New nucleoside phosphoramidate compound compositions and methods for their preparation

Abstract

The present application belongs to the field of pharmaceutical preparations, and relates to an oral preparation comprising (a) a compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal thereof; and (b) an adsorbent. The present application also relates to a preparation method of the oral preparation of the present application and application of the oral preparation of the present application in medicines for preventing and / or treating hepatitis C. The oral preparation of the present application can be completely dissolved, well absorbed in vivo, and has few impurities and good stability.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparations, specifically relating to oral formulations of (2S)-2-((([1,1′-biphenyl]-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoyl)amino)propionate, their preparation methods, and applications. Background Technology

[0002] Hepatitis C virus (HCV) infection is a globally prevalent disease, with over 200 million chronic carriers worldwide. Egypt has a chronic infection rate of 15%, Pakistan 4.8%, and China 3.2%, ranking among the top three globally. The clinical manifestations of HCV infection are diverse, ranging from mild inflammation to severe cirrhosis and liver cancer. Chronic hepatitis C can also lead to various extrahepatic complications, including rheumatoid arthritis, keratoconjunctivitis sicca, lichen planus, glomerulonephritis, mixed cryoglobulinemia, B-cell lymphoma, and porphyria cutanea latentis, possibly due to abnormal immune responses. Furthermore, in the decompensated stage of hepatitis C cirrhosis, various complications can occur, such as ascites and peritoneal infection, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal syndrome, and liver failure.

[0003] HCV belongs to the genus *Hepatitis Virus* of the family Flaviviridae, and its genetic structure is similar to that of two other genera within the Flaviviridae family, namely *Phexavira* and *Flavaviri*. Currently, the standard treatment for HCV infection includes interferon and a combination of interferon and ribavirin. However, only 50% of patients respond to this treatment, and interferon has significant side effects, such as flu-like symptoms, weight loss, and fatigue. The combination of interferon and ribavirin produces considerable side effects, including hemolysis, anemia, and fatigue.

[0004] In addition, developed drugs for treating HCV infection include protease inhibitors, thiazolidinediones, thiazolidinediones and N-benzoylaniline, phenanthrenequinone, helicase inhibitors, nucleoside polymerase inhibitors and mycotoxins, antisense phosphate-thionucleotide oligonucleotides, IRES-dependent translation inhibitors, ribozymes, and nucleoside analogs. Currently, the use of nucleoside phosphate compounds for treating flaviviridae viruses, especially HCV infection, is an important research direction in this field.

[0005] CN104031104A discloses a novel nucleoside aminophosphate compound as shown in formula (1), with the chemical name (2S)-2-((([1,1′-biphenyl]-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl (hereinafter referred to as "compound of formula (1)").

[0006]

[0007] CN104031104A reports that isopropyl (2S)-2-((([1,1′-biphenyl]-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoyl)amino)propionate has excellent antiviral activity and low cytotoxicity, and can be used to treat flaviviridae viruses, especially hepatitis C virus infection.

[0008] To date, no suitable formulations containing this compound or its pharmaceutically acceptable salts, isomers, solvates, hydrates, or crystals have been reported. Isopropyl (2S)-2-((([1,1′-biphenyl]-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phospho)amino)propionate is a prodrug that exerts its antiviral effect by conversion to an active metabolite in the liver; therefore, oral administration may be a more suitable dosage form. However, it is well known in the art that many problems exist in the formulation of such drugs, such as long-term stability, control of related substances, drug absorption, and bioavailability. These problems are determined by a variety of factors, including many factors affecting drug absorption, such as drug dissolution or release from the formulation, drug solubility under physiological conditions, and permeability in the gastrointestinal tract.

[0009] During the initial formulation research, the inventors of this invention discovered that the solubility and permeability of the compound of formula (1) need to be improved, and it is easy to aggregate in water, which brings great challenges to the formulation research of this type of compound.

[0010] Therefore, there is a need to provide oral formulations of isopropyl (2S)-2-((([1,1′-biphenyl]-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoyl)amino)propionate that are completely soluble, well absorbed in vivo, have few impurities, low hygroscopicity, and good stability. Summary of the Invention

[0011] One object of the present invention is to provide an oral formulation comprising a compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal thereof, which is capable of complete dissolution and has good stability.

[0012] Another object of the present invention is to provide a method for preparing the oral formulation of the present invention.

[0013] Another object of the present invention is to provide the use of the oral formulation of the present invention in the preparation of a medicament for the prevention and / or treatment of Flaviviridae viruses, especially HCV infection.

[0014] The above-mentioned objectives are achieved through the following technical solutions.

[0015] The present invention provides an oral formulation comprising (a) a compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal thereof; and (b) an adsorbent.

[0016] This invention provides a compound of formula 1a, which is a stereoisomer of the compound of formula 1, chemically named (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate isopropyl ester, suitable for the treatment of patients with hepatitis C virus infection.

[0017]

[0018] The inventors of this invention used the methods of Experimental Examples 1 and 2 in CN105985355A to determine the inhibitory activity of the compound of Formula 1a against HCV 1b and 1a replicons. The results showed that the compound of Formula 1a had significantly better inhibitory activity against HCV virus than the compound of Formula 1.

[0019] In some embodiments, the oral formulation of the present invention comprises (a) a compound of formula (1a) or a pharmaceutically acceptable salt, solvate, hydrate or crystal thereof; and (b) an adsorbent.

[0020] In the oral formulations of this invention, the combination of any form or amorphous compound of formula (1a) with an adsorbent yields formulations with excellent stability. Although amorphous compounds exhibit metastable properties, limiting their commercial application, the inventors of this invention unexpectedly discovered that after accelerated and long-term testing for 6 months, the formulation properties remained highly stable, with no significant changes observed in any of the indicators. Furthermore, the oral formulations of this invention prepared using the crystalline form of the compound of formula (1a) are also highly stable.

[0021] Therefore, in some specific embodiments, the oral formulation according to the present invention is wherein the compound of formula (1a) is a crystalline or amorphous form.

[0022] Preferably, the oral formulation of the present invention is in solid form. In some embodiments, the oral formulation of the present invention includes tablets, capsules, pills, granules, powders, pellets, lozenges, and flat capsules. In specific embodiments, the oral formulation of the present invention is a tablet, capsule, or granule. In some embodiments, the oral formulation of the present invention further includes a disintegrant, a lubricant, and / or a filler.

[0023] In some embodiments, the disintegrant is selected from one or more of corn starch, hydroxypropyl starch, carboxymethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, crospovidone, sodium carboxymethyl starch, sodium crospovidone carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, and pregelatinized starch. More preferably, the disintegrant is selected from one or more of sodium crospovidone carboxymethyl cellulose, sodium carboxymethyl starch, crospovidone, and low-substituted hydroxypropyl cellulose.

[0024] In some embodiments, the lubricant is selected from one or more of magnesium stearate, stearic acid, calcium stearate, zinc stearate, sodium stearate fumarate, talc, and polyethylene glycol.

[0025] In some embodiments, the filler is selected from mannitol, starches, sugars, celluloses, and inorganic salts.

[0026] In some embodiments, the oral formulation according to the invention comprises, based on the amount of the compound of formula (1), the compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal thereof constitutes about 15% to 85% (weight / weight) of the tablet weight, the adsorbent constitutes about 15% to 85% (weight / weight) of the tablet weight, the disintegrant constitutes about 0% to 20.0% (weight / weight) of the tablet weight, the lubricant constitutes about 0% to 15.0% (weight / weight) of the tablet weight, and the filler constitutes about 0% to 30% (weight / weight) of the tablet weight.

[0027] In some embodiments, the oral formulation according to the invention comprises, based on the amount of the compound of formula (1), the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal, comprising about 20% to 50% (by weight) of the tablet, the adsorbent comprising about 30% to about 60% (by weight) of the tablet, the disintegrant comprising about 5% to 15.0% (by weight) of the tablet, the lubricant comprising about 0.2% to 10.0% (by weight) of the tablet, and the filler comprising about 10% to 20% (by weight) of the tablet.

[0028] In other embodiments, the oral formulation according to the invention comprises, based on the amount of the compound of formula (1), the compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal thereof, comprising about 30% to about 50% (by weight) of the tablet, the adsorbent comprising about 30% to about 50% (by weight) of the tablet, the disintegrant comprising about 5% to 15.0% (by weight) of the tablet, the lubricant comprising about 0.5% to 5.0% (by weight) of the tablet, and the filler comprising about 10% to 20% (by weight) of the tablet.

[0029] The adsorbent of the present invention can be a porous material with adsorption properties. For example, in some embodiments, the adsorbent is selected from crospovidone, microcrystalline cellulose, lactose, cyclodextrin, low-substituted hydroxypropyl cellulose, and silica.

[0030] In one embodiment, crospovidone is used as an adsorbent, preferably crospovidone with a particle size of 10-30 μm.

[0031] In another embodiment, cyclodextrin is used as an adsorbent.

[0032] In another embodiment, low-substituted hydroxypropyl cellulose is used as an adsorbent. Low-substituted hydroxypropyl cellulose can be added internally during wet granulation in tablet production, or externally, or both simultaneously for better results.

[0033] In other embodiments, inorganic acid salts or inorganic salts are used as adsorbents. Inorganic acid salts include sodium phosphate, calcium phosphate, potassium sulfate, sodium dihydrogen phosphate, and disodium hydrogen phosphate; inorganic salts include sodium chloride, calcium chloride, and potassium chloride.

[0034] In addition, diatomaceous earth, porous cellulose and its salts, starch, sugars and other pharmaceutical excipients can also be used as adsorbents in this invention.

[0035] This invention provides a method for preparing an oral formulation of the present invention, comprising the step of combining the components of the oral formulation of the present invention. The inventors of the present invention have discovered that in the formulation of the present invention, dissolving the compound of formula (1) in an easily soluble solvent and then preparing the formulation results in more complete dissolution.

[0036] In some embodiments, according to the preparation method of the formulation of the present invention, the compound of formula (1) is dissolved in an easily soluble solvent, then mixed with an adsorbent, and then granulated, tableted, and coated to prepare an oral formulation. In other embodiments, according to the preparation method of the formulation of the present invention, the compound of formula (1) and the adsorbent are dissolved in an easily soluble solvent, then granulated, tableted, and coated to prepare an oral formulation.

[0037] In some embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0038] a) Mix the adsorbent thoroughly in the device;

[0039] b) Dissolve the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal in a solvent with stirring; and

[0040] c) Add the mixture from step b) to the adsorbent from step a), dry, and then prepare particles.

[0041] In some specific embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0042] a) Mix the adsorbent thoroughly in the device;

[0043] b) Dissolve the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal in a solvent with stirring;

[0044] c) Add the mixture from step b) to the adsorbent from step a), and dry it in a fluidized bed to obtain granules;

[0045] e) Optionally, the dried granules are mixed evenly with the disintegrant, lubricant, and filler;

[0046] f) Compress the mixed granules into tablets; and

[0047] g) Coating.

[0048] In some specific embodiments, according to the above-described method for preparing oral formulations of the present invention, the solvent in step b) is an alcohol solvent, preferably selected from one or more of methanol, ethanol, and isopropanol. In a more preferred embodiment, the compound of formula (1) is dissolved in ethanol.

[0049] In some specific embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0050] a) Mix the prescribed amount of adsorbent evenly in a wet granulator;

[0051] b) Dissolve the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal in an alcohol solvent with stirring;

[0052] c) Add the mixture from step b) to the adsorbent from step a) to prepare wet granules;

[0053] d) The wet particles are dried using a fluidized bed;

[0054] e) Optionally, the dried granules are mixed evenly with the disintegrant, lubricant, and filler;

[0055] f) Compress the mixed granules into tablets; and

[0056] g) Coating.

[0057] In some embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0058] a) Dissolving a compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or crystal and adsorbent thereof in a solvent to form a mixed solution; and

[0059] b) The mixed solution is dried and then granulated.

[0060] In some specific embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0061] a) Dissolve the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal and adsorbent in a solvent to form a uniformly dispersed mixed solution;

[0062] b) The mixed solution is dried into a mixed powder using a spray drying device;

[0063] c) Mixing the powder with disintegrants and lubricants; and

[0064] d) Tableting.

[0065] In some other embodiments, the oral formulation of the present invention is prepared by the following method, the method comprising:

[0066] a) Dissolve the compound of formula (1) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal and adsorbent in a solvent to form a uniformly dispersed mixed solution;

[0067] b) The mixed solution is dried into a mixed powder using a fluidized bed dryer;

[0068] c) Mixing the powder with disintegrants and lubricants; and

[0069] d) Tableting.

[0070] In one specific embodiment of the present invention, the inventors of the present invention added the compound of formula (1a) or its pharmaceutically acceptable salt, isomer, solvate, hydrate or crystal and silica to ethanol and stirred to disperse evenly. Then, the mixed solution was dried to prepare a mixed powder using a spray drying device. Then, it was mixed with a disintegrant and a lubricant and directly compressed into tablets.

[0071] In some embodiments, the solvent of the present invention is preferably selected from alcohol solvents and diethylene glycol monoethyl ether.

[0072] In one embodiment, the oral formulation according to the present invention uses crospovidone and microcrystalline cellulose with a particle size of 3-100 μm as adsorbents, and crospovidone sodium carboxymethyl cellulose or sodium carboxymethyl starch as disintegrants. Preferably, crospovidone sodium carboxymethyl cellulose is used as the disintegrant.

[0073] The inventors of this invention have discovered that when crospovidone is used as an adsorbent in the oral formulation of this invention, the dissolution endpoint changes with variations in the particle size of the crospovidone. Preferably, this invention uses crospovidone with a particle size of 10-30 μm. Preferably, the coating material comprises hydroxypropyl methylcellulose, lactose, and triacetin. According to the formulation of this invention, preferably, the coating accounts for 2-4% (w / w) of the total weight of the formulation.

[0074] In this invention, the effect of pH on the dissolution profile of the formulation was also investigated. The results showed that the formulation of this invention exhibited good dissolution and stability over a relatively wide pH range. At the same pH value, such as in dissolution media of pH 1.2, pH 3.0, pH 4.5, or pH 6.8 buffer solutions, the batch-to-batch dissolution profiles of the formulation of this invention were consistent, with small batch-to-batch differences, indicating a relatively stable process.

[0075] This invention provides the use of the oral formulation of this invention in the preparation of medicaments for the treatment and / or prevention of Flaviviridae viruses, especially HCV infection.

[0076] When the oral formulation of the present invention is administered, the compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or crystal thereof is converted in vivo to its active form to exert its antiviral effect. In one embodiment, the present invention provides a method for treating, preventing, or controlling HCV, the method comprising orally administering to a patient the formulation of the present invention, wherein the amount of the compound of formula (1) or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or crystal thereof is molarly equal to the therapeutically effective amount of the compound of formula (1).

[0077] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0078] In this document, the term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt formed by the compound of the present invention with an acid, wherein the acid may be selected from: phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, citric acid, maleic acid, malonic acid, mandelic acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-toluenesulfonic acid, malic acid, tartaric acid, amino acids, methanesulfonic acid, or analogs thereof.

[0079] The term "solvent" refers to the form of the compounds of this invention that form solid or liquid complexes by coordination with solvent molecules. Hydrates are a special form of solvate in which coordination with water occurs. Within the scope of this invention, hydrates are preferred solvates.

[0080] The term "crystallization" refers to the various solid forms formed by the compounds described in this invention, including crystalline and amorphous forms. The term "uncoated tablet" refers to tablets that are not coated, as described in this invention.

[0081] The oral formulation of the present invention has good stability, can be stored for a long time, dissolves completely, is well absorbed in the body, has few impurities, low hygroscopicity, and simplifies storage and transportation conditions. Attached Figure Description

[0082] Figure 1 Configuration confirmation diagram of isopropyl propionate (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate.

[0083] Figure 2Dissolution curves of the formulation obtained in Example 9 are shown below, where pH 1.2 represents the dissolution curve of the formulation obtained in Example 9 in a pH 1.2 buffer of 0.05% sodium dodecyl sulfate; pH 3.0 represents the dissolution curve of the formulation obtained in Example 9 in a pH 3.0 buffer; pH 4.5 represents the dissolution curve of the formulation obtained in Example 9 in a pH 4.5 buffer; and pH 6.8 represents the dissolution curve of the formulation obtained in Example 9 in a pH 6.8 buffer. Detailed Implementation

[0084] The present invention will be further described in detail below with reference to the embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, all materials used in the following embodiments are commercially available, and the film coating premix is ​​Opadry (295K620013), a gastrointestinal coating premix powder purchased from Calcare Company.

[0085] Example 1: Preparation of isopropyl (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate

[0086]

[0087] Step 1: Synthesis of (S)-2-[[(1,1′-biphenyl-4-oxy)(pentafluorophenoxy)phosphoryl]amino]isopropyl propionate

[0088]

[0089] In a 50L glass reactor under nitrogen protection, phosphorus oxychloride (1.53 kg, 10 mol) and dichloromethane (10 L) were added, stirred, and cooled to below -30°C. A solution of triethylamine (1.01 kg, 10 mol) in dichloromethane (5 L) was added dropwise, maintaining the internal temperature below -30°C during the addition. After the addition was complete, a solution of 4-hydroxybiphenyl (1.7 kg, 10 mol) in tetrahydrofuran (3.4 L) was slowly added dropwise, stirring for 30 min after the addition was finished. Maintaining the internal temperature below -30°C, L-alanine isopropyl hydrochloride (1.68 kg, 10 mol) was added, followed by a solution of triethylamine (2.02 kg, 20 mol) in dichloromethane (8.0 L), stirring for 30 min. The temperature was raised to about 0°C, and a solution of pentafluorophenol (1.84 kg, 10 mol) and triethylamine (1.01 kg, 10 mol) in dichloromethane (7.0 L) was added dropwise. During the dropwise addition, the reaction was exothermic and the temperature rose. After the dropwise addition was completed, the internal temperature was raised to room temperature. After the reaction was complete, the mixture was filtered, and the filter cake was washed with 2.5 L × 4 of dichloromethane. The filtrates were combined and washed with 25 L of purified water. The mixture was separated, and the organic phase was concentrated under reduced pressure. 5 L × 2 of heptane was added as solvent, and the mixture was centrifuged, dried under vacuum, and weighed to obtain the title compound as a white solid.

[0090] Step 2: Synthesis of isopropyl (S)-2-[[(S)-(1,1′-biphenyl-4-oxy)(pentafluorophenoxy)phosphoryl]amino]propionate]

[0091]

[0092] In a 30L reactor under nitrogen protection, isopropyl (S)-2-[[(1,1'-biphenyl-4-oxy)(pentafluorophenoxy)phosphoryl]amino]propionate (3.70 kg, 7.0 mol), methyl tert-butyl ether (2.40 kg), n-heptane (9.0 kg), triethylamine (77 g, 0.7 mol), and pentafluorophenol (43 g, 0.21 mol) were added. The mixture was heated to 40°C and vigorously stirred mechanically. After the reaction was complete, the mixture was centrifuged and dried under vacuum to obtain the title compound.

[0093] Step 3: Synthesis of isopropyl propionate (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate

[0094]

[0095] In a 50L glass reactor, 1.30 kg (5.0 mol) of 1-[(2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-hydroxymethyl-3-methyltetrahydrofuran-2-yl]pyrimidine-2,4-(1H,3H)-dione product and 15.6 L of tetrahydrofuran were added. The mixture was stirred, protected with nitrogen, and cooled to below -10°C. 8.0 L (8.0 mol) of tert-butylmagnesium chloride solution was added dropwise. After the addition was complete, the reaction was continued at 0–5°C for 1 h. Add 2.12 kg (4.0 mol) of isopropyl (S)-2-[[(S)-(1,1'-biphenyl-4-oxy)(pentafluorophenoxy)phosphoryl]amino]propionate product dropwise. After the addition is complete, react at 5–10 °C. Take a sample after 10–11 h and monitor the reaction with HPLC. After the reaction is complete, cool to below 0 °C and add 4.0 L (8.0 mol) of 2N hydrochloric acid to the reaction solution to quench the reaction. Control the temperature during quenching at 0–10 °C. After the addition is complete, stir for 10 min. Concentrate under reduced pressure, evaporate to dryness to obtain a solid-liquid residue, add 40.0 L of ethyl acetate and 20.0 L of purified water, and separate the layers. The organic phase was washed with 10.0 L of 5% sodium carbonate aqueous solution, then washed with water, filtered, and evaporated to dryness. 7.0 L of dichloromethane was added again, and the mixture was concentrated to obtain an oily foaming material. Isopropyl acetate (15.0 L) was added to the oily foaming material, and after dissolution, the mixture was cooled, resulting in the precipitation of a large amount of white precipitate. The precipitate was cooled, filtered, and the filter cake was dried. The mixture was then dissolved in isopropyl acetate and heated with stirring. Activated carbon was added, and the mixture was stirred for 30 min. The mixture was filtered while hot, cooled, filtered again, and the filter cake was dried to obtain a white solid.

[0096] ESI-MS m / z: 604.2 [M+H]-.

[0097] 1 HNMR (300MHz, DMSO-d6) δ: 11.48 (s, 1H), 7.67 (d, 2H), 7.64-7.62 (m, 2H), 7.59 (d, 1H), 7.47 (t, 2H), 7.37 (t, 1H), 7.33 (d, 2H), 6.04 (d, 1H), 6.02 (d, 1H), 5.8 3(d, 1H), 5.57(d, 1H), 4.89-4.84(m, 1H), 4.42-4.39(m, 1H), 4.29-4.24(m, 1H ), 4.05-4.02(m, 1H), 3.88-3.82(m, 2H), 1.29-1.24(m, 6H), 1.16-1.15(m, 6H).

[0098] Example 2: Configuration confirmation of isopropyl (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate

[0099] Weigh 15 mg of the compound of formula 1a prepared in Example 1 into a 3 mL vial, add 5 mL of a mixed solvent system of dichloromethane / n-heptane with a volume ratio of 2:1, shake to obtain a clear solution, cover the vial with sealing film and punch holes in it, and place at room temperature for 6 days to obtain crystals.

[0100] The single-crystal samples obtained by the above method were subjected to X-ray single-crystal diffraction data collection and their single-crystal structures were analyzed. Table 1 lists the single-crystal structure data and structural refinement parameters of the n-heptane solvate of Formula 1a. The single-crystal structure analysis determined the absolute configuration of the chiral center in the compound of Formula 1a, i.e., as shown in Table 1. Figure 2 As shown, P1(S), C20(R), C21(R), C22(R), C24(R), C13(S). Its asymmetric structural unit contains two molecules of compound 1a and two n-heptane molecules.

[0101] Table 1: Single Crystal Structure Data of Compound 1a

[0102]

[0103] Example 3: Solid oral formulation of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]isopropyl propionate

[0104] Table 2. Prescription

[0105] Cross-linked polyvinylpyrrolidone 38.46 microcrystalline cellulose 15.38 lactose 12.82 Cross-linked carboxymethyl cellulose sodium 7.69 total 100.00

[0106] Unit: weight % (w / w).

[0107] Preparation steps:

[0108] a. Mix the prescribed amount of compound (1a) and cross-linked polyvinyl ketone and microcrystalline cellulose evenly in a wet granulator, add an appropriate amount of ethanol, and prepare wet granules;

[0109] b. Dry the wet granules;

[0110] c. Mix the dried granules with lactose and croscarmellose sodium;

[0111] d. Fill the capsules with the mixed granules.

[0112] The formulation of this embodiment can rapidly disintegrate and disperse uniformly in in vitro dissolution experiments.

[0113] Examples 4-6: Solid oral formulations of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]amino]propionate isopropyl ester

[0114] This embodiment examines the in vitro dissolution of formulations prepared with different proportions of adsorbent, using the ability to disperse as an indicator of in vitro dissolution.

[0115] Table 3. Prescription

[0116]

[0117]

[0118] Unit: weight % (w / w)

[0119] Preparation steps:

[0120] a) Mix the compound of formula (1a) with silicon dioxide, microcrystalline cellulose, lactose, croscarmellose sodium, and magnesium stearate until homogeneous;

[0121] b) Tableting.

[0122] Experimental results show that the formulations in Examples 4 to 6 can all rapidly disintegrate and disperse uniformly in in vitro dissolution experiments.

[0123] Example 7: Solid oral formulation of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]isopropyl propionate

[0124] This embodiment mainly examines the in vitro dissolution of formulations prepared by spray drying, with the time required for complete dissolution as the indicator.

[0125] Table 4. Prescription

[0126] silicon dioxide 33.33 microcrystalline cellulose 4.47 lactose 17.87 Cross-linked carboxymethyl cellulose sodium 10.00 magnesium stearate 1.00 total 100.00

[0127] Unit: weight % (w / w)

[0128] Preparation steps:

[0129] a) Add an appropriate amount of ethanol to the prescribed amount of compound (1a) and silicon dioxide and stir to form a uniformly dispersed mixed solution. Use a spray drying device to dry the mixed solution to obtain a mixed powder of compound (1a) and silicon dioxide.

[0130] b) Mix the compound of formula (1a) and silicon dioxide powder with microcrystalline cellulose, lactose, cross-linked sodium carboxymethyl cellulose and magnesium stearate evenly;

[0131] c) Tableting.

[0132] Experimental results show that the formulation prepared by spray drying can be completely dissolved in 10 minutes in the in vitro dissolution test.

[0133] Example 8. Solid oral formulation of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]isopropyl propionate

[0134] This embodiment mainly examines whether the adsorbent, being an inorganic acid salt, can be completely dissolved.

[0135] Table 5. Prescription

[0136] Sodium phosphate 33.33 microcrystalline cellulose 11.67 lactose 10.67 Cross-linked carboxymethyl cellulose sodium 10.00 magnesium stearate 1.00 total 100.00

[0137] Unit: weight % (w / w)

[0138] Preparation steps:

[0139] a) Mix the prescribed amount of compound (1a) with sodium phosphate, microcrystalline cellulose, lactose, croscarmellose sodium, and magnesium stearate until homogeneous;

[0140] b) Tableting.

[0141] The results show that the formulation prepared in this embodiment can rapidly disintegrate and completely dissolve in the in vitro dissolution experiment.

[0142] Example 9: Solid oral formulation of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]isopropyl propionate

[0143] Table 6. Prescription

[0144]

[0145]

[0146] Unit: weight % (w / w)

[0147] Preparation steps:

[0148] a) Mix the adsorbent cross-linked polyvinyl ketone and microcrystalline cellulose evenly;

[0149] b) The compound of formula (1a) was dissolved in an appropriate amount of ethanol by stirring and then slowly added to the adsorbent to obtain wet granules;

[0150] c) The wet particles from step b) are dried using a fluidized bed, with the inlet air temperature controlled at approximately 60℃±5℃ and the material temperature controlled at approximately 50℃.

[0151] d) Mix the dried granules from step c) with croscarmellose sodium, lactose, and magnesium stearate until homogeneous;

[0152] e) Compress the mixed powder into tablets using a tablet press;

[0153] f) Place the uncoated film in a coating machine and spray in a film coating premix for coating.

[0154] Comparative Example 1: Solid oral formulation of (S)-2-[[[(S)-(1,1'-biphenyl-4-oxy)]-[((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]isopropyl propionate

[0155] This embodiment examines the solubility of a formulation prepared using conventional excipients in an in vitro dissolution experiment.

[0156] Table 7. Prescription

[0157] Cross-linked carboxymethyl cellulose sodium 14.00 magnesium stearate 1.00 total 100.00

[0158] Unit: weight % (w / w).

[0159] Preparation steps:

[0160] a) Mix the prescribed amount of compound (1a) with cross-linked sodium carboxymethyl cellulose and magnesium stearate until homogeneous;

[0161] b) Tableting.

[0162] Experimental results show that the formulation prepared in this embodiment disintegrated in the in vitro dissolution experiment and then agglomerated at the bottom of the dissolution vessel, preventing the release of the drug.

[0163] Experimental Example 1: Solubility of the formulation of the present invention

[0164] The formulation prepared according to Example 9 was tested for dissolution using the dissolution assay method (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0931, Method II). The dissolution medium was 1000 ml of pH 1.2, pH 3.0, pH 4.5, and pH 6.8 buffer solutions containing 0.05% sodium dodecyl sulfate, and the rotation speed was 50 rpm. Samples were taken at 5, 10, 15, 20, 30, 45, and 60 minutes for analysis. The analysis was performed using high-performance liquid chromatography (High-Performance Liquid Chromatography, Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0512). An octadecylsilane-bonded silica column was used, with a column temperature of 30°C, a UV / DAD detector at a detection wavelength of 254 nm, a mobile phase of water-methanol (35:65), and a flow rate of 1.0 ml / min. Dissolution curves were plotted based on the experimental data, as shown below. Figure 2 As shown.

[0165] Experiments show that the formulation of the present invention can be completely dissolved in all the above-mentioned dissolution media, which meets the requirements.

[0166] Experimental Example 2: Stability of the Formulation of the Present Invention

[0167] The formulation prepared according to Example 9 was placed under accelerated conditions of 40℃±2℃ and 75%±5% relative humidity, and under long-term conditions of 25℃±2℃ and 60%±10% relative humidity for 6 months. Dissolution was determined according to the dissolution test method (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0931, Method II), using 1000 ml of pH 1.2 buffer containing 0.05% sodium dodecyl sulfate as the dissolution medium, and a rotation speed of 50 rpm. Related substances and contents were determined according to high-performance liquid chromatography (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0512), using an octadecylsilane-bonded silica column at 30℃, a UV / DAD detector at 254 nm, and a water-methanol (35:65) mobile phase. The experimental results are shown in Table 8.

[0168] Table 8. Formulation stability data of the present invention

[0169] 0 days 101.7％ 96.8％ 0.13％ Accelerated testing for 1 month (40℃±2℃, RH 75%±5%) 101.9％ 99.1％ 0.14％ Accelerated testing for 2 months (40℃±2℃, RH 75%±5%) 100.5％ 100.8％ 0.11％ Accelerated testing for 3 months (40℃±2℃, RH 75%±5%) 100.7％ 100.8％ 0.11％ Accelerated testing for 6 months (40℃±2℃, RH 75%±5%) 101.0％ 97.3％ 0.14％ Long-term test for 3 months (25℃±2℃, RH 60%±10%) 100.5％ 98.1％ 0.13％ Long-term test for 6 months (25℃±2℃, RH 60%±10%) 100.6％ 97.3％ 0.18％

[0170] From the above experimental results, it can be seen that the (S)-2-[[[(S)-(1,1'-biphenyl-4-oxo]

[0171] The isopropyl propionate formulation [((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy]phosphoryl]amino]propionate was fully dissolved in four dissolution media containing 0.05% sodium dodecyl sulfate in pH 1.2, pH 3.0, pH 4.5 and pH 6.8 buffers. After accelerated dissolution for 6 months and long-term dissolution for 6 months, the formulation remained stable with few impurities and no significant changes in any of the indicators, meeting the requirements.

[0172] Although the present invention has been described in detail above, those skilled in the art will understand that various modifications and changes can be made to the present invention without departing from its spirit and scope. The scope of the present invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. An oral formulation comprising a compound of formula (1a) or a pharmaceutically acceptable salt thereof, (1a); and an adsorbent, wherein the adsorbent is selected from one or more of crospovidone, microcrystalline cellulose, cyclodextrin, and diatomaceous earth.

2. The formulation according to claim 1, wherein the compound of formula (1a) is a crystalline or amorphous form.

3. The formulation according to claim 1 or 2, wherein the formulation is in solid form.

4. The formulation according to claim 3, wherein the formulation is a tablet or capsule.

5. The formulation according to claim 1, 2 or 4, further comprising a disintegrant, a lubricant and / or a filler.

6. The formulation according to claim 3, further comprising a disintegrant, a lubricant, and / or a filler.

7. The formulation according to claim 1 or 2, wherein, based on the amount of the compound of formula (1a), the compound of formula (1a) or a pharmaceutically acceptable salt thereof accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the adsorbent accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the disintegrant accounts for 0% to 20.0% (weight / weight) of the tablet weight of the formulation, the lubricant accounts for 0% to 15.0% (weight / weight) of the tablet weight of the formulation, and the filler accounts for 0% to 30% (weight / weight) of the tablet weight of the formulation.

8. The formulation according to claim 3, wherein, based on the amount of the compound of formula (1a), the compound of formula (1a) or a pharmaceutically acceptable salt thereof accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the adsorbent accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the disintegrant accounts for 0% to 20.0% (weight / weight) of the tablet weight of the formulation, the lubricant accounts for 0% to 15.0% (weight / weight) of the tablet weight of the formulation, and the filler accounts for 0% to 30% (weight / weight) of the tablet weight of the formulation.

9. The formulation according to claim 4 or 6, wherein, based on the amount of the compound of formula (1a), the compound of formula (1a) or a pharmaceutically acceptable salt thereof accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the adsorbent accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the disintegrant accounts for 0% to 20.0% (weight / weight) of the tablet weight of the formulation, the lubricant accounts for 0% to 15.0% (weight / weight) of the tablet weight of the formulation, and the filler accounts for 0% to 30% (weight / weight) of the tablet weight of the formulation.

10. The formulation according to claim 5, wherein, based on the amount of the compound of formula (1a), the compound of formula (1a) or a pharmaceutically acceptable salt thereof accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the adsorbent accounts for 15% to 85% (weight / weight) of the tablet weight of the formulation, the disintegrant accounts for 0% to 20.0% (weight / weight) of the tablet weight of the formulation, the lubricant accounts for 0% to 15.0% (weight / weight) of the tablet weight of the formulation, and the filler accounts for 0% to 30% (weight / weight) of the tablet weight of the formulation.

11. A method for preparing a formulation according to any one of claims 1-10, comprising the step of combining the components of the formulation according to any one of claims 1-10.

12. The method of claim 11, comprising granulating a compound of formula (1a) or a pharmaceutically acceptable salt thereof and an adsorbent.

13. The method of claim 12, wherein the granulation comprises the following steps: a) Mix the adsorbent thoroughly in the device; b) Dissolve the compound of formula (1a) or a pharmaceutically acceptable salt thereof in a solvent with stirring; and c) Add the mixture from step b) to the adsorbent from step a), dry, and then prepare particles.

14. The method of claim 12, wherein the granulation comprises the following steps: a) Dissolving the compound of formula (1a) or its pharmaceutically acceptable salt and adsorbent in a solvent to form a mixed solution; and b) The mixed solution is dried and then granulated.

15. The method of claim 13 or 14, wherein the solvent is an alcohol solvent.

16. The method of claim 15, wherein the alcohol solvent is selected from one or more of methanol, ethanol and isopropanol.

17. Use of the formulation of any one of claims 1-10 in the preparation of a medicament for the treatment and / or prevention of hepatitis C virus infection.

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