Salts of nucleoside analogs, their crystal forms, pharmaceutical compositions, and uses

Abstract

The present invention relates to a salt of a nucleoside analog, its crystalline form, pharmaceutical composition, and use thereof, which falls within the field of pharmaceutical technology. Specifically, the salt of the nucleoside analog has a structure represented by Formula I (wherein X is hydrogen or deuterium, Y is an acid, preferably hydrogen bromide, hydrogen chloride, nitric acid, methanesulfonic acid, or maleic acid, and n is 0.5 to 2, preferably 1). When X is hydrogen or deuterium, Y is hydrogen bromide, and n is 1, the salt of the nucleoside analog exists in the form of a crystalline or amorphous substance exhibiting Crystal Form I or Crystal Form A, which has advantages such as good solid state properties, high stability, good solubility, and low hygroscopicity, making it useful for producing a medicament for treating and / or alleviating diseases caused by viruses (particularly SARS-CoV-2). [Formula 1] TIFF2023552672000036.tif60114

Description

Technical Field

[0001] [Reference to Related Applications] The present invention claims priority from Chinese Patent Application No. 202011156037.5, entitled "Salt of Nucleoside Analogue, Its Crystal Form, Pharmaceutical Composition and Use", filed in China on October 26, 2020, and Chinese Patent Application No. 202011505962.4, entitled "Salt of Nucleoside Analogue, Its Crystal Form, Pharmaceutical Composition and Use", filed in China on December 18, 2020. The entire contents of these patent applications are incorporated herein by reference. [Technical Field]

[0002] The present invention pertains to the field of pharmaceutical technology, and specifically relates to acid addition salts of nucleoside analogues, salt form crystals, pharmaceutical compositions and pharmaceutical uses.

Background Art

[0003] Viral infectious diseases pose a serious threat to human life and health. Currently, numerous viruses that cause diseases in humans have been discovered. However, as the scope of human social activities expands and globalization intensifies, new and re-emerging viruses continue to appear around the world. For example, SARS-CoV in 2003, H1N1 influenza virus in 2009, H7N9 avian influenza virus in 2013, MERS-CoV in 2012, Ebola virus in 2014, and more recently, the more severe dengue virus, Zika virus, etc. Most of these viruses are characterized by being easy to spread, highly infectious, and highly pathogenic, causing serious problems for the world's medical and health systems.

[0004] Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new coronavirus that has approximately 79% genomic-wide similarity with SARS-CoV in 2003. Since its discovery, this virus has rapidly spread around the world.

[0005] Viruses have a strong growth ability and are prone to mutation during the replication process. Therefore, there are many viral infectious diseases that are difficult to prevent through vaccination, and antiviral drugs are one of the important means to combat viruses. As organisms, the genetic material of viruses is DNA or RNA, and a large number of nucleoside triphosphates and deoxynucleoside triphosphates are required for their replication. Interfering with the replication of viral genetic material by nucleoside analogs is an important strategy in the development of antiviral drugs.

[0006] In short, in this field, there is a strong desire to develop nucleoside analogs with stable physical properties and high drugability for the treatment of diseases caused by viral infections.

Summary of the Invention

Problems to be Solved by the Invention

[0007] The present invention aims to provide an acid addition salt of a nucleoside analog having stable physical properties and high drugability, a salt form crystal obtained from the acid addition salt, a pharmaceutical composition containing the acid addition salt or its salt form crystal, and to provide a method for producing the same and its pharmaceutical use.

Means for Solving the Problems

[0008] In a first aspect, the present invention provides a compound represented by formula I.

Chemical Formula

[0009] Preferably, the compound of formula I is a compound of formula I-1.

Chemical formula

[0010] More preferably, the compound of formula I or formula I-1 is a compound of formula I-1'.

Chemical formula

[0011] Preferably, the compound of formula I-1' is a crystal having the form of crystal form A, and the crystal form A has at least one of the following characteristics. 1) In the XRPD pattern, characteristic peaks are present at least at 3 positions, preferably at least 5 positions, more preferably at least 7 positions, out of the 2θ values of 5.35° ± 0.2°, 8.11° ± 0.2°, 8.46° ± 0.2°, 15.70° ± 0.2°, 18.08° ± 0.2°, 21.09° ± 0.2°, and 21.91° ± 0.2°. 2) In the DSC pattern, it has an absorption peak at 204 ± 5 °C. 3) After storage for 24 hours under the conditions of 25 °C and 80% RH, the increase in the water content is 1% or less, preferably 0.2% or less, more preferably 0.1% or less. 4) The solubility in deionized water at 37 °C is 0.1 mg / mL or more, preferably 0.2 mg / mL or more, more preferably 0.3 mg / mL.

[0012] More preferably, in the XRPD pattern of the crystalline form A, further, at least 3 of the 2θ values of 16.02° ± 0.2°, 16.88° ± 0.2°, 17.22° ± 0.2°, 17.76° ± 0.2°, 20.55° ± 0.2°, 23.25° ± 0.2°, 23.89° ± 0.2° and 26.14° ± 0.2°, preferably at least 5, more preferably at least 8 have characteristic peaks. Most preferably, the XRPD pattern of the crystalline form A is as shown in Figure 2.

[0013] More preferably, the DSC pattern of the crystalline form A is as shown in Figure 1.

[0014] Alternatively, preferably, the compound of formula I-1’ is an amorphous substance.

[0015] More preferably, the XRPD pattern of the form of the amorphous substance is as shown in Figure 6.

[0016] Preferably, the compound of formula I is the compound of formula I-2.

Chemical formula

[0017] More preferably, the compound of formula I or formula I-2 is the compound of formula I-2’.

Chemical formula

[0018] Preferably, the compound of formula I-2’ is a crystal having the form of crystalline form I, and the crystalline form I has at least one of the following characteristics. 1) In the XRPD pattern, it has characteristic peaks at least at 3 positions, preferably at least 5 positions, more preferably at least 6 positions among the 2θ values of 5.36° ± 0.2°, 8.13° ± 0.2°, 8.48° ± 0.2°, 18.16° ± 0.2°, 20.95° ± 0.2° and 21.95° ± 0.2°. 2) In the DSC pattern, it has an absorption peak at 200 ± 5 °C. 3) After storage for 24 hours under the conditions of 25 °C and RH80%, the increase in the water content is 1% or less, preferably 0.2% or less, more preferably 0.1% or less. 4) Its solubility in deionized water at 37 °C is 0.1 mg / mL or more, preferably 0.2 mg / mL or more, more preferably 0.3 mg / mL.

[0019] More preferably, in the XRPD pattern of the crystalline form I, further, it has characteristic peaks at least at 3 positions, preferably at least 5 positions, more preferably at least 7 positions among the 2θ values of 15.71° ± 0.2°, 16.07° ± 0.2°, 16.90° ± 0.2°, 17.26° ± 0.2°, 20.55° ± 0.2°, 23.27° ± 0.2° and 26.08° ± 0.2°. Most preferably, the XRPD pattern of the crystalline form I is as shown in Figure 4.

[0020] More preferably, the DSC pattern of the crystalline form I is as shown in Figure 3.

[0021] Alternatively, preferably, the compound of formula I-2’ is an amorphous substance.

[0022] More preferably, the XRPD pattern of the form of the amorphous substance is as shown in Figure 7.

[0023] In a second aspect, the present invention provides a method for producing a compound represented by Formula I, Formula I-1, Formula I-1', Formula I-2 or Formula I-2'. The production method comprises: 1) dissolving the compound of Formula II in Solvent A to obtain Solution A, and mixing the solution A with an acid or a solution B obtained by dissolving the acid in Solvent B under ice bath conditions to obtain a mixed solution; [Chemical formula] (wherein X is hydrogen or deuterium.) 2) stirring the mixed solution at room temperature and concentrating it to obtain the target product; The acid is maleic acid, succinic acid, citric acid, tartaric acid, fumaric acid, formic acid, acetic acid, propionic acid, malonic acid, oxalic acid, benzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, 1,5-naphthalenedisulfonic acid, camphoric acid, camphorsulfonic acid, salicylic acid, acetylsalicylic acid, aspartic acid, glutamic acid, lactic acid, gluconic acid, ascorbic acid, gallic acid, mandelic acid, malic acid, sorbic acid, trifluoroacetic acid, taurine, homotaurine, 2-hydroxyethanesulfonic acid, cinnamic acid, mucic acid, hydrogen chloride, hydrogen bromide, hydrogen iodide, sulfuric acid, nitric acid, phosphoric acid, perchloric acid or a combination thereof, preferably hydrogen bromide, hydrogen chloride, nitric acid, methanesulfonic acid, maleic acid or a combination thereof, more preferably hydrogen bromide.

[0024] Preferably, in step 1) of the production method, the usage ratio of the compound of Formula II to Solvent A is 1 g: 2-20 mL, preferably 1 g: 5-10 mL, the acid content in Solution B is 40 wt% - 50 wt%, the molar ratio of the compound of Formula II to the acid is 1: 0.9-1, and in step 2) of the production method, the stirring time is 0.5-5 hours, preferably 0.5-1 hour.

[0025] Preferably, the production method further includes: 3) mixing the product obtained in step 2) with solvent C, and stirring under room temperature and / or heating conditions to precipitate a solid, thereby obtaining the target product.

[0026] More preferably, in step 3) of the production method, the usage ratio of the compound of formula II to solvent C is 1 g: 2-20 mL, preferably 1 g: 5-15 mL; the temperature of the heating conditions is 35-60 °C, preferably 50-60 °C; and the stirring time is 0.5-5 hours, preferably 0.5-2 hours.

[0027] Preferably, in the production method, solvent A, solvent B, and solvent C are each independently selected from water, hydrocarbons, alcohols, ethers, ketones, esters, nitriles, and homogeneous mixtures thereof. The hydrocarbon is selected from n-pentane, n-hexane, n-heptane, petroleum ether, dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, benzene, toluene, xylene, chlorobenzene, and dichlorobenzene; the alcohol is selected from methanol, ethanol, n-propanol, isopropanol, n-butanol, ethylene glycol, and propylene glycol; the ether is selected from ethyl ether, n-propyl ether, isopropyl ether, methyl t-butyl ether, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol dimethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and diethylene glycol dimethyl ether; the ketone is selected from acetone, butanone, and diethyl ketone; the ester is selected from methyl formate, ethyl formate, propyl formate, butyl formate, methyl acetate, ethyl acetate, propyl acetate, and butyl acetate; and the nitrile is selected from acetonitrile and propionitrile. Solvent A and solvent B are miscible with each other. More preferably, in step 1) of the manufacturing method, the solvent A is acetonitrile, and the usage ratio of the compound of formula II to acetonitrile is 1 g: 2-20 mL, preferably 1 g: 5-10 mL; the acid is hydrogen bromide; the solvent B is water; the solution B is hydrobromic acid; the hydrogen bromide content in the hydrobromic acid is 40 wt%-50 wt%; the molar ratio of the compound of formula II to hydrogen bromide is 1: 0.9-1; in step 2) of the manufacturing method, the stirring time is 0.5-5 hours, preferably 0.5-1 hour; in step 3) of the manufacturing method, the solvent C is methyl t-butyl ether, and the usage ratio of the compound of formula II to methyl t-butyl ether is 1 g: 2-20 mL, preferably 1 g: 5-15 mL; the temperature of the heating condition is 35-60 °C, preferably 50-60 °C; the stirring time is 0.5-5 hours, preferably 0.5-2 hours.

[0028] In a third aspect, the present invention provides a pharmaceutical composition comprising a compound represented by formula I, formula I-1, formula I-1', formula I-2 or formula I-2', and optionally a pharmaceutically acceptable additive.

[0029] Preferably, the pharmaceutical composition is an oral preparation or a parenteral preparation. More preferably, the oral preparation is selected from tablets, capsules, granules, powders and syrups, and the parenteral preparation is selected from injections, powder injections, sprays and suppositories.

[0030] In a fourth aspect, the present invention provides a compound represented by formula I, formula I-1, formula I-1', formula I-2 or formula I-2', or a pharmaceutical composition comprising the compound, for the treatment and / or alleviation of diseases caused by viruses. Preferably, the virus is SARS-CoV-2.

[0031] In a fifth aspect, the present invention provides the use of a compound represented by formula I, formula I-1, formula I-1', formula I-2 or formula I-2' or a pharmaceutical composition comprising said compound for the manufacture of a medicament for the treatment and / or alleviation of a disease caused by a virus. Preferably, said virus is SARS-CoV-2.

[0032] In a sixth aspect, the present invention provides a method for the treatment and / or alleviation of a disease caused by a virus, comprising administering to a subject in need thereof a therapeutically and / or alleviatingly effective amount of a compound represented by formula I, formula I-1, formula I-1', formula I-2 or formula I-2' or a pharmaceutical composition comprising said compound. Preferably, said virus is SARS-CoV-2.

[0033] In a seventh aspect, the present invention provides a method for inhibiting the replication of a virus, comprising contacting the virus with an inhibitory effective amount of a compound represented by formula I, formula I-1, formula I-1', formula I-2 or formula I-2' or a pharmaceutical composition comprising said compound. Preferably, said virus is SARS-CoV-2.

Advantages of the Invention

[0034] Through extensive and meticulous research, the present invention synthesized and isolated acid addition salts of nucleoside analogs having antiviral activity (especially anti-SARS-CoV-2 activity) and their crystals, and examined their physical and chemical properties. As a result, acid addition salts (for example, hydrobromide, hydrochloride and maleate of compound Z, hydrobromide and maleate of compound W) and their crystal forms (for example, crystal form A and crystal form I) that are useful in the manufacture of medicaments for the treatment and / or alleviation of diseases caused by viruses (especially SARS-CoV-2) were unexpectedly found, by virtue of advantages such as good solid properties, high stability, good solubility, and low hygroscopicity. In addition, the production method of the present invention also has advantages such as high purity of the product, constant composition, easy storage, simple method and easy reproducibility.

Brief Description of the Drawings

[0035]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Embodiments for Carrying Out the Invention

[0036] [Definition of Terms] Nucleoside analogs Nucleoside analogs are one of the most important antiviral drugs and have played an important role in the clinical treatment of viral diseases. Nucleoside drugs can be converted into the triphosphate form in vivo. Especially in the viral replication stage, nucleoside triphosphates "disguise" as substrates and are incorporated into the viral DNA or RNA strands, thereby inhibiting the replication of genetic material and exerting an antiviral effect.

[0037] Compound Z and Compound W

Chemical Formula

[0038] Salts and solvates Unless otherwise specified, the "salt" referred to in the present invention includes both pharmaceutically acceptable salts (also referred to as "pharmaceutical salts") and pharmaceutically unacceptable salts. Pharmaceutically unacceptable salts are not preferred for administration to patients, but can be used to provide drug intermediates and active pharmaceutical ingredients.

[0039] Unless otherwise specified, the "pharmaceutically acceptable salt" or "pharmaceutically acceptable acid addition salt" referred to in the present invention means an acid addition salt prepared using a pharmaceutically acceptable acid, and examples include, but are not limited to, organic acid salts and inorganic acid salts. The acid for forming the salt is preferably hydrobromic acid, hydrochloric acid, sulfuric acid, nitric acid, methanesulfonic acid or maleic acid, more preferably hydrobromic acid or maleic acid, and most preferably hydrobromic acid.

[0040] The acid addition salts of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) (Compound Z) or (2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) (Compound W) absorb moisture and form hydrates with water bound when left in air or recrystallized, and such acid addition salts containing moisture are also included within the scope of the present invention.

[0041] The present invention also includes solvates (also referred to as "solvated compounds") composed of the acid addition salts of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) (Compound Z) or (2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) (Compound W) and a solvent. Any solvent used in the production of the salt and / or crystal may be used, and there is no particular limitation. Specifically, for example, hydrates, alcoholates, acetoneates, esterates, etherates, tolueneates, etc. can be mentioned, but hydrates and alcoholates are preferred.

[0042] Pharmaceutical composition Unless otherwise specified, the "pharmaceutical composition" referred to in the present invention contains at least one kind of the compound according to the present invention and, optionally, a pharmaceutically acceptable additive. Preferably, the pharmaceutical composition of the present invention contains at least one kind of an acid addition salt composed of a free base selected from Compound Z and Compound W and an acid selected from hydrogen bromide, hydrogen chloride, sulfuric acid, nitric acid, methanesulfonic acid and maleic acid, and, optionally, a pharmaceutically acceptable additive. More preferably, the pharmaceutical composition of the present invention contains a hydrobromide of Compound Z having Crystal Form A and / or a hydrobromide of Compound W having Crystal Form I, and, optionally, a pharmaceutically acceptable additive.

[0043] Unless otherwise specified, the "additive" referred to in the present invention includes excipients, binders, lubricants, disintegrants, colorants, flavoring agents, odor-masking agents, emulsifiers, surfactants, solubilizers, suspending agents, isotonic agents, buffers, preservatives, antioxidants, stabilizers, absorption promoters, etc. generally used in the pharmaceutical field, but is not limited thereto. If necessary, the above additives may be used in appropriate combinations as required.

[0044] When used as a therapeutic or prophylactic agent for viral infections, the acid addition salts of Compound Z or Compound W of the present invention (i.e., the compounds of Formula I) can be administered alone or mixed with a suitable pharmaceutically acceptable additive, as oral preparations such as tablets, capsules, granules, powders, syrups, etc., or as parenteral preparations such as injections, powder injections, sprays, suppositories, etc. These dosage forms can be manufactured by general formulation methods in the art.

[0045] The oral pharmaceutical composition is produced by mixing the hydrobromide of Compound W or Compound Z of the present invention with at least one pharmaceutically acceptable additive so as to contain 10 to 2000 mg of a pharmaceutical active ingredient (API) per unit dose. For example, when the oral pharmaceutical composition is a tablet, the API is mixed with at least one pharmaceutical additive (such as starch, lactose, magnesium stearate, etc.) and tabletted, and further, the core tablet can be coated with a sugar coating or other appropriate substances, or processed to have a sustained-release effect or a release control effect. As another example, when the oral pharmaceutical composition is a capsule, the API is mixed with at least one diluent (such as starch), granulated and sized as desired, and the resulting mixture is filled into a capsule shell.

[0046] Pharmaceutical use The dosage of the drug varies depending on symptoms, age, etc. Taking adults as an example, it can be administered about 0.01 to 1000 mg per time, 1 to 7 times a day for 1 to 7 days according to symptoms. The administration method is not limited.

[0047] The compound of the present invention can be used for the treatment and / or alleviation of diseases caused by viral infections, or for the inhibition of viral replication. Examples of the above viruses include coronavirus, influenza virus, respiratory syncytial virus, Flaviviridae, Filoviridae, porcine epidemic diarrhea virus (PEDV), with coronavirus being preferred and SARS-CoV-2 being more preferred. Therefore, the compound of the present invention can be used in the manufacture of antiviral drugs.

Examples

[0048] The present invention will be further described below with specific examples. These examples are only for explaining the present invention and do not limit the scope of the present invention. When detailed conditions are not specified in the following examples, the experimental methods are according to normal conditions, for example, the conditions described in Molecular Cloning: A Laboratory Manual edited by Sambrook et al. (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. Unless otherwise specified, the percentages and parts in the present invention are by weight.

[0049] Description of reagents and consumables In the examples of the present invention, all reagents such as acetonitrile in the production method are of analytical grade manufactured by Sinopharm Chemical Reagent Co., Ltd. Unless otherwise specified, the reagents used have not been specially treated. (2R,3R,4R,5R)-2-(4-Amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diylbis(2-methylpropionate) (Compound Z) and (2R,3R,4R,5R)-2-(4-Aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diylbis(2-methylpropionate) (Compound W) were prepared with reference to the examples of Patent CN202010313870X and have a purity of over 98% and a deuteration rate of over 98%. Triethylamine and phosphoric acid in the high performance liquid chromatography experiment are chromatography reagents manufactured by Sinopharm Chemical Reagent Co., Ltd. The nuclear magnetic resonance spectrum was measured with nuclear magnetic resonance apparatuses Brucker 500Hz and Brucker 600Hz.

[0050] Common measurement methods 1. Measurement method of powder X-ray diffraction (XRPD) Apparatus: X-ray polycrystalline diffractometer Bruker D8 advance Target: Cu-Kα (40 kV, 40 mA) Distance from the sample to the detector: 30 cm Scan type: Two-axis linkage Scan step width: 0.02° Scan range: 3° to 40° Scan step: 0.1 s Generally, in XRPD, the diffraction angle (2θ value) may have an error in the range of ±0.2°. Therefore, the values related to the diffraction angle of the present invention should be understood to include values within the range of about ±0.2°. Therefore, the present invention includes not only crystal forms that exactly match the characteristic peaks in a specific XRPD pattern, but also crystal forms having an error of about ±0.2° from the characteristic peaks in a specific XRPD pattern.

[0051] 2. Measurement method of DSC Apparatus: Differential scanning calorimeter METTLER TOLEDO Temperature range: 50 - 260 °C Scanning speed: 20 °C / min Nitrogen flow rate: 50 mL / min

[0052] 3. Measurement method of hygroscopicity Appropriately collect the test sample and flatten it in a stoppered glass weighing bottle placed in an artificial climate chamber (temperature 25 °C ± 1 °C, relative humidity 80% ± 2%) for 24 hours. After placing the weighing bottle in the open state together with the lid in the artificial climate chamber (temperature 25 °C ± 1 °C, relative humidity 80% ± 2%) for 24 hours, take it out, measure the moisture content of the test sample before and after being placed in the artificial climate chamber, and examine the hygroscopicity by comparing the moisture change.

[0053] 4. Measurement method of stability Place the test sample in an appropriate clean container and place it under the conditions of high temperature (80 °C), high humidity (25 °C, relative humidity 92.5%), and light irradiation (illuminance 4500 ± 500 lx and 90 μw / cm 2 ) for 7 days, sample on the 7th day, and measure according to the stability confirmation items.

[0054] 5. Measurement method of solubility Weigh approximately 10 mg of the test sample into a 1.5 ml sample tube, add 1 ml of deionized water, place it in a thermostatic mixer, shake at 37 °C for 30 minutes. After shaking, put the sample tube into a centrifuge, centrifuge for 2 minutes, then take 500 μl of the supernatant, dilute it to an appropriate concentration with acetonitrile to obtain a test sample solution. Perform HPLC analysis and determine the solubility of the test sample in water (37 °C) by the external standard method.

[0055] (Example 1) Preparation of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) hydrobromide [Chemical formula] Dissolve (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) (Compound Z, the same below) (12 g, 23.90 mmol) in acetonitrile (100 mL), add 40% hydrobromic acid (4.37 g, 21.6 mmol) dropwise under an ice bath. After the addition is complete, return to room temperature and stir. After 30 minutes, concentrate the reaction solution, add methyl t-butyl ether (150 mL), stir for 30 minutes, then a solid precipitates. Heat to 55 °C and stir for another 2 hours. Cool to room temperature, let stand, then filter and dry to obtain the title compound (10.8 g, yield 78%, HPLC purity 99.2%) as a white solid. 1H-NMR(600 MHz, CDCl3): δ 13.05 (s, 1H), 9.73 (s, 1H), 9.46 (s, 1H), 8.00 (s, 1H), 7.04 (s, 1H), 6.02 (d, J = 5.8 Hz, 1H), 5.41 (dd, J = 5.8, 4.0 Hz, 1H), 4.65 (q, J = 4.0 Hz, 1H), 4.40 - 4.33 (m, 2H), 2.71 - 2.60 (m, 2H), 2.58 - 2.52 (m, 1H), 1.26 - 1.22 (m, 6H), 1.21 - 1.18 (m, 6H), 1.17 - 1.13 (m, 6H). 13 C-NMR(150 MHz, DMSO-d6): δ 176.01, 175.35, 174.59, 150.65, 139.98, 125.81, 115.79, 115.43, 112.21, 82.22, 75.64, 73.33, 70.72, 62.99, 33.67, 33.62, 33.56, 19.11, 19.02, 18.92, 18.87, 18.85, 18.70。 MS: m / z 503.1 [M + 1] + 。

[0056] The results of differential scanning calorimetry (DSC) show that, as shown in Figure 1, the obtained solid began to show an endothermic peak at 201.21 °C and reached the peak at 204.26 °C. The powder X-ray diffraction (XRPD) results show that, as shown in Table 1 and Figure 2, the obtained solid has a crystal form corresponding to crystal form A.

[0057]

Table 1

[0058] The solid powder (2.5 g) of the hydrobromide (I-1’) of compound Z having crystal form A was added to ethanol (10 mL), stirred at room temperature, and after all the solids were dissolved, the solvent was removed by concentration and dried with an oil pump to obtain a powdery solid (2.5 g). The results of powder X-ray diffraction (XRPD) show that, as shown in Figure 6, the obtained solid is an amorphous substance.

[0059] (Example 2) Preparation of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) hydrochloride

Chemical formula

[0060] (Example 3) Preparation of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) nitrate

Chemical formula

[0061] (Example 4) Preparation of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) methanesulfonate

Chemical formula

[0062] (Example 5) Preparation of (2R,3R,4R,5R)-2-(4-Amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) maleate [Chemical Formula] Compound Z (2.85 g, 5.67 mmol) was dissolved in anhydrous ethanol (20 mL), maleic acid (0.66 g, 5.67 mmol) was added, and the mixture was heated to reflux. After 30 minutes, the reaction solution was cooled to room temperature, and n-heptane (40 mL) was added, resulting in the precipitation of a large amount of flocculent solid. The temperature was raised to 45 °C, stirred for 1.5 hours, cooled to 0 °C again, stirred for an additional 30 minutes, and filtered. The filtrate was dried to obtain the title compound (2.5 g, yield 71%, HPLC purity 98.9%) as a white solid. 11H-NMR (600 MHz, DMSO-d6): δ 8.11 (s, 1H), 8.01 (s, 1H), 7.94 (s, 1H), 6.76 (s, 1H), 6.26 (s, 2H), 6.07 (d, J = 5.7 Hz, 1H), 5.44 (dd, J = 5.7, 3.7 Hz, 1H), 4.63 (q, J = 3.7 Hz, 1H), 4.33 (dd, J = 12.4, 3.3 Hz, 1H), 4.28 (dd, J = 12.4, 4.1 Hz, 1H), 2.67 - 2.56 (m, 2H), 2.49 - 2.45 (m, 1H), 1.17 (d, J = 7.0 Hz, 3H), 1.15 (d, J = 7.0 Hz, 3H), 1.12 - 1.09 (m, 6H), 1.05 (d, J = 7.0 Hz, 3H), 1.02 (d, J = 7.0 Hz, 3H). MS: m / z 503.1 [M+1] + .

[0063] (Example 6) Preparation of hemisulfate of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate)

Chemical formula

[0064] (Example 7) Preparation of sulfate of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate)

Chemical formula

[0065] (Example 8) Preparation of (2R,3R,4R,5R)-2-(4-amino-5-deuteropyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) phosphate

Chemical formula

[0066] (Example 9) Preparation of (2R,3R,4R,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano-5-(isobutyryloxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropionate) hydrobromide

Chemical formula

[0067] As shown in Figure 3, the results of differential scanning calorimetry (DSC) indicate that the obtained solid began to show an endothermic peak at 195.41 °C and reached the peak at 200.11 °C. As shown in Table 2 and Figure 4, the results of powder X-ray diffraction (XRPD) indicate that the obtained solid has a crystal form corresponding to Crystal Form I.

[0068]

Table 2

[0069] The solid powder (0.17 g) of the hydrobromide (I-2’) of Compound W having Crystal Form I was added to ethanol (10 mL), stirred at room temperature, and after all the solids had dissolved, the solvent was removed by concentration and dried with an oil pump to obtain a powdery solid (0.17 g). As shown in Figure 7, the results of powder X-ray diffraction (XRPD) indicate that the obtained solid is an amorphous substance.

[0070] (Example 10) Comparison of the Properties of Compound Z and Its Acid Addition Salts (1) Comparison of the Physical Properties and Preparation Conditions of Compound Z and Its Salts The physical properties and the difficulty of preparation of Compound Z and its various salts (obtained in Examples 1 to 8) were compared, and the results are shown in Table 3.

[0071]

Table 3

[0072] As is apparent from the above table, the acid addition salts of Compound Z other than the hemisulfate, sulfate, and phosphate have good physical properties. Also, the hydrobromide and nitrate salts of Compound Z are easier to prepare.

[0073] (2) Comparison of the Hygroscopicity and Stability of the Salts of Compound Z The hygroscopicity and stability of some salts of Compound Z (obtained in Examples 1 to 5) were compared, and the results are shown in Table 4.

[0074]

Table 4

[0075] As is apparent from the above table, the hydrobromide, hydrochloride, nitrate, methanesulfonate, and maleate salts of Compound Z were non-hygroscopic under the conditions of 25°C and 80% RH. The hydrobromide, hydrochloride, and maleate salts of Compound Z were stable under the conditions of high temperature, light irradiation, and high humidity, while the nitrate and methanesulfonate salts of Compound Z showed relatively low stability.

[0076] (3) Comparison of the Solubility of Compound Z and Its Salts The solubility of Compound Z and its salts (obtained in Examples 1, 4, and 5) was compared. An appropriate amount of the sample was weighed into a glass sample tube, and the selected solvent was gradually added, and the transparency was observed. The solubility of each acid salt in deionized water was measured, and the results are shown in Table 5.

[0077] [Table 5]

[0078] From the above table, it is clear that the water solubility of the basic compound of Compound Z is significantly lower than that of the salt of Compound Z. Water solubility has a great impact on drug formulation, oral bioavailability, etc. Therefore, converting Compound Z into a salt is advantageous for formulating the drug for use in human medicine.

[0079] (Example 11) Comparison of the properties of Compound W and its acid addition salts The same operations as in Example 10 were carried out for property comparison, except that Compound W and its various salts were used. Similar to the case of Compound Z and its salts, the hydrobromide, hydrochloride, nitrate, methanesulfonate, and maleate salts of Compound W are all white powdery solids, and it is clear that these salts are superior in solid properties to the free base form. Among them, the hydrobromide, nitrate, methanesulfonate, and maleate salts of Compound W are easier to prepare, and the hydrobromide and maleate salts are the most excellent in stability.

[0080] (Example 12) Superiority of crystalline form A of the hydrobromide salt of Compound Z The hygroscopicity and stability of the hydrobromide salt crystals of Compound Z having crystalline form A were examined, and the results are shown in Table 6.

[0081] [Table 6]

[0082] As is clear from the above table, crystalline form A of the hydrobromide salt of Compound Z has no hygroscopicity under high humidity conditions, and the crystal form is stable under high temperature (80 °C) conditions, as shown in Figure 5.

[0083] (Example 13) Superiority of crystalline form I of the hydrobromide salt of Compound W The hygroscopicity and stability of the hydrobromide salt crystals of Compound W having crystalline form I were examined, and the results are shown in Table 7.

[0084]

Table 7

[0085] As is apparent from the above table, crystalline form I of the hydrobromide salt of compound W had no hygroscopicity under high humidity conditions and had good stability under high temperature (80 °C), high humidity, and light irradiation conditions.

[0086] (Example 14) Confirmation of the composition ratio of the hydrobromide salts of compound Z and compound W The composition ratio of the hydrobromide salts of compound Z and compound W was confirmed by titration. Approximately 0.10 g each of the hydrobromide salt of compound Z obtained in Example 1 and the hydrobromide salt of compound W obtained in Example 9 were accurately weighed, an equal-volume mixture of methanol and water (20 ml) was added and dissolved, the end point was determined by potentiometric titration, and titration was carried out with a silver nitrate titrant (0.1 mol / L). The results are shown in Table 8.

[0087]

Table 8

[0088] As is apparent from the above table, compound Z and hydrogen bromide form a salt in a molar ratio of 1:1, and compound W and hydrogen bromide form a salt in a molar ratio of 1:1.

[0089] (Example 15) Pharmacokinetic evaluation of the hydrobromide salt of compound Z in SD rats (Grouping) Thirty SD rats were randomly divided into five groups, with 6 rats in each group, and half males and half females. They were fasted for more than 12 hours before administration and fed simultaneously 4 hours after administration.

[0090] (Administration) The hydrobromide salt of compound Z was administered as a single intravenous dose at 10 mg / kg to the first group, using DMSO / EtOH / PEG300 / 0.9% NaCl (5 / 5 / 40 / 50, v / v / v / v) as the solvent. The hydrobromide salt of compound Z was administered as a single intragastric dose at 10 mg / kg, 30 mg / kg, and 90 mg / kg to the second to fourth groups, respectively, using PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v) as the solvent. The hydrobromide salt of compound Z was administered intragastrically once a day at 30 mg / kg for 7 consecutive days to the fifth group.

[0091] (Blood Sampling) Blood samples (0.2 ml) were collected from the retro-orbital venous plexus or jugular vein (or other blood sampling methods) of each group before administration (0 h) and at 5 min, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 24, and 48 h after administration, placed in EDTA-K2 anticoagulant tubes, centrifuged at 2000 g for 10 min (4 °C) within 30 min to separate plasma, and stored at -70 °C until measurement. The operations were performed under the condition of crushed ice from blood sampling to centrifugation.

[0092] (Analysis) The concentration of the nucleoside metabolite (2R,3R,4S,5R)-2-(4-amino-5-deuterio-pyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile in rat plasma was analyzed by LC-MS / MS, and the pharmacokinetic parameters were calculated.

[0093]

Table 9

[0094]

Table 10

[0095] As is clear from Table 9, after the hydrobromide salt of compound Z (10 mg / kg) was administered to SD rats by intravenous injection, the disappearance of the nucleoside metabolite from plasma was rapid, and the average half-life (t1 / 2) was 1.44 ± 0.53 h.

[0096] As is apparent from Table 10, when the hydrobromide salt of Compound Z was administered intragastrically to SD rats, it was rapidly absorbed, nucleoside metabolites were rapidly generated, and the maximum plasma concentration (Cmax) was reached in about 1 hour. After single intragastric administrations of 10, 30, and 90 mg / kg, the bioavailability (F%) of the hydrobromide salt of Compound Z, calculated from the nucleoside metabolites, was 86.4%, 79.2%, and 79.6%, respectively. After multiple (30 mg / kg, once a day, for 7 consecutive days) intragastric administrations, since the plasma Cmax and AUC0-t of the nucleoside metabolites on the 7th day were 0.84 times and 0.71 times those after single administration, it is considered that the nucleoside metabolites do not accumulate in the rat body.

[0097] (Example 16) Pharmacokinetic evaluation of the hydrobromide salt of Compound Z in beagle dogs (Grouping) Eighteen beagle dogs were randomly divided into three groups (A, B, C), with 6 dogs in each group and an equal number of males and females. They were fasted for more than 12 hours before administration and fed simultaneously 4 hours after administration.

[0098] (Administration) Administration was carried out in two phases, and the drug withdrawal period was 1 week. In the first phase, the hydrobromide salt of Compound Z was administered intragastrically to groups A and B at 10 mg / kg and 20 mg / kg, respectively, and to group C at 20 mg / kg once a day for 7 consecutive days. PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v) was used as the vehicle. In the second phase, the hydrobromide salt of Compound Z at 10 mg / kg and 40 mg / kg was administered to groups A and B by single intravenous administration and intragastric administration, respectively. DMSO / EtOH / PEG300 / 0.9% NaCl (5 / 5 / 40 / 50, v / v / v / v) was used as the solvent for intravenous administration, and the same vehicle as in the first phase was used for intragastric administration.

[0099] (Blood Sampling) For each group, 1.0 mL of blood was collected from the anterior limb vein or other sites into anticoagulant blood collection tubes containing stabilizer at 0 hours (before administration), 5 minutes after administration (only for the intravenous injection group), and 0.25, 0.5, 1, 2, 4, 6, 8, 10, 24, and 48 hours after administration. After processing, plasma was separated and stored in a -70°C refrigerator.

[0100] (Processing) The concentration of the nucleoside metabolite (2R,3R,4S,5R)-2-(4-amino-5-deuterated pyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxyl-5-(hydroxymethyl)tetrahydrofuran-2-carbonitrile in the plasma of beagle dogs was analyzed by LC-MS / MS, and pharmacokinetic parameters were calculated.

[0101]

Table 11

[0102]

Table 12

[0103] As is evident from Table 11, after the hydrobromide salt of Compound Z (10 mg / kg) was administered to beagle dogs by intravenous injection, the average t1 / 2 of the nucleoside metabolite was 3.94 ± 0.85 hours.

[0104] As is evident from Table 12, when the hydrobromide salt of Compound Z was administered intragastrically to beagle dogs, it was rapidly absorbed, and the nucleoside metabolite was rapidly generated, reaching Cmax in about 1 hour. The t1 / 2 at a dose of 20 mg / kg was 4.21 hours. After single intragastric administration of 10, 20, and 40 mg / kg, the F% of the hydrobromide salt of Compound Z, calculated using the nucleoside metabolite, was 87.4%, 101.7%, and 99.9%, respectively. After multiple intragastric administrations (20 mg / kg, once a day, for 7 consecutive days), since the plasma Cmax and AUC0-t of the nucleoside metabolite on the 7th day were 0.95 times and 0.85 times those after single administration, it is considered that the nucleoside metabolite does not accumulate in the body of beagle dogs.

[0105] (Example 17) Safety Evaluation of Hydrobromide Salt of Compound Z in SD Rats and Beagle Dogs In accordance with the "Good Laboratory Practice for Non-clinical Research of Drugs" (2017) of the NMPA, the safety of the hydrobromide salt of Compound Z in SD rats and Beagle dogs was evaluated, and an acute toxicity test and a 14-day long-term toxicity test were respectively conducted.

[0106] 1. Toxicity Test by Single Oral Administration to SD Rats (Method) Forty SD rats were randomly divided into four groups, with 10 rats in each group and half males and half females. For each group, the solvent of PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v) and the hydrobromide salt of Compound Z dissolved in this solvent (200, 600, 2000 mg / kg as free base) were respectively administered by single oral administration. After administration, continuous observation was carried out for 14 days, and planned dissection was performed on the 15th day. During the test period, clinical findings, body weight, food intake, and gross anatomical findings were evaluated.

[0107] (Result) During the test period, there was no premature death or euthanasia in each group. No abnormal clinical symptoms were observed in each administration dose group during the test. Compared with the control group, no changes were observed in the average body weight and food intake of each administration group due to the test article, and no abnormalities were observed in the gross anatomical findings of the dissection. Therefore, under the conditions of this test, as a result of single oral administration of the hydrobromide salt of Compound Z to SD rats at 200, 600, or 2000 mg / kg, all animals showed good tolerance, and the maximum tolerated dose (MTD) is considered to be 2000 mg / kg or more.

[0108] 2. Toxicity Test by Single Oral Administration to Beagle Dogs (Method) Eight beagle dogs were randomly divided into four groups of two dogs each, with one male and one female in each group. The solvent PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v) and the hydrobromide salt of compound Z dissolved in this solvent (50, 250, 1000 mg / kg as the free base) were each administered once by gavage. After administration, the dogs were observed continuously for 14 days, and planned dissection was performed on the 15th day. Clinical findings, body weight, food intake, clinical pathology (blood, blood coagulation, plasma biochemistry), gross findings, etc. were evaluated.

[0109] (Results) During the test period, no unplanned deaths were observed in each group. In the 1000 mg / kg group, vomitus containing a suspicious drug was seen only in the female on the first day, and loose stools and vomitus containing food were seen in the male on the second day. In the 1000 mg / kg group, a significant weight loss was seen only in the male on the second day. Compared with the control group, no significant changes due to the test article were observed in the food intake, clinical pathology, and gross findings at autopsy in each dosing group. Therefore, under the conditions of this test, as a result of single-dose gavage administration of the hydrobromide salt of compound Z at 50, 250, and 1000 mg / kg to beagle dogs, all animals were tolerable, and the maximum tolerated dose (MTD) was considered to be 1000 mg / kg or more.

[0110] 3. Toxicity test by continuous 14-day gavage administration to SD rats and 14-day drug withdrawal (Method) 120 Sprague-Dawley rats were randomly divided into 4 groups of 30 each, with an equal number of males and females. The solvent, PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v), and the hydrobromide salt of compound Z dissolved in this solvent (100, 300, 600 mg / kg) were administered intragastrically once a day for 14 consecutive days. Two-thirds of each group (20 per group, equal number of males and females) were sacrificed after the dosing period (day 15), and the remaining animals in each group (10 per group, equal number of males and females) were sacrificed after a 14-day recovery period (day 29). Clinical observations, body weight, food intake, ophthalmic examination, hematology, blood coagulation, plasma biochemistry and plasma electrolytes, urine examination, gross anatomical findings, organ weights and histopathological examinations, as well as observations, measurements and evaluations of the accompanying micronucleus test in rat bone marrow were performed.

[0111] (Results) During the test period, 2 rats died in the 600 mg / Kg group. The relationship between the cause of death and the drug was unclear. In the 300 mg / kg and 600 mg / Kg groups, adverse effects due to the drug were observed in hematology, plasma biochemistry, urine examination, pathological examination, etc., but after the end of the recovery period, each test index returned to normal or showed a tendency to recover. Therefore, under the conditions of this test, after intragastric administration of the hydrobromide salt of compound Z to Sprague-Dawley rats at 100, 300, 600 mg / kg once a day for 14 consecutive days and a 14-day drug withdrawal for recovery, the no-observed-adverse-effect level (NOAEL) was 100 mg / kg, with no adverse effects observed in both males and females.

[0112] 4. Toxicity Test by 14-Day Intragastric Administration and 14-Day Drug Withdrawal in Beagle Dogs (Method) Forty beagle dogs were randomly divided into four groups of ten each, with an equal number of males and females. The solvent PEG400 / Kolliphor® HS15 / ultrapure water (40 / 10 / 50, v / v / v) and the hydrobromide salt of compound Z dissolved in this solvent (30, 100, 250 mg / kg) were administered intragastrically once a day for 14 consecutive days. Three-fifths of each group (six per group, equal number of males and females) were sacrificed at the end of the dosing period (day 15), and the remaining animals in each group (four per group, equal number of males and females) were sacrificed after a 14-day recovery period (day 29). Clinical observations, ophthalmic examinations, body weight, food intake, body temperature, electrocardiograms (HR, PR interval, QRS interval, T wave amplitude, QT interval, QTcV interval), clinical pathology (hematology, blood coagulation, plasma biochemistry, urine), and pathological examinations (gross findings, organ weights, histopathological examinations) were observed, measured, and evaluated.

[0113] (Results) During the test period, no unplanned deaths were observed in any of the animals. In the 250 mg / kg and 100 mg / kg groups, drug-related adverse effects were observed in ophthalmic examinations, clinical pathology, and pathological examinations, but after the end of the recovery period, each test index either returned to normal or showed a tendency to recover. Therefore, under the conditions of this test, when the hydrobromide salt of compound Z was administered intragastrically once a day at 30, 100, 250 mg / kg for 14 consecutive days to beagle dogs and then allowed to recover after a 14-day drug withdrawal period, the no-observed-adverse-effect level (NOAEL) for both males and females was 30 mg / kg.

[0114] All the documents referred to in the present invention are hereby incorporated by reference into the present application in the same manner as if each document was individually incorporated by reference. After reading the above content of the present invention, those skilled in the art can make various adjustments or modifications to the present invention, and it should be understood that these equivalent forms are also included within the scope defined by the appended claims.

Claims

Claim 1 A compound of formula I-1', which is a crystal having the form of crystalline form A, wherein the crystalline form A has the following feature 1): 1) In the XRPD pattern, it has characteristic peaks at least at three of the 2θ values of 5.35° ± 0.2°, 8.11° ± 0.2°, 8.46° ± 0.2°, 15.70° ± 0.2°, 18.08° ± 0.2°, 21.09° ± 0.2° and 21.91° ± 0.2°, a crystal having the same. 【Chemical Formula 2】 Claim 2 The crystalline form A has the following features 2) to 4): 2) In the DSC pattern, it has an absorption peak at 204 ± 5 °C, 3) After storage for 24 hours under the conditions of 25 °C and 80% RH, the increase in the water content is 1% or less, 4) The solubility in deionized water at 37 °C is 0.1 mg / mL or more, The crystal according to claim 1, having at least one of the above. Claim 3 In the XRPD pattern of the crystalline form A, further, it has characteristic peaks at least at three of the 2θ values of 16.02° ± 0.2°, 16.88° ± 0.2°, 17.22° ± 0.2°, 17.76° ± 0.2°, 20.55° ± 0.2°, 23.25° ± 0.2°, 23.89° ± 0.2° and 26.14° ± 0.2°. The crystal according to claim 1 or 2, characterized in that. Claim 4 A compound of formula I-2', which is a crystal having the form of crystalline form I, wherein the crystalline form I has the following feature 1): 1) In the XRPD pattern, it has characteristic peaks at least at three of the 2θ values of 5.36° ± 0.2°, 8.13° ± 0.2°, 8.48° ± 0.2°, 18.16° ± 0.2°, 20.95° ± 0.2° and 21.95° ± 0.2°, a crystal having the same. 【Chemical Formula 3】 Claim 5 The crystalline form I has the following features 2) to 4): 2) In the DSC pattern, it has an absorption peak at 200 ± 5 °C, 3) After storage for 24 hours under the conditions of 25 °C and 80% RH, the increase in the water content is 1% or less, 4) The solubility in deionized water at 37 °C is 0.1 mg / mL or more, The crystal according to claim 4, having at least one of the above. Claim 6 In the XRPD pattern of the crystalline form I, further, it has characteristic peaks at least at three of the 2θ values of 15.71° ± 0.2°, 16.07° ± 0.2°, 16.90° ± 0.2°, 17.26° ± 0.2°, 20.55° ± 0.2°, 23.27° ± 0.2° and 26.08° ± 0.2°. The crystal according to claim 4 or 5, characterized in that. Claim 7 1) A step of dissolving the compound of formula II in solvent A to obtain solution A, and mixing acid, or solution B obtained by dissolving acid in solvent B, with the solution A under ice bath conditions to obtain a mixed solution, wherein the acid is hydrogen bromide; 【Chemical Formula 5】 (In the formula, X is hydrogen or deuterium.) 2) A step of stirring the mixed solution at room temperature and concentrating it to obtain the target product; The method for producing a crystal according to any one of claims 1 to 6, comprising the above steps.

8. In step 1), The usage ratio of the compound of formula II to solvent A is 1 g: 2 - 20 mL; The acid content in solution B is 40 wt% - 50 wt%; The molar ratio of the compound of formula II to the acid is 1: 0.9 - 1; In step 2), The stirring time is 0.5 - 5 hours. The production method according to claim 7, characterized by the above.

9. 3) A step of mixing the product obtained in step 2) with solvent C, and stirring under room temperature and / or heating conditions to precipitate a solid, thereby obtaining the target product. The production method according to claim 7 or 8, further comprising the above step.

10. In step 3), The usage ratio of the compound of formula II to solvent C is 1 g: 2 - 20 mL; The temperature of the heating condition is 35 - 60 °C; The stirring time is 0.5 - 5 hours. The production method according to claim 9, characterized by the above.

11. Solvent A, solvent B and solvent C are each independently selected from water, hydrocarbons, alcohols, ethers, ketones, esters, nitriles and homogeneous mixtures thereof; The hydrocarbons are selected from n - pentane, n - hexane, n - heptane, petroleum ether, dichloromethane, chloroform, carbon tetrachloride, 1,2 - dichloroethane, benzene, toluene, xylene, chlorobenzene and dichlorobenzene; The alcohols are selected from methanol, ethanol, n - propanol, isopropanol, n - butanol, ethylene glycol and propylene glycol; The ether is selected from ethyl ether, n-propyl ether, isopropyl ether, methyl t-butyl ether, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol dimethyl ether, tetrahydrofuran, dioxane, dimethoxyethane and diethylene glycol dimethyl ether, The ketone is selected from acetone, butanone and diethyl ketone, The ester is selected from methyl formate, ethyl formate, propyl formate, butyl formate, methyl acetate, ethyl acetate, propyl acetate and butyl acetate, The nitrile is selected from acetonitrile and propionitrile, and The solvent A and the solvent B are miscible with each other The production method according to claim 9 or 10, characterized in that.

12. In step 1), The solvent A is acetonitrile, the usage ratio of the compound of formula II to the acetonitrile is 1 g: 2 to 20 mL, the solvent B is water, the solution B is hydrobromic acid, and the content of hydrogen bromide in the hydrobromic acid is 40 wt% to 50 wt%, and the molar ratio of the compound of formula II to the hydrogen bromide is 1: 0.9 to 1, In step 2), The stirring time is 0.5 to 5 hours, In step 3), The solvent C is methyl t-butyl ether, the usage ratio of the compound of formula II to the methyl t-butyl ether is 1 g: 2 to 20 mL, the temperature of the heating condition is 35 to 60 ° C, and the stirring time is 0.5 to 5 hours The production method according to claim 9, characterized in that.

13. 1) The crystal according to any one of claims 1 to 6, 2) Optionally, a pharmaceutically acceptable additive A pharmaceutical composition comprising.

14. The pharmaceutical composition is an oral preparation or a parenteral preparation, The oral preparation is selected from tablets, capsules, granules, powders and syrups, The parenteral preparation is selected from injections, powder injections, sprays and suppositories The pharmaceutical composition according to claim 13, characterized in that.

15. The crystal according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 13 or 14 for use in the treatment and / or alleviation of a disease caused by a virus.

16. The pharmaceutical composition according to claim 15, wherein the virus is SARS-CoV-2.

17. Use of the crystal according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 13 or 14 for the manufacture of a medicament for the treatment and / or alleviation of a disease caused by a virus.

18. The use according to claim 17, wherein the virus is SARS-CoV-2.

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