Pyrimidine-5-carboxamide compounds

Abstract

The present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof, and a method for treating type 2 diabetes using the compound.

Description

[0001] The present invention relates to pyrimidine-5-carboxamide compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions and therapeutic uses of the compounds.

[0002] Nicotinamide N-methyltransferase (NNMT) is an enzyme that catalyzes the transfer of a methyl group from the universal methyl donor S-(5'-adenosyl)-L-methionine (SAM) to nicotinamide (NAM) to form 1-methylnicotinamide (1-MeNAM). NNMT is a potential therapeutic target for the treatment of type 2 diabetes mellitus (T2DM).

[0003] WO2018 / 183668 discloses certain quinoline derivatives which are described as NNMT inhibitors. Bisubstrate small molecule inhibitors of NNMT are also described (Martin, NI et al., "Bisubstrate Inhibitors of Nicotinamide N-Methyltransferase (NNMT) with Enhanced Activity", Journal of Medicinal Chemistry , 2019, 62, 6597-6614). Nicotinamide analogs have also been reported as NNMT inhibitors (see Rajagopal, S.; Ruf, S.; et al., "Novel nicotinamide analog as inhibitor of nicotinamide N-methyltransferase", Bioorganic and Medicinal Chemistry Letters , 2018, 28, 922-925, and Dhakshinamoorthy, S.; Kannt, A.; et al., "A small molecule inhibitor of Nicotinamide N-methyltransferase for the treatment of metabolic disorders"; Scientific Reports , 2018, 8, 3660). To date, no NNMT inhibitors have been approved for therapeutic use.

[0004] There is a need for alternative NNMT inhibitor compounds and therapeutic methods using the same. In particular, there is a need for NNMT inhibitor compounds that are potent and orally bioavailable.

[0005] To this end, in one embodiment, the present invention provides novel compounds that are potent NNMT inhibitors for the treatment of T2DM.

[0006] In one embodiment, the present invention provides a compound of formula I,

[0007]

[0008] in,

[0009] R is selected from cyclopropyl and geminal cyclopropyl;

[0010] or a pharmaceutically acceptable salt thereof. Formula I includes all single enantiomers and diastereomers thereof, as well as mixtures of enantiomers and racemates.

[0011] In one embodiment, the invention provides a compound of formula Ia,

[0012]

[0013] or a pharmaceutically acceptable salt thereof.

[0014] In one embodiment, the present invention provides a compound of formula Ib,

[0015]

[0016] or a pharmaceutically acceptable salt thereof. Formula lb includes all single enantiomers thereof, as well as mixtures of enantiomers and racemates.

[0017] In another embodiment, the present invention provides a pharmaceutical composition comprising a compound according to Formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient. In a preferred embodiment, the pharmaceutically acceptable composition is formulated for oral administration.

[0018] In another embodiment, the present invention provides a method for treating T2DM in a mammal, the method comprising administering a pharmaceutically acceptable composition to a mammal in need of treatment, the composition comprising an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient. In one embodiment, the pharmaceutically acceptable composition is formulated for oral administration. Preferably, the mammal is a human being.

[0019] In another embodiment, the present invention also provides a method for treating T2DM in a mammal, comprising administering to a mammal in need of treatment an effective amount of a compound according to Formula I or a pharmaceutically acceptable salt thereof.

[0020] In another embodiment, the present invention provides a method of treating metabolic syndrome in a mammal, comprising administering to a mammal in need of treatment an effective amount of a compound according to formula I or a pharmaceutically acceptable salt thereof.

[0021] In another embodiment, the present invention also provides a method of treating chronic kidney disease (CKD) in a mammal, comprising administering to a mammal in need of treatment an effective amount of a compound according to Formula I or a pharmaceutically acceptable salt thereof.

[0022] In one embodiment, the invention provides a compound according to formula I or a pharmaceutically acceptable salt thereof for use in therapy.

[0023] In another embodiment, the present invention also provides a compound according to formula I or a pharmaceutically acceptable salt thereof for use in treating T2DM in a mammal.

[0024] In another embodiment, the present invention also provides a compound according to formula I or a pharmaceutically acceptable salt thereof for use in treating metabolic syndrome in a mammal.

[0025] In another embodiment, the present invention also provides a compound according to formula I or a pharmaceutically acceptable salt thereof for use in treating CKD in a mammal.

[0026] In one embodiment, the invention provides the use of a compound according to formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating T2DM in a mammal.

[0027] In one embodiment, the invention provides the use of a compound according to formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating metabolic syndrome in a mammal.

[0028] In one embodiment, the present invention also provides the use of a compound according to formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating CKD in a mammal.

[0029] In a preferred embodiment, the compound according to formula I is administered orally. In another preferred embodiment, the mammal to be treated is a human.

[0030] As used herein, the term "geminal" has the ordinary meaning recognized by skilled chemists. That is, the carbon of the cyclopropyl group is directly linked to the carbon of the substitution point to form a linked cyclopropyl group.

[0031] As used herein, the terms "treating," "treat," or "treatment" refer to reducing, decreasing, or reversing the progression or severity of an existing symptom, disorder, or condition. Such disorders may include T2DM and metabolic syndrome.

[0032] As used herein, the term "patient" refers to a mammal. Preferably, the patient is a human.

[0033] As used herein, the term "effective amount" refers to the amount or dosage of a compound of the present invention or a pharmaceutically acceptable salt thereof, which provides the desired effect in a patient under diagnosis or treatment after single or multiple doses of administration to the patient.

[0034] The effective amount can be determined by one 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 need to be considered, including, but not limited to: the species of the patient; its size, age, and general health; the specific disease or condition involved; the extent or involvement or severity of the disease or condition; the response of the individual patient; the specific compound being administered; the mode of administration; the bioavailability characteristics of the administered formulation; the dosing regimen selected; the use of concomitant medications; and other relevant circumstances.

[0035] The compounds of the invention are formulated as pharmaceutical compositions, which are administered by any route that makes the compounds bioavailable. Preferably, such compositions are for oral administration. Such pharmaceutical compositions and methods for their preparation are well known in the art (see, e.g., Remington, JP, "Remington: The Science and Practice of Pharmacy", LV Allen, Editor, 22nd Edition, Pharmaceutical Press, 2012).

[0036] The compound of formula I is easily converted and separable into a pharmaceutically acceptable salt. The formation of salt can occur when a pharmaceutically acceptable acid is added to form an acid addition salt or when a pharmaceutically acceptable base is added to form a base addition salt. Salt can also be formed simultaneously when nitrogen or oxygen is deprotected (i.e., removing the protecting group). Examples, reactions and conditions for forming salts can be found in the following: Gould, PL, "Salt selection for basic drugs," International Journal of Pharmaceutics, 33: 201-217 (1986); Bastin, RJ, et al., "Salt Selection and Optimization Procedures for Pharmaceutical New Chemical Entities," Organic Process Research and Development, 4: 427-435 (2000); and Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, 66: 1-19, (1977).

[0037] Increased expression and activity of NNMT have been implicated in various disease pathologies, including metabolic syndrome, cardiovascular disease, neurodegeneration, and cancer. Of particular interest is the demonstrated correlation between adipose NNMT activity and insulin resistance. This mechanism appears to be reversible, as adipose NNMT activity is reduced following interventions that improve insulin resistance (i.e., bariatric surgery and lifestyle interventions), see Kannt, A.; Pfenninger, A.; Teichert, L; et al., “Association of nicotinamide-N-methyltransferase mRNA expression in human adipose tissue and the plasma concentration of its product, 1-methylnicotinamide, with insulin resistance,” Diabetologia, 2015, 58, 799-808). Gene knockout of the NNMT gene in mice showed a protective effect against diet-induced obesity, and the animals showed enhanced insulin sensitivity, validating its potential utility as a therapeutic target for metabolic disorders and type 2 diabetes (Kraus D, Yang Q, Kong D, et al., "Nicotinamide N-methyltransferase knockdown protects against diet-induced obesity", Nature, 2014, 508, 258-262).

[0038] Another utility of NNMT inhibitors as therapeutic targets of interest in metabolic disorders is their role in regulating cellular methylation capacity via SAM usage. NAM methylation via NNMT utilizes a methyl group from the universal methyl donor SAM, converting SAM to its demethylated form, S-adenosyl-L-homocysteine ​​(SAH). SAH is the substrate for the enzyme adenosylhomocysteinase, which converts SAH to homocysteine. Elevated homocysteine ​​levels (hyperhomocysteinemia) are observed in patients with chronic and end-stage renal disease and are believed to promote cellular homeostasis and redox imbalance, leading to increased oxidative stress (Ostrakhovitch, EA; Tabibzadeh S. “Homocysteine ​​in Chronic Kidney Disease,” Advances in Clinical Chemistry , 2015, 72, 77-106). Improving hyperhomocysteinemia in these patients via NNMT inhibition may serve as a valuable therapeutic mechanism for the treatment of chronic kidney disease.

[0039] The compounds of the present invention or their salts may be prepared by a variety of methods, some of which are illustrated in the following preparations and examples. The products of each step in the following preparations and examples may be recovered by conventional methods, including extraction, evaporation, precipitation, chromatography, filtration, grinding and crystallization. Single isomers, enantiomers and diastereomers may be separated or resolved at any convenient point in the synthesis by methods such as selective crystallization techniques or chiral chromatography (see, for example, J. Jacques, et al., " Enantiomers, Racemates, and Resolutions ", John Wiley and Sons, Inc., 1981, and EL Eliel and SH Wilen, "Stereochemistry of Organic Compounds", Wiley-Interscience, 1994).

[0040] Certain abbreviations are defined as follows: "EtOAc" means ethyl acetate; "HPLC" means high performance liquid chromatography; "NMP" means N-methyl-2-pyrrolidone; "RT" means room temperature; and "SFC" means supercritical fluid chromatography.

[0041] Preparation 1

[0042] 2-Chloro-4-[[1-(hydroxymethyl)cyclopropyl]amino]pyrimidine-5-carboxamide

[0043]

[0044] To a solution of 2,4-dichloropyrimidine-5-carboxamide (10 g, 52 mmol) in NMP (50 mL) was added N,N-diisopropylethylamine (27 mL, 155 mmol) at room temperature followed by (1-aminocyclopropyl)methanol hydrochloride (6.95 g, 56.2 mmol) in portions, a slight exotherm was observed during the addition (reaching 32 °C in 10 min). The suspension was stirred at room temperature under nitrogen. After 1 h 15 min, the reaction mixture was poured into a mixture of water and ice (500 mL) and stirred for one hour. The solid was filtered and washed 3 times with water and then dried in a vacuum oven at 45 °C overnight to give the title compound (11.05 g, 85%) as a beige solid. ES / MS m / z 243 (M+H).

[0045] Example 1

[0046] 2-[(4-Fluoro-4-methyl-pentyl)amino]-4-[[1-(hydroxymethyl)cyclopropyl]amino]pyrimidine-5-carboxamide

[0047]

[0048] To a solution of 2-chloro-4-[[1-(hydroxymethyl)cyclopropyl]amino]pyrimidine-5-carboxamide (88.3 g, 346 mmol) in NMP (450 mL) was added potassium carbonate (143 g, 1034 mmol) at room temperature, followed by 4-fluoro-4-methyl-pentane-1-amine acetate (68.5 g, 382 mmol). The mixture was heated at 80 °C under nitrogen. After 2 hours, the reaction mixture was cooled to room temperature and poured into an ice / water mixture (2000 mL). The mixture was stirred overnight. The solid was filtered out, washed with water, and dried overnight in a vacuum oven at 45 °C to give a beige solid. The solid was mixed with ethanol (850 mL) and heptane (850 mL), heated in a 95 °C bath until a solution was obtained, and then the mixture was slowly cooled to room temperature overnight. The mixture was cooled in an ice-water bath for 30 minutes, the solid was filtered, and washed with heptane. The solid was dried in a vacuum oven. To remove residual ethanol, EtOAc (850 mL) was added and the mixture was heated in a 75 °C bath for 2 hours. The mixture was allowed to slowly cool to room temperature overnight. Concentrate in vacuo to reduce the mixture to about one-third of its original volume. Heptane (250 mL) was added to the mixture and stirred in an ice-water bath for 30 minutes. The solid was filtered, washed with heptane, and dried in a vacuum oven at 45 °C for 2 days to give the title compound (78.4 g, 68%) as a white solid. ES / MS m / z 326 (M+H).

[0049] Example 2

[0050] 4-[(1-Cyclopropyl-2-hydroxy-ethyl)amino]-2-[(4-fluoro-4-methyl-pentyl)amino]pyrimidine-5-carboxamide

[0051]

[0052] To a solution of 2,4-dichloropyrimidine-5-carboxamide (300 mg, 1.56 mmol) in NMP (1.5 mL, 16 mmol) was added N,N-diisopropylethylamine (0.4 mL, 2 mmol) at room temperature, followed by racemic 2-amino-2-cyclopropyl-ethanol (158 mg, 1.56 mmol). The suspension was stirred at room temperature under nitrogen. After 1 hour, 4-fluoro-4-methylpentane-1-amine acetate (186 mg, 1.56 mmol) was added, and the mixture was heated at 80 ° C under nitrogen for 16 hours. The mixture was cooled to room temperature and water and EtOAc were added. The organic phase was separated and washed with water and saturated NaCl aqueous solution. The organics were dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase HPLC using 25-55% acetonitrile / (20 mM NH4HCO3 aqueous solution) to give the title compound 163 mg (40%) as a white solid. ES / MS m / z 340 (M+H).

[0053] Example 2a

[0054] 4-{[(1S)-1-cyclopropyl-2-hydroxyethyl]amino}-2-[(4-fluoro-4-methylpentyl)amino]pyrimidine-5-carboxamide

[0055]

[0056] Racemic 4-[(1-cyclopropyl-2-hydroxy-ethyl)amino]-2-[(4-fluoro-4-methylpentyl)amino]pyrimidine-5-carboxamide (148 mg, 0.37 mmol) was purified by chiral SFC (column: Chiralpak® IG 25×2 cm, 5 μm particle size; column temperature: 40°C; flow rate: 65 mL / min; eluent: 30% (0.2% dimethylethylamine in isopropanol) / CO2; 10 mg injection every 8 minutes) and the first-eluting isomer provided 52 mg of the title compound as a white solid. ES / MS m / z 340 (M+H).

[0057] Example 2b

[0058] 4-{[(1R)-1-cyclopropyl-2-hydroxyethyl]amino}-2-[(4-fluoro-4-methylpentyl)amino]pyrimidine-5-carboxamide

[0059]

[0060] Chiral SFC elution of the second isomer from Example 2a provided 49 mg of the title compound as a white solid. ES / MS m / z 340 (M+H).

[0061] Biological assays

[0062] Biochemical NNMT inhibition

[0063] The compounds of the present invention inhibit the biochemical activity of human and mouse NNMT enzymes and are quantified in a mass spectrometry-based assay. The test compounds are diluted and acoustically transferred to the assay plate to achieve a final DMSO concentration of 2% in the assay. The assay buffer contains 50 mM Tris-HCl (Invitrogen 15567-027) at pH 7.5 and 0.05 mg / mL fatty acid-free BSA (Sigma A6003). The human NNMT assay is assembled using the Hamilton Nimbus to deliver 10 μM SAM (S-(5'-adenosyl)-L-methionine) (US radiolabeled 0768), 15 μM NAM (nicotinamide) (Sigma 72340) and 10 nM full-length, n-terminal his-tagged human NNMT to the assay plate containing the diluted test compounds. The corresponding mouse NNMT assay includes 20 μM SAM, 20 μM NAM and 10 nM mouse NNMT. Max control wells, reflecting full enzyme activity, contained DMSO without test compound. Min control wells, reflecting no enzyme activity, were identical to the Max control wells but without NNMT. After two hours of incubation at room temperature, the wells were treated with 158.5 nM 13 C-SAH (S-adenosyl-L-homocysteine ​​(adenosine- 13 C 10 The reaction was stopped with 1 aliquot of stop solution containing 50 nM d7-methylnicotinamide (BDGBiosynthesis 140138-25) as an internal standard. MeNAM (1-methylnicotinamide) and SAH (S-adenosylhomocysteine) levels were measured by RapidFire mass spectrometry and converted to area ratios by dividing the product level by the corresponding internal standard level. The product area ratios were converted to percent inhibition using the following equation: % inhibition = 100–[(test compound-minimum median) / (maximum median-minimum median) × 100]. IC 50 By using Next Generation ResultsRel-IC 50 The percent inhibition at each inhibitor concentration was determined by fitting to the following equation: Data were analyzed using the 4-parameter nonlinear logistic equation y = (A+((BA) / (1 + ((C / x)^D)))) where y = % specific inhibition, A = bottom of the curve, B = top of the curve, and C = relative IC 50= the concentration that causes 50% inhibition based on the data range from top to bottom, D = Hill Slope = the slope of the curve.

[0064] In this assay, Example 1 showed a relative IC of 74 nM. 50 Inhibits human NNMT with a relative IC of 21 nM 50 Inhibits mouse NNMT. Example 2a has a relative IC of 385 nM 50 Inhibits human NMMT with a relative IC of 57 nM compared to Example 2b 50 Inhibits human NNMT.

[0065] In vivo PD assay

[0066] C57Bl / 6 male mice were maintained at 22°C on a 12-hour light / dark cycle with ad libitum access to food (Teklad 2014) and water. Animals were orally administered three concentrations (1, 3, and 10 mg / kg) of the test compound prior to subcutaneous injection of 25 mg / kg of deuterated nicotinamide (Cambridge Isotope Laboratories DLM-6883). Plasma was collected at 0, 60, 120, and 240 minutes after injection of deuterated nicotinamide (d4-NAM), and labeled d4-1-methylnicotinamide was quantified by LC / MS according to the following method. The inhibition curve of the test compound was calculated as the area under the curve (AUC) of the plasma d4-1-MeNAM shift measured over 240 minutes, and statistical tests were performed by one-way ANOVA (GraphPad Prism 7.03). The resulting AUC calculations demonstrated a dose-dependent decrease in plasma d4-1-MeNAM in mouse plasma following administration of Example 1 compared to vehicle controls (Table 1).

[0067] Table 1

[0068] .

[0069] LC / MS Method for Quantification of Deuterated 1-Methylnicotinamide as a Product of the NNMT Reaction :

[0070] Five microliters of mouse plasma were mixed with forty-five microliters of an internal standard solution containing 100 ng / mL of d7-1-MeNAM in water and 0.1% formic acid. Next, two hundred microliters of 1% ammonium hydroxide in acetonitrile were added to each sample and centrifuged at 2700 × g (4 °C) for 10 minutes. The upper layer was removed for LC / MS analysis via an isocratic HILIC HPLC method (using a Shimadzu Series 30 LC system) consisting of 72% acetonitrile and 28% of 20 mM ammonium acetate and 1% ammonium hydroxide in 95% water / 5% acetonitrile (v / v). The column was a Waters xBridge Amide, 3.5 µm, 2.1 × 50 mm (part #186004859). The mobile phase flow rate was 0.7 mL / min at ambient room temperature. Six microliters of sample were injected for mass spectrometry analysis (Sciex 6500 triplequad) using a 3.6 minute injection cycle. 1-MeNAM was detected in positive SRM mode using the following transitions: 137.0 / 94.0 for unlabeled 1-MeNAM, 144.1 / 101.0 for d7-1-MeNAM, and 141.1 / 98.1 for d4-1-MeNAM. Analyte peak area determination and quantification were achieved using Sciex MultiQuant version 2.1 software.

Claims

1. A compound of the formula: in, R is selected from cyclopropyl and geminal cyclopropyl; or a pharmaceutically acceptable salt thereof.

2. The compound as claimed in claim 1, wherein the compound is or a pharmaceutically acceptable salt thereof.

3. The compound as claimed in claim 1, wherein the compound is or a pharmaceutically acceptable salt thereof.

4. The compound as claimed in claim 1, wherein the compound is or a pharmaceutically acceptable salt thereof.

5. A pharmaceutical composition comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for inhibiting nicotinamide N-methyltransferase.

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