Preparation method of finerenone key intermediate and finished product

Abstract

The present invention relates to the field of pharmaceutical technologies, and specifically, to a preparation method of finerenone and its key intermediate. The present invention uses enzymatic resolution to prepare the key chiral intermediate (S)-compound of formula I of finerenone. The present invention has the advantages of high chiral selectivity in the reaction, being green and environmentally friendly, high yield, and not requiring the use of chromatographic separation technology, and is suitable for industrial production, etc.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically, to a method for preparing a key intermediate and a finished product of finerenone with high chiral purity using a hydrolase. Background Art

[0002] Finerenone is a non-steroidal selective mineralocorticoid receptor antagonist, which has been shown in preclinical studies to block the harmful effects caused by overactivation of the mineralocorticoid receptor. In diabetic patients, overactivation of the mineralocorticoid receptor is considered to lead to the progression of chronic kidney disease and cardiovascular impairment, which may be driven by factors such as metabolism, hemodynamics, or inflammation and fibrosis.

[0003] On July 9, 2021, based on the positive results of the FIDELIO-DKD Phase III clinical study in adult patients with chronic kidney disease and type 2 diabetes, the US FDA approved the marketing of finerenone ( ).

[0004] On December 22, 2021, the Committee for Medicinal Products for Human Use (CHMP) of the European Medicines Agency (EMA) recommended the approval of the marketing application for the non-steroidal selective mineralocorticoid receptor antagonist finerenone, and recommended finerenone (10 mg or 20 mg) for the treatment of adult patients with chronic kidney disease (stages 3 and 4 with albuminuria) and type 2 diabetes.

[0005] On June 29, 2022, the marketing application for finerenone declared by Bayer was approved for adult patients with chronic kidney disease related to type 2 diabetes, which can reduce the risk of continuous decline in estimated glomerular filtration rate (eGFR) and end-stage renal disease.

[0006] Multiple synthetic routes have been reported in the literature on finerenone. Among them, a compound patent discloses a method for preparing finerenone, and the synthetic route is as follows:

[0007]

[0008] Although the raw materials of this method are easy to prepare and the reaction conditions are relatively mild, the last step uses chromatographic separation technology, which has disadvantages such as low yield and poor atom economy. At the same time, chiral chromatographic separation is costly, difficult to operate, and requires high equipment investment, which is not conducive to cost control and industrial production of the product.

[0009] In view of the good market prospect of finerenone, it is necessary to develop an economical, safe, and more environmentally friendly method for preparing the key intermediate and finished product of finerenone. Summary of the Invention

[0010] The object of the present invention is to solve the deficiencies of the prior art and provide a more economical, environmentally friendly and safe biocatalytic technology for preparing finerenone and its intermediate compounds with high chiral purity.

[0011] The technical solution adopted by the present invention to solve the above technical problems is: A method for preparing finerenone and its key intermediate compound (S)-Formula I, which includes the following steps:

[0012] (1) The compound of Formula I is subjected to enzymatic hydrolysis resolution and then ester hydrolysis to obtain the compound of Formula II;

[0013]

[0014] (2) The compound of Formula II is subjected to ammonolysis to obtain finerenone;

[0015]

[0016] The present invention also relates to another method for preparing finerenone and its key intermediate, including the following steps:

[0017] (1) The compound of Formula Ia is subjected to enzymatic hydrolysis resolution and then ester hydrolysis to obtain the compound of Formula IIa;

[0018]

[0019] (2) The compound of Formula IIa reacts with triethyl orthoformate to obtain the compound of Formula IIIa;

[0020]

[0021] (3) The compound of Formula IIIa is subjected to hydrolysis to obtain the compound of Formula II;

[0022]

[0023] (4) The compound of Formula II is subjected to ammonolysis to obtain finerenone;

[0024]

[0025] In the embodiments of the present invention, R is C1-C8 alkyl, C3-C6 cycloalkyl, benzyl, alkoxy or cyanomethyl, cyanoethyl or cyanopropyl, preferably R is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, more preferably methyl, cyanomethyl, cyanoethyl or cyanopropyl.

[0026] In an embodiment of the present invention, the hydrolase is selected from Pseudomonas fluorescens, Aspergillus niger, Candida rugosa, Pseudomonas cepacia, Candida antarctica A, Candida antarctica B, Mucor miehei, Thermomyces lanuginosus, Humicola, Bacillus subtilis (Alcalase 2.4L), Bacillus subtilis (Savinase 12T), Mucor miehei, Gordonia polyisoprenivorans, Bacillus subtilis (Neutrase 0.8), porcine pancreas, preferably Gordonia polyisoprenivorans.

[0027] In an embodiment of the present invention, the dosage ratio of the compound of formula I to the hydrolase is 1 mmol:(100 - 500 mg), preferably 1 mmol:(200 - 500 mg).

[0028] In an embodiment of the present invention, in the enzymatic resolution reaction, a cosolvent that helps the enzymatic reaction is further added, preferably one or more of methyl tert-butyl ether, ethyl acetate, toluene, dimethyl sulfoxide, methanol, and more preferably methyl tert-butyl ether.

[0029] In an embodiment of the present invention, the temperature of the enzymatic resolution reaction is 0 - 60 °C, and the reaction time is 1 - 36 hours. Preferably, the reaction temperature is 20 - 35 °C, and the reaction time is 24 - 36 hours.

[0030] In an embodiment of the present invention, the form of the hydrolase is immobilized.

[0031] In an embodiment of the present invention, the pH value of the enzymatic resolution reaction is between 6 and 10, preferably 7 - 9.

[0032] Beneficial effects:

[0033] 1. The inventor first selected Gordonia polyisoprenivorans hydrolase to resolve Compound I. Through extensive research by the inventor, it was found that enzymatic resolution is the key step restricting the entire reaction route. Only by ensuring or controlling the purity and ee value of the (S)-Compound I can the purity of the final product finerenone and related substances be controlled, thereby meeting the quality standards for clinical use. The inventor conducted a detailed study on the enzymatic resolution conditions, focusing on factors such as the type of enzyme, conversion time, type of cosolvent, reaction temperature, dosage ratio of substrate to hydrolase, pH, etc. By further screening the commercially available enzymes, such as Pseudomonas fluorecens, Aspergillus niger, Candida rugosa, Pseudomonas cepacia, Candida antarctica A, Candida antarctica B, Mucor miehei, Thermomyces lanuginosus, Humicola, Bacillus subtilis (Alcalase 2.4L), Bacillus subtilis (Savinase 12T), Mucor miehei, Gordonia polyisoprenivorans, Bacillus subtilis (Neutrase 0.8), and porcine pancreas, the inventor surprisingly found that Gordonia polyisoprenivorans is the key factor affecting the resolution yield and ee value. Therefore, Gordonia polyisoprenivorans was selected as the resolving hydrolase.

[0034] 2. The S-Compound I obtained by resolving Compound I using Gordonia polyisoprenivorans has the advantages of high chiral selectivity, high yield, and does not require the use of chromatographic separation technology, making it suitable for industrial production. Description of the Drawings

[0035] Figure 1 1H NMR spectrum of Compound Ia-1

[0036] Figure 2 1H NMR spectrum of Compound IIa

[0037] Figure 3 1H NMR spectrum of Compound I-1

[0038] Figure 4 1H NMR spectrum of Compound II

[0039] Figure 5 Liquid chromatogram of S-form I compound Detailed implementation mode

[0040] The technical solutions of the present invention will be described below through specific embodiments, but the protection scope of the present invention is not limited thereto.

[0041] Example 1: Preparation of 2-cyanoethyl 4-(4-cyano-2-methoxyphenyl)-2,8-dimethyl-5-oxo-1,4,5,6-tetrahydro-1,6-naphthyridine-3-carboxylate (Ia-1):

[0042]

[0043] Add 20.0 g (124 mmol) of 4-formyl-3-methoxybenzonitrile (C), 24.0 g (155 mmol) of 2-cyanoethyl 3-oxobutyrate (A), 2.00 g (23.5 mmol) of piperidine, 1.47 g (24.5 mmol) of glacial acetic acid and 350 mL of dichloromethane to the reaction flask, and heat under reflux to separate water for 6.5 h. Cool the mixture to room temperature, and wash the organic phase successively with water and saturated brine. Dry the organic phase over anhydrous sodium sulfate. Filter, and concentrate the filtrate under reduced pressure to dryness to obtain 43.7 g of a brown-yellow oil. Dissolve the above-obtained residue in 520 mL of isopropanol, and add 11.2 g (90.0 mmol) of 4-amino-5-methylpyridone (D), and reflux the reaction (80 - 85 °C) for 22 h. Cool the mixture to room temperature. Filter with suction, and dry the wet product to obtain 27.2 g of a bright yellow powder.

[0044] Elute and column through with DCM / MeOH = 20:1 - 10:1, collect the positive fraction, and concentrate to dryness under reduced pressure at 45 °C to obtain 22.0 g (54.4 mmol) of a pale yellow solid, with a yield of 43.8%. MS: m / z = 405 [M+H]+

[0045] 1H NMR (400 MHz, DMSO-d6) δ = 10.81 (s, 1H), 8.21 (s, 1H), 7.38–7.30 (m, 2H), 7.24 (dd, 1H), 6.96 (s, 1H), 5.21 (s, 1H), 4.13 (m, 1H), 4.04 (m, 1H), 3.75 (s, 2H), 2.81 (m, 2H), 2.36 (s, 3H), 2.03 (s, 3H).

[0046] Example 2: Preparation of (S)-4-(4-cyano-2-methoxybenzyl)-2,8-dimethyl-5-oxo-1,4,5,6-tetrahydro-1,6-naphthyridine-3-carboxylic acid:

[0047]

[0048] 1.9 g (47 mmol) of compound (Ia-1) was dissolved in 45 ml of methyl tert-butyl ether in a reaction flask, 45 ml of 0.1 M PBS at pH 7.0 was added, 500 mg of Gordonia polyisoprenivorans was added, and the reaction was carried out at 37 °C and pH 7.5. The conversion rate was 51% after 24 hours and 53.6% after 48 hours. After the reaction was completed, the pH was adjusted to 9.0, the reaction solution was filtered, extracted with 400 ml × 2 of ethyl acetate, the organic phases were combined, and concentrated to obtain 0.82 g of a pale yellow solid, yield: 43.1%. The above solid was dissolved in 8 mL of tetrahydrofuran, 8 mL of sodium hydroxide solution (0.3 mol / L) was added dropwise, and the reaction was carried out at room temperature for 40 min. Hydrochloric acid was added to the system to adjust the pH to 4, 10 mL of dichloromethane was added, and the mixture was stirred and crystallized for 0.5 h, and then filtered to obtain a white solid. The solid was washed successively with water and dichloromethane. After filtration, the filter cake was dried to obtain 0.65 g of a white solid, with an overall yield of 39.4% and an ee value of 99.0%.

[0049] Method for measuring the conversion rate in the examples: Agilent ZORBAX 3.5um, SB-C18, 2.1×50mm; 0.8 ml / min; 40 °C; 254 nm, 82% MeOH (0 - 5.3 min), 100%

[0050] MeOH (5.3 min - 6.5 min), 82% MeOH (6.5 min - 10 min), conversion rate = product peak area / (residual compound I or Ia peak area + product peak area) × 100%. Method for measuring the ee value in the examples: Chiralpak IA chiral column, length: 250 nm, inner diameter: 4.6 nm, particle size: 5.0 um, conditions: 40 °C, 0.8 ml / min, 255 nm, eluent: A ethanol; B n-hexane, isogradient: A 10%; B 90%, ee value = (eeR - eeS) / (eeR + eeS) * 100%. MS: m / z = 352 [M + H]+, 1H-NMR (400 MHz, DMSO-d6) δ = 11.41 (s, 1H), 10.86 (s, 1H), 7.97 (s, 1H), 7.33 (s, 1H), 7.28–7.21 (m, 2H), 6.93 (s, 1H), 5.19 (s, 1H), 3.73 (s, 3H), 2.31 (s, 3H), 2.02 (s, 3H).

[0051] Example 3: Preparation of ethyl 2-cyano-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxylate (I-1):

[0052]

[0053] Add 4.04 g (10.0 mmol) of ethyl 2-cyano-4-(4-cyano-2-methoxyphenyl)-2,8-dimethyl-5-oxo-1,4,5,6-tetrahydronaphthalene-1,6-dicarboxylate (Ia-1), 4.40 g (27.2 mmol) of triethyl orthoacetate and 6.0 g of NMP (N-methylpyrrolidone) into a reaction flask. Then add a catalytic amount (0.30 g) of concentrated sulfuric acid. Heat the mixture to 115 °C and react for 3.0 h. Then cool the reaction solution to 50 °C. Slowly add 4 mL of water, stir until a small amount of solid precipitates, and then add 8 mL of water. Stir and cool to 0 °C, and stir at 0 °C for 2 h. Filter, and wash the filter cake with an appropriate amount of water. Dry the filter cake to obtain 3.5 g of a pale yellow solid, with a yield of 81.0%. MS: m / z = 433 [M+H]+, 1H NMR (400 MHz, Chloroform-d) δ 7.65 (s, 1H), 7.41 (d, 1H), 7.15 (m, 1H), 7.03 (d, 1H), 6.17 (s, 1H), 5.46 (s, 1H), 4.28–4.10 (m, 4H), 3.79 (s, 3H), 2.62 (m, 2H), 2.46 (s, 3H), 2.16 (s, 3H), 1.22 (t, 3H).

[0054] Example 4: Preparation of 4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydronaphthalene-1,6-dicarboxylate (II):

[0055]

[0056] Add 2.16 g (5 mmol) of Compound I-1 dissolved in 45 ml of methyl tert-butyl ether and 45 ml of 0.1 M PBS at pH 7.0. Add 500 mg of Gordonia polyisoprenivorans, react at 37 °C and pH 7.5. The conversion rate is 52.0% after 24 hours and 56.3% after 36 hours. Adjust the pH to 9.0 after the reaction ends. Filter the reaction solution, extract with ethyl acetate, combine the organic phases, and concentrate to obtain 0.92 g of a pale yellow solid, with an ee value of 99.5% and a yield of 42.6% (theoretical yield 50%).

[0057] To the above solid, add 6.0 mL of THF and 1.0 mL of water, stir and cool down to 0 °C. Dropwise add an aqueous sodium hydroxide solution. After the addition is complete, react at 0 °C for 4 h. After the reaction is completed, wash the reaction solution with methyl tert-butyl ether and discard the organic phase. Adjust the pH of the aqueous phase to ~7 with dilute hydrochloric acid at 0 °C. Heat the reaction solution to 20 °C, add an ammonium chloride solution and stir for 1 h, filter, wash with an appropriate amount of water, and dry the filter cake to obtain 0.72 g of a pale yellow solid, with an overall yield of 37.9% and an ee value of 99.5%. The calculated catalytic selectivity E value of Gordonia polyisoprenivorans for the substrate is 40. MS: m / z = 380 [M+H]+

[0058] 1H NMR (400 MHz, DMSO-d6) δ = 11.46 (s, 1H), 8.16 (s, 1H), 7.57 (s, 1H), 7.32 (s, 1H), 7.27 (m, 2H), 5.34 (s, 1H), 4.14–3.95 (m, 2H), 3.74 (s, 3H), 2.38 (s, 3H), 2.15 (s, 3H), 1.12 (t, 3H).

[0059] Measurement method for conversion rate in the examples: Agilent ZORBAX 3.5 um, SB-C18, 2.1×50 mm; 0.8 ml / min; 40 °C; 254 nm, 82% MeOH (0 - 5.3 min), 100% MeOH (5.3 min - 6.5 min), 82% MeOH (6.5 min - 10 min), conversion rate = product peak area / (residual compound I or Ia peak area + product peak area) × 100%.

[0060] Measurement method for ee value in the examples: Chiralpak IA chiral column, length: 250 nm, inner diameter: 4.6 nm, particle size: 5.0 um, conditions: 40 °C, 0.8 ml / min, 255 nm, eluent: A ethanol; B n-hexane gradient: A 10%; B 90%, ee value = (eeR - eeS) / (eeR + eeS) * 100%.

[0061] Example 5: Resolution effects of different types of hydrolases

[0062] Adopt the same preparation method as in Example 4 to screen the resolution effects of different hydrolases on the compound of formula -I

[0063]

[0064] Example 6: Comparison of the resolution effects of Gordonia polyisoprenivorans and AK lipase of Pseudomonas fluorescens

[0065] Using the same preparation method as in Example 4, the AK lipase of Pseudomonas fluorescens was used to replace Gordonia polyisoprenivorans for comparison of the resolution effect.

[0066] Types of enzymes Yield ee value of S-form-1 compound Gordonia polyisoprenivorans Yield 42.6% ee value 99.5% AK lipase of Pseudomonas fluorescens Yield 29% ee value 70%

Claims

1. A method for preparing a finerenone intermediate, characterized in that It includes the following steps: (1) The compound of formula I is subjected to enzymatic resolution under the action of a hydrolase to obtain the S-compound of formula I; wherein, R is C1-C8 alkyl, benzyl or cyanomethyl, cyanoethyl or cyanopropyl; wherein the hydrolase is Gordonia polyisoprenivorans.

2. The method according to claim 1, wherein: R is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl.

3. The method according to claim 1, wherein the dosage ratio of the compound of formula I to the hydrolase is 1 mmol:(100-500 mg).

4. The method according to claim 1, wherein: In the enzymatic resolution reaction, a cosolvent that helps the enzymatic catalytic reaction is further added.

5. The method according to claim 4, wherein the cosolvent is selected from one or more of methyl tert-butyl ether, ethyl acetate, toluene, dimethyl sulfoxide, methanol.

6. The method according to any one of claims 1 to 5, characterized in that: The temperature of the enzymatic resolution reaction is 0-60 °C, and the reaction time is 1-36 hours.

7. The method according to claim 6, characterized in that: The reaction temperature is 20-35 °C, and the reaction time is 24-36 hours.

8. The method according to any one of claims 1 to 5, characterized in that: The form of the hydrolase described is immobilized.

9. The method according to any one of claims 1 to 5, characterized in that: The pH value of the enzymatic resolution reaction is between 6 and 10.

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