A composition and its preparation method and use

Abstract

The present invention relates to a composition and a preparation method and use thereof, belonging to the field of pharmaceutical preparations. The composition comprises an ATV014 compound and a pharmaceutically acceptable excipient, has good stability, high solubility, and high bioavailability, and can be used to prevent, alleviate or treat coronavirus infection or the replication or reproduction of its homologous variant virus and the resulting cytopathic effect.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and in particular to a composition and a preparation method and use thereof. Background Art

[0002] The new coronavirus is an enveloped single-stranded RNA virus belonging to the genus β coronavirus. Similar to SARS and MERS, the SARS-CoV-2 genome encodes nonstructural proteins: 3-chymotrypsin-like protease (3CLpro), papain-like protease (PLpro), helicase, and RNA-dependent RNA polymerase (RdRp); structural proteins: such as spike glycoprotein and accessory proteins. The surface spike glycoprotein of the new coronavirus binds to the angiotensin-converting enzyme (ACE2) receptor on the surface of human cells to infect human respiratory epithelial cells. After entering the host cell, the virus disintegrates, releasing the nucleocapsid and viral RNA into the cytoplasm. The open reading frame (ORF1a / b) at the 5ʹ end of the viral RNA will encode polyproteins (pp1a and pp1ab), which play an important role in the processing and maturation of enzymes required for viral replication. Pp1a and pp1ab can be cleaved by papain-like proteases (PLpro) and 3C-like proteases (3CLpro) to produce non-structural proteins, including RNA-dependent RNA polymerase and helicase, which play a key role in the transcription and replication of the new coronavirus. At present, the surface spike glycoprotein of the coronavirus recognition receptor, the important protein 3CLpro involved in the replication and transcription process, PLpro and RdRp are four very attractive targets for the development of antiviral drugs.

[0003] Regarding the development of the COVID-19 vaccine, the UK first approved the emergency use of the Pfizer and BioNTech COVID-19 vaccine on December 2. On the one hand, the general effectiveness of this vaccine is still unknown, and on the other hand, the strict low-temperature storage requirements have brought great inconvenience to its widespread use.

[0004] Regarding the development of new crown drugs, Remdesivir is currently the only new crown drug approved by the US FDA. Remdesivir is a prodrug of aminomethyl monophosphate of an adenosine analog, originally developed by Gilead for the anti-Ebola virus. As an RdRp inhibitor, Remdesivir has shown anti-new crown virus activity at the cellular level, but clinical trials have shown that Remdesivir does not significantly reduce mortality in humans. And because the clinical dosage is close to the safe dosage, some obvious side effects have to attract attention.

[0005] Therefore, there is still a need for a drug with good safety, stability and bioavailability to treat the new coronavirus. Summary of the invention SUMMARY OF THE INVENTION

[0007] To solve the above problems, in a first aspect, the present invention provides a composition comprising an ATV014 compound and a pharmaceutically acceptable excipient.

[0008] . In some embodiments, the pharmaceutically acceptable excipients may include at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients. In some embodiments, the diluent is at least one of microcrystalline cellulose, mannitol and pregelatinized starch. In some embodiments, the diluent is microcrystalline cellulose and mannitol. In some embodiments, the diluent is microcrystalline cellulose or mannitol. Using microcrystalline cellulose or mannitol as a diluent is more conducive to improving the compatibility of raw materials and excipients, preferably microcrystalline cellulose or mannitol as a diluent; more preferably microcrystalline cellulose PH102 or mannitol 50C as a diluent; most preferably mannitol 50C as a diluent.

[0009] In a second aspect, the present invention provides a use of the aforementioned composition in the preparation of a product for preventing, alleviating or treating coronavirus infection or the replication or reproduction of its homologous variant viruses and the cytopathic effects produced therefrom.

[0010] In a third aspect, the present invention provides a method for preparing the aforementioned composition, wherein the method adopts grinding or crushing ATV014 to reduce the particle size of ATV014, which is beneficial to improving the dissolution rate of the product and also beneficial to making the particle hardness moderate during the dry granulation process. The preparation method adopts the ground ATV014 for dry granulation, which is beneficial to obtaining particles with better hardness. The preparation method adopts dry granulation, which is beneficial to improving the disintegration rate of the product. DETAILED DESCRIPTION OF THE INVENTION

[0012] In order to solve the above problems, the present invention provides a composition and a pharmaceutical preparation, use and preparation method thereof.

[0013] In a first aspect, a composition is provided.

[0014] A composition comprising: an ATV014 compound and a pharmaceutically acceptable excipient,

[0015] .

[0016] In some embodiments, the pharmaceutically acceptable excipients may include at least one selected from: diluents, disintegrants, binders, lubricants and other excipients. In some embodiments, the pharmaceutically acceptable excipients include diluents, disintegrants, binders and lubricants.

[0017] In certain embodiments, the diluent may include at least one selected from microcrystalline cellulose, mannitol, calcium hydrogen phosphate and pregelatinized starch. In certain embodiments, the diluent is at least one of microcrystalline cellulose, mannitol and pregelatinized starch. In certain embodiments, the diluent is microcrystalline cellulose and mannitol. In certain embodiments, the diluent is microcrystalline cellulose or mannitol. Using microcrystalline cellulose or mannitol as diluent is conducive to improving the compatibility of raw materials and auxiliary materials. In some more selected embodiments, the dilution is microcrystalline cellulose PH102 or mannitol 50C, which is more conducive to improving the compatibility of raw materials and auxiliary materials. In some most preferred embodiments, the diluent is mannitol 50C, which is most conducive to the compatibility of raw materials and auxiliary materials.

[0018] In some embodiments, the disintegrant may include at least one selected from cross-linked sodium carboxymethylcellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and hydroxypropyl cellulose. In some embodiments, the disintegrant is cross-linked sodium carboxymethylcellulose.

[0019] In some embodiments, the binder may include at least one selected from hydroxypropyl cellulose, povidone and starch. In some embodiments, the binder is hydroxypropyl cellulose.

[0020] In some embodiments, the lubricant may include at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate. In some embodiments, the lubricant is magnesium stearate.

[0021] In some embodiments, the other excipients may include glidants.

[0022] In some embodiments, the glidant may include colloidal silicon dioxide or talc.

[0023] In some embodiments, the content of the ATV014 compound can be 15wt%-70wt% based on the total mass of the composition. In some embodiments, the content of the ATV014 compound is 50wt%-60wt% based on the total mass of the composition. In some embodiments, the content of the ATV014 compound is 15wt%, 20wt%, 25wt%, 30wt%, 35wt%, 40wt%, 45wt%, 50wt%, 55wt%, 56wt%, 60wt%, 65wt% or 70wt% based on the total mass of the composition.

[0024] In some embodiments, the content of the diluent may be 20wt%-70wt% based on the total mass of the composition. In some embodiments, the content of the diluent is 30wt%-40wt% based on the total mass of the composition. In some embodiments, the content of the diluent is 20wt%, 25wt%, 30wt%, 35wt%, 36wt%, 37wt%, 40wt%, 45wt%, 50wt%, 55wt%, 60wt%, 65wt% or 70wt% based on the total mass of the composition.

[0025] In some embodiments, the content of the disintegrant can be 1wt%-10wt% based on the total mass of the composition. In some embodiments, the content of the disintegrant is 2wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the disintegrant is 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt% or 10wt% based on the total mass of the composition.

[0026] In some embodiments, the content of the binder can be 1wt%-10wt% based on the total mass of the composition. In some embodiments, the content of the binder is 2wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the binder is 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt% or 10wt% based on the total mass of the composition.

[0027] In some embodiments, the content of the lubricant can be 0.5wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the lubricant is 1.5wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the lubricant is 0.5wt%, 1wt%, 1.5wt%, 2wt%, 3wt%, 4wt% or 5wt% based on the total mass of the composition.

[0028] In some embodiments, the composition may not include a surfactant. In some embodiments, the composition may not include sodium lauryl sulfate.

[0029] In some embodiments, the composition may further include an additional lubricant.

[0030] In some embodiments, the additional lubricant comprises at least one selected from magnesium stearate, sodium stearate and sodium stearyl fumarate. In some embodiments, the additional lubricant is magnesium stearate.

[0031] In some embodiments, the content of the added lubricant is 0.5wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the added lubricant is 1.5wt%-5wt% based on the total mass of the composition. In some embodiments, the content of the added lubricant is 0.5wt%, 1wt%, 1.5wt%, 2wt%, 3wt%, 4wt% or 5wt% based on the total mass of the composition.

[0032] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent comprises at least one selected from microcrystalline cellulose, mannitol, calcium hydrogen phosphate and pregelatinized starch, the disintegrant comprises at least one selected from cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and hydroxypropyl cellulose, the binder comprises at least one selected from hydroxypropyl cellulose, polyvinylpyrrolidone and starch, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 15wt%-70wt%, the content of the diluent is 20wt%-70wt%, the content of the disintegrant is 1wt%-10wt%, the content of the binder is 1wt%-10wt%, and the content of the lubricant can be 0.5wt%-5wt%.

[0033] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent comprises at least one selected from microcrystalline cellulose, mannitol, calcium hydrogen phosphate and pregelatinized starch, the disintegrant comprises at least one selected from cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and hydroxypropyl cellulose, the binder comprises at least one selected from hydroxypropyl cellulose, polyvinylpyrrolidone and starch, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant can be 1.5%-5wt%.

[0034] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent comprises at least one selected from microcrystalline cellulose and mannitol, the disintegrant comprises cross-linked sodium carboxymethyl cellulose, the binder comprises hydroxypropyl cellulose, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant can be 1.5%-5wt%.

[0035] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent is microcrystalline cellulose PH102 or mannitol 50C, the disintegrant comprises cross-linked sodium carboxymethyl cellulose, the binder comprises hydroxypropyl cellulose, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant can be 1.5%-5wt%.

[0036] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent is microcrystalline cellulose PH102 and mannitol 50C, the disintegrant comprises cross-linked sodium carboxymethyl cellulose, the binder comprises hydroxypropyl cellulose, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant can be 1.5%-5wt%.

[0037] In some embodiments of the present invention, a composition comprises: an ATV014 compound and a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises at least one selected from: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent is mannitol 50C, the disintegrant comprises cross-linked sodium carboxymethyl cellulose, the binder comprises hydroxypropyl cellulose, the lubricant comprises at least one selected from magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant can be 1.5%-5wt%.

[0038] In some embodiments, the dosage form of the composition can be an oral solid preparation or an injection. In some embodiments, the dosage form of the composition is a tablet, a granule or a capsule.

[0039] In some embodiments, the dosage form may be in a strength of 10 mg to 500 mg (e.g., 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg). In some embodiments, the dosage form is in a strength of 15 mg to 300 mg. In some embodiments, the dosage form is in a strength of 20 mg to 200 mg. In some embodiments, the dosage form is in a strength of 50 mg to 200 mg. In some embodiments, the dosage form is in a strength of 50 mg.

[0040] Thirdly, a use is provided.

[0041] A use of the composition of the first aspect in preparing a medicine.

[0042] In some embodiments, a composition described in the first aspect is used in the preparation of a product for preventing, alleviating or treating coronavirus infection or the replication or reproduction of its homologous variant viruses and the cytopathic effects produced therefrom.

[0043] In some embodiments, the infection comprises fever, cough, sore throat, pneumonia, acute respiratory tract infection, severe acute respiratory tract infection, hypoxemic respiratory failure and acute respiratory distress syndrome, sepsis or septic shock.

[0044] In some embodiments, a composition described in the first aspect is used in the preparation of a product for detecting a coronavirus or a homologous variant virus thereof.

[0045] In some embodiments, the coronavirus includes: MHV-A59, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, SARS-CoV-2, mouse hepatitis virus, feline infectious peritonitis virus, canine coronavirus, bovine coronavirus, avian infectious bronchitis virus or porcine coronavirus. Preferably, the SARS-CoV-2 includes a mutant or unmutated strain of SARS-CoV-2; more preferably, the mutant strain of SARS-CoV-2 includes SARS-CoV-2 mutant strain B.1, SARS-CoV-2 mutant strain B.1.351, SARS-CoV-2 mutant strain B.1.617.2, SARS-CoV-2 mutant strain C.37, SARS-CoV-2 mutant strain P.1 family tree, SARS-CoV-2 mutant strain B.1.525, SARS-CoV-2 mutant strain B.1.427 or SARS-CoV-2 mutant strain B.1.429.

[0046] In some embodiments, the composition is suitable for use in humans or animals. In some embodiments, the animal comprises bovine, equine, ovine, porcine, canine, feline, rodent, primate, avian, or piscine.

[0047] In a fourth aspect, a method for preparing the composition of the first aspect is provided.

[0048] A method for preparing the composition of the first aspect comprises: mixing the ATV014 compound and pharmaceutically acceptable excipients, dry granulating, adding an external lubricant, mixing, tableting or capsule filling or bagging to obtain the composition.

[0049] In some embodiments of the present invention, a method for preparing the composition of the first aspect comprises: mixing the ATV014 compound and a pharmaceutically acceptable excipient, wet granulating, grinding, drying, dry grinding, adding an external lubricant, mixing, tableting or capsule filling or bagging to obtain the composition.

[0050] In some embodiments of the present invention, a method for preparing the composition of the first aspect comprises: grinding or pulverizing the ATV014 compound and mixing it with a pharmaceutically acceptable excipient, dry granulating, adding an external lubricant, mixing, tableting or capsule filling or bagging to obtain the composition.

[0051] In some embodiments of the present invention, a method for preparing the composition of the first aspect comprises: grinding or pulverizing the ATV014 compound and mixing it with a pharmaceutically acceptable excipient, wet granulating, grinding, drying, dry grinding, adding an external lubricant, mixing, tableting or capsule filling or bagging to obtain the composition.

[0052] In some embodiments, the particle size D90 of the ATV014 may be ≤500μm. In some embodiments, the particle size D90 of the ATV014 is ≤300μm. In some embodiments, the particle size D90 of the ATV014 is ≤200μm. In some embodiments, the particle size D90 of the ATV014 is ≤100μm. In some embodiments, the particle size D90 of the ATV014 is ≤50μm. In some preferred embodiments, the particle size D90 of the ATV014 is ≤25μm. In some preferred embodiments, the particle size D90 of the ATV014 is ≤20μm. In some more preferred embodiments, the particle size D90 of the ATV014 is ≤10μm.

[0053] Beneficial Effects

[0054] Compared with the prior art, a certain embodiment of the present invention has at least one of the following beneficial technical effects:

[0055] (1) The formulation and formulation ratio adopted in the present invention are beneficial to improving dissolution and bioavailability.

[0056] (2) The prescription and prescription ratio adopted in the present invention are beneficial to improving the stability of the drug under conditions of high temperature, high humidity, light, etc.

[0057] (3) The prescription and prescription ratio adopted by the present invention can be used to prevent, alleviate or treat coronavirus infection or the replication or reproduction of its homologous variant virus and the resulting cytopathic effect.

[0058] (4) Using a method without adding surfactant is beneficial to increasing the dissolution rate of the product.

[0059] (5) The content of lubricant (e.g., magnesium stearate) is preferably 1.5% or more, which is beneficial to the hardness and smoothness of the granules produced by dry granulation.

[0060] (6) The content of the binder (e.g., hydroxypropyl cellulose) is preferably 3% or more, which is beneficial to the hardness and smoothness of the dry granulated particles.

[0061] (7) The content of the binder (e.g., hydroxypropyl cellulose) is preferably about 3%, which is beneficial to the dissolution rate of the product.

[0062] (8) Grinding or crushing ATV014 to reduce the particle size of ATV014 is beneficial to increasing the dissolution rate of the product and also to making the particle hardness moderate during the dry granulation process.

[0063] (9) Using ground ATV014 for dry granulation is beneficial to obtaining particles with better hardness.

[0064] (10) Dry granulation is beneficial to improving the disintegration rate of the product.

[0065] (11) Compared with other diluents, using microcrystalline cellulose or mannitol as a diluent is more conducive to improving the compatibility of raw materials and excipients. Microcrystalline cellulose or mannitol is preferably used as the diluent, microcrystalline cellulose PH102 or mannitol 50C is more preferred, and mannitol 50C is most preferred.

[0066] Definition of terms:

[0067] In the present invention, "room temperature" refers to ambient temperature, which may be 20°C-30°C; in some embodiments, 22°C-28°C; in some embodiments, 24°C-26°C; in some embodiments, 25°C.

[0068] In the above text of the present invention, regardless of whether the words "about" or "approximately" are used, all the numbers disclosed herein are approximate values. Based on the disclosed numbers, the value of each number may vary by less than ±10% or reasonable differences considered by those skilled in the art, such as ±1%, ±2%, ±3%, ±4% or ±5%.

[0069] The term "optional", "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur. For example, "optional surfactant" means that a surfactant may or may not be present.

[0070] The term "external lubricant" refers to a substance added after granulation to reduce the friction between the granules obtained by granulation, prevent the raw materials and excipients from sticking to the punch surface, or reduce the friction between the tablet and the wall of the die hole.

[0071] The term "strength" refers to the weight of active ingredient in one unit of dosage form (a single tablet or capsule).

[0072] The term "weight percent" or "percent by weight" or "wt %" is defined as the weight of a single component in a composition divided by the total weight of all components of the composition multiplied by 100.

[0073] The term "and / or" should be understood to mean any one of the optional items or a combination of any two or more of the optional items.

[0074] The term "wt%" means percent by mass.

[0075] As used herein, the term "treatment" refers to clinical intervention intended to alter the natural course of a disease in the individual being treated. Desired therapeutic effects include, but are not limited to, preventing the onset or recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of a disease, preventing metastasis, reducing the rate of disease progression, improving or alleviating the disease state, and alleviating or improving prognosis.

[0076] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0077] In this application, "composition" can be conveniently presented in unit dosage form and can be prepared by any of the methods well known in the pharmaceutical art. All methods include the step of combining the active ingredient with a carrier that constitutes one or more accessory ingredients. Usually, the composition is prepared by uniformly and thoroughly combining the active compound with a liquid carrier, a finely divided solid carrier, or both.

[0078] In the present invention, the abbreviations of some compounds represent the compound structures:

[0079] Table 1:

[0080] DETAILED DESCRIPTION

[0081] In order to enable those skilled in the art to better understand the technical solution of the present invention, some non-limiting embodiments are further disclosed below to further illustrate the present invention in detail.

[0082] The reagents used in the present invention can be purchased from the market or prepared by the method described in the present invention.

[0083] "#0" indicates capsule shell No. 0. "#4" indicates capsule shell No. 4. "API" indicates API, which refers to compound ATV014 in the following specific examples. "rpm" indicates the speed unit "revolutions per minute". "SDS" indicates sodium dodecyl sulfate. "h" indicates hour. "min" indicates minute. "μM" indicates micromoles per liter, i.e., μmol / L. "mL" or "ml" indicates milliliter. "Compound RDV" or "RDV" indicates Remdesivir.

[0084] Example 1: Preparation of ATV014

[0085]

[0086] 1.50 g of compound 5 was dissolved in 15 ml of dichloromethane, and then 0.42 mL of cyclohexanecarboxylic acid and 55.40 mg of 4-dimethylaminopyridine were added, and after stirring for 10 min, 1.02 g of dicyclohexylcarbodiimide was added, and stirred at room temperature for 24 h. After column chromatography separation (eluent: petroleum ether / ethyl acetate (V / V) = 1 / 1), compound 7 was obtained.

[0087] 1.50 g of compound 7 was dissolved in 3 mL of 37% hydrochloric acid aqueous solution and 15 mL of tetrahydrofuran, stirred for 6 hours, and then sodium carbonate was added to adjust the pH to 8. The organic solvent was removed by rotary evaporation, and separated by column chromatography (eluent: petroleum ether / ethyl acetate (V / V) = 1 / 3) to obtain 0.28 g of compound ATV014 (white solid, yield 45.8%). The obtained compound ATV014 was tested for hydrogen and carbon spectra, and the results were as follows:

[0088] Hydrogen spectrum: 1H NMR (600 MHz, DMSO-d6) δ (ppm): 7.92 (s, 1H), 7.86 (br, 1H), 6.92 (d, J=4.5 Hz, 1H), 6.81 (d, J=4.5 Hz, 1H), 6.33 (d, J=5.9 Hz, 1H), 5.38(d, J=5.9 Hz, 1H), 4.70 (t, J=5.3 Hz, 1H), 4.32-4.29 (dd, J=12.2 Hz, 2.6 Hz,1H), 4.24-4.21 (m, 1H), 4.16-4.13 (dd, J=12.3 Hz, 4.8 Hz, 1H), 3.98-3.95 (q,J=5.9 Hz, 1H), 2.26-2.22 (m, 1H), 1.75-1.72 (m, 2H), 1.64-1.56 (m, 3H), 1.30-1.12 (m, 5H).

[0089] Carbon spectrum: 13 C NMR (150 MHz, DMSO-d6) δ (ppm): 175.34, 156.06, 148.4,124.0, 117.4, 117.0, 110.7, 101.2, 81.7, 79.4, 74.5, 70.6, 63.0, 42.6, 29.0,28.9, 25.7, 25.2, 25.1.

[0090] Example 2: Inhibitory effect of compounds on SARS-CoV-2 replicon in HEK293T cells

[0091] Compound GS-441524, ATV014 or ATV014 intermediate 5 were respectively used as test compounds and the following steps were performed:

[0092] HEK293T cells were seeded in 24-well plates. When the cells grew to a density of 40-50%, 250 ng of SARS-CoV-2 replicon plasmid was transfected by LIPO2000 (liposome 2000). After 6-8 h of transfection, the cell supernatant was discarded, and fresh DMEM medium was replaced. The test compounds were added to a final concentration of 50 μM, 10 μM, 5 μM, 2 μM, 1 μM, 0.1 μM or 0.01 μM, respectively. After 60 h of transfection, the cell supernatant was discarded, and TRIZOL was used to collect cell RNA. Total RNA was extracted and cDNA was obtained by reverse transcriptase. Finally, the internal reference gene Gapdh and SARS-CoV-2 N gene subgenome in cDNA were detected by fluorescent quantitative PCR to reflect the viral replication in the SARS-CoV-2 replicon, and the inhibitory effect of different concentrations of drugs on the virus was calculated, and the IC of the drug was calculated. 50 , the results are shown in Table 2.

[0093] Table 2: Inhibitory effects of compounds on SARS-CoV-2 replicon in HEK293T cells

[0094]

[0095] Conclusion: The test compounds inhibited the replication of SARS-CoV-2 in HEK293T cells to varying degrees. The activity of ATV014 was 3.65 times that of compound GS-441524, and the activity was significantly improved. The inhibitory effects of different compounds on SARS-CoV-2 replicon in HEK293T cells are shown in Table 2.

[0096] Example 3: Inhibitory effect of compounds on SARS-CoV-2 in Vero-E6 cells

[0097] Take compounds RDV, GS-441524, and ATV014 as test compounds, and perform the operation according to the following steps:

[0098] Vero-E6 cells were seeded in 48-well plates. When the cell density was about 70-80%, the supernatant was discarded and replaced with fresh DMEM medium, and then each compound was added to the medium to a final concentration of 50 μM, 10 μM, 5 μM, 2 μM, 1 μM, 0.5μM, 0.25μM, 0.1μM or 0.01μM. Cells were infected with three SARS-CoV-2 mutants (B.1, B.1.351 and B.1.617.2) at a multiplicity of infection (MOI) of 0.05. Antiviral activity was assessed by quantitative real-time polymerase chain reaction (qRT-PCR) to quantify the number of viral copies in the supernatant 48 hours after infection. We calculated the inhibitory effects of different concentrations of the tested drugs on viral replication and calculated their IC50 values. The IC50 of different compounds against SARS-CoV-2 in Vero-E6 cells is shown in Table 3.

[0099] Table 3: Inhibitory effects of different compounds on SARS-CoV-2 in Vero-E6 cells

[0100]

[0101] Example 4: Investigation of surfactants (preparation method: wet granulation)

[0102] Capsules were prepared according to the formulation described in Table 4.

[0103] Table 4: Prescription ratio

[0104]

[0105] Preparation method: Grind ATV014 (prescription 37181-037-B does not require grinding), and sieve ATV014 and other components, mix the internal components according to the prescription ratio, add water and wet granulate, grind, dry, dry grind, add external components, mix, and fill with capsules to obtain capsules.

[0106] Dissolution test: The obtained capsules were taken and 900 mL of 0.1N hydrochloric acid was used as the medium. The dissolution was tested at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes and 75 minutes by using the paddle method at a speed of 75 rpm. The results are shown in Table 5.

[0107] Table 5: Dissolution test results

[0108]

[0109] Result analysis:

[0110] (1) Using a formulation without adding surfactants (such as sodium dodecyl sulfate (SDS)) is more conducive to improving the dissolution rate.

[0111] (2) Grinding the API helps to increase the dissolution rate.

[0112] (3) By grinding the API and not adding surfactants (such as sodium dodecyl sulfate (SDS)), the two can synergize and greatly improve the dissolution rate.

[0113] Example 5: Preparation of API by air flow milling (wet granulation)

[0114] Capsules were prepared according to the formulation described in Table 6.

[0115] Table 6: Prescription ratio

[0116]

[0117] Preparation method: grind ATV014, and sieve ATV014 and other components, mix the internal components according to the prescription ratio, add water and wet granulate, grind, dry, dry grind, add external components, mix, fill with capsules, and obtain capsules.

[0118] Dissolution test: The obtained capsules were taken and 900 mL of 0.1 N hydrochloric acid was used as the medium. The dissolution was tested at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes and 75 minutes by using the paddle method at a speed of 75 rpm. The results are shown in Table 7.

[0119] Table 7: Dissolution test results

[0120]

[0121] Result analysis:

[0122] 1. The formulation containing jet milled API and without SDS has a fast dissolution rate and a high dissolution platform.

[0123] Example 6: Effect of ATV014 grinding on dissolution (preparation method: dry granulation)

[0124] Capsules were prepared according to the formulation described in Table 8.

[0125] Table 8: Prescription ratio

[0126]

[0127] Preparation method: Grind ATV014 (prescription 37181-027B does not require grinding), and sieve ATV014 and other components, mix the internal components according to the prescription ratio, dry granulate, add the external components, mix, and fill with capsules to obtain capsules.

[0128] Dissolution test: The obtained capsules were taken and 900 mL of 0.1 N hydrochloric acid was used as the medium. The dissolution was tested at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes and 75 minutes by using the paddle method at a speed of 75 rpm. The results are shown in Table 9.

[0129] Table 9: Dissolution results

[0130]

[0131] Result analysis:

[0132] (1) Compared with wet granulation, dry granulation is more conducive to improving product dissolution.

[0133] (2) Grinding of API helps improve the dissolution rate of the product.

[0134] Example 7: Effect of ATV014 grinding on dry granulation particles and disintegration time (preparation method is dry granulation or direct filling)

[0135] Capsules were prepared according to the formulation described in Table 10.

[0136] Table 10: Prescription ratio

[0137]

[0138] Preparation method of prescription 37181-020-B: grind ATV014, and sieve ATV014 and other components, mix the internal components according to the prescription ratio, dry granulate, add the external components, mix, fill with capsules, and obtain capsules.

[0139] Preparation method of prescription 37181-019-B: After sieving unground ATV014 and other components separately, the internal components are mixed according to the prescription ratio, and then the external components are added, mixed, and filled into capsules to obtain capsules.

[0140] Preparation method of prescription 37181-021-B: Grind ATV014, and sieve ATV014 and other components, mix the internal components according to the prescription ratio, add the external components, mix, and fill them with capsules to obtain capsules.

[0141] Observation of dry granulation phenomenon: Granulation was carried out using the dry granulation process parameters shown in Table 11. During the granulation process, the material properties were observed. The results are shown in Table 11.

[0142] Table 11: Observation results of dry granulation phenomena

[0143]

[0144] Summary: Using ground ATV014 for dry granulation is beneficial to obtaining particles with better hardness.

[0145] Disintegration time investigation: The capsules obtained in this example were taken, and the disintegration time of the capsules was tested according to General Rules 0921 of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The results are shown in Table 12.

[0146] Table 12: Disintegration time test results

[0147]

[0148] Conclusion: The capsules prepared by grinding API and dry granulation followed by filling have a fast disintegration time, and dry granulation is beneficial to improving the disintegration rate of the product.

[0149] Example 8: Investigation of binder and lubricant content

[0150] Capsules were prepared according to the formulation described in Table 13.

[0151] Table 13: Prescription ratio

[0152]

[0153] Preparation method: grind ATV014, and sieve ATV014 and other components, mix the internal components according to the prescription ratio, dry granulate, add the external components, mix, and fill with capsules to obtain capsules.

[0154] Observation of dry granulation phenomenon: Granulation was carried out using the dry granulation process parameters shown in Table 14. During the granulation process, the material properties were observed. The results are shown in Table 14.

[0155] Table 14: Observation of dry granulation phenomena

[0156]

[0157] summary:

[0158] 1. The magnesium stearate content is preferably 1.5wt% or more, which is beneficial to the hardness and smoothness of the particles in dry granulation.

[0159] 2. The hydroxypropyl cellulose content is preferably 3 wt% or more, which is beneficial to the hardness and smoothness of the particles in dry granulation.

[0160] Dissolution test: Roller pressure is 6-12 kg / cm 2The obtained capsules were tested for dissolution at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes and 75 minutes using a paddle method at a speed of 75 rpm using 900 mL of 0.1 N hydrochloric acid as a medium. The results are shown in Table 15.

[0161] Table 15: Dissolution Results

[0162]

[0163] Conclusion: The capsules obtained from prescriptions 37181-047-B and 37181-051-B can be dissolved quickly and completely, among which the release rate of prescription 37181-051-B is faster.

[0164] Example 9: Capsules of different specifications

[0165] Capsules were prepared according to the formulation described in Table 16.

[0166] Table 16: Prescription ratio

[0167]

[0168] Preparation method: grind ATV014, and sieve ATV014 and other components, mix the internal components according to the prescription ratio, dry granulate, add the external components, mix, and fill with capsules to obtain capsules.

[0169] Dissolution test: The obtained capsules were taken and 900 mL of 0.1N hydrochloric acid was used as the medium. The dissolution was tested at 5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes and 75 minutes by using a paddle method at a speed of 75 rpm. The results are shown in Table 17.

[0170] Table 17: Dissolution Results

[0171]

[0172] Conclusion: Capsules of different strengths all have good dissolution rates, and the 50 mg capsule has a faster dissolution rate.

[0173] Example 10: Stability Study

[0174] The capsules provided by the present invention were taken and placed under the conditions shown in Table 18 to examine their stability. The results showed that the relevant substances in the capsules provided by the present invention had good stability.

[0175] Table 18: Results of stability study on related substances

[0176]

[0177] Example 11: Compatibility study of raw and auxiliary materials

[0178] Excipient Compatibility Study

[0179] The raw and auxiliary materials were mixed in the proportions shown in Table 19, and placed under high temperature conditions (60°C), high humidity conditions (25°C, 92.5%RH), accelerated conditions (40°C±2°C, 75%RH±5%) and light conditions (4500lx±500lx) for 10 days and 30 days respectively. The mixture was taken out and the changes in its content and related substances were tested. The results are shown in Table 20.

[0180] Table 19: Compatibility test of raw materials and auxiliary materials

[0181]

[0182] Table 20: Content test results (the result value unit is %)

[0183]

[0184] N / A: According to the EC protocol, it is an alternative excipient and has not been tested.

[0185] in conclusion:

[0186] Detection of related substances: There is no obvious change in impurities when the auxiliary materials selected for the present invention and ATV014 are placed under accelerated (40℃ / 75%±5%RH), high humidity (25℃ / 90%±5%RH) or high temperature (60℃) conditions for 30 days and under light (visible light not less than 1.2×106 Lux·hr, near ultraviolet not less than 200w·hr / m2) conditions for 10 days. All auxiliary materials have good compatibility with raw materials.

[0187] From the results in Table 20, we can see that:

[0188] In the investigation of diluents (No. 2-6), the use of microcrystalline cellulose PH102 is beneficial to the stability of ATV014 under high temperature (60℃) and light (visible light not less than 1.2×106 Lux·hr, near ultraviolet not less than 200w·hr / m2) conditions. Compared with other diluents, the use of mannitol 50C is more conducive to the stability of ATV014 under high temperature conditions (60℃), high humidity conditions (25℃, 92.5%RH), accelerated conditions (40℃±2℃, 75%RH±5%) and light conditions (4500lx±500lx), with unexpected technical effects.

[0189] In the investigation of disintegrants (No. 7-8), cross-linked sodium carboxymethyl cellulose VIVASOL was used. ®or crospovidone XL-10 are beneficial to the stability of ATV014 under high temperature conditions (60℃), high humidity conditions (25℃, 92.5%RH), accelerated conditions (40℃±2℃, 75%RH±5%) and light conditions (4500lx±500lx).

[0190] In the investigation of adhesives (No. 9-10), the use of hydroxypropyl cellulose-EXF or povidone K30 was beneficial to the stability of ATV014 under high temperature conditions (60°C), high humidity conditions (25°C, 92.5%RH), accelerated conditions (40°C±2°C, 75%RH±5%) and light conditions (4500lx±500lx).

[0191] In the investigation of surfactants (No. 11), the use of sodium dodecyl sulfate is beneficial to the stability under high temperature conditions (60°C), high humidity conditions (25°C, 92.5%RH), accelerated conditions (40°C±2°C, 75%RH±5%) and light conditions (4500lx±500lx).

[0192] In the investigation of lubricants (No. 12~13), the use of magnesium stearate LIGAMED MF-2-V or sodium stearyl fumarate PRUV is beneficial to the stability of ATV014 under high temperature conditions (60℃), high humidity conditions (25℃, 92.5%RH), accelerated conditions (40℃±2℃, 75%RH±5%) and light conditions (4500lx±500lx).

[0193] In the investigation of glidants (No. 14), the use of silicon dioxide 200 Pharma was beneficial to the stability of ATV014 under high temperature conditions (60°C), high humidity conditions (25°C, 92.5%RH), accelerated conditions (40°C±2°C, 75%RH±5%) and light conditions (4500lx±500lx).

[0194] In the investigation of capsule materials and coating materials (No. 15-16), the use of hollow capsules, 0# rich yellowopaque or Opadry® II complete coating system 85F620077-CN, US is beneficial to the stability of ATV014 under high temperature conditions (60°C), high humidity conditions (25°C, 92.5%RH), accelerated conditions (40°C±2°C, 75%RH±5%) and light conditions (4500lx±500lx).

[0195] The method of the present invention has been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein within the content, spirit and scope of the present invention to implement and apply the technology of the present invention. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention.

Claims

1. A composition comprising: ATV014 compound and a pharmaceutically acceptable excipient, ； The pharmaceutically acceptable excipients include diluents, disintegrants, binders and lubricants; The diluent is mannitol 50C; Calculated based on the total mass of the composition, the content of the lubricant is 1.5wt%-5wt%; the content of the binder is 3wt%-5wt%.

2. The composition according to claim 1, wherein the pharmaceutically acceptable excipients further comprise other excipients.

3. The composition according to claim 1, wherein the disintegrant comprises at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and hydroxypropyl cellulose; and / or The binder comprises at least one of hydroxypropyl cellulose, povidone and starch; and / or The lubricant includes at least one of magnesium stearate, stearic acid and sodium stearyl fumarate.

4. The composition according to claim 2, wherein the other excipients include a glidant.

5. The composition according to claim 1, further comprising an external lubricant. The composition according to claim 4 , wherein the glidant comprises colloidal silicon dioxide or talc.

7. The composition according to claim 5, wherein the external lubricant comprises at least one of magnesium stearate, sodium stearate and sodium stearyl fumarate.

8. The composition according to any one of claims 2 to 3, wherein the content of the ATV014 compound is 15 wt% to 70 wt% based on the total mass of the composition; and / or The content of the diluent is 20wt%-70wt% based on the total mass of the composition; and / or Calculated based on the total weight of the composition, the content of the disintegrant is 1 wt%-10 wt%.

9. The composition according to claim 5 or 7, wherein the content of the added lubricant is 0.5wt%-5wt%, calculated based on the total mass of the composition.

10. The composition according to any one of claims 2 to 7, which does not contain a surfactant.

11. The composition according to any one of claims 2 to 7, which does not contain sodium lauryl sulfate.

12. The composition according to any one of claims 2 to 3, characterized in that include: ATV014 compound and pharmaceutically acceptable excipients, the pharmaceutically acceptable excipients comprising: a diluent, a disintegrant, a binder, a lubricant and other excipients, the diluent being mannitol 50C, the disintegrant comprising at least one of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and hydroxypropyl cellulose, the binder comprising at least one of hydroxypropyl cellulose, polyvinylpyrrolidone and starch, the lubricant comprising at least one of magnesium stearate, stearic acid and sodium stearyl fumarate, and calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant is 1.5%-5wt%.

13. The composition according to any one of claims 2 to 3, characterized in that include: ATV014 compound and pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable excipients include: a diluent, a disintegrant, a binder, a lubricant and other excipients, wherein the diluent is mannitol 50C, the disintegrant includes cross-linked sodium carboxymethyl cellulose, the binder includes hydroxypropyl cellulose, and the lubricant includes at least one of magnesium stearate, stearic acid and sodium stearyl fumarate. Calculated based on the total mass of the composition, the content of the ATV014 compound is 50wt%-60wt%, the content of the diluent is 30wt%-40wt%, the content of the disintegrant is 2wt%-5wt%, the content of the binder is 2wt%-5wt%, and the content of the lubricant is 1.5%-5wt%.

14. The composition according to any one of claims 1 to 7, which is in the form of an oral solid preparation. The composition according to claim 14 , wherein the dosage form is tablet, granule or capsule.

16. Use of the composition according to any one of claims 1 to 15 in the preparation of a product for preventing or treating coronavirus infection.

17. The use according to claim 16, wherein the infection comprises fever, cough, pharyngitis, pneumonia, acute respiratory tract infection, severe acute respiratory tract infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis or septic shock.

18. The use according to any one of claims 16-17, characterized in that: The composition is suitable for humans or animals; the animals include bovines, equines, ovines, porcines, canines, felines, rodents, primates, birds or fish.

19. A method for preparing the composition according to any one of claims 1 to 15, comprising: The ATV014 compound and pharmaceutically acceptable excipients are mixed, dry granulated, an external lubricant is added, mixed, tableted or capsule filled or bagged to obtain the composition; or The ATV014 compound and pharmaceutically acceptable excipients are mixed, wet granulated, ground, dried, dry milled, an external lubricant is added, mixed, tableted or capsule filled or bagged to obtain the composition; or Grinding or crushing the ATV014 compound, mixing it with a pharmaceutically acceptable excipient, dry granulating it, adding an external lubricant, mixing it, tableting it or filling it in a capsule or bagging it to obtain the composition; or The ATV014 compound is ground or pulverized, mixed with pharmaceutically acceptable excipients, wet granulated, ground, dried, dry milled, an external lubricant added, mixed, tableted or capsule filled or bagged to obtain the composition.

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