Dual-function tracheodilator and crystallization and preparation methods therefor

Abstract

The main purpose of the present invention is to provide a quaternary ammonium salt structural compound having both an M3 receptor antagonist function and a β2 receptor agonist function, namely (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octylonium hydrochloride chloride, as well as crystallization and preparation methods therefor, and a use thereof. The compound can form a crystalline solid, has better stability and solubility in water, has a faster onset time for airway relaxation, and is more suitable for drug preparation.

Description

A dual-function tracheodilator and its crystallization and preparation method Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a quaternary ammonium salt structure compound having both M3 (muscarinic receptor, 3-subtype abbreviated as M3) receptor antagonism and β2 (adrenergic receptor, 2-subtype receptor abbreviated as β2) receptor agonism, namely, (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium chloride hydrochloride and a preparation method thereof; the present invention also relates to a crystalline form of the quaternary ammonium salt compound suitable for practical application and its application in the preparation of pharmaceutical preparations. Background Art

[0002] Asthma and chronic obstructive pulmonary disease (COPD) are the most common prevalent diseases. Bronchodilators are the first-line treatment for asthma and COPD. Commonly used bronchodilators include M2 ​​receptor antagonists, such as ipratropium bromide and tiotropium bromide, and β2 receptor agonists, such as albuterol, formoterol, and vilanterol. Combinations of M2 receptor antagonists and β2 receptor agonists have shown superior efficacy in treating moderate to severe asthma and COPD compared to single M2 receptor antagonists or β2 receptor agonists. For example, a combined inhalation solution of ipratropium bromide and albuterol has a synergistic effect, resulting in a stronger efficacy than either drug alone. A combined inhalation powder of umeclidinium bromide and vilanterol is more effective than either umeclidinium or vilanterol alone. Tiotropium bromide / olodaterol combination inhalation powder, approved by the FDA in 2015, is also more effective than either drug alone.

[0003] In addition, compounds with both β2 adrenergic receptor agonist activity and M receptor antagonist activity (MABA for short) have been reported in many literatures for the treatment of asthma and COPD. MABA can produce bronchodilatory effects through two independent modes of action. Because it is a single molecule with a single pharmacokinetics, its efficacy is even better than that of compound preparations, which will bring good news to patients with moderate to severe asthma and COPD.

[0004] WO2018108089A1 discloses a class of long-acting MABA compounds, among which Example 93-R bromide (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)-2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium salt (compound of formula (1)) is a preferred compound. Its receptor binding experiment shows that the compound has both β2 adrenergic receptor agonist activity and M3 receptor antagonist activity, and the two functions have a good match. Compared with the existing technology, it has the characteristics of rapid onset and low toxic side effects in the treatment of asthma and COPD.

[0005] The method disclosed in WO2018108089A1 shows that the bromide ions of all compounds (including the compound of formula (1)) can be replaced by other acid radicals in a molar ratio of 1:1. There is no mention of the product obtained by the formation of salts between the amino group and the acid. We found that after the above-mentioned compound was dissolved in a benign solvent and the solvent was removed under vacuum and reduced pressure, a solid substance was obtained. However, it was always impossible to obtain a crystalline solid. The solid had poor stability and low solubility in water, and could not be made into a qualified pharmaceutical preparation. Such solids were tested by liquid chromatography and found that the content of 5 single impurities exceeded 0.1% (Figure 1, HPLC chart of the compound of formula (1)). Therefore, a crystallization purification method must be found to solve this technical problem and provide technical guarantees for its commercial application. The inventors tried to crystallize and purify the compound of formula (1) using a variety of single solvents and mixed solvents, such as methanol, ethanol, isopropanol, isobutanol, 2-butanone, tetrahydrofuran, acetonitrile, methyl tert-butyl ether, water, toluene, ethyl acetate, isopropyl acetate, n-heptane and other single solvents and mixed solvents composed of the above solvents, but all were unsuccessful. In view of this, without changing the free base structure of the compound of formula (1), the bromide ion of the compound of formula (1) was replaced with other anions to change the physical and chemical properties of the product, so that the product formed by the compound has stable and consistent crystals and controllable quality, which can be truly put into commercial application. During the continuous experimental process, it was extremely accidental and fortunate to obtain the product crystal formed by the base of the compound of formula (1): HCl = 1:2 (Figure 2, HPLC chart of the compound of formula (2)), which greatly reduced the impurity level of the product and improved the quality of the product. The crystals were dissolved in ethanol and single crystals were prepared to obtain single crystals of the compound of formula (2). X-ray diffraction results showed the structural formula of the single crystals, and the chemical name was (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)-2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium chloride hydrochloride. More unexpectedly, the use of other anions such as bromine, iodine, sulfate, nitrate in different proportions and various but not limited to organic acids such as acetic acid, propionic acid, tartaric acid, citric acid, methanesulfonic acid, furoic acid, etc. did not produce corresponding crystalline products. Even the product generated by using the compound of formula (1) with base:HCl in a ratio of 1:1 could not be crystallized. The difference between the compound of formula (2) and the compound of formula (1) is that the bromide ion in formula (1) is replaced by a chloride ion, and the amine group in the compound structure forms a hydrochloride. In other words, the free base of the compound of formula (1) can only be crystallized when it is the chloride hydrochloride (the compound of formula (2)), which proves the uniqueness of this compound's structure.The crystalline form I, crystalline form II and amorphous form of the compound of formula (2) show more stable physical and chemical properties and higher water solubility than the compound of formula (1). The three crystals of the compound of formula (1) and the compound of formula (2) all have strong airway dilation effects, but the crystalline form I, crystalline form II and amorphous form of the compound of formula (2) have an onset time that is significantly faster than that of the compound of formula (1), laying a solid foundation for improving the quality of the compound and broadening its practical application range.

[0006] The production process of the compound disclosed in WO2018108089A1 is as follows: (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-azabicyclo[2,2,2]octane free base is reacted with 3-[4-(2-oxopropyl)phenoxy]propane bromide to generate brominated (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-[3-(4-acetonylphenoxy)propyl]-1- Azabicyclo[2,2,2]octanium salt, which reacts with (R)-2-amino-1-[(4-hydroxy-3-formamido)phenyl]ethanol to form (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-(2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium bromide. The target product obtained in the above process is a pair of diastereomers. To obtain a single stereoisomer sample, preparative HPLC separation and preparation are required.

[0007] After continuous research by the inventors, the process route in synthetic route 1 was adopted, that is, intermediate I was subjected to a nucleophilic substitution reaction with 1,3-dibromopropane to generate intermediate II, intermediate II was subjected to a quaternization reaction with (2R,3R)-3-[(2-cyclopentyl-2-hydroxy-2-phenyl)ethoxy]-1-azabicyclo[2,2,2]octane free base to obtain intermediate III, the nitro group in the structure of intermediate III was reduced to amino group by iron powder and ammonium chloride solution, and then the intermediate was treated with 20% sodium chloride aqueous solution to displace the bromide ion in the structure to obtain intermediate IV, intermediate IV was then reacted with mixed acid anhydride in dichloromethane to formylate the amino group in the structure to prepare intermediate V, and finally intermediate V was subjected to hydrogenation reduction to prepare target compound VI, i.e., compound of formula (2).

[0008] In the synthesis route of the compound of formula (2), the same effect can be achieved by using a chloride anion resin exchange process for ion exchange. However, the method of synthetic route 1 is adopted to wash with a 20% sodium chloride aqueous solution for ion replacement, which is conducive to large-scale commercial production and greatly reduces production costs. At the same time, the solvent anhydrous formic acid in the mixed anhydride formylation method reported in the literature is replaced with dichloromethane. Formylation can be carried out by adding only a small amount of anhydrous formic acid and acetic anhydride (mixed anhydride method), which greatly reduces the content of impurities in the final product, is conducive to the recrystallization and purification of the product, is also conducive to the safety and environmental protection of production, and reduces production costs. It is more conducive to commercial production, which is both economical and environmentally friendly.

[0009] After extensive creative research, the inventors successfully solved the above problems.

[0010] The reaction formula is as follows:

[0011] Synthesis Route 1 Summary of the Invention

[0012] One of the purposes of the present invention is to provide a compound as shown in formula (2):

[0013] Another object of the present invention is to provide a method for preparing the compound of formula (2), such as synthetic route 1, comprising the following steps:

[0014] Step 1, preparing intermediate II by reacting intermediate I with 1,3-dibromopropane;

[0015] The reaction solvent is selected from methanol, ethanol, N,N-dimethylformamide and N,N-dimethylacetamide, preferably methanol and ethanol; the molar ratio of 1,3-dibromopropane to intermediate I is 5-10:1, preferably 8-10:1; the base used in the reaction is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, etc., and the molar ratio of the base to intermediate I is 2-5:1, preferably 3:1.

[0016] Step 2: Intermediate II is reacted with (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-azabicyclo[2,2,2]octane to prepare intermediate III:

[0017] The molar ratio of intermediate II to (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-azabicyclo[2,2,2]octane (prepared according to the "Preparation 2" method in the specific embodiment of WO2018108089) is 1-3:1, preferably 1.1:1; the reaction solvent is selected from methanol, ethanol, N,N-dimethylformamide and N,N-dimethylacetamide, etc., preferably methanol, ethanol, etc.

[0018] Step 3: Using iron powder to reduce the nitro group of intermediate III to an amino group, the molar ratio of intermediate III to iron powder is 1:3-8, preferably 1:5; the solvent used in the reaction is selected from a 5%-10% aqueous solution of methanol, ethanol, tetrahydrofuran and acetone, preferably an aqueous solution of ethanol and methanol.

[0019] Step 4: Intermediate IV reacts with a mixed acid anhydride for formylation, and then is treated with hydrochloric acid to prepare intermediate V. The molar ratio of intermediate IV to formic acid and acetic anhydride is 1:1-3:3-8, preferably 1:1.2:6. The solvent is selected from formic acid and dichloromethane, preferably dichloromethane.

[0020] Step 5. The intermediate V is hydrogenated under Pd-C catalysis to remove the benzyl protecting groups on the amino and hydroxyl groups to obtain the target product, (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)-2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium chloride hydrochloride. The reaction solvent is selected from formic acid, acetic acid, methanol and ethanol, preferably formic acid.

[0021] Another object of the present invention is to provide three different crystal forms of the compound of formula (2) and a preparation method thereof.

[0022] The crystal form analysis method for studying the crystallization of the present invention is as follows:

[0023] X-ray powder diffraction (XRPD)

[0024] All solid samples were analyzed using a powder X-ray diffraction analyzer (Bruker D8 advance) equipped with a LynxEye detector. The 2θ scan angle was from 3° to 40°, with a scan step size of 0.02°. The tube voltage and current were 40 kV and 40 mA, respectively. A zero-background sample pan was used for the measurements.

[0025] The pharmaceutical compositions of the present invention generally contain a therapeutically effective dose of a compound of the present invention. Typically, such pharmaceutical compositions contain about 0.001% to 100% by weight of the active ingredient.

[0026] Any conventional carrier or excipient may be used in the present invention. The selection of a particular carrier or excipient, or combination of carrier and excipient, will depend on the mode of administration or the medical condition or disease type being treated for a particular patient. Techniques for preparing pharmaceutical compositions for specific modes are within the knowledge of those skilled in the art. In addition, carriers or excipients, or combinations of carriers and excipients, are commercially available.

[0027] Representative examples of pharmaceutically acceptable carriers include, but are not limited to, the following: (1) sugars, such as glucose, lactose, sucrose, etc.; (2) starches, such as corn starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, cellulose acetate, etc.; (4) talc; (5) excipients, such as cocoa butter, waxes; (6) oils, such as olive oil, soybean oil, etc.; (7) alcohols, such as ethanol, propylene glycol, glycerol, sorbitol, polyethylene glycol, mannitol, etc.; (8) esters, such as ethyl oleate, ethyl laurate; (9) pyrogen-free water; (10) isotonic saline; (11) phosphate buffer; (12) compressed propellant gases, such as chlorofluorocarbons, hydrofluorocarbons, etc.; (13) other non-toxic miscible substances used in pharmaceutical compositions.

[0028] The compositions of the present invention are usually prepared by thoroughly mixing the compound of the present invention with one or more optional carriers. If necessary, the homogeneous mixture obtained in the present invention can be shaped or loaded into tablets, capsules, pills, cans or cartridges using conventional equipment and methods.

[0029] Pharmaceutical composition of the present invention is suitable for inhalation administration. Compositions for inhalation administration are typically in the form of atomized inhalation, aerosol or powder spray. Such compositions are usually administered with known dosing devices, such as a nebulizer, metered dose inhaler (MDI) or dry powder inhaler (DPI) or other similar dosing devices.

[0030] The compositions containing the active ingredient of the present invention are administered by atomization via a nebulizer. A nebulizer typically generates a high-speed airflow, atomizing the pharmaceutical composition containing the active ingredient for inhalation into the patient's respiratory tract. Therefore, the active ingredient is typically dissolved in a suitable solvent to form a solution that is placed in the nebulizer. Alternatively, the active ingredient is micronized and combined with a suitable carrier to form a suspension of micronized particles suitable for inhalation. Micronization is generally defined as reducing the solid particles to a diameter of less than 10 μm or more. Suitable nebulizers are commercially available.

[0031] Representative pharmaceutical compositions for use with a nebulizer inhaler include those containing 5 μg / ml to 10 mg / ml of the compound of formula (2) or a pharmaceutically acceptable solvate thereof.

[0032] The present invention encompasses pharmaceutical compositions for inhalation administration using a dry powder inhaler. Dry powder inhalers typically form a free-flowing powder of the active ingredient in the patient's airstream during inhalation. Therefore, the active ingredient is typically formulated with a suitable excipient, such as lactose, to provide a free-flowing powder.

[0033] A representative pharmaceutical composition for use in a dry powder inhaler comprises micronized particles of dry lactose having a particle size between about 1 μm and about 100 μm and a compound of formula (2).

[0034] Dry powder formulations can be prepared by dry mixing the active ingredient with excipients, or without excipients, and then loading the pharmaceutical composition into a dry powder dispenser, or into inhaler cartridges or capsules for use with dry powder delivery devices. Dry powder delivery devices are commercially available.

[0035] The compositions of the present invention comprising the compound of formula (2) are administered by inhalation using a metered dose inhaler. Such a metered dose inhaler utilizes a compressed propellant gas to release a measured amount of the compound of formula (2). Thus, the pharmaceutical composition administered using a metered dose inhaler is contained in a liquefied propellant solution or suspension.

[0036] Representative pharmaceutical compositions for metered dose inhalers comprise from 0.001% to about 3% by weight of a compound of formula (2) or a pharmaceutically acceptable solvate thereof; from about 0% to about 40% of a co-solvent ethanol or glycol, preferably from 5% to about 30%; from about 0% to 3% by weight of a surfactant; and the remainder being a hydrofluoroalkane (HFA) propellant.

[0037] Such compositions are typically prepared by adding ice-cold or pressurized hydrofluoroalkanes to a suitable container containing the active ingredient, ethanol (if present), and a surfactant (if present). To prepare a suspension, the active ingredient is micronized and then mixed with a propellant. The formulation is then placed in an aerosol canister, forming part of a metered dose inhaler device.

[0038] Methods for preparing inhalable particles and formulations and other examples suitable for administration by inhalation are described in the literature.

[0039] The present invention relates to a method for treating a patient's pulmonary disease such as COPD or asthma, comprising administering to the patient a compound of formula (2), or a combination thereof with a steroidal anti-inflammatory agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 HPLC spectrum of pure compound of formula (1)

[0041] Figure 2 HPLC spectrum of pure compound of formula (2)

[0042] Figure 3 XRPD spectrum of the compound of Example 2 (Form I)

[0043] Figure 4 XRPD spectrum of the compound of Example 3 (Form II)

[0044] FIG5 XRPD spectrum of the sample of the compound of Example 4 (Form III) Specific implementation method:

[0045] Preparation Example

[0046] Intermediate I: 4-((R)-2-(Benzyl((R)-2-(4-(Benzyloxy)-3-nitrophenyl)-2-hydroxyethyl)amine)propyl)phenol

[0047] To a 500 mL reactor, 128.0 g (0.530 mol) of (R)-4-(2-(benzylamino)propyl)phenol (prepared according to the method of US9029421), 146.0 g (0.538 mol) of (R)-2-(4-(benzyloxy)-3-nitrophenyl)oxirane (prepared according to the method of US6268533), and 250 mL of 1,4-dioxane were added. The mixture was stirred, heated to reflux (approximately 114°C), and reacted for 8 hours. After completion of the reaction, the reaction mixture was cooled to an internal temperature of approximately 55°C. The reaction mixture was evaporated under reduced pressure at 55°C until no liquid was distilled out. The residue was dissolved in 300 mL of ethyl acetate and washed with water three times (3 x 50 mL). The organic phase was dried over anhydrous magnesium sulfate, the desiccant was filtered, and the solution was evaporated under reduced pressure to dryness to obtain 258.1 g of a light reddish-brown oil (95.0% yield).

[0048] Example 1

[0049] Preparation of (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-{2-[4-((R)-2-{(R)-[2-hydroxy-2-(3-formamido-4-hydroxy)phenyl]ethylamino}propyl)phenoxy]propyl}-1-azabicyclo[2,2,2]octanium chloride hydrochloride (steps described in Synthetic Scheme 1)

[0050] Step 1: Preparation of Intermediate II

[0051] To a 5L reactor, add 148.1g (0.289mol) of Intermediate I. Add 1000mL of methanol in portions and heat to approximately 55°C with stirring to dissolve. Then, add 583.2g (2.889mol) of 1,3-dibromopropane, 0.613kg (0.5784mol) of anhydrous sodium carbonate, and 200mL of purified water. Continue stirring and raise the temperature to reflux. Stir and react for 10 hours. Samples are taken and tested for endpoint by HPLC. The residual amount of Intermediate I should not exceed 2%. After the reaction, cool the reaction mixture to an internal temperature of 25°C, add 500mL of ethyl acetate, filter to remove inorganic salts, rinse the solid with 1000mL of ethyl acetate three times, combine the filtrates, and remove the solvent under reduced pressure to dryness. Dissolve the residue in 900mL of dichloromethane and wash with 200mL of purified water three times with stirring. Allow the layers to separate, and dry the organic phase over 60g of anhydrous magnesium sulfate. Filter, add 2500 mL of n-heptane to the filtrate, stir thoroughly, pour off the upper solvent, remove excess 1,3-dibromopropane, dissolve the residue in 500 mL of water, and remove the solvent under reduced pressure to dryness to obtain 145.3 g of a light yellow oil (Intermediate II), which was used directly in the next step. Yield: 79.4%. m / z 632.48, 634.49 (M-1), 1HNMR(DMSO-d6)δppm 8.228 (d, 1H, J = 3.9Hz), 7.672 (d, 1H, J = 7.6Hz), 7.484 (m, 2H), 7.403 (m, 2H) ,7.324(m,1H),7.293(m,1H),7.13-7.25(m,7H),6.865(d,J=7.8Hz,2H),5.5 74(s,1H),5.158(s,2H),4.872(m,1H),4.082(m,2H),3.621(m,2H),3.508( m, 2H), 3.118 (m, 1H), 2.711 (m, 2H), 2.511 (m, 1H), 2.13 (m, 2H), 1.11 (d, 3H).

[0052] Step 2: Preparation of Intermediate III

[0053] To a 2L reaction flask, add 145.1g (0.229mol) of Intermediate II, 66.3g (0.210mol) of (R)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-azabicyclo[2,2,2]octane, and 500mL of anhydrous ethanol. Heat with stirring to dissolve. Raise the temperature to reflux and stir for 3 hours. After completion of the reaction, TLC analysis showed that the (R)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-azabicyclo[2,2,2]octane concentration was less than 1%. Evaporate to dryness under reduced pressure. The residue was dissolved in 300 mL of dichloromethane, and 300 mL of isopropyl ether was added with stirring. Stirring was continued for 15 minutes, and the mixture was allowed to stand. The supernatant was discarded, and the lower oily material was dissolved in 700 mL of dichloromethane, and washed with 300 mL of purified water × 3 times with stirring. The mixture was allowed to stand for stratification, and the organic phase was separated. 100.00 g of anhydrous magnesium sulfate was added, and the mixture was allowed to stand for drying. The desiccant was removed by filtration under reduced pressure, and the desiccant was rinsed with 200 ml of dichloromethane × 3 times. The filtrate was transferred to a rotary evaporator in portions and evaporated under reduced pressure at 37°C until no liquid was distilled out. The temperature was raised to 50°C and rotary evaporation was continued for 1 hour. The oil pump was used to reduce the pressure and rotary evaporation (rotary evaporation speed was 100 r / min) until a foamy solid was obtained, and 179.29 g of intermediate III was obtained as a yellow foamy solid with a yield of 90.0%.

[0054] m / z 869.01(M + -Br), 1HNMR(DMSO-d6)δppm 8.218 (d, 1H, J = 3.9Hz), 7.66 (d, 1H, J = 7.6Hz), 7.540 (m, 2H), 7.491 (m, 2H), 7.4 03(m,2H),7.324(m,1H),7.293(m,3H),7.13-7.25(m,8H),6.865(d,J=7.8Hz,2H ), 5.574 (s, 1H), 5.520 (s, 1H), 5.149 (s, 2H), 4.807 (m, 1H), 4.050 (m, 2H), 3.793 (m, 1H), 3.611 (m, 2H), 3.222 (m, 4H), 3.102 (m, 1H), 2.711 (m, 2H), 2.601 (m, 1H), 2.511(m,1H),2.47(m,1H),2.310(m,2H),2.26(m,2H),2.15(m,2H),1.902(m,2H),1.770- 1.790 (m, 3H), 1.731 (m, 2H), 1.602-1.682 (m, 3H), 1.561 (m, 2H), 1.44 (m, 2H), 1.11 (d, 3H).

[0055] Step 3, preparation of intermediate IV

[0056] In a 5L reactor, add 177.6g (0.1871mol) of Intermediate III and 1500mL of anhydrous ethanol. Heat and stir to dissolve. Then add 41.7g (0.747mol) of activated iron powder and rinse the mixture several times with 400mL of purified water. Then add 40g (0.749mol) of ammonium chloride and 260g of purified water. Stir and heat to reflux for 8h. After the reaction, cool the reaction mixture to the internal temperature, filter under reduced pressure to remove iron sludge, and concentrate the filtrate to dryness under reduced pressure. The residue was dissolved in 1000 mL of dichloromethane and washed with 300 mL of purified water. The organic phase was separated and washed three times with 400 mL of 20% sodium chloride aqueous solution. The separated organic phase was dried over 60 g of anhydrous magnesium sulfate and filtered under reduced pressure. The filtrate was concentrated to dryness to a foamy solid. The solid was dissolved in 400 mL of anhydrous ethanol and the pH was adjusted to approximately 5 with an appropriate amount of hydrochloric acid. Activated carbon was added for decolorization. The filtrate was concentrated under reduced pressure to dryness to obtain 163.3 g of Intermediate IV as a light brown solid, with a yield of 96.5%. m / z 839.1 (M-Cl). 1HNMR(DMSO-d6)δppm 7.393-7.601(m,6H),7.290-7.324(m,5H),7.13-7.25(m,6H)6.991(s,1H), 6.930(m,1H),6.859(m,2H),6.674(m,1H),5.674(s,1H),5.510(s,1H),5.27 1(s,2H),5.161(s,1H),4.868(m,1H),4.048(m,3H),3.920(m,1H),3.793(m, 2H), 3.721 (m, 2H), 3.665 (m, 1H), 3.620 (m, 2H), 3.481 (m, 1H), 3.35 (m, 1H), 3 .272(m,4H),3.223(m,5H),3.112(m,1H),2.961(m,1H),2.770(m,1H),2.84 3(m,1H),2.715(m,1H),2.601(m,1H),2.640(m,1H),2.515(m,1H),2.47(m,1 H),2.310(m,2H),2.261(m,2H),2.15(m,2H),1.891(m,2H),1.770(m,2H),1. 729 (m, 2H), 1.602-1.682 (m, 8H), 1.525 (m, 2H), 1.441 (m, 2H), 1.120 (d, 3H).

[0057] Step 4. Preparation of Intermediate V

[0058] In a 5L three-necked reaction flask, add 437g (0.95mol) of anhydrous formic acid, stir, cool to 0-5°C, add 311.0g (0.305mol) of acetic anhydride dropwise below 15°C, and keep the temperature below 15°C for 30 minutes. Under stirring, add a solution containing 162.8g (0.179mol) of intermediate IV dissolved in 2000mL of dichloromethane dropwise, keeping the internal temperature at 0-10°C. After the addition is complete, control the internal temperature at 10-15°C and continue stirring for 6 hours. After the reaction is completed, add 2000mL of dichloromethane to dilute the reaction mixture, wash the reaction mixture with 800mL of 10% sodium chloride aqueous solution × 5, add 160g of anhydrous magnesium sulfate, and stir and dry for 2 hours. Filter under reduced pressure to remove the desiccant, and concentrate the filtrate under reduced pressure to a foamy solid to obtain 155.8g of intermediate V as a light brown foamy solid, with a yield of 93.3%. m / z 839.1 (M-Cl), 1HNMR(DMSO-d6)δppm 8.56(s,1H),7.393-7.541(m,6H),7.290-7.324(m,5H),7.13-7.25(m,8H),6.860(m,2H ), 6.674(m,1H), 5.674(s, 1H), 5.510(s, 1H), 5.158(s,1H), 4.862(m, 1H), 4.050(m, 3H), 3.923(m,1H), 3.790(m,2H), 3.720(m,2H), 3.656(m,1H), 3.618(m,2H), 3.480(m,1H), 3.347(m,1H), 3.271 (m, 4H), 3.220 (m, 5H), 3.110 (m, 1H), 2.959 (m, 1H), 2.768 (m, 1H), 2.840 (m, 1H), 2.695 (m, 1H), 2 .629(m,1H),2.60(m,1H),2.512(m,1H),2.468(m,1H),2.308(m,2H),2.260(m,2H),2.15(m,2H),1.88 0 (m, 2H), 1.770 (m, 2H), 1.726 (m, 2H), 1.60-1.680 (m, 8H), 1.515 (m, 2H), 1.438 (m, 2H), 1.110 (d, 3H).

[0059] Step 5: Preparation of target compound

[0060] In a 5L hydrogenation kettle, add 1300mL of methanol and 155.6g (0.166mol) of intermediate V. Stir at approximately 25°C for 10 minutes. Then add 35.1g of 10% palladium on carbon (pre-washed with purified water) and 1800mL of purified water. Nitrogen is introduced to an internal pressure of approximately 0.4MPa to displace the air. This operation is repeated twice. Hydrogen is introduced to an internal pressure of approximately 0.4MPa to displace the nitrogen. This operation is repeated twice. Hydrogen is introduced to an internal pressure of approximately 0.4MPa to displace the nitrogen. This operation is repeated twice. Hydrogen is introduced to an internal pressure of approximately 0.4MPa. The internal temperature is maintained at 25°C for 5 hours. After the reaction is completed, the pressure is released and the reaction kettle is rinsed with 1300mL of methanol, which is then added to the reaction solution. The reaction solution is filtered under reduced pressure (0.45μm filter membrane) to remove the palladium on carbon. The palladium on carbon is rinsed with 300mL of methanol four times. The filtrate was transferred to a rotary evaporator and evaporated under reduced pressure at 50°C until no liquid was distilled out. The evaporation was continued under reduced pressure until dryness was achieved. The evaporation was then continued under reduced pressure using an oil pump (evaporation speed of about 100 r / min) until a foamy solid formed. 116.3 kg of crude compound of formula (2) was obtained as a pale yellow solid, with a yield of 92.5%. Samples were taken for water content testing, and the water content was controlled to be no more than 1.5%.

[0061] Example 2. Preparation of Crystalline Form I of the Compound of Formula (2)

[0062] In a 10mL reaction flask, add 1.311g of the crude compound of formula (2) and 3mL of anhydrous methanol, stir, and heat to 50°C to dissolve. Add 7mL of isobutyl acetate and continue stirring for 5 minutes. Cool to 5°C and stir to crystallize. After a large amount of solid precipitates, maintain the temperature at around 25°C and continue crystallization for 1.5 hours. Cool to 5±2°C and stir to crystallize for 3.5 hours. Filter under reduced pressure and rinse the filter cake with 0.159kg of a 50% anhydrous ethanol and isopropanol mixture x 3. The solid is vacuum dried at 80-85°C for 4 hours to obtain 0.817g of an off-white powdery solid with a yield of 62.3%. Elemental analysis: C 29 H 40 BrNO3, calculated value C65.65, H7.60, Br15.06, N2.64; found value C65.60, H7.58, Br15.0, N2.62. m / z 687.5 (M-Cl-HCl), elemental analysis 1HNMR (DMSO-d6) δppm10.197 (1H), 9.644 (s, 1H), 9.498 (m, 1H), 8.758 (m, 1H), 8.292 (d, 1H), 8.145 (s, 1H), 7.484 (d, J = 7.6 Hz, 2H), 7.283 (t, J = 7.6 Hz, 2H), 7.13-7.22 (m, 3H), 6.92-6.99 (m, 2H), 6.895 ( d, 2H), 6.084 (s, 1H), 4.938 (m, 1H), 4.802 (s, 1H), 3.988 (t, 2H), 3.795 (m, 1H), 3.718 (d, 1H), 3.653 (m, 1H), 3.4 80 (d, 1H), 3.07-3.16 (m, 7H), 2.91-3.07 (m, 2H), 2.611 (t, 1H), 2.399 (m, 1H), 2.217 (m, 1H), 2.065 (m, 2H), 1.893 (m, 1H), 1.65-1.77 (m, 2H), 1.57-1.65 (m, 3H), 1.44-1.57 (m, 2H), 1.29-1.44 (m, 2H), 1.231 (m, 1H), 1.087 (m, 4H).

[0063] The results showed that the product did not contain water of crystallization or other crystallization solvents, and was consistent with the molecular formula of the compound of formula (2); XRPD results showed that the product was crystalline, named Form I, and its X-ray powder diffraction pattern was shown in Figure 3, and the data were shown in Table 1.

[0064] Table 1 Characteristic X-ray powder diffraction data of target compound Form I

[0065] The crystal characteristics of the compound form I are X-ray powder diffraction pattern (CuKα, at about 25° C.), and can be further characterized by data including 2θ values ​​selected from the group consisting of: 9.4±0.3, 14.7±0.3, 15.3±0.3, 18.6±0.3, 19.8±0.3, 20.8±0.3, 25.1±0.3.

[0066] Example 3: Preparation of Crystalline Form II of the Compound of Formula (2)

[0067] In a 5L reactor, add 1.50g of the crude compound of formula (2) and 3mL of anhydrous ethanol, stir, and heat to approximately 50°C to dissolve. Then cool to approximately 25°C and stir to crystallize. After a large amount of solid precipitates, maintain the temperature at approximately 25°C for 1.5h of crystallization. Cool to 5±2°C and stir to crystallize for 3.5h. Filter under reduced pressure, rinse the filter cake with 3mL of a 50% anhydrous ethanol and isopropanol mixture three times. Dry the solid in a vacuum at 80-85°C for 4h to obtain 1.17g of an off-white powder, designated Form II, with a yield of 78.0%. The X-ray powder diffraction pattern of Form II is shown in Figure 4, and the X-ray powder diffraction data of the target compound are shown in Table 2.

[0068] Table 2 Characteristic X-ray powder diffraction data of target compound crystal form II

[0069] The crystal characteristics of the compound crystal form II are as follows: X-ray powder diffraction pattern (CuKα, at about 25° C.), and can be further characterized by data including 2θ values ​​selected from the group consisting of: 3.5±0.3, 7.1±0.3, 10.7±0.3, 17.1±0.3, 18.8±0.3.

[0070] Example 4: Preparation of amorphous crystal form of the compound of formula (2)

[0071] In a 50 ml single-necked bottle, add 2.00 g of the compound of formula (2) (crystal form I) and 20.00 g of isopropyl acetate containing 1.0% water. Maintain the internal temperature at about 35°C and stir magnetically for 1 hour. Then, filter out the solid and repeat the above process once. The solid is vacuum-dried at 80-85°C for 4 hours to obtain an amorphous sample. The sample appears as an off-white loose solid. The X-ray powder diffraction pattern of this product is shown in Figure 5.

[0072] Preparation Example 1

[0073] The dry powder formulation for inhalation administration was prepared by the following method

[0074] Components and dosages per dose in the composition

[0075] Example 2 compound 1.0 mg

[0076] Lactose 25mg

[0077] The compound of Example 3 of the present invention was micronized to an average particle size of 1 to 10 μm, and thoroughly mixed with lactose. The mixture was placed in capsules and administered using a powder inhaler.

[0078] Preparation Example 2

[0079] The dry powder formulation for inhalation administration was prepared by the following method

[0080] Components and dosages per dose in the composition

[0081] Example 3 compound 0.30 mg

[0082] Lactose 25mg

[0083] Magnesium stearate 0.04mg

[0084] 1.19% by weight of the compound of Example 4 of the present invention was micronized to an average particle size of 1-10 μm, thoroughly mixed with 0.16% by weight of micronized magnesium stearate, and then thoroughly mixed with 98.65% by weight of lactose to prepare a 25.34 mg / capsule capsule preparation for administration using a powder inhaler.

[0085] Preparation Example 3

[0086] The dry powder formulation for inhalation administration was prepared by the following method.

[0087] Components and dosages per dose in the composition

[0088] Example 4 compound 0.05 mg

[0089] Lactose 25mg

[0090] The compound of Example 4 of the present invention was micronized to an average particle size of 1 to 10 μm, and thoroughly mixed with lactose. The mixture was placed in capsules and administered using a powder inhaler.

[0091] Preparation Example 4

[0092] The inhalation solution is contained in a low-density polyethylene ampoule for inhalation solution and is administered by aerosol inhalation.

[0093] Components and dosages per dose in the composition

[0094] Preparation Example 5

[0095] The inhalation solution is contained in a low-density polyethylene ampoule for inhalation solution and is administered by aerosol inhalation.

[0096] Components and dosages per dose in the composition

[0097] Preparation Example 6

[0098] The aerosol liquid is contained in a pressure-resistant container consisting of an aluminum can and a metered-dose valve; it is sprayed out in a mist-like form by an actuator and inhaled through the mouth.

[0099] Specifications: 20 μg / press (calculated as anhydrous form of formula (2), 60 presses / bottle (actual dosage is 150%). The formula composition of the unit dose product is as follows:

[0100] Preparation Example 7

[0101] The aerosol liquid is contained in a pressure-resistant container consisting of an aluminum can and a metered-dose valve; it is sprayed out in a mist-like form by an actuator and inhaled through the mouth.

[0102] Specifications: 200 μg / press (calculated as anhydrous form according to formula (2), 60 presses / bottle (actual dosage is 150%). The formula composition of the unit dose product is as follows:

[0103] Experimental Example 1: Impurity Removal Effect of Recrystallization

[0104] The following high performance phase chromatography conditions were used to determine the impurity removal effect of the recrystallization process of the crystals of the present invention.

[0105] Instrument: High performance liquid chromatography equipped with UV detector

[0106] Solvent: Mobile phase A

[0107] First, the purity (%) of the crude product of the compound of formula (2), the three crystals, and the compound of formula (1) was calculated based on HPLC chromatography using the following formula: Purity of the active ingredient (%) = (peak area of ​​the active ingredient) / (sum of all peak areas) × 100%. The impurity removal rate (%) in the crystals of the compound of formula (2) based on the solid of the compound of formula (2) = [{(purity of the active ingredient in each crystal) - (purity of the active ingredient of the compound of formula (2))} / {100% - (purity of the active ingredient of the crude product of the compound of formula (2))}] × 100%

[0108] The results are shown in Table 3:

[0109] Table 3. Results of Recrystallization to Remove Impurities from API

[0110] The results show that the crystalline forms I, II, and amorphous forms of the compound of formula (2) of the present invention can remove most impurities from the raw drug substance compared to the crude compound of formula (2). The purity of the raw drug substance of the crystalline forms I, II, and amorphous forms of the compound of formula (2) is not much different. Furthermore, the impurity contents of the crude compounds of formula (1) and formula (2) are relatively similar, fully demonstrating the important role of the crystallization process in removing impurities.

[0111] Experimental Example 2: Destruction Test of Influencing Factors (HPLC Method See Experimental Example 1)

[0112] The crude products of the compound of formula (1), the compound of formula (2), and the crystal form I, crystal form II, and amorphous form were subjected to high temperature, high humidity, and light damage tests. The main peak content (stability), total impurities (total impurities %), and main peak purity of each test sample were analyzed.

[0113] (1) Take 10 mg of each undamaged test sample, accurately weigh it, place it in a 20 ml volumetric flask, add solvent to dissolve and dilute to the scale, and shake well.

[0114] (2) High temperature destruction samples: Take 10 mg of each test sample, weigh accurately, place in a 20 ml volumetric flask, place at 80 ° C for 12 days, add solvent to dissolve and dilute to the scale, and shake well.

[0115] (3) High temperature destruction samples: Take 10 mg of each test sample, weigh accurately, place in a 20 ml volumetric flask, place at 105 ° C for 24 hours, add solvent to dissolve and adjust the volume to the scale, and shake well.

[0116] (4) High humidity damage samples: Take 10 mg of each test sample, weigh accurately, place in a weighing bottle, and leave it open at a humidity of 75% for 24 hours. Then add solvent to dissolve it and transfer it to a 20 ml volumetric flask, dilute to the scale, and shake well.

[0117] (5) Light-damaged samples: Take 10 mg of each test sample, weigh accurately, and place in a 20 ml transparent volumetric flask. After placing it at 4500 Lx ± 500 Lx for 12 days, add solvent to dissolve and dilute to the scale, and shake well.

[0118] Take the above test solution and inject it into the liquid chromatograph respectively, and record the chromatogram.

[0119] The test results are shown in Table 4:

[0120] Table 4. Results of relevant substance inspection and destruction test

[0121] Result analysis: The test sample compound (1) produced 0.3% of single impurity in 5.4 min after being placed at high temperature of 80°C for 12 days, and the crude product of compound (2) produced 0.21% of single impurity in 5.4 min after being placed at high temperature of 80°C for 12 days. The crystal form I, crystal form II and amorphous form of compound (2) did not produce more than 0.2% of impurities, and had good stability; after being placed at high temperature of 105°C for 24 hours, each test sample produced impurities in 5.4 min and 42.5 min, but the total impurity and single impurity of compound (1) were significantly higher than those of the crude product of compound (2), and the crude product of compound (2) was higher than the three crystals; in the stability test under high humidity and light conditions, each test sample did not produce more than 0.2% of single impurity, but the total impurity and single impurity of compound (1) were significantly higher than those of the crude product of compound (2), and the crude product of compound (2) was higher than the three crystals.

[0122] Conclusion: Under high temperature, light and high humidity conditions, the stability of the compound of formula (1) is worse than that of the crude compound of formula (2), and the crude compound of formula (2) is worse than the three crystals of the compound of formula (2). The stability of the three crystals of the compound of formula (2) under the above conditions is basically the same, with no obvious difference.

[0123] Experimental Example 3: Solubility of the crude compound of formula (2), three crystals thereof, and the compound of formula (1)

[0124] According to the determination method under the general rules of Part IV of the Chinese Pharmacopoeia: shake vigorously for 30 seconds every 5 minutes in water at 25°C ± 2°C; observe the dissolution within 30 minutes: Results: The crude product of the compound of formula (2) and its three crystalline forms are easily soluble in water, with a solubility of more than 700 mg per 1 ml of water; the compound of formula (1) is slightly soluble in water, with a solubility of less than 1.0 mg per 1 ml of water. The results are shown in Table 5.

[0125] Table 5. Solubility results of three crystal forms of the compound of formula (1) and the compound of formula (2)

[0126] Conclusion: The solubility of the crude product and three crystals of the compound of formula (1) and formula (2) in water differs significantly. The solubility of the compound of formula (1) in water is slightly soluble, which is not conducive to the preparation of aqueous solution preparations (such as inhalation solutions). On the other hand, the solubility of the crude product and three crystals of the compound of formula (2) in water is freely soluble.

[0127] Experimental Example 4: Relaxation effect of three crystals of the compound of formula (1) and the compound of formula (2) on the contraction reaction of guinea pig isolated tracheal smooth muscle induced by carbachol (CCh)

[0128] Preparation of isolated guinea pig tracheal smooth muscle specimens: Guinea pigs were anesthetized by intraperitoneal injection of 1.5 g / kg of ulanose (15% ulanose, 10 mL / kg intraperitoneally). After anesthesia, the trachea between the larynx and the carina was rapidly removed and placed in a mixture of 5% CO₂ and 95% O₂ in KH₄ solution. The loose connective tissue surrounding the trachea was separated and cut into tracheal rings approximately 3 mm wide. The cartilage was cut open and the ends were ligated with thread. The rings were placed in a McFadden bath containing 5 mL of KH₄ solution at 37°C and continuously aerated with a mixture of 5% CO₂ and 95% O₂. A muscle tension transducer was connected to the upper end of the tube. The initial resting tension of the tracheal segment was set at 1.0 g. Changes in muscle tension were recorded. The KH₄ solution (5 mL) was changed every 15-30 minutes. Experiments were initiated after the muscle tension of the tracheal segment stabilized.

[0129] Dosing method: After the muscle tension of the tracheal piece is stable, add a final concentration of 3×10 -6 After the contraction tension of the tracheal piece reached the plateau phase, the drug was added with a final concentration of 10 -11 , 10 -10 , 10 -9 , 3×10 -9 , 10 -8 , 3×10 -8 , 10 -7 , 10 -6 , 10 -5 , 10 -4 The three crystals of the compound of formula (1) and the compound of formula (2) (crystal I, crystal II and amorphous form) of M were prepared. The positive control was a mixture of ipratropium bromide (IPR) and salbutamol (SAL). The mixture IPR:SAL was 1:6. The concentration was calculated based on IPR. The highest concentration was only added to 10 -5 M, blank control group was added with solvent. The MedLab biological signal acquisition system was used to record the tension of the tracheal piece. If no reaction occurred (subthreshold concentration), the next dose was added in sequence. If a reaction occurred, the next dose was added after the relaxation plateau was reached. The final concentration was 3×10 -5 M isoproterenol (Iso) was added to achieve maximum relaxation and the curve was recorded, with the relaxation effect of Iso being regarded as 100% relaxation.

[0130] Data entry and statistical analysis: Excel and SPSS (Version 20) software package were used for statistical processing and EC was calculated. 50Data were expressed as mean ± standard error. Data were tested for homogeneity of variance. If the variances were homogeneous (P>0.05), a one-way analysis of variance was performed, and the Dunnet test was performed between each dose group and the control group. If the variances were unequal (P≤0.05), a nonparametric test was performed, and the Mann-Whitney U test was performed between each dose group and the control group. A P<0.05 was considered statistically significant when compared with the control group. When comparing two groups, an independent sample t-test was performed, and a P<0.05 was considered statistically significant.

[0131] Experimental results: Relaxation effect of compound of formula (1), crystal I of compound of formula (2), crystal II of compound of formula (2), amorphous form of compound of formula (2), and IPR+SAL mixture on tracheal smooth muscle EC 50 (95% confidence interval) The results are shown in Table 6.

[0132] Table 6. Relaxation effect of the example compounds on isolated tracheal smooth muscle EC 50

[0133] Conclusion: The compound of formula (1), three crystals of the compound of formula (2) (crystal I, crystal II and amorphous form) and IPR+SAL (mixture) all have a strong relaxing effect on the contraction of tracheal smooth muscle caused by CCh. There is no significant difference in the intensity of the effect of the compound of formula (1) and three crystals of the compound of formula (2) (crystal I, crystal II and amorphous form), and all are stronger than the positive control drug IPR+SAL mixture.

[0134] Experimental Example 5: Start-up time of the relaxation effect of the compound of formula (1), three crystals of the compound of formula (2) (crystal I, crystal II, and amorphous form), and the IPR+SAL mixture on the contractile response of isolated guinea pig tracheal smooth muscle induced by CCh

[0135] For the preparation of isolated guinea pig tracheal smooth muscle, see Experimental Example 4.

[0136] Dosing method: Prepared guinea pig tracheal smooth muscle samples, using the MedLab biological signal acquisition system to record changes in muscle tension, after the tracheal muscle tension stabilized, respectively, with a final concentration of 3×10 -6 M CCh induced contraction of tracheal smooth muscle samples. After the contraction tension of the tracheal slices reached a plateau, the EC values ​​of the three crystals (crystal I, crystal II and amorphous form) of the compound of formula (1), the compound of formula (2) and the IPR+SAL mixture (calculated in IPR) were obtained with reference to Experimental Example 4. 80 The concentrations are: 1.73×10 -8 , 1.69×10 -8 , 1.72×10 -8 , 1.75×10 -8 and 1.01×10-8 M. The final concentration was 3×10 -6 After M CCh induced contraction of tracheal smooth muscle samples and tension reached a plateau, the test drug was added to each test group to a concentration of EC 80 The control group was treated with KH solution. The tracheal segment tension was recorded using a MedLab biological signal acquisition system, with the plateau phase of maximum relaxation being taken as 100% relaxation, and the time to 10% relaxation being recorded and calculated as the start-up time.

[0137] For data entry and statistical analysis, see Experimental Example 4.

[0138] The test results are shown in Table 7.

[0139] Table 7. Onset time of relaxation of isolated tracheal smooth muscle by the example compounds

[0140] Conclusion: The compound of formula (1), the three crystals of the compound of formula (2) (crystal I, crystal II and amorphous form) and the IPR+SAL mixture all have a strong relaxing effect on the contraction of tracheal smooth muscle caused by CCh. The compound of formula (1), the three crystals of the compound of formula (2) (crystal I, crystal II and amorphous form) and the positive control drug IPR+SAL mixture all have a fast onset time of less than 5 minutes. There is no significant difference between the compound of formula (2) and the IPR+SAL mixture, but both are significantly faster than the compound of formula (1) (P < 0.05).

Claims

1. A compound represented by the structure of formula (2):

2. A method for preparing the compound of claim 1, comprising the following steps: (1) The reaction solvent in step 1 is selected from methanol, ethanol, N,N-dimethylformamide and N,N-dimethylacetamide, etc., preferably methanol, ethanol, etc.; the molar ratio of 1,3-dibromopropane to intermediate I is 5-10:1, preferably 8-10:1; the base used in the reaction is sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, etc., and the molar ratio of the base to intermediate I is 2-5:1, preferably 3:1; (2) The molar ratio of intermediate II in step 2 to (R)-(-)-3-[(R)-2-hydroxy-2-cyclopentyl-2-phenyl]ethoxy-1-azabicyclo[2,2,2]octane is 1-3:1, preferably 1.1:1; the reaction solvent is selected from methanol, ethanol, N,N- Dimethylformamide and N,N-dimethylacetamide, etc., preferably methanol, ethanol, etc.; (3) The molar ratio of intermediate III in step 3 to iron powder is 1:3-8, preferably 1:5; the solvent used in the reaction is selected from a 5%-10% aqueous solution of methanol, ethanol, tetrahydrofuran and acetone, preferably an aqueous solution of ethanol and methanol; (4) The molar ratio of the intermediate IV in step 4 to formic acid and acetic anhydride is 1:1-3:3-8, preferably 1:1.2:6, and the solvent is selected from formic acid and dichloromethane, preferably dichloromethane; (5) The reaction solvent in step 5 is selected from formic acid, acetic acid, methanol and ethanol, preferably formic acid.

3. The crystalline form I of the compound of claim 1, characterized in that: In the X-ray powder diffraction pattern, there are diffraction peaks at 2θ values ​​of 9.4±0.3, 14.7±0.3, 15.3±0.3, 18.6±0.3, 19.8±0.3, 20.8±0.3, and 25.1±0.

3.

4. Crystalline Form II of the compound of claim 1, characterized in that: In the X-ray powder diffraction pattern, there are diffraction peaks at 2θ values ​​of 3.5±0.3, 7.1±0.3, 10.7±0.3, 17.1±0.3, and 18.8±0.

3.

5. The amorphous form of the compound according to claim 1, characterized in that: It has the X-ray powder diffraction pattern characteristics shown in Figure 5.

6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 as an active ingredient.

7. Use of any compound according to claim 1 to 5 or any pharmaceutical composition according to claim 6 in the preparation of a medicament as a bronchodilator.

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