HK40038962A - Solid form of dihydropyrimidine compound and preparation method therefor and use thereof - Google Patents

Description

Solid form of dihydropyrimidine compound, preparation method and application thereof Technical Field

The present invention relates to solid forms of (E) -3- ((R) -4- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) morpholin-2-yl) acrylic acid (hereinafter referred to as "compound of formula (I)"), processes for preparing said solid forms, pharmaceutical compositions comprising said solid forms, and the use of said solid forms for the prevention or treatment of viral diseases including but not limited to viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes and Acquired Immune Deficiency Syndrome (AIDS).

Background

Hepatitis B Virus (HBV) is a common hepadnatropic viral pathogen. The virus can cause diseases such as acute hepatitis, chronic hepatitis, hepatic fibrosis, liver cirrhosis, liver cancer and the like.

Drugs for the treatment of hepatitis B include interferons and nucleoside analogs (e.g., lamivudine and adefovir dipivoxil). Wherein the interferon causes cells to produce antiviral protein by reacting with cell surface receptor, thereby inhibiting replication of hepatitis B virus. Its disadvantages are low effective response rate and long-term injection administration. Nucleoside analogs function primarily by inhibiting replication of the viral polymerase (reverse transcriptase). The medicine has the defect that the long-term administration of the medicine is easy to cause virus variation and generate drug resistance.

In addition, viral hepatitis B can be treated with non-nucleoside analogs. Research by Deres et al found that heteroaryl dihydropyrimidines (Bay41-4109) could prevent HBV viral replication by inhibiting viral capsid protein assembly (Science, 2003, 299, 893-896). The specific mechanism of action is that dihydropyrimidine compounds induce core protein misassembly, thereby forming unstable capsid protein and accelerating the degradation of core protein (biochem. Pharmacol., 2003, 66, 2273-2279). The heteroaryl dihydropyrimidines HAP1(Proc. Natl. Acad. Sci., 2005, 102, 8138-.

Although all of the above compounds exhibit antiviral activity to varying degrees, their activity has not been satisfactory and some compounds also exhibit significant toxic effects (e.g., GLS4 exhibits significant hERG cardiotoxicity).

The compound of formula (I) developed recently not only shows high antiviral property, but also no cardiotoxicity, has good pharmacokinetic properties, and is further beneficial to improve the therapeutic effect on viral diseases:

different crystal forms of the same drug may have obvious differences in the aspects of stability, bioavailability and the like, thereby affecting the curative effect of the drug. Therefore, the discovery and the acquisition of a stable crystal form of a compound are more beneficial to the processing of medicaments and the use in pharmaceutical compositions, and have very important significance in providing more qualitative and quantitative information for the curative effect research of solid medicaments, and are urgent requirements of the medicament development process.

Summary of The Invention

The present invention provides solid forms of the compound of formula (I) having the chemical name: (E) -3- ((R) -4- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) morpholin-2-yl) acrylic acid:

in one aspect, the invention provides a crystal a of the compound of formula (I) having an X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at diffraction angles (2 θ) of 8.7 ± 0.2 °, 17.5 ± 0.2 °, 19.3 ± 0.2 °, 20.3 ± 0.2 ° and 21.4 ± 0.2 °.

Another aspect of the present invention provides a method for preparing the crystalline a or amorphous form of the present invention.

Another aspect of the present invention provides a pharmaceutical composition comprising crystalline a and/or amorphous form of the present invention, and one or more pharmaceutically acceptable carriers or one or more other therapeutic agents.

Another aspect of the present invention provides a pharmaceutical formulation comprising crystalline a and/or amorphous form of the present invention, and one or more pharmaceutically acceptable carriers.

Another aspect of the present invention provides a use of the crystalline a and/or amorphous form of the present invention, the pharmaceutical composition of the present invention, and/or the pharmaceutical preparation of the present invention for the preparation of a medicament for the prevention or treatment of a viral disease.

Another aspect of the present invention provides the crystalline a and/or amorphous form of the present invention, the pharmaceutical composition of the present invention, and/or the pharmaceutical preparation of the present invention, which is used for preventing or treating viral diseases.

Another aspect of the present invention provides a method for preventing or treating a viral disease, which comprises administering an effective amount of the crystalline a and/or amorphous form of the present invention, the pharmaceutical composition of the present invention and/or the pharmaceutical preparation of the present invention to a subject in need thereof.

Detailed Description

Definition of

Unless defined otherwise below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. Reference to the techniques used herein is intended to refer to those techniques commonly understood in the art, including those variations of or alternatives to those techniques that would be apparent to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

As used herein, the terms "comprising," "including," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps, although not necessarily present (i.e., these terms also encompass the terms "consisting essentially of … …" and "consisting of … …").

The word "about" as used herein refers to a value that one of ordinary skill in the art would consider to be within an acceptable standard error of the stated value, e.g., ± 0.05, ± 0.1, ± 0.2, ± 0.3, ± 0.5, ± 1, ± 2 or ± 3, etc.

The term "solid form" as used herein includes all solid state forms of the compound of formula (I), e.g. crystalline forms or amorphous forms.

The term "amorphous" as used herein refers to any solid substance that is not ordered in three dimensions. In some cases, amorphous solids can be characterized by known techniques including XRPD crystal diffraction analysis, solid-state nuclear magnetic resonance (ssNMR) spectroscopy, Differential Scanning Calorimetry (DSC), or some combination of these techniques. As demonstrated below, amorphous solids produce XRPD patterns without distinct diffraction signature peaks.

The term "crystalline form" or "crystalline" as used herein refers to any solid substance exhibiting a three-dimensional ordering, as opposed to an amorphous solid substance, which results in a characteristic XRPD pattern having well-defined peaks.

The term "X-ray powder diffraction pattern (XRPD pattern)" as used herein refers to an experimentally observed diffraction pattern or a parameter, data or value derived therefrom. XRPD patterns are generally characterized by peak position (abscissa) and/or peak intensity (ordinate).

The term "2 θ" as used herein refers to the peak position in degrees (°) set based on X-ray diffraction experiments, and is typically the abscissa unit in the diffraction pattern. If the reflection is diffracted when the incident beam makes an angle theta with a certain lattice plane, the experimental setup requires recording the reflected beam at an angle of 2 theta. It should be understood that reference herein to particular 2 theta values for particular crystalline forms is intended to refer to 2 theta values (in degrees) measured using the X-ray diffraction experimental conditions described herein.

The term "Differential Scanning Calorimetry (DSC) profile" as used herein refers to the curve recorded by a differential scanning calorimeter.

The term "thermogravimetric analysis (TGA) profile" as used herein refers to the curve recorded by a thermogravimetric analyzer.

As used herein, the term "substantially the same" means that representative peak position and/or intensity variations are taken into account. For example, for X-ray diffraction peaks, one skilled in the art will appreciate that the peak position (2 θ) will show some variation, typically as much as 0.1-0.2 degrees, and that the instruments used to measure diffraction will also cause some variation. In addition, those skilled in the art will appreciate that relative peak intensities will vary due to inter-instrument variation as well as the degree of crystallinity, preferred orientation, prepared sample surface, and other factors known to those skilled in the art.

Crystal and preparation method

In some embodiments, the present invention provides a crystal A of the compound of formula (I), characterized in that said crystal A has an X-ray powder diffraction pattern having characteristic peaks at diffraction angles of 8.7 + -0.2 °, 17.5 + -0.2 °, 19.3 + -0.2 °, 20.3 + -0.2 ° and 21.4 + -0.2 °,

in a preferred embodiment, the XRPD pattern of crystal a of said compound of formula (I) comprises characteristic peaks at diffraction angles (2 θ) of 8.7 ± 0.2 °, 16.0 ± 0.2 °, 17.5 ± 0.2 °, 17.8 ± 0.2 °, 19.3 ± 0.2 °, 20.3 ± 0.2 °, 21.4 ± 0.2 °, 22.3 ± 0.2 ° and 23.1 ± 0.2 °.

In a more preferred embodiment, the XRPD pattern of crystal a of said compound of formula (I) comprises characteristic peaks at diffraction angles (2 θ) of 8.7 ± 0.2 °, 10.8 ± 0.2 °, 15.8 ± 0.2 °, 16.0 ± 0.2 °, 17.5 ± 0.2 °, 17.8 ± 0.2 °, 19.3 ± 0.2 °, 19.5 ± 0.2 °, 20.3 ± 0.2 °, 21.1 ± 0.2 °, 21.4 ± 0.2 °, 22.3 ± 0.2 °, 23.1 ± 0.2 ° and 27.0 ± 0.2 °.

In a particularly preferred embodiment, the XRPD pattern of crystal a of said compound of formula (I) comprises characteristic peaks at the following diffraction angles (2 Θ), with the 2 Θ values having a range of error of ± 0.2 °:

in a more preferred embodiment, the XRPD pattern of crystal a of said compound of formula (I) comprises peaks at diffraction angles (2 θ) substantially the same as shown in figure 1. In a most preferred embodiment, the XRPD pattern of crystal a of said compound of formula (I) is as shown in figure 1.

In some embodiments, the compound of formula (I) has a Differential Scanning Calorimetry (DSC) profile of crystal a that exhibits an endothermic peak at 173 ± 2 ℃.

In a more preferred embodiment, the DSC pattern of said crystal a is as shown in figure 2.

In some embodiments, the thermogravimetric analysis (TGA) profile of the crystal a of the compound of formula (I) begins to decompose at 190 ± 2 ℃.

In a preferred embodiment, the TGA profile of crystal a is shown in figure 3.

Another aspect of the present invention provides a process for preparing crystals a of the compound of formula (I) above, including but not limited to slow volatilization, suspension stirring, osmosis or recrystallization.

Wherein, the solvent adopted in the method is selected from: methanol, ethanol, n-propanol, isopropanol, n-butanol, acetone, butanone, ethyl acetate, butyl acetate, isopropyl acetate, dimethyl carbonate, tetrahydrofuran, dichloromethane, trichloromethane, methyl tert-butyl ether, acetonitrile, anisole, toluene, diethyl ether, water, isopropyl ether, n-hexane, n-heptane, cyclohexane, petroleum ether.

In some embodiments of the invention, crystal a is prepared using a slow volatilization method comprising the steps of: dissolving a compound of formula (I) in a proper solvent 1, standing at room temperature, volatilizing, removing the solvent, and collecting a solid to obtain a crystal A;

wherein the 1 st suitable solvent is selected from: one or more of methanol, ethanol, n-propanol, isopropanol, n-butanol, acetone, butanone, ethyl acetate, butyl acetate, isopropyl acetate, dimethyl carbonate, tetrahydrofuran, dichloromethane, trichloromethane, methyl tert-butyl ether, acetonitrile, anisole, toluene and diethyl ether; the solvent is used in an amount that the compound of formula (I) is dissolved out.

Preferably, the mass to volume ratio (mg/ml) of said compound of formula (I) to the 1 st suitable solvent is from 100: 1 to 10: 1.

Alternatively, the 1 st suitable solvent is a mixed solvent of a solvent a and a solvent B, wherein the solvent a is selected from: one or more of acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane, water, methyl tert-butyl ether; the solvent B is selected from one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene and dichloromethane;

preferably, the volume ratio of the solvent A to the solvent B is 1: 2-2: 1; preferably, the volume ratio of the solvent A to the solvent B is 1: 1;

preferably, the solvent A or the solvent B is used in such an amount that the compound of formula (I) is completely dissolved.

In some embodiments of the invention, crystal a is prepared using a slow volatilization method comprising the steps of: dissolving a compound shown in a formula (I) in a solvent A, adding a solvent B, mixing, standing at room temperature, volatilizing, removing the solvent, and collecting a solid to obtain a crystal A; preferably, the a solvent is selected from: one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane, water and methyl tert-butyl ether; the solvent B is selected from one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane and water;

preferably, the volume ratio of the solvent A to the solvent B is 1: 2-2: 1; preferably, the volume ratio of the solvent A to the solvent B is 1: 1.

In other embodiments of the present invention, crystal a is prepared using a recrystallization process comprising the steps of: heating and stirring the compound of the formula (I) in a proper solvent of the formula 2, and dissolving; slowly cooling to room temperature, filtering, and collecting solid to obtain crystal A;

preferably, the 2 nd suitable solvent is selected from isopropanol, acetonitrile, toluene, n-propanol; preferably, the mass-to-volume ratio (g/ml) of the compound of formula (I) to the solvent is 1: 5-10;

alternatively, the suitable solvent of formula 2 is diethyl ether, and the mass to volume ratio (g/ml) of the compound of formula (I) to diethyl ether is 1: 60.

In other embodiments of the present invention, crystal a is prepared using a suspension stirring method, comprising the steps of: dispersing and suspending the compound of the formula (I) or amorphous substances thereof by using a proper solvent of the No. 3, suspending and stirring at room temperature or high temperature, and collecting solids to obtain crystals A;

preferably, said 3 rd suitable solvent is selected from: one or more of ethanol, n-propanol, n-butanol, diethyl ether, isopropyl ether, acetonitrile, toluene, n-hexane, n-heptane, cyclohexane, methyl tert-butyl ether, water and petroleum ether; preferably, the 3 rd appropriate solvent is used in an amount that the solute is suspended therein (mass to volume ratio (mg/ml) of solute to solvent is preferably 150: 1 to 10: 1); preferably, the high temperature condition is 60 ℃.

In still other embodiments of the present invention, crystals a are prepared using an infiltration process comprising the steps of: placing the compound of formula (I) in a container A, placing in a container B containing an appropriate amount of a 4 th appropriate solvent, sealing the container B, standing at room temperature, and collecting the solid to obtain a crystal A;

preferably, said 4 th suitable solvent is selected from: one or more of methanol, ethanol, isopropyl acetate, n-hexane, acetonitrile, diethyl ether, methyl tert-butyl ether and toluene; preferably, the standing time is not less than 8 days.

Another aspect of the invention provides an amorphous form of the compound of formula (I) having an XRPD pattern without sharp distinct diffraction peaks.

In a preferred embodiment, the amorphous form has an XRPD pattern as shown in figure 4.

Another aspect of the present invention provides a process for preparing an amorphous form of the compound of formula (I), said process comprising the steps of: dissolving the compound of the formula (I) in a proper solvent of 5 th for clarification, and then carrying out reduced pressure rotary evaporation to obtain the amorphous substance;

preferably, the 5 th suitable solvent is selected from one or more of dichloromethane, chloroform;

preferably, the temperature of the water bath is 40-50 ℃; more preferably 45 deg.c.

In yet another aspect, the compounds of formula (I) of the present invention may form corresponding salts of compounds of formula (I) with organic or inorganic acids, including but not limited to: fumarate, citrate, tartrate, phosphate, maleate, succinate, adipate, sulfate, hydrochloride, carbonate, phosphate, hydrobromide, nitrate, malate, glycolate, mucate, lactate, gentisate, methanesulfonate, camphorsulfonate, benzenesulfonate, p-toluenesulfonate, ethanedisulfonate, naphthalenedisulfonate, hippurate, nicotinate, oxalate, malonate, L-arginine, lysine;

wherein the molar ratio of the compound of formula (I) to the organic or inorganic acid is preferably 1: 1 or 2: 1 or 3: 1.

In another aspect, the compounds of formula (I) of the present invention may form a salt with an organic or inorganic base of the corresponding compound of formula (I).

Preferably, the salts of the compounds of formula (I) with inorganic bases include, but are not limited to, ammonium, magnesium, potassium, sodium, calcium, lithium salts, and the like;

alternatively, the organic base is selected from: meglumine, benzylamine, betaine, dimethylethanolamine, diethylaminoethanol, tromethamine, diethanolamine, ethylenediamine, imidazole, piperazine, tromethamine, triethylamine, choline and the like;

wherein the molar ratio of the compound of formula (I) to the organic or inorganic base is preferably 1: 1 or 2: 1 or 3: 1.

Pharmaceutical compositions and methods of treatment

A further aspect of the invention provides a pharmaceutical composition comprising crystalline a and/or amorphous form of a compound of formula (I) of the invention, together with one or more pharmaceutically acceptable carriers or one or more other therapeutic agents.

The "other therapeutic agent" means other substances having pharmacological activity than the crystalline A and/or amorphous form of the compound of formula (I) of the present invention, for example, other antiviral agents capable of exerting a synergistic therapeutic effect with the compound of formula (I).

A further aspect of the invention provides a pharmaceutical formulation comprising crystalline a and/or amorphous form of a compound of formula (I) of the invention, together with one or more pharmaceutically acceptable carriers.

The "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic agent is administered and which is, within the scope of sound medical judgment, suitable for contact with the tissues of humans and/or other animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.

The pharmaceutical formulations of the present invention may act systemically and/or locally. For this purpose, they may be administered by a suitable route, for example by injection, intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or transdermal administration; or by oral, buccal, nasal, transmucosal, topical, in the form of ophthalmic preparations or by inhalation.

For these routes of administration, the formulations of the present invention may be administered in suitable dosage forms.

The dosage form may be a solid, semi-solid, liquid, or gaseous formulation, including, but not limited to, tablets, capsules, powders, granules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, suspensions, elixirs, syrups.

The pharmaceutical formulations of the present invention may be prepared by any method known in the art, for example, by mixing, dissolving, granulating, sugar-coating, milling, emulsifying, lyophilizing, and the like.

The amount or amount of a compound of the invention in a pharmaceutical formulation may be from about 0.01mg to about 1000mg, suitably 0.1-500mg, preferably 0.5-300mg etc.

A further aspect of the present invention provides the use of a crystalline a and/or amorphous form of a compound of formula (I), a pharmaceutical composition of the invention and/or a pharmaceutical formulation of the invention in the manufacture of a medicament for the prevention or treatment of a viral disease; preferably, the viral disease includes, but is not limited to, viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes and Acquired Immune Deficiency Syndrome (AIDS).

A further aspect of the present invention provides a method for preventing or treating a viral disease, which comprises administering to a subject in need thereof an effective amount of crystalline a of a compound of formula (I) of the present invention, an amorphous form, a pharmaceutical composition of the present invention and/or a pharmaceutical formulation of the present invention. Preferably, the viral disease includes, but is not limited to, viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes and Acquired Immune Deficiency Syndrome (AIDS).

A further aspect of the present invention provides a crystalline a and/or amorphous form of the compound of formula (I), a pharmaceutical composition of the present invention or a pharmaceutical formulation of the present invention for use in the prevention or treatment of a viral disease. Preferably, the viral disease includes, but is not limited to, viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes and Acquired Immune Deficiency Syndrome (AIDS).

The crystal of the compound of formula (I) provided by the invention has excellent effect in preventing or treating viral diseases, and also shows good chemical stability, physical stability and pharmacokinetic properties. For example, the crystalline form of the compound of formula (I) of the present invention has excellent solubility, low hygroscopicity, and the like, and thus is more advantageous for sufficient dissolution upon administration and formulation, maintaining sufficient biological activity. Meanwhile, the high-temperature-resistant high-humidity-resistant high-; and, it has good light stability, and does not need special packaging treatment for preventing the influence of light, thereby reducing the cost. The degradation caused by the influence of illumination can not be generated, and the safety of the medicine and the effectiveness after long-term storage are improved.

Drawings

FIG. 1: an XRPD pattern for crystal A of the compound of formula (I).

FIG. 2: DSC profile of crystal a of compound of formula (I).

FIG. 3: a TGA profile of a crystal a of the compound of formula (I).

FIG. 4: an XRPD pattern of an amorphous form of the compound of formula (I).

FIG. 5: a DVS profile of crystal a of the compound of formula (I).

FIG. 6: XRPD comparison pattern of crystal a of compound of formula (I) after 0 and 7 days standing at 60 ℃.

FIG. 7: the XRPD pattern of the compound of formula (I) prepared in example 1.

Examples

The present invention is further illustrated by the following examples, which are only for illustrating the technical solutions of the present invention and are not intended to limit the scope of the present invention, and those skilled in the art can make some insubstantial modifications and adjustments, and still fall within the scope of the present invention.

Test instrument information and methods used for the experiment:

x-ray powder diffraction (XRPD):

an X' Pert3 Powder diffractometer was used, which was irradiated with a Cu target and detected using an Absolute scan at room temperature. The detection range was 3.5 ° to 40 °, the step size was 0.013 °, the dwell time was 50s, and 1 scan.

Differential Scanning Calorimetry (DSC) test apparatus was: DSC1(METTLER TOLEDO).

The thermogravimetric analysis (TGA) test instrument was: METTLER TOLEDO.

The heating rates of the DSC and TGA instruments were 10K/min.

The dynamic moisture sorption instrument (DVS) experimental conditions were as follows:

detection was performed using DVS Intrasic (SMS) in cycle-DMDT mode at 25 ℃.

Example 1 preparation of (E) -3- ((R) -4- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) morpholin-2-yl) acrylic acid (compound of formula (I))

Methyl (R) -6- (bromomethyl) -4- (2-chloro-4-fluorophenyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylate (400mg, 0.90mmol) and trifluoroacetic acid salt of (R, E) -3- (morpholin-2-yl) acrylic acid (488mg, 1.80mmol) were dissolved in dichloromethane (10mL) at room temperature, followed by addition of N, N-diisopropylethylamine (696mg, 5.40mmol) and reaction at room temperature overnight. And concentrating the reaction solution to obtain a crude product. The crude product was purified by preparative liquid chromatography to give 205mg of the compound of formula (I). The resulting sample was subjected to XRPD analysis, the XRPD pattern being shown in figure 7, which indicated that the resulting solid was an amorphous form of the compound of formula (I).

The structure is characterized as follows:

¹H NMR(400MHz，DMSO-d₆)12.44(s，1H)，9.68(s，1H)，7.98(dd，J＝27.6，3.1Hz，2H)，7.48-7.36(m，2H)，7.18(td，J＝8.5，2.6Hz，1H)，6.73(dd，J＝15.8，4.1Hz，1H)，6.04(s，1H)，5.93(dd，J＝15.8，1.6Hz，1H)，4.23(d，J＝9.3Hz，1H)，4.01-3.90(m，3H)，3.68(t，J＝10.2Hz，1H)，3.52(s，3H)，2.94(d，J＝11.0Hz，1H)，2.82(d，J＝11.1Hz，1H)，2.41(dd，J＝11.0，8.6Hz，1H)，2.08(t，J＝10.7Hz，1H)。ESI-MS(m/z)：521.1[M+H]⁺。

EXAMPLE 2 preparation of Crystal A of the Compound of formula (I)

30mg of the compound of formula (I) prepared in example 1 was weighed, dissolved in 0.9ml of methanol, and then allowed to stand at room temperature, evaporated, the solvent removed, crystallized, the solid collected, and the resulting sample was subjected to XRPD detection, the XRPD pattern is shown in FIG. 1, and the relevant data is shown in Table 1, i.e. the crystal A of the compound of formula (I).

TABLE 1

Performing DSC measurement on the obtained sample, wherein the DSC spectrum is shown in figure 2, and the peak value of an endothermic peak appears at 173.18 ℃;

the resulting samples were subjected to TGA measurements and the resulting TGA profile is shown in FIG. 3, with decomposition beginning at about 190 ℃.

EXAMPLE 3 preparation of Crystal A of the Compound of formula (I)

Following the same procedure as in example 2, wherein the methanol in example 2 was replaced with the solvent and the amount in table 2 below, crystal a of the compound of formula (I) was prepared and XRPD examination of the obtained sample showed that the obtained product was the same as crystal a obtained in example 2.

TABLE 2

EXAMPLE 4 preparation of Crystal A of the Compound of formula (I)

The compound of formula (I) prepared in example 1 was dissolved in solvent A to prepare a solution containing 30mg/ml of the compound of formula (I), and then solvent B was added in an amount equal to the volume of solvent A (the types and volumes of solvent A and solvent B are shown in Table 3), and the mixture was allowed to stand at room temperature, evaporated, removed, crystallized, and collected as a solid, and XRPD detection showed that the obtained crystals were the same as those of crystal A of example 2.

TABLE 3

EXAMPLE 5 preparation of Crystal A of the Compound of formula (I)

500mg of the compound of formula (I) prepared in example 1 was weighed, and a certain amount of solvent (see Table 4 for the kind of solvent and the addition ratio) was added thereto, heated and stirred, and dissolved to be clear, and then slowly cooled to room temperature, filtered, and the solid was collected, and XRPD detection showed that the obtained crystals were the same as the crystals A of example 2.

TABLE 4

Solvent(s)	Ratio (solute: solvent)
N-propanol	0.5g/4.0ml
Acetonitrile	0.5g/4.0ml
Ether (A)	0.5g/30.0ml
Isopropanol (I-propanol)	0.5g/4.0ml
Toluene	0.5g/4.0ml

EXAMPLE 6 suspension crystallization of amorphous form of Compound of formula (I)

50mg of the compound of the formula (I) prepared in example 1 was weighed, and 0.5ml of a solvent (the kind of the solvent and the addition ratio are shown in Table 5) was added thereto to disperse and suspend, followed by hermetically stirring at room temperature for 72 hours, filtration, collection of a solid, and XRPD detection showed that the obtained crystals were the same as the crystals A of example 2.

TABLE 5

Solvent(s)	Ratio (solute: solvent)
Ethanol	50mg/0.5ml
N-propanol	50mg/0.5ml
N-butanol	50mg/0.5ml
N-hexane	50mg/2.0ml
N-heptane	50mg/2.0ml
Cyclohexane	50mg/2.0ml
Acetonitrile	50mg/0.5ml
Ether (A)	50mg/1.0ml
Methyl tert-butyl ether	50mg/0.5ml
Isopropyl ether	50mg/1.0ml
Toluene	50mg/0.5ml
Water (W)	50mg/2.0ml
Petroleum ether	50mg/2.0ml

EXAMPLE 7 preparation of Crystal A of the Compound of formula (I)

200mg of a compound of the formula (I) prepared in example 1 was weighed out, dispersed and suspended by adding 4ml of a solvent (see Table 6 for the kind of the solvent and the addition ratio), heated and stirred at 60 ℃ for 8 hours, filtered, and the solid was collected and the XRPD test showed that the obtained crystals were the same as the crystals A of example 2.

TABLE 6

Solvent(s)	Ratio (solute: solvent)
N-hexane	0.2g/4.0ml
Cyclohexane	0.2g/4.0ml
N-heptane	0.2g/4.0ml
Isopropyl ether	0.2g/4.0ml
Petroleum ether	0.2g/4.0ml
Water (W)	0.2g/4.0ml

EXAMPLE 8 suspension Trans-crystallization of a Compound of formula (I)

A sample of 150mg of the compound of formula (I) prepared in example 1 was weighed, dispersed and suspended by adding an appropriate amount of solvent (see table 7 for the kind of solvent and the addition ratio), sealed and stirred at room temperature for 72 hours, filtered, and the solid was collected and XRPD detected to show that the obtained crystals were the same as crystal a of example 2.

TABLE 7

EXAMPLE 9 preparation of Crystal A of the Compound of formula (I)

A sample of 30mg of the compound of formula (I) prepared in example 1 was weighed into a vial, placed open in a 50ml beaker containing the appropriate amount of solvent (see table 8 for the type and amount of solvent) and sealed, left to stand at room temperature for about 8 days, and the solid was collected and detected by XRPD to obtain the same crystals as crystal a of example 2.

TABLE 8

Solvent(s)	Volume (ml)
Methanol	6ml
Ethanol	6ml
Acetic acid isopropyl ester	6ml
N-hexane	6ml
Acetonitrile	6ml
Ether (A)	12ml
Methyl tert-butyl ether	12ml
Toluene	6ml

EXAMPLE 10 preparation of amorphous form of Compound of formula (I)

3g of a compound of formula (I) prepared in example 1 was weighed, dissolved in 50ml of a solvent (see Table 9 for the kind of solvent and the addition ratio), filtered, and then dried by rotary evaporation in a water bath at 45 ℃ under reduced pressure to obtain a solid, and the obtained sample was subjected to XRPD analysis, the XRPD pattern of which is shown in FIG. 4, to obtain an amorphous form of the compound of formula (I).

TABLE 9

Solvent(s)	Ratio (solute: solvent)
Methylene dichloride	3g/50ml
Chloroform	3g/50ml

Examples of the experiments

Experimental example 1 stability investigation of Crystal A at Room temperature

A crystal A sample of the compound of the formula (I) is placed at room temperature, and is sampled and detected for XRPD on the fifth day and the fifteenth day respectively.

And (3) detection results: the XRPD pattern showed that after fifteen days at room temperature the crystals were the same as in example 2 and crystal a had not been transformed.

Experimental example 2 stability examination of Crystal A under high temperature conditions

A crystal A sample of the compound of formula (I) is placed under vacuum at 60 ℃ and sampled for XRPD detection on the next, fifth and seventh days.

And (3) detection results: the XRPD detection result shows that after the crystal is placed under vacuum at 60 ℃ for seven days, the crystal A is the same as the crystal A in the example 2, and the crystal A is not transformed (the XRPD pattern of the crystal A after the crystal A is placed at 60 ℃ for 7 days and the comparison pattern of the crystal A at 0 day are shown in figure 6).

Experimental example 3 stability examination of Crystal A under grinding conditions

A crystal A sample of the compound of the formula (I) is uniformly ground in a mortar, and samples are taken for detection of XRPD after 2min and 5min of grinding respectively.

And (3) detection results: XRPD detection showed that after 5min of milling, the crystals were the same as in example 2, with no transformation of crystal a.

Experimental example 4 DVS examination of Crystal A

A sample of crystal a of the compound of formula (I) is tested for hygroscopicity by means of a dynamic moisture sorption instrument (DVS): the hygroscopicity is measured at 25 ℃ and 10% step humidity in the range of 0% -90% -0%.

The DVS assay profile is shown in fig. 5, with the results showing: the moisture-attracting weight gain of the sample is 0.28% under the humidity condition of 80%, and the moisture-attracting weight gain is in the range of 0.2% -2% according to the regulation of 'Chinese pharmacopoeia' 2015 edition, belonging to 'slight moisture-attracting'.

Experimental example 5 pharmacokinetic experiment of Crystal A in rat

1. Sample to be tested

Preparing an intravenous administration test article: a sample of example 5 (the crystal preparation solvent is isopropanol) is dissolved by using 5% DMSO + 5% Solutol (HS15) + 90% normal saline to be used as a test solution for intravenous administration;

preparing a gastric lavage administration test sample: a sample of example 5 (isopropanol as the crystal preparation solvent) was prepared as a suspension using 97.5% 0.5% Methylcellulose (MC) + 2.5% Solutol (HS15) and used as a test for intragastric administration.

2. Test methods and results

Randomly dividing 12 SD rats into A, B groups with 6 rats each group and half each animal, and respectively performing single intravenous administration on group A to obtain test drugs(ii) a And B, respectively intragastrically administering the test sample medicines with the administration dose of 3 mg/kg. The blood sampling time points for intravenous and intragastric administration were 0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 10 and 24h before and after administration. Pharmacokinetic parameters were calculated from the blood concentrations and the results are shown in table 10. As can be seen from the data in Table 10, the absolute bioavailability of Crystal A in rats was 62.4% (as AUC)_0-∞Calculated) which is well absorbed orally in rats.

Table 10 pharmacokinetic parameters after single administration of crystal a in rats (n ═ 6, male and female halves)

Remarking: t is_maxData are median (range).

EXPERIMENTAL EXAMPLE 6 safety test of Crystal A of Compound represented by formula (I)

Preparing a test article: the required amount of the crystal a sample prepared in example 5 (the crystal preparation solvent was isopropanol) was weighed, put in a mortar, added with polyethylene glycol-15 hydroxystearate (HS15) in an amount of 2.5% by volume of the total volume, and ground uniformly, and then washed with 0.5% methylcellulose solution (MC) until no visible residue remained, and transferred to a calibrated container, and then the volume was fixed with 0.5% methylcellulose solution, and stirred uniformly with a magnetic stirrer to prepare a suspension as a test sample.

Single dose toxicity in SD rats

SD rats were subjected to single-dose toxicity test in 4 groups, namely a vehicle control group and low, medium and high dose groups of the compound crystal A represented by the formula (I), wherein each group comprises 10 rats and each half of the rats are male and female. The administration was continued for 14 days after oral gavage. The animal condition is found to be good in the observation period, the weight and the food intake do not change obviously, the hematology and blood biochemical indexes are normal, and no abnormal condition is found in the general anatomical observation. At the end of the test, it was found that SD rats had better tolerance to the crystal A of the compound represented by the formula (I) under the test conditions, and no abnormality was found within 14 days after the administration.

Single dose toxicity in Beagle dogs

The single-dose toxicity test of Beagle dogs sets 3 groups, namely a solvent control group and a low-dose group and a high-dose group of the compound crystal A shown in the formula (I), wherein each group comprises 4 animals and half of animals. Each group of dogs was observed for 14 consecutive days after oral gavage. The animal condition is good, the weight is not obviously changed, the hematology and blood biochemical indexes are normal, the II-lead electrocardiogram, the respiratory frequency and the blood pressure are not obviously abnormal, and the abnormal condition is not found in the general anatomical observation in the observation period.

The test result shows that the Beagle dog has better tolerance to the crystal A of the compound shown in the formula (I) by single administration, and no abnormality exists within 14 days after the administration.

Experimental example 7 solubility measurement of Crystal A

Testing the solvent: methanol, acetonitrile, ethanol, isopropanol, 0.1mol/L hydrochloric acid aqueous solution and 0.1mol/L sodium hydroxide aqueous solution.

The operation method comprises the following steps: the examination is carried out according to the fifteenth (2) item of the four general examples in the Chinese pharmacopoeia 2015 edition. Weighing a proper amount of the crystal A sample prepared in example 5 (the crystal preparation solvent is isopropanol), adding the crystal A sample into a certain volume of solvent at 25 +/-2 ℃, and strongly shaking for 30 seconds every 5 minutes; dissolution was observed within 30 minutes, as no visible solute particles were present, i.e., complete dissolution was observed.

And (3) test results: the crystal A sample was dissolved in methanol, acetonitrile, ethanol, slightly soluble in isopropanol, and slightly soluble in 0.1mol/L aqueous hydrochloric acid and 0.1mol/L aqueous sodium hydroxide.

Experimental example 8 determination of logP value of Crystal A

The logP value of the sample of crystal a prepared in example 5 (the crystal preparation solvent was isopropanol) was measured using a Sirius T3 physico-chemical constant measuring instrument. The examination method was a Sirius logP test (pH-metric medium logP), and the pH-metric medium logP assay mode was selected, with titration sequences ranging from low to high pH.

And (3) measuring results: the logP value of the product is 2.4-2.6. The crystal A has excellent membrane permeability, and is beneficial to ADME process in vivo and receptor affinity.

The solid forms of the compounds of formula (I) and the methods for their preparation disclosed herein can be prepared by those skilled in the art by appropriate modification of the starting materials, process parameters, and the like, in view of the disclosure herein. While the methods and products of the present application have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that the techniques of the present application can be practiced with modification, or with appropriate modification, and combinations of the methods and products described herein without departing from the spirit and scope of the present application. It is expressly intended that all such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and content of this application.

Claims (18)

1. A crystal A of the compound of formula (I), characterized in that it has an X-ray powder diffraction pattern having characteristic peaks at diffraction angles of 8.7 + -0.2 °, 17.5 + -0.2 °, 19.3 + -0.2 °, 20.3 + -0.2 ° and 21.4 + -0.2 °,
2. the crystal A according to claim 1, characterized in that the crystal A has an X-ray powder diffraction pattern having characteristic peaks at diffraction angles of 8.7 ± 0.2 °, 16.0 ± 0.2 °, 17.5 ± 0.2 °, 17.8 ± 0.2 °, 19.3 ± 0.2 °, 20.3 ± 0.2 °, 21.4 ± 0.2 °, 22.3 ± 0.2 ° and 23.1 ± 0.2 °; preferably, the crystal A has an X-ray powder diffraction pattern as shown in figure 1.
3. Crystal a according to claim 1 or 2, characterized in that the peak of the endothermic peak of the DSC profile of crystal a of the compound of formula (I) occurs at 173 ± 2 ℃; preferably, the DSC pattern of said crystal a is as shown in figure 2.
4. The crystal a according to claim 1 or 2, characterized in that the TGA profile of the crystal a of the compound of formula (I) starts to decompose at 190 ± 2 ℃; preferably, the TGA profile of crystal a is as shown in figure 3.
5. A process for the preparation of a crystal a of the compound of formula (I) according to any one of claims 1 to 4, characterized in that it is a slow volatilization process, a suspension stirring process, an osmosis process or a recrystallization process.
6. The method for preparing crystal A according to claim 5, characterized in that the slow volatilization method comprises the following steps: the compound of formula (I) is dissolved in an appropriate solvent of formula 1, then allowed to stand at room temperature, volatilized, the solvent is removed, and the solid is collected to give crystal a.
7. The method for preparing crystal a according to claim 6, characterized in that the 1 st suitable solvent is selected from: one or more of methanol, ethanol, n-propanol, isopropanol, n-butanol, acetone, butanone, ethyl acetate, butyl acetate, isopropyl acetate, dimethyl carbonate, tetrahydrofuran, dichloromethane, chloroform, methyl tert-butyl ether, acetonitrile, anisole, toluene and diethyl ether; the amount of the appropriate solvent 1 is such that the compound of formula (I) is soluble.
8. The method for producing crystal a according to claim 6, characterized in that the 1 st appropriate solvent is a mixed solvent of an a solvent and a B solvent, wherein the a solvent is selected from: one or more of acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane, water, methyl tert-butyl ether; the solvent B is selected from one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene and dichloromethane;

   preferably, the volume ratio of the solvent A to the solvent B is 1: 2-2: 1; preferably, the volume ratio of the solvent A to the solvent B is 1: 1.
9. The method for preparing crystal a according to claim 5, characterized in that said recrystallization method comprises the following steps: heating and stirring the compound of the formula (I) in a proper solvent of the formula 2, and dissolving; slowly cooling to room temperature, filtering, and collecting solid to obtain crystal A;

   preferably, the 2 nd suitable solvent is selected from isopropanol, acetonitrile, toluene, n-propanol; preferably, the mass-to-volume ratio (g/ml) of the compound of formula (I) to the 2 nd appropriate solvent is 1: 5-10;

   alternatively, the suitable solvent of formula 2 is diethyl ether, and the mass to volume ratio (g/ml) of the compound of formula (I) to diethyl ether is 1: 60.
10. The method for producing crystal a according to claim 5, characterized in that the suspension stirring method comprises the steps of: dispersing and suspending the compound of the formula (I) or amorphous substances thereof by using a proper solvent of the No. 3, suspending and stirring at room temperature or high temperature, and collecting solids to obtain crystals A;

    preferably, said 3 rd suitable solvent is selected from: one or more of ethanol, n-propanol, n-butanol, diethyl ether, isopropyl ether, acetonitrile, toluene, n-hexane, n-heptane, cyclohexane, methyl tert-butyl ether, water and petroleum ether; preferably, the 3 rd suitable solvent is used in an amount such that the solute is suspended therein; preferably, the high temperature condition is 60 ℃.
11. The method for preparing crystal a according to claim 5, characterized in that the infiltration method comprises the following steps: placing the compound of formula (I) in a container A, placing in a container B containing an appropriate amount of a 4 th appropriate solvent, sealing the container B, standing at room temperature, and collecting the solid to obtain a crystal A;

    preferably, said 4 th suitable solvent is selected from: one or more of methanol, ethanol, isopropyl acetate, n-hexane, acetonitrile, diethyl ether, methyl tert-butyl ether and toluene; preferably, the standing time is not less than 8 days.
12. The method for preparing crystal A according to claim 5, characterized in that the slow volatilization method comprises the following steps: dissolving a compound shown in a formula (I) in a solvent A, adding a solvent B, mixing, standing at room temperature, volatilizing, removing the solvent, and collecting a solid to obtain a crystal A; preferably, the a solvent is selected from: one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane, water and methyl tert-butyl ether; the solvent B is selected from one or more of methanol, acetone, tetrahydrofuran, ethyl acetate, acetonitrile, toluene, dichloromethane and water;

    preferably, the volume ratio of the solvent A to the solvent B is 1: 2-2: 1; preferably, the volume ratio of the solvent A to the solvent B is 1: 1.
13. A process for preparing an amorphous form of the compound of formula (I), characterized in that it comprises the following steps: dissolving the compound of the formula (I) in a proper solvent of 5 th for clarification, and then carrying out reduced pressure rotary evaporation to obtain the amorphous substance;

    preferably, the 5 th suitable solvent is selected from one or more of dichloromethane, chloroform; preferably, the temperature of the water bath is 40-50 ℃; more preferably 45 deg.c.
14. A pharmaceutical composition comprising crystalline a according to any one of claims 1 to 4 and/or amorphous form prepared according to the process of claim 13, together with one or more pharmaceutically acceptable carriers or one or more other therapeutic agents.
15. A pharmaceutical formulation comprising crystalline a according to any one of claims 1 to 4 and/or amorphous form prepared according to the process of claim 13, together with one or more pharmaceutically acceptable carriers.
16. Use of the crystal a of any one of claims 1 to 4, the amorphous form prepared by the process of claim 13, the pharmaceutical composition of claim 14 or the pharmaceutical formulation of claim 15 for the manufacture of a medicament for the prophylaxis or treatment of a viral disease, preferably viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes or acquired immunodeficiency syndrome (AIDS).
17. A method for preventing or treating a viral disease, comprising administering to a subject in need thereof an effective amount of the crystalline a of any one of claims 1 to 4, the amorphous form prepared according to the process of claim 13, the pharmaceutical composition of claim 14, or the pharmaceutical formulation of claim 15; preferably, the viral disease is viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes or Acquired Immune Deficiency Syndrome (AIDS).
18. The crystal a of any one of claims 1 to 4, the amorphous form prepared by the process of claim 13, the pharmaceutical composition of claim 14 or the pharmaceutical formulation of claim 15, for use in the prevention or treatment of a viral disease, preferably viral hepatitis a, viral hepatitis b, viral hepatitis c, influenza, herpes or Acquired Immune Deficiency Syndrome (AIDS).