Aminopyrimidine compounds suitable for use as SSAO inhibitors and methods for preparing the same

Abstract

The present application relates to aminopyrimidine compounds useful as SSAO inhibitors and methods for preparing the same. The present invention provides a pharmaceutically acceptable salt of the compound of the formula: a method for treating liver disease in a patient and a method for preparing the compound.

Description

[0001] This application is a divisional application of the invention patent application filed on February 7, 2018, with application number 201880000825.X and titled “Aminopyrimidine compounds suitable for use as SSAO inhibitors”. Technical Field

[0002] The present invention relates to aminopyrimidine compounds, pharmaceutically acceptable salts of said compounds and therapeutic uses of said compounds and their salts. Background Art

[0003] Semicarbazide-sensitive aminooxidase / vascular adhesion protein-1 (SSAO / VAP-1) exists in two forms: a membrane-bound isoform and a plasma-soluble isoform. It is mainly expressed from the surface of endothelial cells, vascular smooth muscle, and adipocytes. SSAO / VAP-1 is involved in many cellular processes, including glucose disposal, inflammatory responses and related pain, and leukocyte recruitment. High activity levels of this enzyme are associated with diabetes, atherosclerosis, stroke, chronic kidney disease, and Alzheimer's disease, among other conditions. SSAO / VAP-1 has been implicated in the pathogenesis of liver diseases, such as fatty liver. (Weston CJ et al., J. Neural. Transm., 2011, 118, 1055.) Fatty liver disease (FLD) covers a range of disease conditions characterized by excessive accumulation of fat in the liver, often accompanied by inflammation. FLD can cause non-alcoholic fatty liver disease (NAFLD), which is characterized by insulin resistance. If left untreated, NAFLD can progress to persistent inflammation or non-alcoholic steatohepatitis (NASH), progressive liver fibrosis, and ultimately cirrhosis. There is a need to provide alternative treatments for liver diseases such as NAFLD and / or NASH.

[0004] It is believed that SSAO / VAP-1 inhibitors will reduce liver inflammation and fibrosis and thus provide treatment for liver disease, especially for the treatment of NAFLD and / or NASH. In addition, because the activation of SSAO / VAP-1 is involved in inflammation and related pain, the inhibition of SSAA / VAP-1 enzymes may be useful for the treatment of pain and pain associated with osteoarthritis in particular. (Luis M. et al., J of Pharm and Experimental Therapeutics, 2005, 315, 553).

[0005] US Patent No. 8,426,587 discloses haloallylamine compounds useful as SSAO / VAP1 inhibitors.

[0006] Currently, there are no approved drugs for the treatment of NASH; the standard of care for NASH consists of dietary control and / or lifestyle changes. In addition, the current standard of care for pain is dominated by nonsteroidal anti-inflammatory drugs (NSAIDS) and opiates. Both drug classes are recommended for short-term use only. It is desirable to have more treatment options to control pain, especially chronic pain. The present invention provides compounds that inhibit the SSAO / VAP-1 enzyme and can address one or more of these needs. Summary of the invention

[0007] The present invention provides a compound of the following formula 1:

[0008]

[0009] Wherein R1 is selected from: as well as R2 is selected from: H, -C(O)NH2, -C(O)NH(CH3), -C(O)NH(CH3)2, as well as R3 is H or CH3, R4 is H or R5 is selected from: H, -C 1-4 Alkyl, -C 3-4 Cycloalkyl, -CH2-C 3-4 cycloalkyl; and n is 1 or 2; or a pharmaceutically acceptable salt thereof.

[0010] Description The bond to fluorine indicates that the fluorine atom and the methoxypyrimidine group can be Z (same side, together) or E (different side, opposite) relative to each other. (See Brecher J. et al., "Graphical Representation of Stereochemical Configuration", Pure and Appl. Chem, 2006, 78 (10) 1897, p. 1959). The structure illustrated by Formula 1 includes compounds that exhibit Z stereochemical configuration or E stereochemical configuration around the double bond; or mixtures of compounds that exhibit Z or E stereochemical configuration alone. Preferred compounds of the present invention have E stereochemical configuration around the double bond.

[0011] The present invention also provides a compound of formula 2:

[0012]

[0013] Wherein R1 is selected from: as well as R2 is selected from: H, -C(O)NH2, -C(O)NH(CH3), -C(O)NH(CH3)2, as well as R3 is H or CH3; R4 is H or R5 is selected from: H, -C 1-4 Alkyl, -C 3-4 Cycloalkyl and -CH2-C 3-4 cycloalkyl; and n is 1 or 2; or a pharmaceutically acceptable salt thereof.

[0014] In another form, the invention provides compounds according to formula 1 or 2, wherein R1 is selected from: as well as and wherein R2, R3, R4, R5 and n are as provided above; or a pharmaceutically acceptable salt thereof. In a preferred embodiment, R1 is and R5 and n are as provided above. In another preferred embodiment, R1 is and R5 is selected from: H, -CH3, More preferably R5 is -CH3.

[0015] In another form, the present invention provides compounds according to formula 1 or 2, wherein R1 is R2 is selected from: H, -C(O)NH2, -C(O)NH(CH3), -C(O)NH(CH3)2, as well as and R3 and R4 are as provided above, or a pharmaceutically acceptable salt thereof. In a preferred embodiment, R3 is H and R4 is as provided above, or a pharmaceutically acceptable salt thereof. In another preferred embodiment, R3 is H and R4 is H.

[0016] In another form, the present invention provides compounds according to formula 1 or 2, wherein R1 is R4 is and R2 and R3 are as provided above, or a pharmaceutically acceptable salt thereof. More preferably, R2 and R3 are both H.

[0017] In another form, the invention provides a compound according to the following Formula 3:

[0018]

[0019] or a pharmaceutically acceptable salt thereof.

[0020] In another form, the present invention provides a compound of formula 4:

[0021]

[0022] or a pharmaceutically acceptable salt thereof. In one form, the compound of formula 4 is provided as a free base.

[0023] In another form, the compound of formula 4 is provided in the form of a pharmaceutically acceptable salt. Preferably, the compound of formula 4 is provided in the form of a mono- or dihydrochloric acid addition salt.

[0024] In another form, the invention provides a pharmaceutical composition comprising a compound according to any one of Formulae 1 to 4, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent or excipient. The pharmaceutical composition can be used to treat a patient suffering from a liver disorder.

[0025] In another form, the present invention provides a method for treating a liver disorder in a patient in need thereof. The method comprises administering to the patient an effective amount of a compound according to Formula 1 to 4, or a pharmaceutically acceptable salt thereof. In certain embodiments, the method comprises treating a liver disorder in a patient in need of treatment, wherein the liver disorder is selected from: liver fibrosis, alcohol-induced fibrosis, alcohol steatosis, non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). In a particularly preferred embodiment, the method comprises treating a patient in need of NASH treatment. Preferably, the method comprises administering an effective amount of a compound of Formula 4 or a pharmaceutically acceptable salt thereof for the treatment of NASH.

[0026] In another form, the present invention provides a compound according to any one of formulas 1 to 4, or a pharmaceutically acceptable salt thereof, for use in therapy. In a preferred embodiment, the present invention provides a compound according to any one of formulas 1 to 4, or a pharmaceutically acceptable salt thereof, for use in treating a liver disorder. The liver disorder may be selected from: liver fibrosis, alcohol-induced fibrosis, alcohol steatosis, NAFLD, and NASH. In one embodiment, the therapy is for treating liver fibrosis. In another embodiment, the therapy is for NAFLD. In yet another embodiment, the therapy is for NASH.

[0027] In yet another form, the invention provides the use of a compound according to Formula 1 to 4 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a liver disorder. In a preferred embodiment, the liver disorder is selected from: liver fibrosis, alcohol-induced fibrosis, alcohol steatosis NAFLD and NASH.

[0028] The term "pharmaceutically acceptable salt" as used herein refers to salts of compounds of the present invention that are considered to be acceptable for clinical and / or veterinary use. Examples of pharmaceutically acceptable salts and common methods of preparing them can be found in "Handbook of Pharmaceutical Salts: Properties, Selection and Use" P. Stahl et al., 2nd Revised Edition, Wiley-VCH, 2011 and SM Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Sciences, 1977, 66 (1), 1-19.

[0029] The pharmaceutical composition used in the present invention can be prepared using pharmaceutically acceptable additives. The term "pharmaceutically acceptable additive" used in pharmaceutical compositions as used herein refers to one or more carriers, diluents and excipients that are compatible with other additives of the composition or formulation and harmless to the patient. Examples of pharmaceutical compositions and methods of preparing the same can be found in "Remington: The Science and Practice of Pharmacy", edited by Lloyd V. et al., 22nd edition, Mack Publishing Co., 2012.

[0030] As used herein, the term "effective amount" refers to an amount as a dosage effective for treating a condition, such as liver diseases including liver inflammation, fibrosis, and steatohepatitis. The effective amount can be readily determined by the care physician as a skilled artisan by using conventional techniques and by observing the results obtained under similar circumstances. Factors considered in determining the effective amount or dosage of a compound include: whether the compound or its salt will be administered; co-administration of other agents (if used); the species of the mammal; its size, age, and general health; the extent or severity of the condition; the response of the individual patient; the mode of administration; the bioavailability characteristics of the administered formulation; the dosage regimen selected; the use of other concomitant agents; and other relevant circumstances.

[0031] As used herein, the terms "treating", "to treat" or "treatment" include slowing, reducing or reversing the progression or severity of an existing symptom, disorder, condition or disease, which may include treating liver disease, such as liver inflammation, fibrosis and steatohepatitis.

[0032] As used herein, the term "patient" refers to a mammal, preferably the patient is a human or a companion mammal, such as a dog or cat.

[0033] The treating physician, veterinarian or other medical personnel will be able to determine the effective amount of the compound for treating a patient in need. Preferred pharmaceutical compositions can be formulated as tablets or capsules for oral administration, solutions for oral administration or injectable solutions. Tablets, capsules or solutions can include the compound of the present invention in an amount that is effective for treating a patient in need of treatment.

[0034] Abbreviations used herein are defined according to Daub GH et al., "The Use of Acronyms in Organic Chemistry," Aldrichimica Acta, 1984, 17(1), 6-23. Other abbreviations are defined as follows: “Boc” refers to tert-butyloxycarbonyl; “DBAD” refers to dibenzyl azodicarboxylate; “DCM” refers to dichloromethane; “DIPEA” refers to N,N-diisopropylethylamine; “DMF” refers to dimethylformamide; “DMSO” refers to dimethyl sulfoxide; “EDTA” refers to ethylenediaminetetraacetic acid; “EGTA” refers to ethylene glycoltetraacetic acid; “ES / MS” refers to electrospray mass spectrometry; “EtOH” refers to ethyl acetate; “EtOH” refers to ethanol or ethyl alcohol; “HATU” refers to (dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methanimine hexafluorophosphate; “HEPES” refers to 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; “hr or hrs” refers to hour / hour; “IC 50 ” refers to the concentration of an agent that produces 50% of the maximum possible inhibitory response of that agent (relative IC 50 ), or the concentration of the agent that produces 50% inhibition of the target activity compared to placebo control (absolute IC 50 ); “IU” refers to international units; “LCMS” refers to liquid chromatography mass spectrometry; “MAOa and MAOb” refer to monoamine oxidase a and b isoforms, respectively; “MeOH” refers to methyl alcohol or methanol; “min” or mins refers to minutes; “MTBE” refers to methyl tert-butyl ether; “NASH” refers to nonalcoholic steatohepatitis; “NMP” refers to N-methyl-pyrrolidone or 1-methyl-2-pyrrolidone; PE refers to petroleum ether; t (R) = retention time; "sat" refers to saturated solution; "SSAO" refers to semicarbazide-sensitive amine oxidase; "hSSAO" refers to human SSAO; and "TG" refers to triglyceride; "THF" refers to tetrahydrofuran.

[0035] In the preparation described herein, hydroxyl and amino functional groups can be protected to facilitate the synthesis of compounds described herein. Examples of protected functional groups can be found in "Greene's Protective Groups in Organic Synthesis," Wuts PGM et al., 5th edition, John Wiley and Sons, 2014. Other functional groups that can be easily converted to hydroxyl or amino groups can be used. Such functional groups, preparations and transformations of these groups can be found in "Comprehensive Organic Transformations: A Guide to Functional Group Preparations," by Larock. RC, Wiley VCH, 1999, and in "March's Advanced Organic Chemistry: Reactions, Mechanisms and Structure," ed. Smith MB, 7th edition, Wiley-Interscience, 2013. DETAILED DESCRIPTION

[0036] The compound of the present invention or its salt can be prepared by a variety of procedures, some of which are described in the following flow, preparation and example. The product of each step in the following flow can be recovered by a conventional method, and the method includes extraction, evaporation, precipitation, chromatography, filtration, grinding and crystallization. In addition, individual isomers, enantiomers and diastereomers can be separated or dissolved by such as selective crystallization techniques or chiral chromatography (see, for example, J. Jacques et al., " Enantiomers, racemates and resolution (Enantiomers, Racemates, and Resolutions) ", John Wiley and Sons, 1981 and EL Eliel (EL Eliel) and SH Wilen (SH Wilen), " Stereochemistry of Organic Compounds ", " Wiley-International Science ", 1994). In the following flow, unless otherwise indicated, all substituents are as previously defined. Reagents and starting materials are easily available to those skilled in the art.

[0037] Process 1

[0038]

[0039] Scheme 1 depicts a general synthesis of compounds of formula 1, wherein "PG" is a protecting group for the hydroxyl group. Specifically, in step 1, sub-step 1, the chlorine of compound A can be replaced by the nitrogen of R1 substituted with a cyclic amine to obtain compound B. In step 1, sub-step 2, deprotection of the hydroxyl group can be achieved by a variety of methods depending on the specific protecting group. In step 2, sub-step 1, the hydroxyl group of the resulting compound B can be alkylated with a suitably protected amine 2-bromo-3-fluoro-propyl-2-ene-amine to obtain a compound of formula 1. Alkylation can generally be achieved under alkaline conditions. The protected amine can be deprotected to obtain a compound of formula 1.

[0040] Preparation and Examples

[0041] The following Preparations and Examples further illustrate the invention and represent typical syntheses of compounds of the invention.

[0042] Preparation 1

[0043] 4-(5-Bromo-2-pyridinyl)-2-oxo-piperazine-1-carboxylic acid tert-butyl ester

[0044]

[0045] Triethylamine (2.4mL, 17mmol), DMAP (160mg, 1.30mmol) and tert-butyl carbonate (1.87g, 1.1 equivalents, 8.59mmol) are added to a solution of 4-(5-bromo-2-pyridyl)piperazine-2-one (2.00g, 7.81mmol) in DCM (45mL). The reaction mixture is stirred for 3 days at 25°C to give a yellow suspension. DCM (50mL) is added to the reactant and the mixture is washed with brine (5×50mL). The organic layer is concentrated to give a crude product in the form of a yellow solid. The material is subjected to silica gel chromatography eluted with a gradient of 0 to 50% EtOAc / PE to give the title compound (1.57g, 55%) in the form of a white solid.

[0046] Preparation 2

[0047] 2-Oxo-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]piperazine-1-carboxylic acid tert-butyl ester

[0048]

[0049] To a suspension of tert-butyl 4-(5-bromo-2-pyridyl)-2-oxo-piperazine-1-carboxylate (1.57 g, 4.41 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (1.23 g, 1.1 eq., 4.85 mmol) and potassium acetate (1.29 g, 3.00 eq., 13.2 mmol) in 1,4-dioxane (30 mL) was added Pd(dppf)Cl2) (0.32 g, 0.1 eq., 0.441 mmol) at 25°C. The reaction mixture was degassed under N2 for 1 hr while stirring at 100°C. The reaction mixture was filtered and the filtrate was concentrated to give the crude product as a yellow oil. The material was subjected to silica gel chromatography eluting with a gradient of 0 to 50% EtOAc / PE to give the title compound as a light yellow solid (1.70 g, 96%).

[0050] Preparation 3

[0051] 4-(5-Hydroxy-2-pyridinyl)-2-oxo-piperazine-1-carboxylic acid tert-butyl ester

[0052]

[0053] Sodium hydroxide (aqueous solution 2mL, 1M) and hydrogen peroxide (aqueous solution 2mL, 30 mass%) were added to a suspension of tert-butyl 2-oxo-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazine-1-carboxylate (500mg, 1.24mmol) in THF (5mL, 61.6mmol) at 0°C. The reaction mixture was stirred for 2 hours at 25°C. The reactant was quenched with saturated Na2SO3 solution and the pH value was adjusted to 6-7 with 1M HCl. The mixture was extracted with EtOAc (3×20mL) and the combined organic extracts were concentrated to give a crude product as a yellow oil. The crude product was subjected to silica gel chromatography eluted with a gradient of 0 to 70% EtOAc in PE to give the title compound (120mg, 33%) as a yellow solid. LCMS (m / z): 294.0 [M+H] + .

[0054] Preparation 4

[0055] 1-Methyl-3-(4-pyridyloxy)pyrrolidin-2-one

[0056]

[0057] A solution of DBAD (3.06 g, 13.0 mmol) in THF (10 mL) was added dropwise to 3-hydroxy-1-methyl-pyrrolidin-2-one (500 mg, 4.34 mmol), pyridine-4-ol (0.826 g, 8.69 mmol) and (n-butyl) 3P (2.72 g, 13.0 mmol) in DCM (9 mL) and THF (15 mL) at 0 ° C. The reaction mixture was stirred for 4 hours at room temperature. The reaction mixture was concentrated under vacuum and the residue was purified by flash chromatography on silica gel eluted with a gradient of 0.5% to 10% MeOH in DCM to give the title compound (300 mg, 32% yield) as a light yellow oil. LCMS (m / z): 193.0 [M+H] + , 1 HNMR(400MHz, CDCl3)δ8.50-8.35(m,2H),7.00-6.87(m,2H),4.94(dd,J=6.0,7.6Hz,1H), 3.56-3.46(m,1H),3.44-3.32(m,1H),2.93(s,3H),2.63-2.50(m,1H),2.21-2.09(m,1H).

[0058] Preparation 5

[0059] 3-(1-Benzylpyridin-1-ium-4-yl)oxy-1-methyl-pyrrolidin-2-one bromide

[0060]

[0061] Bromomethylbenzene (0.534 g, 3.12 mmol) was added to a mixture of 1-methyl-3-(4-pyridyloxy)pyrrolidin-2-one (5,300 mg, 1.56 mmol) in DMF (8.0 mL) and the reaction mixture was warmed to 55 °C for 16 hours. The reaction mixture was concentrated under vacuum, diluted with water (20 mL) and extracted with EtOAc (2 x 10 mL). The aqueous layer was concentrated under vacuum to give the title compound (450 mg, 71.4%) as a white solid. 1 H NMR(400MHz,d6-DMSO)δ9.04(d,J=7.2Hz,2H),7.75(d,J=7.6Hz,2H),7.56-7.48(m,2H),7.47-7.33 (m,3H),5.72(s,2H),5.61(t,J=8.0Hz,1H),3.42-3.37(m,5H),2.70-2.58(m,1H),2.17-2.00(m,1H)

[0062] Preparation 6

[0063] 3-[(1-Benzyl-3,6-dihydro-2H-pyridin-4-yl)oxy]-1-methyl-pyrrolidin-2-one

[0064]

[0065] NaBH4 (212 mg, 5.58 mmol) was added to a solution of 3-(1-benzylpyridin-1-ium-4-yl)oxy-1-methyl-pyrrolidin-2-one bromide (450 mg, 1.12 mmol) in MeOH (8.0 mL) at 0°C and the reaction mixture was stirred at 0°C for 10 min. The reaction mixture was diluted with EtOAc (40 mL) and washed with NaHCO3 (saturated aqueous solution) (30 mL) and brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The crude product was subjected to silica gel flash chromatography eluted with a gradient of 0-1% MeOH / DCM to give the title compound (120 mg, 36%) as a colorless gum. LCMS (m / z): 287.1 [M+H] + , 1 H NMR (400MHz, CDCl3) δ7.45-7.14(m,5H),4.71(t,J=3.2Hz,1H),4.62-4.50(m,1H),3.74-3.50(m,2H),3.50-3.40(m,1H),3.40-3.25(m,1H),3. 18-3.06(m,1H),3.05-2.95(m,1H),2.89(s,3H),2.75-2.65(m,1H),2.5 5-2.47(m,1H),2.43-2.30(m,1H),2.25-2.15(m,2H),2.06-1.94(m,1H)

[0066] Preparation 7

[0067] 1-Methyl-3-(4-piperidinyloxy)pyrrolidin-2-one

[0068]

[0069] Palladium on carbon (50% water, 10% w, 20.0 mg) was added to a solution of 3-[(1-benzyl-3,6-dihydro-2H-pyridin-4-yl)oxy]-1-methyl-pyrrolidin-2-one (120 mg, 0.40 mmol) in EtOH (6.0 mL) and the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 5 hours. The reaction mixture was filtered through celite and the filtrate was concentrated under vacuum to give the title compound (85.0 mg, 96.9%) as a colorless gum. 1H NMR (400MHz, CDCl3) δ4.20-4.10(m,1H),3.90-3.80(m,1H),3.45-3.35(m,1H),3.30-3.18(m,1H),3.14 -3.04(m,2H),2.85(s,3H),2.66-2.55(m,2H),2.36-2.28(m,1H),1.98-1.90(m,2H),1.55-1.35(m,2H)

[0070] Preparation 8

[0071] 2-Cyclopropyl-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester

[0072]

[0073] Combine tert-butyl 1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (161 mg, 0.60 mmol), copper(II) acetate (110 mg, 0.61 mmol) and cesium carbonate (98 mg, 0.30 mmol). Pyridine (145 mg, 1.83 mmol), 2-cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (203 mg, 1.21 mmol) and toluene (1.2 mL) are then added via a syringe. The mixture is heated to 110 ° C for 64 hours. The mixture is washed with EtOAc and filtered through celite. The filtrate is concentrated and the residue is subjected to silica gel flash chromatography eluted with 50% EtOAc / hexanes to obtain the title compound as a light yellow oil. LCMS (m / z): 317.3 [M+Na] +

[0074] Preparation 9

[0075] 2-Cyclopropyl-2,8-diazaspiro[4.5]decan-1-one hydrochloride

[0076]

[0077] Tert-butyl 2-cyclopropyl-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (172 mg, 0.555 mmol) in HCl was dissolved in MeOH (6.0 mL, 3 mmol, 0.5 mmol / mL) and heated to 80 °C for 50 min. The mixture was concentrated under vacuum to give the title compound as a light brown oil which was used without further purification. LCMS (m / z): 195.3 [M+H] +

[0078] Preparation 10

[0079] 4-(2-Bromoethyl)piperidine-1,4-dicarboxylic acid O1-tert-butyl O4-methyl ester

[0080]

[0081] Lithium diisopropylamine in hexane (9.70 mL, 19.396 mmol, 2 mol / L) was added dropwise to a solution of piperidine-1,4-dicarboxylic acid O1-tert-butyl O4-methyl ester (3.146 g, 12.93 mmol) in THF (45 mL) at -78 ° C under N2. The resulting mixture was stirred for 30 min at -78 ° C and then 1,2-dibromoethane (2.23 mL, 25.861 mmol) was added. The resulting mixture was warmed to room temperature and stirred for 1 hr. The reactant was quenched with saturated NH4Cl aqueous solution (20 mL) and extracted with EtOAc (2 × 30 mL). The organic extracts were combined, dried over Na2SO4, filtered and concentrated to dryness. The crude material was purified by flash chromatography on silica gel eluted with 20% EtOAc / hexane to give the title compound (0.698 g, 15%) as a slightly yellow oil. LCMS (m / z): ( 79 Br / 81 Br)372.2 / 374.2[M+Na] +

[0082] Preparation 11

[0083] 2-tert-Butyl-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester

[0084]

[0085] 4-(2-Bromoethyl)piperidine-1,4-dicarboxylic acid O1-tert-butyl O4-methyl ester (698 mg, 1.89 mmol) was dissolved in MeOH (5 mL) and 2-methylpropan-2-amine (1.59 mL, 15.1 mmol) was added. The solution was heated to 120 °C for 16 hours via microwave irradiation. The solution was concentrated and the residue was then purified via silica gel flash chromatography eluting with 25% EtOAc / hexanes to give the title compound as a light yellow oil (74 mg, 11%). LCMS (m / z): 333.3 [M+Na] +

[0086] Preparation 12

[0087] 2-tert-Butyl-2,8-diazaspiro[4.5]decan-1-one hydrochloride

[0088]

[0089] Tert-butyl 2-tert-butyl-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (74 mg, 0.21 mmol) in HCl was dissolved in MeOH (5 mL, 2.5 mmol, 0.50 mmol / mL) and heated to 80 °C for 5 min via microwave irradiation. The solution was concentrated to give the title compound (56 mg, 95%) as a light yellow oil. LCMS (m / z): 211.2 [M+H] +

[0090] Preparation 13

[0091] 2-Methyl-2,8-diazaspiro[4.5]decan-1-one hydrochloride

[0092]

[0093] Tert-butyl 2-methyl-1-oxo-2,8-diazaspiro[4.5]decane-8-carboxylate (45.2 g, 168 mmol) was cooled to 0°C. HCl in MeOH (250 mL, 4.0 M) was added and the solution was stirred vigorously for 30 minutes. The mixture was warmed to room temperature, stirred for 5 hours, and then concentrated to dryness to give the title compound (34.6 g, 98.4%) as a pale yellow solid. ES / MS (m / z) 169.2 (M+H).

[0094] Preparation 14

[0095] N-[(E)-2-[(2-Chloropyrimidin-5-yl)oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0096]

[0097] Potassium carbonate (1.126 g, 8.14 mmol) was added to a solution of 2-chloropyrimidine-5-ol (501 mg, 3.84 mmol) and tert-butyl N-[(E)-2-(bromomethyl)-3-fluoro-allyl]carbamate (507 mg, 1.89 mmol) in DMF (10 mL) and the resulting mixture was stirred at room temperature overnight. The reactants were quenched by adding water and EtOAc and the aqueous phase was extracted with EtOAc (3×50 mL). The organic extracts were combined, the solution was dried over Na2SO4, filtered and the filtrate was concentrated under vacuum. The material was subjected to silica gel flash chromatography with EtOAc / hexanes to give the title compound (658 mg, 87%) as a white solid. LCMS (ESI): m / s 340.2 [M+Na] + .

[0098] Preparation 15

[0099] 8-(5-Benzyloxypyrimidin-2-yl)-2,8-diazaspiro[4.5]decan-1-one

[0100]

[0101] DIPEA (1.7 g, 13 mmol) and 5-benzyloxy-2-chloro-pyrimidine (0.61 g, 2.8 mmol) were added to a mixture of 2-methyl-2,8-diazaspiro[4.5]dec-1-one hydrochloride (0.50 g, 2.6 mmol) in NMP (10 mL). The reaction mixture was stirred at 100 °C for 20 hours. The reaction mixture was then diluted with water (40 mL) and extracted with EtOAc (2 × 20 mL). The organic extracts were combined; washed with brine (3 × 20 mL); dried over Na2SO4; filtered; and the filtrate was concentrated under vacuum to give a residue. The residue was subjected to flash chromatography on silica gel eluted with a gradient of 0-1% MeOH / DCM to give the title compound (0.39 g, 42%) as a light yellow solid. 1 H NMR(400MHz, CDCl3)δ8.11(s,2H),7.45-7.30(m,5H),5.79(s,1H),5.02(s,2H),4.55-4.45(m,2H),3 .38(t,J=6.8Hz,2H),3.21-3.10(m,2H),2.15(t,J=6.8Hz,2H),1.98-1.85(m,2H),1.55-1.43(m,2H).

[0102] Preparation 16

[0103] 1-(5-Benzyloxypyrimidin-2-yl)piperidine-4-carboxamide

[0104]

[0105] A mixture of 5-benzyloxy-2-chloro-pyrimidine (400 mg, 1.81 mmol), piperidine-4-carboxamide (0.28 g, 1.2 eq., 2.18 mmol) and DIPEA (2.0 eq., 3.63 mmol) in DMF (6 mL) was stirred at 100 °C under N2 for 17 hours. The reaction mixture was poured into water (60 mL) and filtered. The filter cake was washed with EtOAc (30 mL) and the material was stirred for 0.5 hr. The solution was filtered and the filter cake was dried under reduced pressure to give the title compound (310 mg, 49%) as a pink solid. 1H NMR: (400MHz, CD3OD) δ8.13(s,2H),7.47-7.28(m,5H),5.07(s,2H),4.67-4.60(m ,2H),2.95-2.84(m,2H),2.50-2.40(m,1H),1.85-1.75(m,2H),1.68-1.60(m,2H).

[0106] Preparation 17

[0107] 8-(5-Benzyloxypyrimidin-2-yl)-2-cyclopropyl-2,8-diazaspiro[4.5]decan-1-one

[0108]

[0109] 5-Benzyloxy-2-chloro-pyrimidine (79 mg, 0.36 mmol) and potassium carbonate (164 mg, 1.19 mmol) were added together. 2-Cyclopropyl-2,8-diazaspiro[4.5]dec-1-one; hydrochloride (72 mg, 0.30 mmol) was dissolved in EtOH (3.0 mL) and added to the reaction mixture. The reactants were heated to 110 ° C for 62 hours via microwave irradiation. The mixture was diluted with EtOAc and the slurry was filtered through celite. The filtrate was concentrated and the crude mixture was purified by flash chromatography on silica gel eluted with 55% EtOAc / hexanes to give the title compound (40 mg, 35%) as a white solid. LCMS (m / z): 379.2 [M+H] + .

[0110] Preparation 18

[0111] 8-(5-Benzyloxypyrimidin-2-yl)-2-tert-butyl-2,8-diazaspiro[4.5]decan-1-one

[0112]

[0113] NMP (1 mL) was added to 2-tert-butyl-2,8-diazaspiro[4.5]dec-1-one; hydrochloride (56 mg, 0.20 mmol), 5-benzyloxy-2-chloro-pyrimidine (54 mg, 0.25 mmol) and potassium carbonate (113 mg, 0.82 mmol). The mixture was heated to 120 ° C for 16 hours via microwave irradiation. The mixture was diluted with EtOAc and filtered through celite. The filtrate was concentrated to give a residue and the residue was subjected to silica gel flash chromatography eluted with 25% EtOAc / hexanes to give the title compound as a white solid (16 mg, 19% yield). LCMS (m / z): 395.3 [M+H] +

[0114] Preparation 19

[0115] 9-(5-Benzyloxypyrimidin-2-yl)-2,9-diazaspiro[5.5]undecan-1-one

[0116]

[0117] 1-Oxo-2,9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.907g, 6.96mmol) was dissolved in DCM (50mL) and trifluoroacetic acid (10mL) was added at room temperature under stirring. The resulting solution was stirred for 4 hours. The solution was concentrated to give the intermediate product 2,9-diazaspiro [5.5] undecane-1-one as a light yellow oil; 2,2,2-trifluoroacetic acid (2.096g, 6.683mmol). DMF (10mL) was added to the crude mixture 5-benzyloxy-2-chloro-pyrimidine (880mg, 3.99mmol), cuprous iodide (158mg, 0.830mmol), N,N'-bis (2-phenoxyphenyl) oxamide (220mg, 0.804mmol) and tribasic potassium phosphate (2.612g, 12.06mmol). The resulting mixture was stirred under N2 and heated to 100°C for 6 hours. The mixture was diluted with EtOAc and filtered through celite. The filtrate was concentrated and the resulting crude material was subjected to silica gel flash chromatography eluting with 7% MeOH / DCM to give the title compound (869 mg, 61%) as a yellow solid. LCMS (m / z): 353.2 [M+H] +

[0118] Preparation 20

[0119] 8-(5-Benzyloxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]decan-1-one

[0120]

[0121] Sodium hydride (60% by mass, 66 mg, 1.6 mmol) in mineral oil was added to a solution of 8-(5-benzyloxypyrimidin-2-yl)-2,8-diazaspiro[4.5]dec-1-one (0.39 g, 1.1 mmol) in DMF (8.0 mL); the mixture was stirred at 0 ° C for 20 min. Iodomethane (0.31 g, 2.2 mmol) was added to the cold (0 ° C) mixture. The mixture was allowed to warm to ambient temperature and the mixture was stirred for 1 hr. The reactants were quenched with water (30 mL). The resulting mixture was extracted with EtOAc (2×15 mL). The organic extracts were combined and washed with brine (2×20 mL); dried over anhydrous Na2SO4; filtered; and the filtrate was concentrated under vacuum to give the title compound (0.42 g, 98%) as a yellow solid, which can be used without further purification. 1 H NMR (400MHz, CDCl3) δ8.04(s,2H),7.38-7.24(m,5H),4.95(s,2H),4.42(dt,J=4.0,13.6Hz,2H),3.27(t,J =6.8Hz,2H),3.15-3.02(m,2H),2.79(s,3H),1.96(t,J=6.8Hz,2H),1.90-1.78(m,2H),1.42-1.34(m,2H).

[0122] Alternative preparation 20

[0123] 8-(5-Benzyloxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]decan-1-one

[0124] 5-Benzyloxy-2-chloro-pyrimidine (24.824 g, 112.50 mmol), 2-methyl-2,8-diazaspiro[4.5]dec-1-one hydrochloride (29.736 g, 145.27 mmol) and potassium carbonate (46.643 g, 337.50 mmol) were combined in NMP. NMP (170 mL) and triethylamine (23.5 mL, 168.75 mmol) were added and the mixture was heated to 130 ° C for 30 hours. The mixture was cooled, filtered to collect the solid, and then the solid was washed with EtOAc. The filtrate was concentrated and the concentrated solution was poured into crushed ice (about 1.2 L). A light brown solid precipitate appeared immediately. The mixture was stirred for 30 min and then the mixture was allowed to stand at room temperature overnight. The mixture was filtered to collect the solid and the solid was washed with MTBE (400 mL). The solid was dried under vacuum at 50 °C for 1.5 days to give the title compound (36.924 g, 88.48%) as a light brown solid which was used without further purification. ES / MS (m / z) 353.3 (M+H).

[0125] Preparation 21

[0126] 8-(5-Benzyloxypyrimidin-2-yl)-2-(cyclopropylmethyl)-2,8-diazaspiro[4.5]decan-1-one

[0127]

[0128] Sodium hydride (22 mg, 0.55 mmol, 60% by mass) in mineral oil was added to a stirred solution of 8-(5-benzyloxypyrimidin-2-yl)-2,8-diazaspiro[4.5]dec-1-one (60 mg, 0.18 mmol) in DMF (5.0 mL) at 0 ° C. The mixture was allowed to warm to room temperature and stirred for 5 min. (Bromomethyl)cyclopropane (100 mg, 0.71 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The solvent was evaporated under reduced pressure to give a crude product. The material was subjected to silica gel flash chromatography eluted with 40% EtOAc / hexanes to give the title compound (40 mg, 55%) as a white solid. LCMS (m / z): 392.2 [M+H] +

[0129] Preparation 22

[0130] 9-(5-Benzyloxypyrimidin-2-yl)-2-methyl-2,9-diazaspiro[5.5]undecan-1-one

[0131]

[0132] 9-(5-Benzyloxypyrimidin-2-yl)-2,9-diazaspiro[5.5]undecane-1-one (183 mg, 0.51 mmol) was dissolved in THF (8 mL) and the solution was cooled to 0 ° C. Sodium hydride (41 mg, 1.018 mmol, 60% by mass) was added in one portion. The solution was stirred at 0 ° C for 15 min. Iodomethane (0.064 mL, 1.02 mmol) was added at 0 ° C, the mixture was warmed to room temperature and stirred for 30 min. The mixture was cooled to 0 ° C, sodium hydride (20 mg, 0.51 mmol) was further added and stirred for 15 min. Iodomethane (0.032 mL, 0.51 mmol) was added, the mixture was warmed to room temperature, and stirred for 30 min. Saturated NH4Cl (aqueous solution) was added to quench the reactants and diluted with EtOAc. The organic phase was separated and the aqueous phase was extracted with EtOAc (2×). The organic extracts were combined, dried over Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was subjected to silica gel flash chromatography eluting with 75% EtOAc / hexanes to give the title compound (158 mg, 83%). LCMS (m / z): 367.2 [M+H] +

[0133] Preparation 23

[0134] 8-(5-Hydroxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]decan-1-one

[0135]

[0136] Palladium on carbon (50%, 10% w in water, 42 mg) was added to a solution of 8-(5-benzyloxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]dec-1-one (0.42 g, 1.1 mmol) in MeOH (40 mL). The mixture was stirred at room temperature for 4 hours under a hydrogen atmosphere and then at 40 ° C for 6 hours. The mixture was filtered through a pad of celite. The filtrate was concentrated under vacuum to give the title compound (0.34 g, 97%) as a pale yellow solid, which was used without further purification. 1 H NMR (400MHz, CDCl3) δ8.05 (s, 2H), 4.54-4.44 (m, 2H), 3.37 (t, J = 7.2Hz, 2H), 3.15-3.05 (m, 2H), 2 .88(s,3H),2.07(t,J=7.2Hz,2H),1.96-1.88(m,2H),1.45-1.40(m,2H), ES / MS(m / z)262.9(M+H).

[0137] Alternative preparation 23

[0138] 8-(5-Hydroxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]decan-1-one

[0139] A suspension of 8-(5-benzyloxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]decan-1-one (18.190 g, 49.04 mmol) in EtOAc (100 mL) and MeOH (100 mL) was transferred to a high pressure reactor to which a suspension of 5% palladium on activated carbon (1.84 g, 0.865 mmol) in MeOH (40 mL) was added. TM The mixture was hydrogenated in a shaker under 310 kPa hydrogen for 4 hours. The mixture was diluted with MeOH and filtered through celite. The filtrate was concentrated to dryness to give the title compound (12.9 g, 46.7 mmol, 95.3%) as a yellow solid, which was used without further purification. ES / MS (m / z) 263.3 (M+H).

[0140] Preparation 24

[0141] 2-Cyclopropyl-8-(5-hydroxypyrimidin-2-yl)-2,8-diazaspiro[4.5]decan-1-one

[0142]

[0143] 8-(5-Benzyloxypyrimidin-2-yl)-2-cyclopropyl-2,8-diazaspiro[4.5]dec-1-one (39 mg, 0.10 mmol) was dissolved in EtOAc (2.0 mL). Palladium on activated carbon (10 mg, 0.005 mmol, 5 mass%) was suspended in MeOH (1.0 mL) and transferred to the above solution. 1,4-cyclohexadiene (0.096 mL, 1.01 mmol) was added and the resulting mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc and filtered through celite. The filtrate was concentrated to give the title compound (29 mg, 98%) as a white solid, which was used directly without further purification. LCMS (m / z): 289.1 [M+H] +

[0144] Preparation 25

[0145] 2-(Cyclopropylmethyl)-8-(5-hydroxypyrimidin-2-yl)-2,8-diazaspiro[4.5]decan-1-one

[0146]

[0147] 8-(5-Benzyloxypyrimidin-2-yl)-2-(cyclopropylmethyl)-2,8-diazaspiro[4.5]dec-1-one (40 mg, 0.097 mmol) and palladium / carbon (20 mg, 5 mass%) were stirred together in MeOH (10 mL) under H2 atmosphere at room temperature for 16 hours. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give a crude product, which was subjected to silica gel flash chromatography eluted with 80% EtOAc and 20% hexanes to give the title compound (17 mg, 55%) as a white solid. LCMS (m / z): 303.3 [M+H] +

[0148] Preparation 26

[0149] 9-(5-Hydroxypyrimidin-2-yl)-2,9-diazaspiro[5.5]undecane-1-one

[0150]

[0151] Palladium on activated carbon (50 mg, 0.023 mmol) was suspended in MeOH (15 mL) and added to 9-(5-benzyloxypyrimidin-2-yl)-2,9-diazaspiro[5.5]undecan-1-one (97 mg, 0.27 mmol). A hydrogen balloon was equipped, evacuated 3 times with hydrogen, and the resulting mixture was stirred at room temperature for 4 hours. The mixture was diluted with MeOH and the solution was filtered through celite. The filtrate was concentrated to give the title compound (72 mg, 99%) as a white solid, which was used directly without further purification. LCMS (m / z): 263.1 [M+H] +

[0152] The following compound was prepared essentially according to the procedure of Preparation 26 and the mixture was stirred for 1-5 hours.

[0153]

[0154]

[0155] Preparation 30

[0156] tert-Butyl N-[(E)-3-fluoro-2-[[2-(2-methyl-1-oxo-2,8-diazaspiro[4.5]dec-8-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0157]

[0158] 8-(5-Hydroxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]dec-1-one (0.14 g, 0.49 mmol), tert-butyl N-[(E)-2-(bromomethyl)-3-fluoro-allyl]carbamate (0.11 g, 0.41 mmol) and potassium carbonate (0.17 g, 1.2 mmol) were combined in anhydrous DMF (5.0 mL). The resulting mixture was warmed to 50 °C and stirred for 1.5 hours. The mixture was diluted with water (30 mL) and extracted with EtOAc (2×15 mL). The organic extracts were combined; washed with brine (2×20 mL); dried over Na2SO4; filtered; and the filtrate was concentrated under vacuum to give a residue. The residue was subjected to flash chromatography on silica gel eluted with 0-0.5% MeOH / DCM to give the title compound (0.16 g, 82%) as a white solid. 1H NMR (400MHz, CDCl3) δ8.09 (s, 2H), 6.70 (d, J = 82.0Hz, 2H), 4.78 (br s,1H),4.60-4.45(m,2H),4.39(s,2H),4.00(s,2H),3.41-3.28(m,2H),3.25-3.18( m,2H),2.94-2.76(m,3H),2.12-2.01(m,2H),2.00-1.85(m,2H),1.50-1.38(m,12H).

[0159] Alternative preparation 30

[0160] tert-Butyl N-[(E)-3-fluoro-2-[[2-(2-methyl-1-oxo-2,8-diazaspiro[4.5]dec-8-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0161] 8-(5-Hydroxypyrimidin-2-yl)-2-methyl-2,8-diazaspiro[4.5]dec-1-one (12.9 g, 46.7 mmol), tert-butyl N-[(E)-2-(bromomethyl)-3-fluoro-allyl]carbamate (13.2 g, 49.1 mmol) and potassium carbonate (19.4 g, 140 mmol) were combined with DMF (73 mL). The resulting mixture was stirred at room temperature for 5 hours. The mixture was diluted with EtOAc and filtered through celite. The filtrate was concentrated to give a residue. The residue was subjected to flash chromatography on silica gel eluted with 3% MeOH / DCM to give the title compound (18.6 g, 40.6 mmol, 86.8%) as a yellow oil. ES / MS (m / z) 450.3 (M+H).

[0162] The following compounds were prepared essentially according to the procedure of Alternative Preparation 30 and the reactions were stirred for 3 to 64 hours.

[0163]

[0164] a Stir at room temperature and heat the reaction to 80 °C for 1.5 hours.

[0165] Preparation 37

[0166] 4-[5-[(E)-2-[(tert-Butoxycarbonylamino)methyl]-3-fluoro-allyloxy]-2-pyridinyl]-2-oxo-piperazine-1-carboxylic acid tert-butyl ester

[0167]

[0168] N-[(E)-2-(bromomethyl)-3-fluoro-allyl]carbamic acid tert-butyl ester (0.12 g, 1.1 eq., 0.450 mmol) and potassium carbonate (0.17 g, 3 eq., 1.23 mmol) were added to a solution of tert-butyl 4-(5-hydroxy-2-pyridyl)-2-oxo-piperazine-1-carboxylate (120 mg, 0.409 mmol) in DMF (3 mL, 39 mmol). The mixture was stirred at 70 ° C for 2 hours to give a yellow suspension. Water (20 mL) was added to the mixture and the mixture was extracted with EtOAc (3×20 mL). The extracts were combined and the combined organic extracts were washed with brine (2×20 mL). The organic extracts were concentrated to give the title compound (180 mg) as a yellow oil, which was used directly without further purification. LCMS (ESI): m / s 381.2 [M+H] + .

[0169] Preparation 39

[0170] N-[(E)-2-[[2-(4-Carbamoyl-1-piperidinyl)pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0171]

[0172] The procedure was essentially according to that of Preparation 38, except that 2 equivalents of potassium carbonate were used. ES / MS (m / z) [M+H] + 410.2

[0173] Preparation 40

[0174] tert-Butyl N-[(E)-3-fluoro-2-[[2-(3-methyl-2-oxo-imidazolidin-1-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0175]

[0176] A mixture of tert-butyl N-[(E)-2-[(2-chloropyrimidin-5-yl)oxymethyl]-3-fluoro-allyl]carbamate (114 mg, 0.36 mmol), methylimidazolidinone (115 mg, 1.151 mmol), cuprous iodide (40.1 mg, 0.211 mmol), trans-N,N'-dimethylcyclohexane-1,2-diamine (40 μL), cesium carbonate (351 mg, 1.077 mmol) and 1,4-dioxane (10 mL) was heated to 160° C. for 3 hours under microwave conditions under N2 atmosphere. The reaction mixture was filtered and concentrated under vacuum. The residue was subjected to preparative HPLC with the following conditions: LC column: SunFire C18 30×100 mm 5 μm; A: H 2 O (0.1% FA); B: ACN (0.1% FA), gradient 24-39% ACN in 11 min, stop at 17 min; column room temperature; flow rate 30 mL / min.; t (R) = 10.0 min (UV). The title product was obtained as a white solid. (137 mg, 27%). LCMS (ESI): m / s 382.2 [M+H] + .

[0177] Preparation 41

[0178] tert-Butyl N-[(E)-3-fluoro-2-[[2-(3-oxo-2,8-diazaspiro[4.5]dec-8-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0179]

[0180] A mixture of tert-butyl N-[(E)-2-[(2-chloropyrimidin-5-yl)oxymethyl]-3-fluoro-allyl]carbamate (65 mg, 0.20 mmol), 2,8-diazaspiro[4.5]decan-3-one (67 mg, 0.41 mmol), DIPEA (0.11 mL, 0.63 mmol) and 1,4-dioxane (5.0 mL) was stirred at 120° C. under microwave irradiation for 12 hours. The solvent was evaporated under reduced pressure to give the crude product, which was subjected to silica gel flash chromatography eluting with a gradient of 0-5% MeOH / DCM to give the title compound (61 mg, 68.46%) as a white solid. LCMS (ESI): m / s 436.3 [M+H].

[0181] The following compound was prepared essentially according to the procedure of Preparation 41, heating from 110 to 120°C for 3-12 hours.

[0182]

[0183]

[0184]

[0185] a See below for purification procedures.

[0186] Purification of Preparation 43

[0187] tert-Butyl N-[(E)-3-fluoro-2-[[2-[4-(2-oxopyrrolidin-1-yl)-1-piperidinyl]pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0188] The crude material was subjected to preparative HPLC with the following conditions: Column: SunFire C18 30×100 mm 5 μm; A: H 2 O (0.1% FA); B: ACN (0.1% FA), gradient: 33-48% ACN in 11 min, stop at 18 min; Column temperature: room temperature; Flow rate 30 mL / min.; t (R) = 8.7 min (UV) to give the title compound (16 mg, 21%) as a white solid.

[0189] Purification of Preparation 45

[0190] N-[(E)-2-[[2-[(3S)-3-(Cyclopropylcarbamoyl)-1-piperidinyl]pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0191] The crude material was subjected to preparative HPLC with the following conditions: Column: C18 30×150mm 5μm; A: H2O 10mM NH4HCO3; B: ACN, gradient: 35-40% ACN in 11min, stop at 17min; column temperature: room temperature; flow rate 35mL / min.; t (R) = 9.8 min (UV) to give the title compound (120 mg, 53%).

[0192] Purification of Preparation 47

[0193] N-[(E)-2-[[2-(3,3-Dimethyl-5-oxo-piperazin-1-yl)pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0194] The crude material was subjected to preparative HPLC with the following conditions: Column: SunFire C18 30×100 mm 5 μm; A: H 2 O (0.1% FA); B: ACN (0.1% FA), gradient: 29-44% ACN in 11 min, stop at 17 min; Column temperature: room temperature; Flow rate: 30 mL / min.; t (R) = 10.1 min (UV) to give the title compound (15 mg, 6%) as a white solid.

[0195] Preparation 51

[0196] N-[(E)-2-[[2-(4,4-dimethyl-2-oxo-imidazolidin-1-yl)pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0197]

[0198] Tert-butyl N-[(E)-2-[[2-[(2-amino-2-methyl-propyl)amino]pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamate (151 mg, 0.41 mmol) was dissolved in THF (10 mL) and 1,1'-carbonyldiimidazole (122 mg, 0.73 mmol) was added in one portion. The mixture was stirred at room temperature overnight and then heated to 60°C for 5 hours. The reaction mixture was concentrated under vacuum and the crude mixture was used without further purification.

[0199] Preparation 52

[0200] tert-Butyl N-[(E)-3-fluoro-2-[[2-[4-(1-methyl-2-oxo-pyrrolidin-3-yl)oxy-1-piperidinyl]pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0201]

[0202] A mixture of 1-methyl-3-(4-piperidinyloxy)pyrrolidin-2-one (85 mg, 0.386 mmol), tert-butyl N-[(E)-2-[(2-chloropyrimidin-5-yl)oxymethyl]-3-fluoro-allyl]carbamate (0.123 g, 0.386 mmol) and DIPEA (0.102 g, 0.772 mmol, 0.135 mL) was stirred in NMP (2.0 mL) at 100 °C for 16 hours. The reaction mixture was diluted with water (30 mL) and the mixture was extracted with EtOAc (3×15 mL). The combined organic extracts were washed with brine (3×30 mL), dried over anhydrous Na2SO4, filtered and the filtrate concentrated under vacuum. The residue was subjected to flash chromatography on silica gel eluted with 0-0.5% MeOH / DCM to give the title compound (85 mg, 41%) as a colorless gum. LCMS (m / z): 480.2 [M+H] + ,

[0203] 1 H NMR (400MHz, CDCl3) δ8.08 (s, 2H), 6.70 (d, J = 81.6Hz, 1H), 4.77 (br s,1H),4.39(d,J=3.6Hz,2H),4.38-4.25(m,2H),4.20-4.14(m,1H),4.13-4.05(m,1H),4.04-3.94(m,2H),3.45-3 .36(m,1H),3.35-3.20(m,3H),2.87(s,3H),2.42-2.28(m,1H),2.06-1.94(m,3H),1.62-1.50(m,2H),1.43(s,9H)

[0204] Preparation 53

[0205] 1-[5-[(E)-2-[(tert-Butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]piperidine-4-carboxylic acid

[0206]

[0207] Lithium hydroxide (300 mg, 12.5 mmol) was added to a stirred solution of 1-[5-[(E)-2-[(tert-butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]piperidine-4-carboxylic acid methyl ester (260 mg, 0.55 mmol) in a mixture of THF (8.0 mL) and water (4.0 mL). The resulting mixture was stirred at room temperature for 16 hours. Filter to remove solids and concentrate the filtrate to give the crude title product, which can be used directly without further purification. LCMS (ESI): m / s 411.3 [M+H].

[0208] Preparation 54

[0209] 1-[5-[(E)-2-[(tert-Butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]-4-methyl-piperidine-4-carboxylic acid

[0210]

[0211] Lithium hydroxide (19 mg, 0.79 mmol) was added to a stirred solution of 1-[5-[(E)-2-[(tert-butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]-4-methyl-piperidine-4-carboxylic acid ethyl ester (35 mg, 0.077 mmol) in a mixture of THF (4.0 mL) and water (2.0 mL). The resulting mixture was stirred at room temperature for 16 hours. The mixture was heated to 100 ° C for 2 hours under microwave irradiation. 10% HCl was added to adjust the pH value to about 3 and the mixture was evaporated under reduced pressure to dryness to give the crude title product as a white solid, which was used directly without further purification. LCMS (ESI): m / s 425.3 [M+H].

[0212] Preparation 55

[0213] N-[(E)-2-[[2-[4-(Dimethylcarbamoyl)-1-piperidinyl]pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamic acid tert-butyl ester

[0214]

[0215] Dimethylamine (0.10 mL, 0.20 mmol, 2 mol / L) in THF was added to a stirred solution of 1-[5-[(E)-2-[(tert-butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]piperidine-4-carboxylic acid (60 mg, 0.14 mmol) in DMF (4.0 mL), followed by HATU (0.10 g, 0.26 mmol) and DIPEA (0.05 mL, 0.3 mmol). The mixture was stirred at room temperature for 16 h. The solvent was removed under reduced pressure to give the crude product, which was purified by preparative HPLC using the following conditions: Column: SunFire C18 30×100 mm 5 μm; A: H2O (0.1% FA); B: ACN (0.1% FA), gradient: 31-46% ACN in 11 min, stop at 18 min; Column temperature: room temperature; Flow rate: 30 mL / min.; t (R) = 9.2 min (UV). The title product was isolated as a white solid (23 mg, 37%). LCMS (ESI): m / s 438.4 [M+H].

[0216] Preparation 56

[0217] tert-Butyl N-[(E)-3-fluoro-2-[[2-[4-[(3S)-3-hydroxypyrrolidine-1-carbonyl]-1-piperidinyl]pyrimidin-5-yl]oxymethyl]allyl]carbamate

[0218]

[0219] The procedure was essentially according to the method of Preparation 55 using the appropriate amine, pyrrolidin-3-ol.

[0220] Example 1

[0221] 8-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2-methyl-2,8-diazaspiro[4.5]decan-1-one dihydrochloride

[0222]

[0223] Tert-butyl N-[(E)-3-fluoro-2-[[2-(2-methyl-1-oxo-2,8-diazaspiro[4.5]dec-8-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate (0.16 g, 0.34 mmol) was dissolved in 3M HCl (5.0 mL) in MeOH. The resulting mixture was stirred at room temperature for 1 hr. The mixture was concentrated under vacuum to give a residue. The residue was dissolved in water (5 mL); the mixture was lyophilized to give the title compound (0.13 g, 86%) as a light yellow gum. ES / MS (m / z) 350.2 (M+H).

[0224] Example 2

[0225] 8-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2-methyl-2,8-diazaspiro[4.5]decan-1-one

[0226]

[0227] A saturated aqueous solution of potassium carbonate (100 mL) and 2-methyl-THF (200 mL) was added to 8-[5-[(E)-2-(aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2-methyl-2,8-diazaspiro[4.5]dec-1-one dihydrochloride (9.17 g, 21.2 mmol). The aqueous layer was separated from the organic layer. The aqueous layer was washed with 2-methyl-THF (200 mL) and the organic extract was added to the organic layer. The organic layer was dried over NaSO4; filtered; and the filtrate was concentrated under vacuum to give the title compound (85% by mass, 7.47 g, 18.2 mmol, 85.9%). ES / MS (m / z) 350.2 (M+H).

[0228] Example 3

[0229] 4-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]-2-pyridyl]piperazin-2-one

[0230]

[0231] HCl (3 mL, 4 M in 1,4-dioxane) was added to a solution of 4-[5-[(E)-2-[(tert-butoxycarbonylamino)methyl]-3-fluoro-allyloxy]-2-pyridinyl]-2-oxo-piperazine-1-carboxylic acid tert-butyl ester (180 mg, 0.364 mmol) in 1,4-dioxane (2 mL). The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure to give a crude product in the form of a yellow solid. The material was subjected to preparative HPLC using the following conditions: Column: Kromasil C18 250*50mm*10μm, 1-30% B and A: water / 0.05% NH4OH, B: ACN, flow rate: 25 mL / min to give the title compound (20.2 mg, 19%). LCMS (ESI): m / s 280.9 [M+H] + , 1 H NMR(400MHz,d4-MeOH 7.95(d,J=3.2Hz,1H),7.37(dd,J=9.2,3.2Hz,1H),6.97(d,J=84Hz,1H),6.83(d,J=8.0Hz,1H),4. 57(d,J=3.2Hz,2H),4.03(s,2H),3.71(t,J=4.0Hz,2H),3.62-3.55(m,2H),3.44(t,J=4.0Hz,2H).

[0232] Example 4

[0233] 3-[[1-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-4-piperidinyl]oxy]-1-methyl-pyrrolidin-2-one dihydrochloride

[0234]

[0235] A mixture of tert-butyl N-[(E)-3-fluoro-2-[[2-[4-(1-methyl-2-oxo-pyrrolidin-3-yl)oxy-1-piperidinyl]pyrimidin-5-yl]oxymethyl]allyl]carbamate (85 mg, 0.160 mmol) in MeOH (4.0 mL, 4 M) in HCl was stirred at room temperature for 1 hr and then the reaction mixture was concentrated under vacuum. The residue was subjected to preparative HPLC using the following conditions: column: YMC-Actus Triart C18 150*30mm*5μm, 0-30% B and A: water / 0.05% HCl, B: ACN, flow rate: 25 mL / min to give the title compound (45 mg, 61%) as a yellow gum. LCMS (m / z): 380.3[M+H] + , 1H NMR (400MHz, d4-MeOH) δ6.86 (s, 2H), 5.68 (d, J = 80.4Hz, 1H), 3.13 (d, J = 2.8Hz, 2H), 2.73 (t, J = 7.8Hz, 1H), 2.55-2.41(m,3H),2.29-2.14(m,4H),1.88-1.76(m,2H),1.29(s,3H),0.95-0.81(m,1H),0.52-0.15(m,5H)

[0236] Example 5

[0237] 1-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-3-methyl-imidazolidin-2-one hydrochloride

[0238]

[0239] Tert-butyl N-[(E)-3-fluoro-2-[[2-(3-methyl-2-oxo-imidazolidin-1-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate (35.1 mg, 0.0920 mmol) was dissolved in HCl (10 mL, 1 M in EtOAc) and the resulting solution was stirred overnight. The suspension was concentrated under vacuum, dissolved in water and the solution was lyophilized to give the title compound as an off-white solid. (24 mg, 63%). LCMS (ESI): m / s 282.2 [M+H] + .

[0240] The following compounds were prepared essentially analogously to the procedure of Example 5 using the appropriate BOC protected allylmethylamine.

[0241]

[0242]

[0243] a Premix with 1 mL 0.5 mol / L HCl in MeOH

[0244] b The mixture was heated to 80 °C for 1 hr.

[0245] Example 8

[0246] 1-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]piperidine-4-carboxamide hydrochloride

[0247]

[0248] A mixture of tert-butyl N-[(E)-2-[[2-(4-carbamoyl-1-piperidinyl)pyrimidin-5-yl]oxymethyl]-3-fluoro-allyl]carbamate (80.0 mg, 0.195 mmol) in HCl (8 mL, 4 M in MeOH) was stirred at 10 °C for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC eluting with 0.05% HCl to give the title compound (43.0 mg, 62%) as a yellow solid. LCMS (ESI): m / s 309.9 [M+H], 1 H NMR(400MHz,d4-MeOH)δ8.49(s,2H),7.28(d,J=80.4Hz,1H),4.74(d,J=2.8Hz,2H),4.60-4.47( m,2H),3.86(s,2H),3.41-3.33(m,2H),2.78-2.60(m,1H),2.08-1.95(m,2H),1.87-1.72(m,2H).

[0249] Example 9

[0250] 8-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2,8-diazaspiro[4.5]decan-3-one dihydrochloride

[0251]

[0252] Tert-butyl N-[(E)-3-fluoro-2-[[2-(3-oxo-2,8-diazaspiro[4.5]dec-8-yl)pyrimidin-5-yl]oxymethyl]allyl]carbamate (61 mg, 0.1401 mmol, 100 mass%) and HCl (4.0 mL, 0.39 mol / L) in MeOH were stirred for 2 hours at 60 ° C under microwave irradiation. The solvent was evaporated under reduced pressure to give the title compound (60 mg, 99%) as a yellow solid. LCMS (ESI): m / s 336.3 [M+H].

[0253] The following compounds were prepared substantially similarly to the procedure of Example 9, heating the reaction to between about 60 and 80°C for 5 minutes to 2 hours.

[0254]

[0255]

[0256]

[0257] a The crude material was dissolved in water and lyophilized to a solid.

[0258] b See below for purification methods.

[0259] Purification of Example 12

[0260] 7-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2,7-diazaspiro[4.5]decan-3-one

[0261] The crude material was subjected to preparative HPLC with the following conditions: Column: C18 30×150mm 5μm; A: H2O 10mM NH4HCO3; B: ACN, gradient: 0-5% ACN in 0-2min, 10-20% ACN in 2-12min, stop at 18min; column temperature: room temperature; flow rate 35mL / min.; t (R) = 10.7 min. (UV) to give the title product (8.5 mg, 36%) as a white solid.

[0262] Purification of Example 20

[0263] 9-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-2-methyl-2,9-diazaspiro[5.5]undecan-1-one

[0264] The crude material was dissolved in water and made basic with NaHCO3. The resulting material was subjected to preparative HPLC with the following conditions: Column: C18 30×100mm 5μm; A: H2O (10mM NH4HCO3); B: ACN, gradient: 10-25% B in 9min, stop at 14min; column temperature: room temperature; flow rate 35mL / min.; t (R) = 8.7 min. (UV) to give the title compound (21 mg, 52%) as a light yellow oil.

[0265] Example 22

[0266] 1-[5-[(E)-2-(Aminomethyl)-3-fluoro-allyloxy]pyrimidin-2-yl]-N,N,4-trimethyl-piperidine-4-carboxamide dicarboxylic acid

[0267]

[0268] Dimethylamine (0.20 mL, 0.40 mmol, 2 mol / L) in THF was added to a stirred solution of 1-[5-[(E)-2-[(tert-butoxycarbonylamino)methyl]-3-fluoro-allyloxy]pyrimidin-2-yl]-4-methyl-piperidine-4-carboxylic acid (32 mg, 0.075 mmol) in DMF (2.0 mL), followed by HATU (60 mg, 0.16 mmol) and DIPEA (0.065 mL, 0.37 mmol). The mixture was stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure to give a crude product. The crude material was subjected to preparative HPLC using the following conditions: LC column: SunFire C18 30×100 mm 5 μm; A: H2O (0.1% FA); B: ACN (0.1% FA), gradient: 5-5% ACN in 0-3 min, 5-10% ACN in 3-13 min, stop at 19 min; column temperature: room temperature; flow rate 30 mL / min.; t (R) = 8.5 min. (UV) to give the title compound (15 mg, 43%) as a white solid. LCMS (ESI): m / s 352.2 [M+H].

[0269] Biological analysis

[0270] SSAO / VAP-1 activity in vitro

[0271] MAO-Glo from Promega (V1402) was used TM The assay kit measures the amine oxidase activity of recombinant hSSAO, hMAOa and hMAOb isoforms. The test compound (using DMSO as a vehicle, SSAO is 0.5% v / v) and enzyme are incubated at room temperature for 10 minutes, followed by the addition of the luminescent substrate. The substrate concentration for human recombinant SSAO is 10 μM. The assay is performed in a well plate in pH 7.4 buffer (50 mM HEPES, 120 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1.4 mM MgCl2, 0.001% Tween-20). Substrate oxidation is performed for 2 hours, followed by the addition of the detection reagent according to the manufacturer's protocol. IC of the test compound 50 Values ​​were calculated by fitting dose response curves using a 4-parameter nonlinear regression program. Compounds of the Examples exhibited hSSAO inhibition IC values ​​less than 100 nM 50 IC of the compound of Example 1 50 The value was 19.36±5.68, n=5 (data are presented as mean±standard deviation).

[0272] The compounds of the tested examples showed IC values ​​exceeding 15 μM and 180 μM, respectively. 50hMAOa and hMAOb, indicating that the compounds of the examples have selectivity for hSSAO compared to hMAOa or hMAOb.

[0273] SSAO target bonding

[0274] Use MAO-Glo from Promega (V1402) TM The assay kit measures SSAO activity in rat plasma and liver tissue. The residual SSAO activity in rats after compound treatment is estimated by measuring the total amine oxidase activity in plasma or liver lysates that are insensitive to the presence of MAO inhibitors clogyline and pargyline. The compound of Example 1 was administered to rats at a dose of 15, 3, 0.6, 0.12, 0.025, 0.005 mg / kg. The same volume (2 ml / kg) of dosing vehicle (hydroxyethyl cellulose 1% w / v, 0.25% Tween 80) was administered to the control group. The plasma and liver after 2 or 24 hours of compound treatment were collected and stored at -78°C until analysis. Tissue lysates were prepared by homogenization in lysis buffer (20 mM HEPES, pH 7.4; 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100 and 1× Roche Complete protease inhibitor tablet). Tissue pellets were removed by centrifugation at 12,000 rpm for 30 min at 4°C. 40 μl of plasma or liver lysate was incubated with clorgyline (10 μM) and pargyline (10 μM) at room temperature for 20 minutes, followed by the addition of luminescent substrate (50 μM) for 60 min. The products produced were quantified according to the manufacturer's procedure. The active fraction, which is insensitive to the presence of MAO inhibitors, was used as a surrogate for residual SSAO activity. The compounds of Example 1 administered at various doses were evaluated in a protocol essentially as described above. The results are listed in the table below.

[0275] SSAO target bonding of Example 1

[0276]

[0277] Data are presented as mean ± SEM, n = 6.

[0278] The results showed that the compound of Example 1 dose-dependently inhibited SSAO activity in rat plasma and liver tissue.

Claims

1. A method for preparing a compound of formula 1 or a pharmaceutically acceptable salt thereof: in: R1 is selected from the group consisting of: R2 is selected from the group consisting of: H, -C(O)NH2, -C(O)NH(CH3) and -C(O)N(CH3)2; R3 is H; R4 is H; R5 is selected from the group consisting of: H, -C 1-4 Alkyl, -C 3-4 Cycloalkyl and -CH2-C 3-4 cycloalkyl; and n is 1 or 2, The method comprises the following steps: a) the compound of formula A: Where Pg is a protecting group, Reaction with a substituted cyclic amine followed by a deprotection step to form a compound of formula B: Wherein R1 is as defined in Formula 1; b) reacting the compound of formula B with an appropriate amine protected 2-bromomethyl-3-fluoro-propyl-2-en-amine followed by a deprotection step to form the compound of formula 1; and c) optionally converting the compound of formula 1 into a pharmaceutically acceptable salt.

2. The method according to claim 1, wherein the compound of formula 1 or a pharmaceutically acceptable salt thereof is a compound of formula 2: or a pharmaceutically acceptable salt thereof.

3. The method according to claim 2, wherein R1 is 4. The method of claim 3, wherein R5 is selected from the group consisting of: H, -CH3, as well as 5. The method according to claim 1, wherein: R1 is and R2 is -C(O)NH2.

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

7. The method according to claim 1, wherein the compound of formula 1 is: or a pharmaceutically acceptable salt thereof.

8. The method of claim 7, wherein the compound is a mono- or di-hydrochloride salt.

9. The method of claim 7, wherein the compound is:

10. The process according to claim 7, wherein step a) is carried out in the presence of a base during the reaction of the compound of formula A with the substituted cyclic amine.

11. The method according to claim 10, wherein the base is N,N-diisopropylethylamine or potassium carbonate.

12. The method according to claim 1, wherein the protecting group of the compound of formula A in step a) is benzyl.

13. The method according to claim 12, wherein the deprotection step of step a) uses palladium catalyzed hydrogenation.

14. The process of claim 1, wherein the suitable amine protected 2-bromomethyl-3-fluoro-propyl-2-en-amine of step b) is tert-butyl N-[(E)-2-(bromomethyl)-3-fluoro-allyl]carbamate.

15. The method according to claim 14, wherein the deprotection step of step b) uses an acid.

16. The method of claim 15, wherein the acid is HCl.

17. The method of claim 1, wherein the compound of formula 1 is selected from the group consisting of: as well as or a pharmaceutically acceptable salt thereof.

18. The method of claim 1, wherein the compound of formula 1 is selected from the group consisting of: as well as or a pharmaceutically acceptable salt thereof.

19. The method of claim 1, wherein the compound of formula 1 is selected from the group consisting of: as well as or a pharmaceutically acceptable salt thereof.

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