl)-4-pyrimidinyl]-1H-pyrazol-1-yl]-1-(1-piperazinyl)-, succinate (1:1.5), or succinic acid-2-(4-{2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl}-1H-pyrazol-1-yl)-1-(piperazin-1-yl)ethan-1-one (1.5 / 1).

[0105] Formula I includes Formulas Ia, Ib, Ic, Id, Ie, If, Ig and Ih, and reference to Formula I hereinafter, for example in methods of treatment and therapeutic uses, is to be understood as reference to each and all of these subformulae.

[0106] In one embodiment, a method of treating T2DM in a patient in need of such treatment is provided, comprising administering to the patient an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof.

[0107] In one embodiment, a method of treating heart failure in a patient in need of such treatment is provided, comprising administering to the patient an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof.

[0108] In one embodiment, a method of treating diabetic nephropathy in a patient in need of such treatment is provided, comprising administering to the patient an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof.

[0109] In one embodiment, a method of treating NASH in a patient in need of such treatment is provided, comprising administering to the patient an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof.

[0110] In one embodiment, a method of treating a disease selected from metabolic syndrome, NAFLD, obesity, cardiovascular disease, coronary artery disease, chronic kidney disease, dyslipidemia, and diabetic complications (e.g., diabetic retinopathy) in a patient in need of such treatment is provided, comprising administering to the patient an effective amount of Formula I, or a pharmaceutically acceptable salt thereof.

[0111] In addition, in one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment. In one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment of T2DM. In one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment of heart failure. In one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment of diabetic nephropathy. In one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment of NASH. In one embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof is provided for treatment of metabolic syndrome, NAFLD, obesity, cardiovascular disease, coronary artery disease, chronic kidney disease, dyslipidemia or diabetic complications (e.g., diabetic retinopathy).

[0112] In addition, in one embodiment, there is provided the use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating T2DM. In one embodiment, there is provided the use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating heart failure. In one embodiment, there is provided the use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating diabetic nephropathy. In one embodiment, there is provided the use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating NASH. In one embodiment, there is provided the use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating metabolic syndrome, NAFLD, obesity, cardiovascular disease, coronary artery disease, chronic kidney disease, dyslipidemia, or diabetic complications (e.g., diabetic retinopathy).

[0113] In one embodiment, a pharmaceutical composition is provided, comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients. In one embodiment, a method for preparing a pharmaceutical composition is provided, comprising mixing a compound of formula I or a pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable carriers, diluents or excipients.

[0114] As used herein, the term "treating" includes inhibiting, slowing, stopping or reversing the progression or severity of an existing symptom or disorder.

[0115] As used herein, the term "patient" refers to a mammal. Preferably, the patient is a human.

[0116] As used herein, the term "effective amount" refers to the amount or dosage of a compound of Formula I or a pharmaceutically acceptable salt thereof, which provides the desired effect in the patient being diagnosed or treated after single or multiple doses of administration to the patient.

[0117] The effective amount can be determined by those skilled in the art by using known techniques and by observing the results obtained under similar circumstances. In determining the effective amount for a patient, many factors are considered, including but not limited to: the species of the patient; its body size, age and general health; the specific disease or disorder involved; the extent or involvement or severity of the disease or disorder; the response of the individual patient; the specific compound administered; the mode of administration; the bioavailability characteristics of the administered formulation; the selected dosing regimen; the use of concomitant medications; and other relevant circumstances. The compound of formula I is effective in a daily dose in the range of about 0.1 to about 15 mg / kg body weight.

[0118] The compound of formula I is formulated into a pharmaceutical composition, which is administered by any route that makes the compound bioavailable. Preferably, such compositions are for oral administration. Such pharmaceutical compositions and methods for preparing them are well known in the art (see, for example, Remington, JP, "Remington: The Science and Practice of Pharmacy", LV Allen, ed., 22nd ed., Pharmaceutical Press, 2012).

[0119] The compounds of formula I and their pharmaceutically acceptable salts can be used in the methods of treatment and therapeutic uses of the present invention, wherein certain configurations are preferred. It should be understood that the following preferences apply to the methods of treatment, therapeutic uses and compounds of the present invention.

[0120] The compounds of the present invention include:

[0121]

[0122]

[0123]

[0124] and pharmaceutically acceptable salts thereof.

[0125] While the present invention contemplates all individual enantiomers and diastereomers as well as mixtures of the compounds, including racemates, compounds of Formula Ia, Formula Ic and Formula Ie and pharmaceutically acceptable salts thereof are particularly preferred.

[0126] Individual enantiomers may be separated or resolved by one of ordinary skill in the art at any convenient point in the synthesis of compounds of Formula I by methods such as selective crystallization techniques, chiral chromatography (see, e.g., J. Jacques, et al., "Enantiomers, Racemates, and Resolutions", John Wiley and Sons, Inc., 1981, and E.L. Eliel and S.H. Wilen, "Stereochemistry of Organic Compounds", Wiley-Interscience, 1994), or supercritical fluid chromatography (SFC) (see, e.g., T.A. Berger; "Supercritical Fluid Chromatography Primer," Agilent Technologies, July 2015).

[0127] Pharmaceutically acceptable salts of compounds of formula I can be formed, for example, by reacting a suitable free base of a compound of formula I with a suitable pharmaceutically acceptable acid in a suitable solvent under standard conditions known in the art (see, for example, Bastin, RJ, et al.; Org. Process. Res. Dev., 4, 427-435, 2000 and Berge, SM, et al.; J. Pharm. Sci., 66, 1-19, 1977). A preferred salt is succinate. A particularly preferred salt is sesquisuccinate. In sesquisuccinate, the ratio of free base to succinic acid is 1:1.5. The succinate is also known as succinate.

[0128] Formula I compound or its salt can be prepared by various methods known to those of ordinary skill in the art, some of which are described in the following flow, preparation and examples. The product of each step in the following flow can be recovered by conventional methods known in the art, including extraction, evaporation, precipitation, chromatography, filtration, grinding and crystallization. In the following flow, unless otherwise indicated, all substituents are as previously defined. Reagents and starting materials are easily available to those of ordinary skill in the art. Without limiting the scope of the invention, the following flow, preparation and examples are provided to further illustrate the present invention. In addition, it is understood by those of ordinary skill in the art that the compound of formula I can be prepared by using starting materials or intermediates with corresponding desired stereochemical configurations that can be prepared by those skilled in the art.

[0129] Certain abbreviations are defined as follows: “ACN” refers to acetonitrile; “BOC” refers to tert-butyloxycarbonyl; “DCM” refers to dichloromethane; “DIPEA” refers to N,N-diisopropylethylamine; “DMF” refers to N,N-dimethylformamide; “DMSO” refers to dimethyl sulfoxide; “ELSD” refers to evaporative light scattering detector; “ES / MS” refers to electrospray mass spectrometry; “EtOAc” refers to ethyl acetate; “EtOH” refers to ethanol; “h” refers to hour; “HATU” refers to 1-[[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4, 5-b] pyridinium 3-oxide hexafluorophosphate; "HPLC" means high performance liquid chromatography; "IPA" means isopropyl alcohol; "Me" means methyl; "MeOH" means methanol; "MTBE" means methyl tert-butyl ether; "min" means minute; "m / z" means mass-to-charge ratio; "Ph" means phenyl; "RBF" means round-bottom flask; "RT" means room temperature; "SCX" means selective cation exchange; "SEM" means standard error of the mean; "SFC" means supercritical fluid chromatography; "TFA" means trifluoroacetic acid; and "THF" means tetrahydrofuran.

[0130] Process 1

[0131]

[0132] Scheme 1 describes a general method for the preparation of compounds of formula I. 1a It may be related to R in the final compound of formula I 1 or may need to be converted to obtain R of Formula I 1 The group R 1a Conventional synthetic transformations (e.g., BOC deprotection, ester hydrolysis, and amide coupling reactions) of can be performed before or after step B. In step A, a Suzuki cross-coupling reaction between a heteroaryl dichloride 1 and a pyrazole borate ester 2 generates a substituted pyrazole compound 3. The reaction is carried out in an organic solvent (e.g., 1,4-dioxane) using a base (e.g., 2M Na2CO3 aqueous solution) in the presence of a palladium catalyst (e.g., tetrakis(triphenylphosphine)palladium(0) or bis(triphenylphosphine)palladium(II) dichloride) at elevated temperatures. In step B, compound 3 is subjected to a nucleophilic aromatic substitution reaction with a substituted azetidine 4 to obtain a compound of formula I. The reaction is carried out in an organic solvent (e.g., 1,4-dioxane) at elevated temperatures in the presence of an organic base (e.g., DIPEA).

[0133] Process 2

[0134]

[0135] Scheme 2 describes a method for preparing a group of compounds of Formula I, wherein the pyrazole is substituted with a nitrogen-containing heterocycle (Formula I'). Pyrazole boronate ester 5 is substituted with a nitrogen-containing heterocycle as described in Scheme 2, wherein "Y" and "Z" can each independently be -CH2- or -CH2-CH2-, and "PG" is a nitrogen protecting group, such as BOC. In step C, a Suzuki cross-coupling reaction between heteroaryl dichloride 1 and pyrazole boronate ester 5 produces substituted pyrazole compound 6. As described in step A of Scheme 1, the reaction is carried out in an organic solvent (such as 1,4-dioxane) using a base (such as 2M aqueous Na2CO3) in the presence of a palladium catalyst (such as tetrakis(triphenylphosphine)palladium(0) or bis(triphenylphosphine)palladium(II) dichloride) at elevated temperature. As described in Scheme 2, two different routes can be used, Route A and Route B.

[0136] In route A, substituted pyrazole compound 6 undergoes deprotection in step D to give compound 7. For example, if "PG" is BOC, deprotection can be accomplished using TFA. In step E, the heterocyclic nitrogen of compound 7 can undergo a substitution reaction, such as reductive amination, acylation, or amide coupling to give compound 8. In this process, such substitutions are described as "R". In step F, compound 8 is subjected to a nucleophilic aromatic substitution reaction with a substituted azetidine 4 to give a compound of formula I'. As described in step B of process 1, the reaction is carried out in an organic solvent (e.g., 1,4-dioxane) in the presence of an organic base (e.g., DIPEA) at elevated temperature. This may be the last step, or the R group may undergo other conventional synthetic transformations, such as removal of protecting groups.

[0137] In route B, substituted pyrazole compound 6 undergoes a nucleophilic aromatic substitution reaction with substituted azetidine 4 in step G to provide compound 9. This can be achieved by a nucleophilic aromatic substitution reaction with substituted azetidine 4 in an organic solvent (e.g., 1,4-dioxane) at elevated temperature in the presence of an organic base (e.g., DIPEA). Alternatively, steps C and G can be accomplished in a one-pot process, where the Suzuki cross-coupling reaction of step C is first accomplished, and then azetidine 4 is added to the reaction along with an organic base (e.g., DIPEA), followed by step G at elevated temperature. The protecting group "PG" is removed in step H (e.g., using TFA if PG is BOC) to provide compound 10. In step I, the nitrogen group of compound 10 may undergo a substitution reaction, such as reductive amination, acylation, or amide coupling, to provide a compound of formula I'. This may be the last step, or the R group may undergo other conventional synthetic transformations, such as removal of the protecting group.

[0138] Process 3

[0139]

[0140] Scheme 3 describes the preparation of a group of compounds of Formula I, wherein the pyrazole is substituted with an acetamide group (Formula I"). In step J, a Suzuki cross-coupling reaction between heteroaryl dichloride 1 and pyrazole boronate ester 11 produces substituted pyrazole compound 12. As described in Schemes 1 and 2, the reaction is carried out in an organic solvent (e.g., 1,4-dioxane) using a base (e.g., 2 M Na2CO3 aqueous solution) in the presence of a palladium catalyst (e.g., tetrakis(triphenylphosphine)palladium(0) or bis(triphenylphosphine)palladium(II) dichloride) at elevated temperature. In step K, heteroaryl chloride 12 undergoes nucleophilic aromatic substitution with azetidine 4 in an organic solvent (e.g., 1,4-dioxane) in the presence of an organic base (e.g., DIPEA) at elevated temperature to give compound 13. In step L, ester hydrolysis is performed using a base (e.g., sodium hydroxide) to give acid 14. In step M, acid 14 is subjected to an amide coupling reaction with an amine of formula HNR'R" to give an amide compound of formula I". The amine HNR'R" may be cyclic (e.g., optionally substituted piperazine). Step M may be the last step, and there may be other conventional synthetic transformations, such as removal of protecting groups.

[0141] Preparation method and examples

[0142] The following preparation methods and examples further illustrate various embodiments of the present invention and represent typical synthesis of compounds of the present invention. Reagents and starting materials are readily available or can be easily synthesized by those of ordinary skill in the art. It should be understood that the preparation methods and examples are illustrated by way of example and not limitation, and various modifications may be made by those of ordinary skill in the art.

[0143] LC-ES / MS HP1200 liquid chromatography system. Electrospray mass spectrometry measurements (acquired in positive ion mode and / or negative ion mode) were performed on a mass selective detector quadrupole mass spectrometer interfaced with an HPLC, which may or may not have an ELSD. LC-MS conditions (low pH): Column: NX C18 2.0×50mm 3.0μm, Gradient: 5-95% B in 1.5 min, then 95% B for 0.5 min Column temperature: 50°C + / - 10°C; Flow rate: 1.2 mL / min; 1 μL injection volume; Solvent A: 0.1% HCOOH in deionized water; Solvent B: 0.1% formic acid in ACN; Wavelengths 200-400 nm and 212-216 nm. If the HPLC is equipped with an ELSD, set to 45°C evaporator temperature, 40°C nebulizer temperature and 1.6 SLM gas flow rate. Alternative LC-MS conditions (high pH): Column: Waters C18 column 2.1×50 mm, 3.5 μm; gradient: 5-95% B in 1.5 min, then 95% B for 0.50 min; column temperature: 50°C+ / -10°C; flow rate: 1.2 mL / min; 1 μL injection volume; solvent A: 10 mM NH4HCO3 pH 9; solvent B: ACN; wavelengths: 200-400 nm and 212-216 nm; if there is ELSD, 45°C evaporator temperature, 40°C nebulizer temperature and 1.6 SLM gas flow rate.

[0144] The XRPD pattern of the crystalline solid was obtained on a Bruker D4Endeavor X-ray powder diffractometer equipped with a CuKα source and a Vantec detector, operating at 35kV and 50mA. The sample was scanned between 4-40 2θ°, with a step size of 0.008 2θ°, a scan rate of 0.5 seconds / step, using a 1.0mm divergence, a 6.6mm fixed anti-scattering, and an 11.3mm detector slit. The dried powder was packed in a quartz sample holder and a glass slide was used to obtain a smooth surface. The diffraction pattern of the crystalline form was collected at ambient temperature and relative humidity. Based on the internal NIST 675 standard, the crystal peak position in MDI Jade was determined after the entire pattern was moved, with peaks at 8.853 and 26.774 2θ°. It is well known in the field of crystallography that for any given crystalline form, the relative intensity of the diffraction peaks may vary due to preferred orientation caused by factors such as crystal morphology and habit. When there is a preferred orientation effect, the peak intensity changes, but the characteristic peak position of the polymorph remains unchanged. See, for example, The United States Pharmacopeia #23, National Formulary #18, pp. 1843-1844, 1995. In addition, it is also well known in the art of crystallography that for any given crystalline form, the angular peak position may vary slightly. For example, the peak position may shift due to temperature changes, sample replacement, or the presence or absence of an internal standard when analyzing the sample. In the present case, a peak position variation of ±0.2 2θ° is assumed to account for these potential variations without hindering the clear identification of the crystalline form shown. The crystalline form can be confirmed based on any unique combination of distinguishing peaks.

[0145] Preparation 1

[0146] (2S)-1-Benzhydryl-2-methyl-azetidine [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl] methanesulfonate

[0147]

[0148] Assemble a 2000mL three-neck RBF equipped with an addition funnel, nitrogen inlet, and thermometer adapter. Purge the container with nitrogen and add (3R)-butane-1,3-diol (25g, 277mmol), DIPEA (127mL, 731mmol), and ACN (556mL). The mixture is cooled to -30°C. Trifluoromethanesulfonic anhydride (101mL, 601mmol) is added dropwise over 3 hours, keeping the internal temperature between -35 and -30°C. After addition, stir at -35 to -30°C for 10 minutes. Trifluoromethanesulfonic anhydride (1.9mL, 11mmol) is added dropwise over 5 minutes, keeping the internal temperature between -35 to -30°C. After addition, stir at -35 to -30°C for 10 minutes. DIPEA (127mL, 731mmol) was added dropwise over 15 minutes, and the internal temperature was maintained between -35 and -30°C. After completion, stirred at -35 to -30°C for 10 minutes. Under nitrogen, aminodiphenylmethane (48.0mL, 270mmol) was dissolved in ACN (49mL) in a separate flask, and the resulting solution was transferred to an addition funnel. The amine solution was added dropwise to the cooling triflate over 40 minutes, and the internal temperature was maintained between -20 and -35°C. After completion, stirred at a temperature of -35 to -30°C for 30 minutes. The reactant was transferred to a water bath, and allowed to heat slowly for 30 minutes. The water bath was removed, and the reactant was heated to RT in 30 minutes. The container was transferred to a heating mantle, and the reaction was heated to 45°C for 30 minutes, and then cooled to RT. The resulting mixture was poured into 1200mL water and extracted with toluene (400mL×3). The extracts were combined and washed with water and saturated aqueous NaCl solution. The organic matter was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was dried overnight under vacuum and then dissolved in DCM (400 mL). A silica gel pad was prepared on a sinter funnel and balanced with 1:1 heptane / EtOAc. The product solution was loaded onto a silica gel pad and washed with 1600 mL 1:1 heptane / EtOAc. The filtrate was concentrated to give a red oil. The oil was dissolved in MeOH (250 mL) and the flask was placed in a water bath (~10 ° C). L(-)-camphorsulfonic acid (61.6 g, 265 mmol) was added in batches, keeping the internal temperature below 20 ° C. The resulting mixture was stirred for 15 minutes and then concentrated in vacuo to give a brown foam. The foam was dried on a vacuum pump for 2 hours. The foam was dissolved in DCM (130 mL) and EtOAc (1100 mL) was slowly added to the stirring solution through an addition funnel. The resulting mixture was transferred to a 4000 mL beaker and stirred overnight in the atmosphere. The beaker was cooled in an ice bath for 10 minutes. The precipitate was collected in a sintered funnel via vacuum filtration and washed with a minimum of ice-cooled EtOAc. The solid was dried on the sintered funnel for 2 hours.The gained white solid is dissolved in a small amount of DCM, transferred in a 2000mL beaker, then slowly diluted with EtOAc, until the clear solution begins to become turbid. Stir the suspension 4 hours in atmosphere. Use sinter funnel to collect solid by vacuum filtration, dry overnight on sinter funnel to obtain title compound (111.8g, 238.06mmol, 86% productive rate), as white solid. 1 H NMR (400MHz, DMSO-d6) δ10.54-10.47(m,1H),7.61(d,J=7.3Hz,5H),7.47-7.37(m,7H) ,5.85(d,J=10.3Hz,1H),4.68-4.61(m,1H),3.91-3.83(m,2H),3.37(s,8H),2.99(d,J =14.6Hz,1H),2.77-2.68(m,1H),2.51-2.44(m,4H),2.30-2.16(m,2H),1.91-1.81(m, 2H),1.42-1.28(m,3H),1.08(s,3H),1.01(d,J=6.6Hz,3H),0.77(s,4H);>98%ee[HPLC: OJ (10 cm × 4.6 mm, 5 μm), 5 mL / min, 40 ° C isocratic 10% EtOH (0.2% i PrNH2) / CO2].

[0149] Preparation 2

[0150] [(1R,4S)-7,7-Dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (2S)-2-methylazetidin-1-ium salt

[0151]

[0152] To a 2250 mL Parr bottle was added 20 wt % Pd(OH)2 / carbon (6.62 g). The bottle was purged with nitrogen and MeOH (250 mL) was added. To the resulting suspension was slowly added (2S)-1-benzhydryl-2-methyl-azetidine [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonate (111 g, 236 mmol) dissolved in MeOH (250 mL). The container was sealed. Purge first with nitrogen, then with hydrogen, and pressurized to 60 PSI. The reaction vessel was vigorously shaken in a Parr shaker at RT for 15 h. The container was purged with nitrogen and then by The reaction mixture was filtered through a pad and washed with MeOH. The filtrate was concentrated to give a white solid and dried under vacuum. The solid was suspended in 780 mL 1:1 MTBE / EtOAc, and the mixture was heated to 65 ° C for 20 hours, then cooled to RT and stirred overnight. The solid was collected by filtration. The solid was suspended in 380 mL MTBE and stirred at RT for 24 hours. The white solid was collected by filtration to give the title compound (41.5 g, 137 mmol, 58% yield). 1 H NMR(400MHz,DMSO-d6)δ8.68-8.55(m,1H),4.51-4.42(m,1H),3.91-3.75(m ,1H),3.36(s,3H),2.91(d,J=14.6Hz,1H),2.69-2.61(m,1H),2.52-2.46(m, 2H),2.28-2.22(m,1H),2.17-2.10(m,1H),1.96(t,J=4.5Hz,1H),1.89-1.79 (m,1H),1.43(d,J=6.7Hz,2H),1.36-1.26(m,1H),1.05(s,2H),0.75(s,2H).

[0153] Preparation 3

[0154] (R)-2-Azetidine methanol hydrochloride

[0155]

[0156] To a 2-neck RBF equipped with a nitrogen inlet was added: (R)-1-(tert-butoxycarbonyl)azetidine-2-carboxylic acid (30 g, 146 mmol), THF (300 ml) and 4-methylmorpholine (17.7 mL, 161 mmol). The mixture was cooled to -10 ° C, and isobutyl chloroformate (21 mL, 161 mmol) was added dropwise. The mixture was stirred for 30 minutes and then heated to RT. The resulting solid was removed by filtration. The filtrate was cooled to 0 ° C, and sodium borohydride (11.1 g, 292 mmol) aqueous solution (90 mL) was added dropwise (note: gas is generated). After addition, the mixture was heated to RT and stirred for 30 minutes. The mixture was diluted with MTBE (300 mL) and water (100 mL). The mixture was washed with saturated NaHCO3 aqueous solution (200 mL) and then with saturated NaCl aqueous solution (200 mL). The organic phase was dried over MgSO4, filtered, and concentrated to dryness to give an oil (27 g). HCl (4.0 M) was carefully added to 1,4-dioxane (110 mL) [Caution: gas is produced], and the resulting mixture was stirred at RT for 3 h. The solvent was evaporated in vacuo to give the title compound as an oil (16 g, 89%). This material was used directly in the preparations of 9, 11, 16, and 36.

[0157] Preparation 4

[0158] (2S,3R)-2-Methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid

[0159]

[0160] Step 1

[0161] Assemble a 3-neck 500mL RBF equipped with an addition funnel and temperature probe. Add but-2-en-1-ol (cis / trans mixture) (23.7mL, 267mmol) and chloroform (200mL) to the flask. Cool the solution in an ice bath until the internal temperature reaches 1.2°C. Add bromine (13.7mL, 267mmol) dropwise via the addition funnel at a rate of ~1 drop / 6 seconds over 2h. After addition, heat the reaction to RT and stir for 30 minutes. Turn off stirring and let the reaction stand for 3 days. Quench with saturated Na2S2O3 solution and stir vigorously for 10 minutes. Let the mixture stand for 3 days. Remove the organic layer and extract the aqueous portion with DCM (x3). Combine the organics and dry over Na2SO4, filter and concentrate in vacuo to give 2,3-dibromobutan-1-ol (62.3g, 269mmol).

[0162] Step 2

[0163] Method A

[0164] Assemble a 3-neck 1L RBF equipped with an addition funnel and a temperature probe. Add 2,3-dibromobutan-1-ol (62.3 g, 269 mmol) and THF (180 mL) to the flask. Place the flask in a water bath at RT. Add a solution of KOH (15.1 g, 269 mmol) in water (135 mL) dropwise over 10 minutes via an addition funnel. Stir for 2 hours at RT. Separate the organic phase. Extract the water portion with 3x150 mL EtOAc. Combine the organics, wash once with ~200 mL brine, dry over Na2SO4 and filter. The organics were carefully concentrated (100 mbar, 30°C until the volume was minimal, then 10 mbar, 30°C for 10 min) to give 37 g of a mixture of 2-(1-bromoethyl)oxirane (60% mass), 2,3-dibromobutan-1-ol (36% mass) and EtOAc (4% mass), as 1 1H NMR. To the mixture, EtOH (100 mL), aminodiphenylmethane (36 mL, 208.6 mmol) and NaHCO3 (26 g, 309 mmol) were added. The reaction mixture was heated to 65°C overnight and cooled to RT.

[0165] Method B

[0166] In the first flask, add 2,3-dibromobutan-1-ol (72.6 g, 313 mmol) and THF (200 mL). Place the flask in a water bath at RT. Add a solution of KOH (17.6 g, 314 mmol) in water (150 mL). Stir overnight at RT. Separate the organic phase. Extract the aqueous portion with EtOAc (3×150 mL). Combine the organics, wash once with ~200 mL of saturated aqueous NaCl solution, dry over Na2SO4 and filter. Concentrate the organic layer in vacuo (100 mbar, 30°C until the volume is minimum, then 10 mbar, 30°C for 10 minutes) to give 41.1 g of a mixture of 2-(1-bromoethyl)oxirane (75% by mass), 2,3-dibromobutan-1-ol (22% by mass) and EtOAc (3% by mass), as 1 Detected by H NMR.

[0167] In a second flask, add 2,3-dibromobutan-1-ol (10 g, 43 mmol) and THF (30 mL). Place the flask in a water bath at RT. Add a solution of KOH (2.42 g, 43.1 mmol) in water (20 mL). Stir overnight at RT. Separate the organic phase. Extract the aqueous portion with 3x50 mL EtOAc. Combine the organics, wash once with ~200 mL saturated aqueous NaCl solution, dry over Na2SO4 and filter. Concentrate the organic layer in vacuo (100 mbar, 30°C until the volume is minimum, then 10 mbar, 30°C for 10 minutes) to give a mixture of 5.4 g of 2-(1-bromoethyl)oxirane (66% mass), 2,3-dibromobutan-1-ol (32% mass) and EtOAc (2% mass), as 1 Detected by H NMR.

[0168] In a third flask, 2,3-dibromobutan-1-ol (10 g, 43 mmol) and THF (30 mL) were added. The flask was placed in a water bath at RT. A solution of KOH (2.42 g, 43.1 mmol) in water (20 mL) was added. The mixture was heated to 50 °C for 2 hours. Cooled to RT. The organic phase was separated. The aqueous portion was extracted with 3x50 mL EtOAc. The organics were combined, washed once with ~200 mL brine, dried over Na2SO4 and filtered. The organic layer was concentrated in vacuo (100 mbar, 30 °C until the volume was minimum, then 10 mbar, 30 °C for 10 minutes) to give a mixture of 5.4 g of 2-(1-bromoethyl)oxirane (70% mass), 2,3-dibromobutan-1-ol (28% mass) and EtOAc (2% mass), as 1 Detected by H NMR.

[0169] The mixtures from the three reactions were combined in a RBF to give 51.9 g of 2-(1-bromoethyl)oxirane (73% mass). EtOH (100 mL), aminodiphenylmethane (44 mL, 255.0 mmol) and NaHCO3 (32 g, 380.926 mmol) were added. The resulting mixture was stirred at RT for 2 hours, then heated to 65 ° C, and continued to stir overnight. Cooled to RT. The crude reaction mixtures from methods A and B were mixed together for purification. The solids were removed by filtration and washed with EtOH. The filtrate was concentrated to dryness. The resulting oil was dissolved in DCM. The resulting solution was washed twice with NH4Cl solution, dried with Na2SO4, filtered and concentrated to a volume of ~150 mL. The mixture was left overnight. The solids were removed by filtration. The filtrate was purified by normal phase silica gel chromatography (70% MTBE:hexanes) to give crude 1-benzhydryl-2-methyl-azetidin-3-ol (66.8 g). ES / MS (m / z): 254 (M+H).

[0170] Step 3

[0171] 1-Benzhydryl-2-methyl-azetidin-3-ol (66.8 g) was dissolved in MeOH (608 mL). The resulting solution was purified using a 5×25 cm Lux i-Cellulose 5 column with a solvent system of 85 / 15 CO2 / EtOH (containing 0.5% dimethylethylamine) at a flow rate of 300 mL / min to give (2S, 3R)-1-benzhydryl-2-methyl-azetidin-3-ol (19.2 g).

[0172] Step 4

[0173] A RBF was charged with (2S,3R)-1-benzhydryl-2-methyl-azetidine-3-ol (19.2 g, 75.8 mmol), L(-)-camphorsulfonic acid (19 g, 80.2 mmol), EtOH (100 mL), and Et2O (50 mL). The mixture was heated until nearly all of the solids dissolved and then sonicated briefly. The mixture was heated to reflux and then cooled to RT and stored in a refrigerator overnight. The solids were collected by filtration, washed with a large volume of Et2O, and dried under reduced pressure to give (2S,3R)-1-benzhydryl-2-methyl-azetidine-3-ol; [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (31.9 g, 65.7 mmol).

[0174] Step 5

[0175] RBF was charged with 20% palladium hydroxide on carbon (50 wt% water) (2 g). The catalyst was wetted with a small amount of EtOH. To the suspension was added a partial solution of (2S,3R)-1-benzhydryl-2-methyl-azetidine-3-ol; [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (20 g, 41.2 mmol) in EtOH (400 mL). The suspension was sparged with nitrogen for 5 minutes and then briefly sparged with hydrogen. The reaction mixture was stirred under a hydrogen balloon until all the starting material was consumed by LC-MS analysis. The reaction mixture was filtered through a Celite pad.

[0176] In a second flask, using the same procedure, (2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol; [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (10 g, 20.6 mmol) was hydrogenated with a solution of 20% palladium hydroxide on carbon (50 wt% water) (1 g) in EtOH (200 mL).

[0177] In a third flask, using the same procedure, (2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol; [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (2 g, 4.12 mmol) was hydrogenated with a solution of 20% palladium hydroxide on carbon (50 wt% water) (0.2 g) in EtOH (40 mL).

[0178] The three filtrates were mixed together and concentrated in vacuo. The material was suspended in n-heptane and sonicated for 10 minutes, then the solid was collected by filtration. The sonication-filtration sequence was repeated four more times. The solid was dried in vacuo overnight to give the title compound (20.11 g; 53% yield over 5 steps). 1 H NMR (399.85MHz, MeOD): 4.35-4.25 (m, 2H), 4.07 (dd, J = 6.9, 10.6Hz, 1H), 3.7 8(dd,J=7.0,10.6Hz,1H),3.30(m,1H),2.79(d,J=14.9Hz,1H),2.70-2.63(m, 1H),2.40-2.33(m,1H),2.08-2.03(m,2H),1.92(d,J=18.4Hz,1H),1.69-1.6 2(m,1H),1.54(d,J=6.6Hz,3H),1.47-1.41(m,1H),1.14(s,3H),0.88(s,3H).

[0179] Preparation 5

[0180] Ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate

[0181]

[0182] 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (15 g, 77.32 mmol), ACN (150 mL), potassium carbonate (32.06 g, 232.0 mmol) and ethyl bromoacetate (9.09 mL, 81.2 mmol) were loaded into a 500 mL RBF. The slurry was stirred overnight at RT. The reaction mixture was partitioned between EtOAc (300 mL) and water (250 mL). The layers were separated and the aqueous layer was extracted with EtOAc. The organics were combined and washed with saturated aqueous sodium chloride solution (500 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give a light yellow oil. Purification by silica gel chromatography with a gradient elution of 0-5% MeOH in DCM gave the title compound (16.8 g, 76%). 1 H NMR (400MHz, DMSO-d6) δ7.94 (s, 1H), 7.60 (s, 1H), 5.08 (s, 2H), 4.14 (q, J = 7.1Hz, 2H), 1.26 (m, 12H), 1.2 (t, J = 7.2Hz, 3H).

[0183] Preparation 6

[0184] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetic acid

[0185]

[0186] A 2000 mL RBF was charged with ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate (39.15 g, 132.8 mmol), 2,4-dichloro-6-(trifluoromethyl)pyrimidine (27.9 g, 126 mmol), 1,4-dioxane (800 mL), aqueous Na2CO3 solution (2M, 200 mL, 400 mmol) and bis(triphenylphosphine)palladium(II) dichloride (2.8 g, 4.0 mmol). The mixture was heated to 85°C. After 2 hours, the mixture was cooled to RT. The reaction mixture was divided into two portions and subjected to the following procedure:

[0187] Method A:

[0188] To the first reaction mixture was added [(1R, 4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (18.69 g, 59.75 mmol) and heated to 70 ° C for 2.5 hours. Cool to RT. Add NaOH aqueous solution (2M, 166 mL, 332 mmol) and stir overnight at RT. Add EtOAc (500 mL) and stir the mixture for 30 minutes. Acidify the mixture to pH = 7 with HCl (5M) aqueous solution and extract with EtOAc (4×250 mL). Combine the organics, wash with saturated NaCl aqueous solution, dry with Na2SO4, filter and concentrate in vacuo. The residue was dissolved in DCM (125 mL) and heptane (125 mL) was added dropwise. Stir the mixture for 30 minutes. The solid was collected by filtration and washed with 1:1 DCM / heptane (50 mL). The solid was air dried.

[0189] Method B:

[0190] The second part of the reaction mixture was concentrated in vacuo to remove 1,4-dioxane. The mixture was distributed between EtOAc and water, the aqueous layer was separated and the organic layer was concentrated to dryness. 1,4-dioxane (420 mL), Na2CO3 aqueous solution (2M, 75 mL, 150 mmol) and [(1R, 4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (18.69 g, 59.75 mmol) were added to the flask containing the residue. The mixture was heated to 70 ° C and stirred for 1.5 hours. The mixture was cooled to RT. NaOH aqueous solution (2M, 125 mL, 250 mmol) was added and stirred overnight. EtOAc (500 mL) was added and stirred for 30 minutes. The aqueous phase was separated and left overnight. The aqueous mixture was acidified to pH = 7 with aqueous HCl (5 M) and extracted with EtOAc (2 x 250 mL). The organics were combined, washed with saturated aqueous NaCl, dried over Na2SO4, filtered and concentrated in vacuo to give a solid.

[0191] Purification method:

[0192] The products from methods A and B were combined and dissolved in THF (485 mL). SiliaMetS Thiol resin (32 g) was added. The mixture was stirred for 1 hour and then filtered. The filtrate was concentrated in vacuo to give the title compound as a white powder (31.7 g, 72%). ES / MS (m / z): 342 (M+H); 340 (MH).

[0193] Preparation 7

[0194] tert-Butyl 4-[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]piperazine-1-carboxylate

[0195]

[0196] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetic acid (31.7 g, 91.0 mmol), tert-butyl piperazine-1-carboxylate (20.5 g, 109 mmol) and DCM (320 mL) were loaded into a 1 L RBF. Et3N (25.6 mL, 182 mmol) was added to the resulting solution, and then 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphane-2,4,6-trioxide anhydride solution (50 wt % solution in EtOAc, 68 mL, 114.2 mmol) was added dropwise. Stir at RT for 1 hour. The reactants were washed with water (500 mL) and then with saturated NaCl aqueous solution. The organics were dried over Na2SO4, filtered, and concentrated under high vacuum to give the title compound (52.5 g, estimated purity 88 wt% based on theoretical quantitative yield of the product). ES / MS (m / z): 510 (M+H); 508 (MH).

[0197] Preparation 8

[0198] tert-Butyl 4-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

[0199]

[0200] 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.7 g, 3.05 mmol) and 1,4-dioxane (15 mL) were added to a microwave vial. To the solution was added tert-butyl 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.55 g, 4.0 mmol), tetrakis(triphenylphosphine)palladium(0) (0.195 g, 0.17 mmol) and aqueous Na2CO3 (2M, 5.5 mL, 11 mmol). The vial was sealed and heated at 85°C in a microwave reactor for 1 hour. In a second vial, the same reaction was carried out on the same scale. The reaction mixtures were mixed in a separatory funnel. The reaction mixture was diluted with saturated aqueous sodium bicarbonate and extracted twice with EtOAc. The extracts were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography eluting with a 0-50% EtOAc / hexanes gradient to afford the title compound as a white solid (2.33 g, 88%). ES / MS (m / z): 432, 434 (M+H); 430, 432 (MH).

[0201] Preparation 9

[0202] tert-Butyl 4-[4-[2-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

[0203]

[0204] In three separate microwave reaction bottles, tert-butyl 4-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate (1 g, 2.3 mmol) and 1,4-dioxane (16 mL) were mixed. (R)-2-azetidine methanol hydrochloride (0.52 g, 4.3 mmol) and DIPEA (1.6 mL, 9.3 mmol) were added. The bottles were sealed and heated at 130 ° C for 2.5 hours in a microwave reactor. The resulting reaction mixtures were combined, then the mixture was diluted with saturated aqueous NaHCO3 and extracted twice with EtOAc. The extracts were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography with a 10-80% EtOAc / hexane gradient to give the title compound as a white foam (3.2 g, 95%). ES / MS (m / z): 483 (M+H).

[0205] Preparation 10

[0206] tert-Butyl 3-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

[0207]

[0208] 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.50 g, 2.30 mmol) and 1,4-dioxane (22 mL) and water (2 mL) were added to a microwave vial. 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylic acid tert-butyl ester (0.630 g, 1.71 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.20 g, 0.26 mmol) and K2CO3 (500 mg, 3.67 mmol) were added to the solution. The vial was sealed and stirred overnight at RT. The crude reaction was loaded onto a silica gel column, dried in a vacuum oven, and the residue was purified by flash chromatography (0-45% EtOAc / hexanes elution) to afford the title compound as a light brown oil (0.375 g, 49%). ES / MS (m / z): 348 (M+H- t Bu); 402(MH).

[0209] Preparation 11

[0210] tert-Butyl 3-[4-[2-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

[0211]

[0212] Mix tert-butyl 3-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate (190 mg, 0.47 mmol) and THF (18 mL) in a bottle. Add (R)-2-azetidine methanol hydrochloride (150 mg, 1.23 mmol) and DIPEA (1.0 mL, 5.7 mmol). Seal the bottle and heat to 100 ° C in a microwave reactor for 1 hour and 40 minutes. After cooling to RT, the crude reaction is loaded onto a silica gel column, dried in a vacuum oven, and the residue is purified by silica gel chromatography with a 0-80% (5% MeOH / EtOAc) / hexane gradient elution to give the title compound as a colorless oil (153 mg, 72%). ES / MS (m / z): 455 (M+H).

[0213] Preparation 12

[0214] 3-[4-[2-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylic acid tert-butyl ester

[0215]

[0216] The title compound was prepared essentially as described in Preparation 11 using (2S,3R)-2-methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid. ES / MS (m / z): 455 (M+H).

[0217] Preparation 13

[0218] tert-Butyl 3-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

[0219]

[0220] The title compound was prepared essentially as described in Preparation 11 using [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (2S)-2-methylazetidin-1-ium salt. The reaction was purified by silica gel chromatography using 45% EtOAc / hexanes. ES / MS (m / z): 439 (M+H).

[0221] Preparation 14

[0222] 2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]acetic acid

[0223]

[0224] A RBF was charged with ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate (2.95 g, 10.5 mmol), 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (2 g, 8.3 mmol), 1,4-dioxane (52.4 mL) and aqueous Na2CO3 (2 M, 13.3 mL, 26.6 mmol). The mixture was sparged with nitrogen for 10 minutes. Bis(triphenylphosphine)palladium(II)dichloride (191 mg, 0.27 mmol) was added. The mixture was stirred at RT for 16.5 hours. The flask was equipped with a reflux condenser and [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (2.73 g, 9.10 mmol) was added. The mixture was heated to 70°C and stirred for 3 hours, then cooled to RT. Aqueous NaOH solution (2M, 21 mL, 42 mmol) was added and the mixture was stirred at RT for 15 minutes. The organic solvent was evaporated in vacuo. The residue was diluted with water (50 mL) and the water was poured off the solid. This operation was repeated twice with water (50 mL). The solid was collected by vacuum filtration and air dried overnight. 2-Methyltetrahydrofuran (30.4 mL) and aqueous citric acid solution (6.5 wt%, 30.4 mL) were added. Stir for 5 minutes and then the layers were separated. The organic layer was washed with aqueous citric acid solution (6.5 wt%, 30.4 mL). The organic solution was dried over MgSO4, filtered, and concentrated to give the title compound (3.14 g, estimated purity 97 wt % based on theoretical quantitative yield of the product). ES / MS (m / z): 366 (M+H); 364 (MH).

[0225] Preparation 15

[0226] tert-Butyl 4-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate

[0227]

[0228] Method A:

[0229] To a microwave reaction bottle was added 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (497 mg, 2.063 mmol), tert-butyl 4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylate (867.2 mg, 2.3 mmol), 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II) DCM complex (91.2 mg, 0.109 mmol), aqueous potassium carbonate (3M solution, 2.1 mL, 6.3 mmol) and 1,4-dioxane (10.5 mL). The mixture was sparged with nitrogen for 5 minutes, sealed, and heated to 80°C. After 1 hour, cooled to RT. The mixture was filtered through a column, rinsed with EtOAc, concentrated and purified by silica gel chromatography using a 15-40% EtOAc in hexane gradient, then flash dried under vacuum at 35°C to give the title compound (705 mg, 64%). ES / MS (m / z): 454, 456 (MH).

[0230] Method B:

[0231] To a RBF was added 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (1.07 g, 4.44 mmol) and 1,4-dioxane (25 mL). 4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylic acid tert-butyl ester (2.08 g, 5.51 mmol), tetrakis(triphenylphosphine)palladium(0) (200 mg, 0.17 mmol) and aqueous Na2CO3 solution (2M, 8.5 mL, 17 mmol) were added. The reaction was degassed with nitrogen and heated at 85°C for 1.5 hours. The organic phase was decanted from the reaction and the crude reaction was divided into two equal amounts for the preparation of 16 and 17. ES / MS (m / z): 454 (MH).

[0232] Preparation 16

[0233] tert-Butyl 4-[4-[5-cyano-6-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate

[0234]

[0235] In a microwave vial was added 1 / 2 of the crude reaction product from Method B of Preparation 15, (R)-2-azetidine methanol hydrochloride (0.5 g, 4 mmol), DIPEA (1.6 mL, 9.2 mmol) and an additional amount of 1,4-dioxane (16 mL). The reaction was heated at 120 °C for 2 hours. The reaction was cooled and concentrated to a crude product. The residue was purified by silica gel chromatography with a 50-80% EtOAc / hexane gradient elution and purified again by silica gel chromatography with a 2-5% IPA / DCM gradient elution to give the title compound as a tan solid (264 mg, 26%). ES / MS (m / z): 507 (M+H).

[0236] Preparation 17

[0237] 4-[4-[5-cyano-6-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylic acid tert-butyl ester

[0238]

[0239] The title compound was prepared essentially as described in Preparation 16 using a second 1 / 2 portion of the crude reaction product from Preparation 15 and (2S,3R)-2-methylazetidin-3-ol [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl] methanesulfonic acid. ES / MS (m / z): 507 (M+H).

[0240] Preparation 18

[0241] 2-Chloro-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile hydrochloride

[0242]

[0243] To the bottle was added tert-butyl 4-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate (705 mg, 1.5 mmol), 4M HCl in 1,4-dioxane (1.5 mL, 6.0 mmol) and DCM (1.5 mL). The mixture was stirred at RT. After 1 hour, the mixture was concentrated in vacuo and the residue was dried in vacuo overnight to give the title compound (608 mg), which was used directly in the preparation of 19 and 20 without further purification. ES / MS (m / z): 356, 358 (free base M+H).

[0244] Preparation 19

[0245] 2-Chloro-6-[1-(1-methyl-4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0246]

[0247] To the bottle was added 2-chloro-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile hydrochloride (303.3 mg, 0.74 mmol), 13.3 M aqueous formaldehyde solution (0.2 mL, 3 mmol), sodium triacetoxyborohydride (378 mg, 1.73 mmol) and 1,2-dichloroethane (6.0 mL). The mixture was stirred at RT for 30 minutes. Saturated aqueous sodium bicarbonate solution was added and extracted twice with DCM. The extracts were combined and concentrated in vacuo to give the title compound (285 mg, 99% yield). ES / MS (m / z): 370, 372 (M+H).

[0248] Preparation 20

[0249] tert-Butyl 4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate

[0250]

[0251] The title compound was prepared from 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile and tert-butyl 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate using essentially the same procedure as described in Example 10 below. ES / MS (m / z): 491 (M+H)

[0252] Preparation 21

[0253] tert-Butyl 4-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

[0254]

[0255] 2,4-Dichloro-6-(trifluoromethyl)pyrimidine (0.258 g, 1.1 mmol) was dissolved in 1,4-dioxane (6 mL), and tert-butyl 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (0.591 g, 1.52 mmol), tetrakis(triphenylphosphine)palladium(0) (68 mg, 0.059 mmol) and 2M aqueous sodium carbonate solution (2.2 mL, 4.4 mmol) were added. The mixture was heated to 85° C. in a microwave reactor for 1 hour. Cool to RT and add [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (0.414 g, 1.36 mmol) and DIPEA (0.6 mL, 3.4 mmol). The mixture is heated at 110°C for 2 hours. The mixture is diluted with saturated aqueous sodium bicarbonate solution and extracted twice with EtOAc. The extracts are combined, dried over sodium sulfate, then filtered and evaporated. The residue is purified by silica gel chromatography with a 0-50% EtOAc / hexane gradient to give the title compound (215 mg, 41%) as a white solid. ES / MS (m / z): 467 (M+H).

[0256] Preparation 22

[0257] tert-Butyl 3-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]azetidine-1-carboxylate

[0258]

[0259] To the bottle was added 3-cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (0.360 g, 1.49 mmol), 1,4-dioxane (7.5 ml), tert-butyl 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate (0.727 g, 1.98 mmol), tetrakis(triphenylphosphine)palladium(0) (95 mg, 0.08 mmol) and 2M aqueous sodium carbonate solution (2.8 mL, 5.6 mmol, 2 mol / L). The bottle was sealed and heated at 85 °C in a microwave reaction bottle for 1 hour. Diluted with saturated sodium bicarbonate solution and extracted twice with EtOAc. The extracts were combined, dried over sodium sulfate, filtered and evaporated. The residue was purified by silica gel chromatography eluting with a 0-90% EtOAc / hexanes gradient to afford the title compound (0.609 g, 95%). ES / MS (m / z): 426, 428 (MH).

[0260] Preparation 23

[0261] 2-Chloro-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

[0262]

[0263] The title compound was prepared from 1-tetrahydropyran-4-yl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole using essentially the same procedure as described for Preparation 22. ES / MS (m / z): 357, 359 (M+H).

[0264] Preparation 24

[0265] tert-Butyl 3-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]azetidine-1-carboxylate

[0266]

[0267] Tert-butyl 3-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]azetidine-1-carboxylate (603 mg, 1.410 mmol) was dissolved in THF (18 mL). (2S)-2-methylazetidine-1-ium salt of [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate (519 mg, 1.711 mmol) and DIPEA (0.75 mL, 4.3 mmol) were added and the mixture was heated to 130°C for 2.5 hours. The solvent was evaporated and the residue was purified by silica gel chromatography eluting with a 0-50% EtOAc / hexanes gradient to give the title compound (498 mg, 76%) as a colorless oil. ES / MS(m / z): 463(M+H), 461(MH).

[0268] Preparation 25

[0269] tert-Butyl N-[2-[4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-1-piperidinyl]-2-oxo-ethyl]carbamate

[0270]

[0271] 2-[(2S)-2-Methylazetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (0.129 g, 0.330 mmol) was dissolved in DCM (1 mL). 2-(tert-Butoxycarbonylamino)acetic acid (69 mg, 0.39 mmol), HATU (0.175 g, 0.460 mmol) and DIPEA (0.18 mL, 1.0 mmol) were added. The mixture was stirred at RT for 3 hours, then diluted with saturated aqueous sodium bicarbonate solution and extracted twice with DCM. The organics were combined, dried over sodium sulfate, then filtered and evaporated. The residue was purified by reverse phase chromatography on a C18 bonded silica gel column (solvent A = 10 mM aqueous ammonium bicarbonate solution, solvent B = ACN; gradient 10-91% B) to give the title compound (129 mg, 71%). ES / MS (m / z): 548 (M+H).

[0272] Preparation 26

[0273] 1-(2,2-Dimethyl-[1,3]dioxolan-4-ylmethyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole

[0274]

[0275] 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.500 g, 2.50 mmol) was suspended in DMF (10 mL), and then 4-(chloromethyl)-2,2-dimethyl-1,3-dioxolane (0.73 mL, 5.0 mmol) and cesium carbonate (1.64 g, 5.03 mmol) were added. The mixture was heated to 75 ° C overnight. The reactants were diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc. The organics were combined and washed four times with saturated aqueous NaCl solution, then dried over sodium sulfate, filtered and evaporated to give the title compound (780 mg), which was used without further purification. ES / MS (m / z): 309 (M+H).

[0276] Preparation 27

[0277] 2-Chloro-6-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0278]

[0279] 3-Cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (0.352 g, 1.46 mmol) was dissolved in 1,4-dioxane (7.3 mL). 1-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (0.602 g, 1.89 mmol), tetrakis(triphenylphosphine)palladium(0) (0.715 g, 0.619 mmol) and 2M aqueous sodium carbonate solution (2.8 mL, 5.6 mmol) were added. The mixture was sparged with nitrogen for 15 minutes and then heated to 85°C for 1.5 hours. The reaction was diluted with saturated aqueous sodium bicarbonate solution and extracted twice with EtOAc. The organics were combined and dried over sodium sulfate, then filtered and evaporated. The residue was purified by silica gel chromatography eluting with a 0-40% EtOAc / hexanes gradient to afford the title compound (207 mg, 36%) as a colorless oil. ES / MS (m / z): 387, 389 (M+H).

[0280] Preparation 28

[0281] tert-Butyl 4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazole-1-carboxylate

[0282]

[0283] Mix 3-cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (195 mg, 0.785 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (250 mg, 0.85 mmol), 2M aqueous sodium carbonate (2.5 mL, 5.0 mmol) and 1,4-dioxane (4 mL) in a bottle. Degas the reactants at RT by bubbling nitrogen through them while stirring for 5 minutes. Add [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (185 mg, 0.240 mmol) and heat the reactants to 100 °C for 2 hours. Add water and extract with EtOAc. Combine the extracts and concentrate in vacuo. The residue was purified by silica gel chromatography eluting with a 25-50% EtOAc / hexanes gradient to afford the title compound (40 mg, 14%) as an orange solid. ES / MS (m / z): 271, 273 (MH-BOC).

[0284] Preparation 29

[0285] tert-Butyl N-[2-[[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]amino]ethyl]carbamate

[0286]

[0287] Add 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetic acid (200 mg, 0.586 mmol), DMF (2 mL), DIPEA (0.31 mL, 1.76 mmol), HATU (0.267 g, 0.703 mmol) and N-BOC-ethylenediamine (0.102 mL, 0.644 mmol) to the bottle. Stir at RT for 8 hours and then purify by preparative HPLC [parameters: solvent-10 mM aqueous ammonium bicarbonate pH 10 / 5% MeOH (solvent A) and ACN (solvent B); pre-column-Waters BEH HILIC 100×30 mm5μm, With 15×30mm BEH HILIC guard column; Column: EVO C18,100×30mm,5μm, Equipped with a 15×30 mm EVO guard column (using an online heater at 50° C.); 33-67% B gradient] to give the title compound (179 mg, 63%). ES / MS (m / z): 484 (M+H).

[0288] Prepare the compounds shown in Table 1 using essentially the same procedure as described for Preparation 29 and the appropriate commercially available amines.

[0289] Table 1

[0290]

[0291]

[0292] Preparation 34

[0293] (3R)-3-[4-[2-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylic acid tert-butyl ester

[0294]

[0295] In a reaction vessel, 2,4-dichloro-6-(trifluoromethyl)pyrimidine (600 mg, 2.71 mmol) was mixed with (R)-tert-butyl 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate (1 g, 2.67 mmol), 2M aqueous sodium carbonate solution (3 mL, 6 mmol) and bis(triphenylphosphine)palladium(II) dichloride (50 mg, 0.068 mmol) in 1,4-dioxane (6 mL). Degassed with nitrogen and the mixture was heated to 80 °C for 2 hours. Diluted with EtOAc (75 mL), washed with water and saturated aqueous NaCl solution. The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was mixed with a solution of (2S,3R)-2-methylazetidine-3-ol [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (420 mg, 1.32 mmol) in 1,4-dioxane (4 mL) and DIPEA (0.7 mL, 4 mmol) in a vial. The bottle was sealed and heated to 120 ° C in a microwave reactor for 1 hour. The reaction mixture was directly loaded onto silica gel and purified by silica gel chromatography with a 0-80% EtOAc / hexane gradient elution to give the title compound (570 mg, 92%) as an off-white foam. ES / MS (m / z): 469 [M+H].

[0296] Preparation 35

[0297] (3S)-3-[4-[2-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylic acid tert-butyl ester

[0298]

[0299] The title compound was prepared essentially as described in Preparation 34 using (S)-tert-butyl 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate. ES / MS (m / z): 469 [M+H].

[0300] Preparation 36

[0301] tert-Butyl 3-[(1R)-4-[2-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylate

[0302]

[0303] Prepare the title compound essentially as described in Preparation 34 using (R)-2-azetidinemethanol hydrochloride. ES / MS (m / z): 469 [M+H].

[0304] Preparation 37

[0305] 4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-3-hydroxy-piperidine-1-carboxylic acid tert-butyl ester

[0306]

[0307] 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (560 mg, 2.9 mmol), 7-oxa-4-azabicyclo[4.1.0]heptane-4-carboxylic acid tert-butyl ester (580 mg, 2.9 mmol,), DMF (10 mL) and cesium carbonate (1.7 g, 5.2 mmol) were added to the bottle. The mixture was heated to 80 ° C for 6 hours. The reactant was diluted with water and extracted with EtOAc. The extracts were combined, dried over sodium sulfate, filtered and concentrated to give crude tert-butyl 3-hydroxy-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylate (650 mg). The material was placed in a RBF and 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (300 mg, 1.2 mmol), 1,4-dioxane (5 mL) and 2M aqueous sodium carbonate solution (1.3 mL, 2.6 mmol) were added. The reactants were degassed at RT by bubbling nitrogen through the reactants while stirring for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (80 mg, 0.06 mmol) was added and degassed for another 3 minutes. The reactants were heated to 80 °C for 4 hours and then cooled to RT. DIPEA (0.6 mL, 3 mmol) and [(1R, 4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (410 mg, 1.4 mmol) were added. Stirred for 30 minutes at RT and then heated to 80 °C for 1 hour. The mixture was diluted with water and extracted with EtOAc. The extracts were combined, dried over sodium sulfate, filtered and concentrated. The residue was purified by reverse phase chromatography (C18, gradient 10-100% ACN / 10 mM aqueous ammonium carbonate + 5% methanol) to give the title compound (95 mg, 15%). ES / MS (m / z): 507 (M+H).

[0308] Example 1

[0309] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone

[0310]

[0311] A 2L RBF was charged with tert-butyl 4-[2-[4-[2-[(2S)-2-methylazetidine-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]piperazine-1-carboxylate (47.0 g, 89.5 mmol) and DCM (470 mL) was added. TFA (60 mL, 778 mmol) was added dropwise to the reaction mixture and stirred at RT for 19 hours. More TFA (8 mL, 110 mmol) was added and stirring was continued for 18 hours. The reaction mixture was slowly added to a flask containing cooled NH4OH aqueous solution (35 wt%, 150 mL, 1300 mmol). The layers were separated. The organic layer was washed with saturated NaCl aqueous solution, dried over Na2SO4, filtered, and concentrated. The residue was dissolved in EtOAc (400 mL) and concentrated in vacuo. The residue was dissolved in IPA (400 mL) and concentrated in vacuo to afford the title compound as an off-white foam (32 g, 84%). ES / MS (m / z): 410 (M+H).

[0312] Example 1a

[0313] 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone sesquisuccinate

[0314]

[0315] To the reactor was added 2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone (959 g, 2.34 mol) and then IPA (11.5 L). Succinic acid (550 g, 4.69 mol) was added and the mixture was heated to 70-80°C to give a solution. The mixture was stirred between 70-80°C for 2 hours and then cooled to 25°C for 6 hours. The mixture was filtered and rinsed with IPA (1 L). The resulting solid was dried at 40-50°C for 6 hours to give the title compound (1070 g, 78%) as a white solid. 1H-NMR (400MHz, DMSO-d6) δ8.41 (s, 1H), 8.18 (s, 1H), 7.33 (s, 1H), 5.24 (s, 2H), 4.53-4.43 (m, 1H), 4.06-3.93 (m, 2H), 3.58-3.47 (m, 4H), 2.95-2.82 (m, 4H), 2.47-2.40 (m, 1H), 2.37 (s, 6H-succinic acid methylene, 1.5 equivalents), 2.02-1.92 (m, 1H), 1.50 (d, J = 4 Hz, 3H). High resolution ES-MS (m / z): theoretical value 410.1911 (free base M+H), found value 410.1916.

[0316] The XRPD peaks of Example 1a are listed in Table 2.

[0317] Table 2: X-ray powder diffraction peaks of Example 1a

[0318] peak Angle (°2θ) Relative intensity (% of strongest peak) 1 12.1 27.5％ 2 14.1 22.7％ 3 16.3 33.3％ 4 17.7 30.5％ 5 18.9 100.0％ 6 20.0 22.1％ 7 20.5 53.8％ 8 21.3 44.9％ 9 22.0 36.6％ 10 24.3 23.2％

[0319] Example 2

[0320] [(2R)-1-[4-[1-(4-piperidinyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

[0321]

[0322] Tert-butyl 4-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate (2.55 g, 5.28 mmol) was dissolved in DCM (50 mL) and TFA (10 mL, 132 mmol) was added. The mixture was stirred at RT for 30 minutes and then applied directly to SCX resin. The resin was washed with MeOH and then with a solution of ammonia in MeOH (7 M). The fractions containing the desired product were combined and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a 0-9% (7 M ammonia / MeOH) / DCM gradient to give the title compound as a white powder (2.02 g, 94%). ES / MS (m / z): 383 (M+H); 381 (MH).

[0323] Example 3

[0324] [(2R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

[0325]

[0326] Tert-butyl 3-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate (153 mg, 0.336 mmol) was dissolved in DCM (15 mL) and TFA (5 mL, 66 mmol) was added. The mixture was stirred at RT for 1 hour and then concentrated under reduced pressure at RT. The residue was co-evaporated with DCM and dried in vacuo. The residue was purified by preparative HPLC (parameters: solvent A = 10 mM aqueous ammonium bicarbonate pH 10 / 5% MeOH, solvent B = ACN; column - EVO C18,100×30mm,5um, With 15×30mm BEHHILIC guard column; Column: EVO C18,100×30mm,5μm, EVO guard column with 15 x 30 mm) to give the title compound (67 mg, 40%) as a white powder. ES / MS (m / z): 355 (M+H); 353 (MH).

[0327] Example 4

[0328] (2S,3R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol

[0329]

[0330] tert-Butyl 3-[4-[2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate (153 mg, 0.336 mmol) was dissolved in DCM (15 mL) and TFA (5 mL, 66 mmol) was added. The mixture was stirred at RT for 1 hour and then concentrated under reduced pressure at RT. The residue was co-evaporated with DCM and dried in vacuo. The residue was purified by preparative HPLC (parameters: solvent A = 10 mM NH4HCO3 aqueous solution with 5% MeOH, pH 10, solvent B = ACN; column - EVO C18,100×30mm,5μm, EVO guard column with 15 x 30 mm) to give the title compound (79 mg, 57%) as a white powder. ES / MS (m / z): 355 (M+H); 353 (MH).

[0331] Example 5

[0332] 2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-N-(2-hydroxyethyl)acetamide

[0333]

[0334] 2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]acetic acid (351 mg, 0.96 mmol) was dissolved in DMF (2 mL), and HATU (482 mg, 1.24 mmol), ethanolamine (0.1 mL, 2 mmol) and DIPEA (0.5 mL, 3 mmol) were added. The mixture was stirred at RT for 2 days. The reaction mixture was directly purified by reverse phase chromatography (C18-bonded silica gel) with a gradient elution of 5-95% MeCN / (NH4)2CO3 aqueous solution (10 mM) to give the title compound (110 mg, 28%). ES / MS (m / z): 409 (M+H); 407 (MH).

[0335] Example 6

[0336] 2-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0337]

[0338] Tert-butyl 4-[4-[5-cyano-6-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate (264 mg, 0.52 mmol) was dissolved in DCM (2 mL). TFA (2 mL) was added to the reaction in one portion and stirred for 1 hour. The reaction was concentrated, diluted with DCM, and the reaction mixture was concentrated. The reaction mass was diluted with MeOH (1 mL) and saturated aqueous NaHCO3 was added to pH ~8. The reaction mixture was purified by reverse phase chromatography (40 g, C18, gradient of 20-100% ACN / 10 mM (NH4)2CO3 aqueous solution with 5% MeOH) to give the title compound (135.5 mg, 64%). ES / MS (m / z): 407 (M+H).

[0339] Example 7

[0340] 2-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0341]

[0342] Dissolve tert-butyl 4-[4-[5-cyano-6-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate (220 mg, 0.43 mmol) in DCM (2 mL). Add TFA (2 mL) to the reaction in one portion and stir for 1 hour. Concentrate the reaction, dilute with DCM, and concentrate the reaction mixture. Dilute the reaction mass with MeOH (1 mL) and add saturated aqueous NaHCO3 to pH ~2. The reaction mixture is purified by reverse phase chromatography (40 g, C18, gradient 20-100% ACN / 10 mM (NH4)2CO3 aqueous solution, containing 5% MeOH) to give the title compound (127 mg, 72%). ES / MS (m / z): 407 (M+H).

[0343] Example 8

[0344] 2-[(2S)-2-Methylazetidin-1-yl]-6-[1-(1-methyl-4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0345]

[0346] 2-Chloro-6-[1-(1-methyl-4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (261 mg, 0.68 mmol), [(1R, 4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (338 mg, 1.1 mmol), DIPEA (0.4 mL, 2 mmol) and THF (3.7 mL) were added to the bottle. The bottle was sealed and heated to 130 ° C for 2 hours. The reaction mixture was distributed between DCM and 1N HCl aqueous solution. The aqueous phase was neutralized with 1N NaOH and then extracted three times with DCM. The organic extracts were combined. Dry with sodium sulfate, filter, and concentrate in vacuo. The material was dried in a vacuum oven at 50 ° C for 1 hour. The residue was purified by silica gel chromatography eluting with a 5-10% MeOH / DCM gradient, and the fractions containing the title compound were combined, concentrated and dried in a vacuum oven at 50°C overnight. The residue was purified by preparative HPLC (parameters: solvent A = 10 mM aqueous ammonium bicarbonate solution containing 5% MeOH, solvent B = ACN; column - XbridgeTM 30 mm×75 mm 5 μm, 45 mL / min; gradient 5-100% B) to obtain the title compound (73 mg, 26%). ES / MS (m / z): 405 (M+H).

[0347] Example 9

[0348] 6-[1-[1-(2-hydroxyethyl)-4-piperidinyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0349]

[0350] To a 20 mL bottle was added 2-chloro-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile hydrochloride (304.3 mg, 0.63 mmol), 2-[tert-butyl(dimethyl)silyl]oxyacetaldehyde (219 mg, 1.19 mmol), DIPEA (0.52 mL, 3.1 mmol), sodium triacetoxyborohydride (277.2 mg, 1.3 mmol) and DCM (2.0 mL). The reaction mixture was stirred at RT for 2 hours. 2-[tert-butyl(dimethyl))silyl]oxyacetaldehyde (219 mg, 1.2 mmol) was added and stirred overnight at RT. A saturated aqueous solution of sodium bicarbonate was added and extracted twice with DCM. The organic phases were combined, washed with a saturated aqueous solution of NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a 40-70% EtOAc in hexanes gradient to give 6-[1-[1-[2-[tert-butyl(dimethyl)silanyl]oxyethyl]-4-piperidinyl]pyrazol-4-yl]-2-chloro-4-(trifluoromethyl)pyridine-3-carbonitrile (220 mg, 58%).

[0351] 6-[1-[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidinyl]pyrazol-4-yl]-2-chloro-4-(trifluoromethyl)pyridine-3-carbonitrile (175 mg, 0.289 mmol), [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (136 mg, 0.45 mmol), DIPEA (0.15 mL, 0.85 mmol) and THF (1.5 mL, 18 mmol) were added to a vial. The bottle was sealed and heated to 130 ° C for 60 minutes. The reaction mixture was partitioned between EtOAc and 1N HCl, the organic layer was removed, and the aqueous layer was extracted three times with EtOAc. The organics were combined, washed with saturated NaCl aqueous solution, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was dissolved in THF (1.3 mL), and the mixture was cooled to 0 ° C, and then a 1M solution of tetrabutylammonium fluoride in THF (0.37 mL, 0.37 mmol) was added dropwise. The reaction was warmed to RT and stirred overnight. The reaction mixture was concentrated in vacuo. The residue was purified by silica gel chromatography using 100% hexane and then 1-10% (0.7N ammonia in MeOH) / DCM gradient elution to give the title compound (93 mg, 73%). ES / MS (m / z): 435 (M+H).

[0352] Example 10

[0353] 6-[1-[2-(Dimethylamino)ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0354]

[0355] To the reaction bottle was added 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (250 mg, 1.04 mmol), N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanamine (271 mg, 1.07 mmol), 1,4-dioxane (10 mL), aqueous potassium carbonate (3M, 1.04 mL, 3.12 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) (40 mg, 0.052 mmol). The bottle was sealed and heated to 80°C for 2 hours, then the mixture was cooled to RT. After 3 g The column was filtered and eluted with EtOAc. The filtrate was concentrated by evaporation. To the residue were added [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (378 mg, 1.24 mmol), DIPEA (0.9 mL, 5.2 mmol) and DMSO (10 mL). The mixture was heated to 120 ° C for 6 hours. The reaction mixture was diluted with water and added to The product was washed with MeOH, then with DCM and finally with a 1:1 mixture of DCM / MeOH, and the eluted fractions were combined. The combined fractions containing the title compound were concentrated. The residue was purified by preparative HPLC (parameters: solvent A = 10 mM aqueous ammonium bicarbonate solution containing 5% MeOH, pH 10, solvent B = ACN; pre-column - Waters BEH HILIC 100×30 mm 5 μm, With 15×30mm BEHHILIC guard column; Column: EVO C18,100×30mm,5μm, Equipped with a 15×30 mm EVO guard column (with an online heater at 50° C.); 33-100% B gradient) to give the title compound (53 mg, 13%). ES / MS (m / z): 379 (M+H).

[0356] Embodiment 11

[0357] 6-[1-(2-Hydroxyethyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0358]

[0359] The title compound was prepared essentially using the procedure described in Example 10 using 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanol. ES / MS (m / z): 352 (M+H).

[0360] Example 12

[0361] 2-[(2S)-2-Methylazetidin-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0362]

[0363] Tert-butyl 4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]piperidine-1-carboxylate (1.99 g, 4.06 mmol) was dissolved in DCM (50 mL) and TFA (10 mL, 132.3 mmol) was slowly added. Stirred for 30 minutes at RT and then the reaction mixture was loaded onto four 10 g SCX columns. The column was washed with MeOH and then with 7N ammonia in MeOH. The basic washings were concentrated in vacuo. The residue was purified by reverse phase chromatography on C18-bound silica gel (Solvent A: 10 mM ammonium bicarbonate with 5% MeOH; Solvent B: ACN; Gradient: 10-71% Solvent B) to give the title compound (709 mg, 45%) as a white solid. ES / MS (m / z): 391 (M+H).

[0364] Example 13

[0365] 2-[(2S)-2-Methylazetidin-1-yl]-4-[1-(4-piperidinyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine

[0366]

[0367] Tert-butyl 4-[4-[2-[(2S)-2-methylazetidine-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate (0.213 g, 0.457 mmol) was dissolved in DCM (5 mL) and TFA (1 mL) was added. The mixture was stirred for 25 minutes and then the reaction mixture was directly loaded onto a 10 g SCX column. The column was washed with MeOH and then eluted with 7N ammonia in MeOH. The alkaline washings were concentrated to give the title compound (129 mg, 77%). ES / MS (m / z): 367 (M+H).

[0368] Prepare the examples in Table 3 using essentially the procedure described in Example 13 and the appropriate protected amine.

[0369] Table 3

[0370]

[0371] Embodiment 17

[0372] 6-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0373]

[0374] The title compound was prepared using essentially the same procedure as described in Example 12, starting with tert-butyl 3-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]azetidine-1-carboxylate. Purification by preparative HPLC (parameters: solvent A - 10 mM aqueous ammonium bicarbonate solution containing 5% MeOH; solvent B - ACN; gradient 35-59% B; column: EVO C18, 100×30 mm, 5 μm) to obtain the title compound. ES / MS (m / z): 363 (M+H).

[0375] Embodiment 18

[0376] 6-[1-[1-(2-aminoacetyl)-4-piperidinyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0377]

[0378] The title compound was prepared using essentially the same procedure as described in Example 12, starting with tert-butyl N-[2-[4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-1-piperidinyl]-2-oxo-ethyl]carbamate. Purification by preparative HPLC (parameters: solvent A - 10 mM aqueous ammonium bicarbonate solution containing 5% MeOH; solvent B - MeOH; gradient 20-50% B, 55 mL / min; column - EVOC18, 30mm×250mm, 5μm). ES / MS(m / z):448(M+H).

[0379] Embodiment 19

[0380] 6-[1-(1-acetyl-4-piperidinyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0381]

[0382] 2-[(2S)-2-methylazetidine-1-yl]-6-[1-(4-piperidinyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (Example 12) (0.111 g, 0.284 mmol) was dissolved in DCM (1 mL) and pyridine (0.026 mL, 0.32 mmol) was added. The mixture was cooled to 0 ° C and acetyl chloride (0.024 mL, 0.34 mmol) was added. The mixture was warmed to RT and stirred for 30 minutes. The mixture was cooled to 0 ° C and pyridine (0.026 mL, 0.32 mmol) and acetyl chloride (0.024 mL, 0.34 mmol) were added. It was heated to RT and stirred for another 30 minutes. The reactant was diluted with saturated sodium bicarbonate aqueous solution and extracted twice with DCM. The organics were combined, dried over sodium sulfate, filtered and evaporated. The residue was purified by preparative HPLC (parameters: solvent A-10 mM aqueous ammonium bicarbonate solution, containing 5% MeOH; solvent B-MeOH; gradient 25-55% B, 55 mL / min; column- EVO C18, 30 mm×250 mm, 5 μm) to give the title compound (67 mg, 55%). ES / MS (m / z): 433 (M+H).

[0383] Examples 20-23

[0384] The examples in Table 4 were prepared using essentially the procedure of Example 5 and the appropriate carboxylic acids and commercially available amines.

[0385] Table 4

[0386]

[0387]

[0388] Embodiment 24

[0389] 6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0390]

[0391] 2-Chloro-6-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (0.204 g, 0.527 mmol) was dissolved in THF (6 mL). To the mixture was added aqueous hydrochloric acid (2M, 3 mL, 6 mmol) and stirred at RT for 1.5 hours. The mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted twice with EtOAc. The organics were combined and dried over sodium sulfate, then filtered and evaporated to give crude 2-chloro-6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (183 mg). The crude material was dissolved in DMF (2 mL), and then [(1R, 4S)-7,7-dimethyl-2-oxo-norbornane-1-yl] methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (222 mg, 0.73 mmol) and DIPEA (0.37 mL, 2.1 mmol) were added. The mixture was heated at 130 ° C for 2.5 hours in a microwave reactor. The reaction mixture was purified by preparative HPLC (parameters: solvent A-10 mM aqueous ammonium bicarbonate solution, containing 5% MeOH; solvent B-ACN; gradient 20-50% B, 60 mL / min; column- EVOC18, 30 mm×250 mm, 5 μm) to obtain the title compound (102 mg, 51%). ES / MS (m / z): 382 (M+H).

[0392] Examples 24a and 24b

[0393] Example 24a: 6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile - Isomer 1

[0394] Example 24b: 6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile - Isomer 2

[0395] The isomers of 6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile (86 mg) were separated using chiral SFC (parameters: column - Cellulose-2, 21×250 mm; column temperature -40°C; solvent -15% EtOH / CO2, 80 mL / min) to obtain the title compound [isomer 1, first eluting isomer: 35 mg, ES / MS (m / z): 382 (M+H); isomer 2, second eluting isomer: 39 mg, ES / MS (m / z): 382 (M+H)]. Analytical chiral SFC (parameters: column - Cellulose-2, 4.6×150 mm; solvent-15% EtOH / CO2, 5 mL / min): Isomer 1-retention time 3.10 min, 95.6% ee; Isomer 2-retention time 3.52 min, 94.4% ee.

[0396] Embodiment 25

[0397] 2-[(2S)-2-Methylazetidin-1-yl]-6-(1H-pyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

[0398]

[0399] Combine tert-butyl 4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridinyl]pyrazole-1-carboxylate (75 mg, 0.20 mmol), 1,4-dioxane (1.5 mL) and EtOH (1 mL) in a bottle. Add DIPEA (130 mg, 1.0 mmol) and (2S)-2-methylazetidin-1-ium salt of [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate (117 mg, 0.39 mmol). Heat the reaction at 200 °C in a microwave reactor for 2 hours. Concentrate the reaction in vacuo. The residue is purified by reverse phase chromatography on C18 bonded silica gel using a gradient elution of 10-100% ACN / water (containing 0.1% formic acid) to give the title compound (35 mg, 41%) as a light yellow solid. ES / MS(m / z):308(M+H),306(MH).

[0400] Embodiment 26

[0401] 2-[(2S)-2-Methylazetidin-1-yl]-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

[0402]

[0403] Combine 2-chloro-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile (105 mg, 0.29 mmol), 1,4-dioxane (1 mL), EtOH (0.5 mL), DIPEA (0.2 mL, 1 mmol) and [(1R,4S)-7,7-dimethyl-2-oxo-norbornane-1-yl]methanesulfonic acid (2S)-2-methylazetidine-1-ium salt (100 mg, 0.33 mmol) in a bottle. Heat the reaction at 150 °C in a microwave reactor for 2 hours. Concentrate the reaction in vacuo. The residue is purified by silica gel chromatography using a 40-50% EtOAc / hexanes gradient to give the title compound (88 mg, 76%) as a light yellow solid. ES / MS (m / z): 392 (M+H).

[0404] Embodiment 27

[0405] 6-[1-(3-Hydroxy-4-piperidinyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

[0406]

[0407] 4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridinyl]pyrazol-1-yl]-3-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (85 mg, 0.17 mmol) was dissolved in DCM (1 mL), then TFA (0.5 mL) was added and stirred for 5 minutes. The reaction was concentrated in vacuo. The residue was purified by reverse phase chromatography (C18, gradient 20-100% ACN / 10 mM aqueous ammonium carbonate + 5% methanol gradient to give the title compound (27 mg, 40%) as a white solid. ES / MS (m / z): 407 (M+H).

[0408] Embodiment 28

[0409] N-(2-aminoethyl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetamide

[0410]

[0411] Tert-butyl N-[2-[[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]amino]ethyl]carbamate (340 mg) was dissolved in neat TFA (5 mL) and stirred at ambient temperature. After 2 minutes, quenched with aqueous NaOH until basic. Extracted with DCM and EtOAc. The extracts were combined, dried over sodium sulfate, filtered, and concentrated. The residue was purified by preparative HPLC [parameters: solvents - 10 mM aqueous ammonium bicarbonate pH 10 / 5% MeOH (solvent A) and ACN (solvent B); pre-column - Waters BEH HILIC 100×30 mm 5μm, With 15×30mm BEH HILIC guard column; Column: EVO C18,100×30mm,5μm, Equipped with a 15×30 mm EVO guard column (using an online heater at 50° C.); 14-48% B gradient] to give the title compound (45 mg, 17%). ES / MS (m / z): 384 (M+H).

[0412] Using essentially the same procedure as described in Example 28 and the appropriate protected amine, prepare the compounds shown in Table 5.

[0413] Table 5

[0414]

[0415]

[0416] Embodiment 33

[0417] 4-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidine

[0418]

[0419] tert-Butyl 3-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate (530 mg, 1.21 mmol) was dissolved in DCM (50 mL). TFA (6 mL) was added and the mixture was stirred at RT for 1 hour. The mixture was concentrated in vacuo to give the crude title compound. Half of the material was purified by preparative HPLC [parameters: solvents - 10 mM aqueous ammonium bicarbonate pH 10 / 5% MeOH (solvent A) and ACN (solvent B); pre-column - Waters BEH HILIC 100×30 mm 5μm, With 15×30mm BEH HILIC guard column; Column: EVO C18,100×30mm,5μm, Equipped with a 15 x 30 mm EVO guard column (with an inline heater at 50°C); 23-58% B gradient] to give the title compound (162 mg, 80% yield from half the starting material) as a white solid. ES / MS (m / z): 339 (M+H).

[0420] Embodiment 34

[0421] 2-[(2S)-2-methylazetidin-1-yl]-4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine

[0422]

[0423] Half of the crude 4-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidine (0.6 mmol) prepared in Example 33 was dissolved in MeOH (6 mL). Acetic acid (0.1 mL) and 3.45 M aqueous formaldehyde solution (1 mL) were added, and the mixture was stirred at RT for 30 minutes. Sodium triacetoxyborohydride (300 mg, 1.4 mmol) was added, and the mixture was stirred for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC [Parameters: Solvent-10 mM aqueous ammonium bicarbonate pH 10 / 5% MeOH (solvent A) and ACN (solvent B); Precolumn-Waters BEHHILIC 100×30 mm 5μm, With 15×30mm BEH HILIC guard column; Column: EVO C18,100×30mm,5μm, With 15 x 30 mm EVO guard column (using online heater at 50°C); 33-67% B gradient] to give the title compound (128 mg, 61%, yield from half the starting material in Example 33) as a white solid. ES / MS (m / z): 353 (M+H).

[0424] Embodiment 35

[0425] (2S,3R)-2-Methyl-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol

[0426]

[0427] The title compound was prepared using essentially the same procedure as described in Example 34, starting from (2S,3R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol (Example 4). ES / MS (m / z): 369 (M+H), 367 (MH).

[0428] Embodiment 36

[0429] [(2R)-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

[0430]

[0431] The title compound was prepared using essentially the same procedure as described in Example 34 starting from [(2R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol (Example 3). ES / MS (m / z): 369 (M+H), 367 (MH).

[0432] Determination

[0433] Assay for KHK enzyme activity against human KHK-C and human KHK-A

[0434] The intrinsic potency of inhibition of KHK C or A activity can be measured using an enzyme assay that measures F1P production. Compounds were prepared in DMSO and tested in a 10-point concentration curve to generate 3-fold serial dilutions of compounds ranging from 20 μM to 1.02 nM in a 96-well plate. The enzyme was prepared in assay buffer [50 mM 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), 10 mM potassium chloride, 100 mM magnesium chloride, 2 mM tris(2-carboxyethyl)phosphine (TCEP), 0.01% n-octyl glucoside] and incubated with the compound for 15 min at RT. The reaction was performed in a volume of 100 μL, containing substrate concentrations of fructose (250 μM for KHK-C assay and 1.25 mM for KHK-A assay) and ATP (150 μM for both isoforms); it was further incubated for 20 min at RT. The reaction was then stopped by adding stop buffer containing 0.2% formic acid and 1 μg / ml 13 C6-fructose-6-phosphate ( 13C6-F6P) internal standard. The plates were stored at -20°C until RapidFire MS analysis.

[0435] RapidFire MS analysis for F1P quantification

[0436] An Agilent 300 RapidFire automated extraction system (Agilent, Santa Clara, CA) with three HPLC quaternary pumps was coupled to an Agilent 6495 triple quadrupole mass spectrometer (Agilent Technologies, Santa Clara, CA) equipped with an electrospray ionization (ESI) interface source. The RapidFire Mass Spec system was equipped with a reusable RapidFire C18 (type C) solid phase extraction (SPE) cartridge (G9205A).

[0437] The solvent A for sample loading and washing is 6mM octylamine (Acros Organics 129495000), which is adjusted to pH 5.0 using acetic acid. The solvent B for sample elution is 20% water in ACN, which contains 0.1% formic acid. Analyze samples successively by directly aspirating 10 μ L from porous plates to the collecting loop under vacuum. 10 μ L samples are loaded onto the C18 short column, and solvent A is used to wash 5000 ms with a flow rate of 1.25 mL / min. The analyte retained is then eluted with solvent B for 5000 ms to a mass spectrometer with a flow rate of 1.25 mL / min. The system is rebalanced for 2000 ms with a flow rate of 1.25 mL / min using solvent A.

[0438] The triple quadrupole mass spectrometer was equipped with an ESI source and the analytes were monitored in negative ion mode [MH]-using selected reaction monitoring (SRM). F1P was monitored at m / z 259.02 / 96.9 and d-P at m / z 264.99 / 97. 13 C6-fructose-6-phosphate. Use 13 The area ratio value to F1P was calculated using C6-fructose-6-phosphate as an internal standard.

[0439] The compounds of Examples 1-36 were tested essentially as described above:

[0440] Table 6

[0441]

[0442]

[0443]

[0444] Data are presented as mean ± SEM (n)

[0445] The results shown in Table 6 above indicate that the compounds of Examples 1-36 inhibited the enzyme activities of both KHK-C and KHK-A.

[0446] KHK cell activity assay

[0447] The efficacy of cellular KHK in inhibiting the conversion of fructose to F1P was measured using a cell-based assay. HepG2 hepatocytes were plated in growth medium [high glucose Dulbecco's Modified Eagle's medium (DMEM), 10% heat-inactivated fetal bovine serum (HI FBS), 1x penicillin / streptomycin] on 96-well cell culture plates and allowed to adhere overnight in a 37°C incubator. The growth medium was washed and replaced with an assay medium containing Gibco OptiMEM 1 Reduced Serum Medium, 0.1% casein, 8.33 mM D-fructose- 13 C6 and compounds ranging in concentration from 100 μM to 0.0051 μM (10-point concentration curve). The plates were incubated at 37°C for 3 h, and then the assay medium was aspirated from the cell wells. Stop solution containing 80% methanol, 2 mM ammonium acetate, and 50 ng / mL fructose-6-phosphate- 13 C6. The plates were stored at -20°C until RapidFireMS analysis (as described above).

[0448] The compounds of Examples 1-36 were tested essentially as described above:

[0449] Table 7

[0450]

[0451]

[0452]

[0453] Data are presented as mean ± SEM (n)

[0454] The results shown in Table 7 above indicate that the compounds of Examples 1-36 inhibited the metabolism of fructose to F1P in HepG2 cells.

[0455] Liquid chromatography-tandem mass spectrometry (LC-MS / MS) method for pharmacokinetic determination: by adding 180 μL MeOH:ACN (1:1, v / v) containing internal standard to 50 μL plasma, protein precipitation is used to extract the sample. The sample is then diluted with MeOH:water (1:1, v / v) to make its concentration within the standard curve range. The Sciex API4000 triple quadrupole mass spectrometer (Applied Biosystems / MDS; Foster City, CA) equipped with a TurboIonSpray interface and run in positive ion mode to analyze the diluted sample by LC-MS / MS. Analytes are chromatographically separated using ECHIELON C18 4um 20X2.1mm columns. LC conditions are water / 1M ammonium bicarbonate (2000:10, v / v) (mobile phase A) and MeOH / 1M ammonium bicarbonate (2000:10, v / v) (mobile phase B).

[0456] Pharmacokinetics in Sprague Dawley rats

[0457] The in vivo pharmacokinetic properties of Example 1 and Example 2 were demonstrated using Sprague-Dawley rats (fasted; n=3 for each route of administration). Administration was by a single oral (PO; 2 or 3 mg / kg; volume of 10 mL / kg) or intravenous (IV; 1 mg / kg; volume of 1 mL / kg) dose in vehicle. Blood was collected from each animal at multiple time points between 0 and 48 hours after administration. The plasma concentrations of Example 1 and Example 2 were determined by the LC-MS / MS method as described above.

[0458] The mean half-life of Example 1 was 12.9 hours and the bioavailability was 83% as determined by PO administration, while the mean half-life of Example 1 was 12.8 hours and the mean clearance was 5.86 mL / min / kg as determined by IV administration. The mean half-life of Example 2 was 5.12 hours and the bioavailability was 95% as determined by PO administration, while the mean half-life of Example 2 was 4.29 hours and the mean clearance was 56.4 mL / min / kg as determined by IV administration. The data show that Examples 1 and 2 have different levels of clearance, but both have high oral bioavailability and long elimination times, as evidenced by adequate mean half-lives.

[0459] Pharmacokinetics in Dogs

[0460] Beagle dogs (fed, n=3) were used to demonstrate the in vivo pharmacokinetic properties of Example 1 and Example 2. The compounds were administered by a single oral (PO; 2 or 3 mg / kg; volume 2 mL / kg) or intravenous (IV; 1 mg / kg; volume 1 mL / kg) dose in vehicle. Blood was collected from each animal at multiple time points between 0-72 hours after dosing. The plasma concentrations of Example 1 and Example 2 were determined by the LC-MS / MS method described above.

[0461] Example 1 had a mean half-life of 36.6 hours and a bioavailability of 87% as determined by PO administration, while Example 1 had a mean half-life of 28 hours and a mean clearance of 3.41 mL / min / kg as determined by IV administration. Example 2 had a mean half-life of 9.79 hours and a bioavailability of -100% as determined by PO administration, while Example 2 had a mean half-life of 10.3 hours and a mean clearance of 19.6 mL / min / kg as determined by IV administration. The data show that Examples 1 and 2 have different levels of clearance, but both have high oral bioavailability and long elimination times, as evidenced by adequate mean half-lives.

Claims

1. The compound of the formula: in X is N or C substituted by CN; R 1 Selected from: H. R 2 and R 3 Both are H, or one is H and the other is OH; R 4 , R 5 , R 6 , R 7 and R 9 are independently H or CH3; R 8 is H, CH3, CH2CH2OH, C(=O)CH2NH2 or C(=O)CH3; and R 10 It is OH or NH2; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein R 1 Selected from: or a pharmaceutically acceptable salt thereof.

3. The compound of claim 1 or claim 2, wherein the compound is: or a pharmaceutically acceptable salt thereof.

4. The compound of claim 3, wherein X is N, or a pharmaceutically acceptable salt thereof.

5. The compound of claim 3, wherein X is C substituted with CN, or a pharmaceutically acceptable salt thereof.

6. The compound of claim 1, wherein the compound is: or a pharmaceutically acceptable salt thereof.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, which is a succinate salt.

8. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating type 2 diabetes.

9. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating heart failure.

10. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diabetic nephropathy.

11. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating non-alcoholic steatohepatitis.

12. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating chronic kidney disease.

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers, diluents or excipients.

14. A method for preparing a pharmaceutical composition, comprising mixing a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable carriers, diluents or excipients.

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