Pharmaceutical product comprising a muscarinic receptor antagonist and a beta2-adrenoceptor agonist

[0164] Preparation of muscarinic antagonist

[0165] The muscarinic antagonist of the present invention can be prepared as follows. Alternative salts of those described in this application can be prepared by conventional chemical action using methods similar to those described.

[0166] General experimental details for dry preparation of muscarinic antagonists

[0167] Unless otherwise specified, the following general conditions are used in the preparation of muscarinic antagonists.

[0168] Unless otherwise specified, all reactions were performed under a nitrogen atmosphere.

[0169] In the embodiment, the NMR spectrum is measured on a Varian Unity Inova spectrometer with a proton frequency of 300 or 400 or 500MHz, or on a Bruker DRX spectrometer with a proton frequency of 400 or 500MHz, or on a BrukerAvance spectrometer with a proton frequency of 600MHz The frequency is measured, or on a Bruker Avance DPX 300 spectrometer at a proton frequency of 300MHz. The MS spectrum was measured on an Agilent 1100MSD G1946D spectrometer or Hewlett Packard HP1100MSD G1946A spectrometer or Waters Micromass ZQ2000 spectrometer. The name is generated using Autonom 2000 (version 4.01.305) software provided by MDL.

[0170] Use PANalytical CubiX PRO instrument or PANalytical X-Pert instrument to collect XRPD data.

[0171] X-ray powder diffraction-XR PD-PANalytical CubiX PRO

[0172] The data was collected with a PANalytical CubiX PRO instrument, which under the θ-θ configuration, the scanning range was 2° to 40° 2θ, and the exposure time was 100 seconds/0.02° increments. X-rays are generated by a long thin focus tube made of copper, which operates at 45kV and 40mA. The wavelength of copper X-ray is The data was collected on a zero background container, and ~2 mg of compound was placed on the container. The container is made of single crystal silicon, which is cut along a non-diffractive plane and then polished on an optical plane refiner. The incidence of X-rays on this plane is offset by Bragg extinction.

[0173] X-ray powder diffraction-PANalytical X-Pert

[0174] The data was collected using a PANalytical X-Pert instrument, which was in the 2θ-θ configuration, with a scanning range of 2° to 40° 2θ, 100 seconds exposure time/0.02° increments. X-rays are generated by a long thin focus tube made of copper, which operates at 45kV and 40mA. The wavelength of copper X-ray is The data was collected on a zero background container, and ~2 mg of compound was placed on the container. The container is made of single crystal silicon, which is cut along a non-diffraction plane and then polished on an optical plane refiner. The incidence of X-rays on this plane is offset by Bragg extinction.

[0175] Differential scanning calorimetry (DSC) thermograms are measured using TA Instruments Q 1000 DSC Differential Scanning Calorimeter with aluminum pan and perforated lid. The sample weight varies between 0.5 and 5 mg. The operation is carried out under the following conditions: the nitrogen flow rate is 50 mL/min and the temperature under study is between 25 and 300°C at a constant rate of 10°C per minute.

[0176] Thermogravimetric analysis (TGA) thermograms are measured using TA Instruments Q500TGAThermogravimetric Analyser with platinum discs. The sample weight varies from 1 to 5 mg. The operation was performed under the following conditions: the nitrogen flow rate was 60 mL/min and the temperature under study was increased from 25°C to 200-300°C at a constant rate of 10°C per minute.

[0177] The gravimetric vapor absorption (GVS) distribution is measured using Surface Measurements Systems Dynamic Vapour Sorption DVS-1 or DVS Advantage GVS instruments. Put about 1-5 mg of solid sample in a glass tube or wire screen and in a two-cycle step method (40 to 90 or 0 to 90 or 0% relative humidity (RH) in a 10% RH step) Record the sample weight in.

[0178] Abbreviations used in the experimental section:

[0179] Aq = water or water solution

[0180] DCE = 1,2-Dichloroethane

[0181] DCM=dichloromethane

[0182] DMF=Dimethylformamide

[0183] DMSO = dimethyl sulfoxide

[0184] EtOAc = ethyl acetate

[0185] EtOH=ethanol

[0186] DSC = Differential Scanning Calorimeter

[0187] GVS=Gravimetric analysis steam absorption

[0188] TGA = thermogravimetric analysis

[0189] XRPD = X-ray powder diffraction

[0190] HATU=O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyl Hexafluorophosphate

[0191] MeCN = Acetonitrile

[0192] MeOH = methanol

[0193] RT = room temperature

[0194] Rt = retention time

[0195] THF=tetrahydrofuran

[0196] Satd = saturated

[0197] Based on the structure depicted in the examples and the stereochemistry specified according to the Cahn-Ingold-Prelog system, the muscarinic antagonists described in this application and the intermediates used in their preparation have been developed by MDL Information Systems IUPAC name generated by Beilstein Autonom2000naming package provided by Inc.

[0198] Specific implementation plan

[0199] Example 1: (R)-3-(1-Phenyl-cycloheptanecarbonyloxy)-1-(pyrazin-2-ylcarbamoylmethyl)-1-nitrogen -Bicyclo[2.2.2]octane bromide

[0200] a) 1-Phenyl-cycloheptanol

[0201]

[0202] To a solution of magnesium (1.2 g) in anhydrous tetrahydrofuran (60 mL), crystals of iodine were added to a solution of magnesium (1.2 g) in anhydrous tetrahydrofuran (60 mL) under a nitrogen atmosphere at a rate at which the reaction maintained stable reflux, followed by bromobenzene (7.85 g). The reaction mixture was stirred for 20 minutes and then cycloheptanone (4.48 g) was carefully added. After stirring for 10 minutes, saturated aqueous ammonium chloride (10 mL) was added and the reaction mixture was partitioned between water (100 mL) and isohexane (100 mL). Dry the organic layer (MgSO 4 ) And evaporation to give the subtitle compound (7.6g) as an oil.

[0203] 1 H NMR(299.946MHz, CDCl 3 )δ7.53-7.47 (m, 2H), 7.36-7.29 (m, 2H), 7.26-7.19 (m, 1H), 2.07 (ddd, 2H), 1.97-1.50 (m, 11H).

[0204] b) 1-Methoxy-1-phenyl-cycloheptane

[0205]

[0206] 1-Phenyl-cycloheptanol (Example 1a) (7.6 g) was dissolved in tetrahydrofuran (100 mL) and sodium hydride (60% in oil, 2.0 g) was added. The reaction mixture was stirred at 60°C for 5 minutes and methyl iodide (7.1 g) was added. The mixture was kept at 60°C overnight and then a certain amount of sodium hydride (60% in oil, 2.0 g) and methyl iodide (7.1 g) were added and the reaction mixture was refluxed for 70 hours. The reaction mixture was partitioned between water (100 mL) and isohexane (100 mL) and the organic layer was separated and dried (MgSO 4 ) And evaporation to obtain the subtitle compound (11.31 g).

[0207] 1 H NMR(300MHz, CDCl 3 )δ7.43-7.37(m, 2H), 7.37-7.30(m, 2H), 7.24-7.19(m, 1H), 2.98(s, 3H), 2.12-1.88(m, 4H), 1.88-1.45( m, 8H).

[0208] c) 1-Phenyl-cycloheptane carboxylic acid

[0209]

[0210] Potassium (2.62g) and sodium (0.52g) were heated together in mineral oil at 120°C for 30 minutes under a nitrogen atmosphere and then cooled to room temperature. The oil was removed and replaced with ether (100 mL) and 1-methoxy-1-phenyl-cycloheptane (Example 1b) (4.9 g) was added and the reaction mixture was stirred at room temperature overnight under nitrogen. The reaction mixture was cooled to -78°C and solid carbon dioxide (~20 g) was added while stirring. The reaction mixture was warmed to room temperature and water (150 mL) was carefully added under nitrogen. The aqueous layer was separated, neutralized with concentrated hydrochloric acid and extracted with ether (150 mL). Dry the organic layer (MgSO 4 ) And evaporation to give the subtitle compound (4.15 g) as an oil.

[0211] 1 H NMR(300MHz, CDCl 3 )δ7.40-7.20 (m, 5H), 2.49-2.35 (m, 2H), 2.16-2.03 (m, 2H), 1.76-1.47 (m, 8H).

[0212] d) Methyl 1-phenyl-cycloheptane carboxylate

[0213]

[0214] 1-Phenyl-cycloheptane carboxylic acid (Example 1c) (4.15 g) was refluxed in methanol (150 mL) and concentrated hydrochloric acid (5 mL) for 24 hours. The solvent was evaporated and the residue was dissolved in ether (100 mL), washed with water (100 mL), saturated sodium bicarbonate (50 mL) and water (100 mL), and dried (MgSO 4 ) And evaporation to give the subtitle compound (3.5g) as an oil.

[0215] 1 H NMR(300MHz, CDCl 3 )δ 7.37-7.18 (m, 5H), 3.63 (s, 3H), 2.47-2.35 (m, 2H), 2.08-1.97 (m, 2H), 1.70-1.48 (m, 8H).

[0216] e) 1-Phenyl-cycloheptane carboxylic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester

[0217]

[0218] Mix 1-phenyl-cycloheptane carboxylate (Example 1d) (1.0g) and (R)-quinuclidin-3-ol (0.39g) in heptane (50mL) containing sodium (~5mg) ) And reflux in Dean and Stark apparatus for 24 hours. The heptane (20 mL) was replaced with toluene (20 mL) and refluxing was continued for 3 days. The reaction mixture was partitioned between water (50mL) and ether (50mL) and the ether layer was separated and dried (MgSO 4 ) And evaporate. The crude product was purified by silica gel column chromatography (eluted with ethyl acetate/triethylamine (99/1)) to give the title compound as an oil (0.83 g).

[0219] m/e 328[M+H] +

[0220] 1 H NMR(300MHz, CDCl 3 )δ7.35-7.27 (m, 4H), 7.23-7.16 (m, 1H), 4.78-4.71 (m, 1H), 3.12 (ddd, 1H), 2.79-2.32 (m, 7H), 2.16-1.98 ( m, 2H), 1.91-1.80 (m, 1H), 1.70-1.34 (m, 12H).

[0221] f) 2-Bromo-N-(pyrazin-2-yl)-acetamide

[0222]

[0223] To a stirred suspension of pyrazin-2-ylamine (1.878 g) and potassium carbonate (8.19 g) in dichloromethane (25 mL) was added 2-bromoacetyl bromide (1.72 mL). The reaction mixture was stirred overnight and then washed with water (2x 50 mL). Dry the organic layer (MgSO 4 ) And concentrated to obtain the subtitle compound as a solid (0.700 g).

[0224] 1 H NMR(400MHz, CDCl 3 )δ9.51(d, 1H), 8.63(s, 1H), 8.42(d, 1H), 8.30(dd, 1H), 4.06(s, 2H).

[0225] Example 1: (R)-3-(1-Phenyl-cycloheptanecarbonyloxy)-1-(pyrazin-2-ylcarbamoylmethyl)-1-nitrogen -Bicyclo[2.2.2]octane bromide form A

[0226]

[0227] Combine 1-phenyl-cycloheptane carboxylic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 1e) (0.200g) and 2-bromo-N -(Pyrazin-2-yl)-acetamide (Example 1f) (0.132g) was dissolved in acetonitrile (1mL) and left to stand overnight. The resulting solid was filtered and washed with acetonitrile (2×1 mL) and ether (3 mL). The dried solid was recrystallized from acetone (15 mL) and ether (10 mL) to obtain the title compound (0.240 g).

[0228] m/e 463[M] +

[0229] 1 H NMR(400MHz, DMSO-D 6 )δ11.37 (s, 1H), 9.28 (s, 1H), 8.50-8.46 (m, 2H), 7.39-7.30 (m, 4H), 7.27-7.21 (m, 1H), 5.16-5.08 (m, 1H), 4.33(s, 2H), 4.17-4.07(m, 1H), 3.69-3.56(m, 4H), 3.48-3.38(m, 1H), 2.44-2.26(m, 3H), 2.25-2.04( m, 2H), 2.03-1.87 (m, 3H), 1.85-1.71 (m, 1H), 1.68-1.45 (m, 8H).

[0230] For Example 1: (R)-3-(1-phenyl-cycloheptanecarbonyloxy)-1-(pyrazin-2-ylcarbamoylmethyl)-1-nitrogen -Analysis of Bicyclo[2.2.2]octane Bromide Form A

[0231] The sample of crystal form A of Example 1 obtained by the above operation was analyzed by XRPD (PANalytical X'Pert or CubiX system), DSC and TGA.

[0232] The melting temperature of the bromide crystal form A of Example 1 determined by DSC was actually 202°C (initial) (±2°C). The weight loss observed by TGA before melting was 2.7%. GVS determined to obtain a 3% weight gain (%w/w) (±0.2%) at 80%RH.

[0233] Example 1 The XRPD spectrum of bromide crystal form A is in figure 1 Show in.