Crystal form of ethanesulfonyl phenyl acetamide compound, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a crystal form of an ethanesulfonyl phenyl acetamide compound, a preparation method therefor and a use thereof. A crystal form 1 of a compound shown in formula I provided in the present invention has an X-ray powder diffraction pattern expressed by 2θ angles and having diffraction peaks at 862±0.2°, 15.446±0.2°, 19.406±0.2°, and 23.809±0.2°. The crystal form 1 of the present invention has good stability.

Description

A crystal form of ethylsulfonylphenylacetamide compound, preparation method and application thereof

[0001] This application claims the benefit of Chinese patent application No. 2024101185507, filed on January 26, 2024. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0002] The present invention relates to a crystal form of an ethylsulfonylphenylacetamide compound, a preparation method and application thereof. Background Art

[0003] Retinoic acid receptor-related orphan receptors (RORs) belong to the ligand-dependent nuclear receptor superfamily of transcription factors and play important roles in a range of physiological and pathological processes, including reproductive development, circadian rhythm regulation, metabolic disorders, inflammation, and immune system regulation. RORs primarily include RORα, RORβ, and RORγ. RORα is primarily distributed in the liver, skeletal muscle, skin, lung, adipose tissue, kidney, thymus, brain, and blood. RORβ is primarily distributed in the central nervous system, including the brain, retina, and pineal gland. RORγ is highly expressed in the thymus and also in the kidney, liver, heart, skeletal muscle, adipose tissue, testis, prostate, and pancreas. RORγ is divided into two subtypes, RORγ1 and RORγt (also known as RORγ2), based on the site of transcriptional splicing. RORγ1 is primarily expressed in the thymus, testis, pancreas, heart, liver, skeletal muscle, and kidney, while RORγt is exclusively expressed in immune organs.

[0004] Th17 cells are a subtype of T helper cells characterized by the secretion of the cytokine interleukin-17 (IL-17). Initially, they were thought to primarily contribute to immune responses against bacterial and fungal infections by recruiting neutrophils. Subsequent studies have revealed that Th17 cells play a key role in numerous mouse models of autoimmune diseases. Increased IL-17 levels have also been detected in several human autoimmune diseases, including psoriasis, multiple sclerosis (MS), rheumatoid arthritis (RA), and inflammatory bowel disease (IBD). Increased numbers of Th17 cells have been found in tissues and peripheral blood samples from patients with autoimmune diseases. Therefore, Th17 cells, or the cytokine IL-17 they produce, are closely linked to the pathogenesis of autoimmune diseases and inflammation. Inhibiting Th17 cell differentiation could be used to treat these conditions.

[0005] Studies have shown that RORγt is a key regulator of Th17 cell differentiation. Littman et al. were the first to report that RORγt is essential for the differentiation of initial CD4+ T cells into Th17 cells. Mice lacking RORγt lack lymph nodes and Peyer's patches, and the maturation of T cells is also affected, with the number of various T cells being lower than that of normal mice. Modulating RORγt activity through small molecule compounds can directly affect the differentiation of Th17 cells. Inhibiting RORγt significantly reduces the level of IL-17, a cytokine secreted by Th17. Therefore, RORγt can serve as a new target for the treatment of autoimmune diseases. The development of small molecule RORγt regulators for the treatment of RORγt-mediated diseases such as autoimmune diseases and inflammatory diseases is of great significance.

[0006] Chinese patent CN114685363A discloses an ethylsulfonylphenylacetamide compound, 2-(4-(ethylsulfonyl)phenyl)-N-(6-(2-methyl-2-(pyridin-2-yl)propionyl)pyridin-3-yl)acetamide, whose molecular formula is C 24 H 25 N3O4S, whose structural formula is shown in Formula I: The solid obtained by the method of Example 7 is the following crystal form 2;

[0007] Compound I has inhibitory activity against RORγt and can effectively inhibit the RORγt protein receptor, thereby regulating the differentiation of Th17 cells, inhibiting the production of IL-17, and then treating RORγt-mediated related autoimmune diseases. It is particularly suitable for psoriasis, multiple sclerosis, atopic dermatitis, inflammatory bowel disease and other diseases.

[0008] Given the importance of solid drug crystal forms and their stability in clinical treatment, in-depth research on the crystal form of the above-mentioned compound I is of great significance for the development of drugs suitable for industrial production and with good biological activity. Summary of the Invention

[0009] The present invention provides a crystalline form 1 of a compound as shown in Formula I, which has an X-ray powder diffraction pattern expressed in 2θ angles at diffraction peaks at 9.862±0.2°, 15.446±0.2°, 19.406±0.2°, and 23.809±0.2°;

[0010] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed in 2θ angles further has diffraction peaks at one or more (e.g., 2, 3, 4, or 5) of 16.802±0.2°, 18.167±0.2°, 19.705±0.2°, 27.070±0.2°, and 27.629±0.2°.

[0011] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed in 2θ angles has diffraction peaks at 9.862±0.2°, 15.446±0.2°, 16.802±0.2°, 18.167±0.2°, 19.406±0.2°, 19.705±0.2°, 23.809±0.2°, 27.070±0.2° and 27.629±0.2°.

[0012] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed in 2θ angles also has diffraction peaks at one or more (e.g., 2, 3, or 4) of 9.091±0.2°, 11.502±0.2°, 15.285±0.2°, and 17.729±0.2°.

[0013] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed in 2θ angle is 15.884±0.2°, 18.773±0.2°, 20.414±0.2°, 20.922±0.2°, 22.881±0.2°, 23.202±0.2°, 24.050±0.2°, 24.568±0.2°, 25.49 There are diffraction peaks at one or more of 0±0.2°, 25.769±0.2°, 26.491±0.2°, 27.852±0.2°, 30.331±0.2°, 30.477±0.2°, 33.450±0.2°, 34.311±0.2°, 34.813±0.2°, 37.472±0.2° and 38.460±0.2°.

[0014] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed in 2θ angles is also 4.718±0.2°, 10.719±0.2°, 15.064±0.2°, 17.148±0.2°, 18.595±0.2°, 22.180±0.2°, 22.399±0.2°, 22.682±0.2°, 23.526±0.2°, 24.310±0.2°, 25.232±0.2°, 28.218±0.2°, 28.650 There are diffraction peaks at one or more of the following positions: 29.689±0.2°, 30.731±0.2°, 31.170±0.2°, 31.649±0.2°, 31.975±0.2°, 32.612±0.2°, 32.975±0.2°, 33.954±0.2°, 34.592±0.2°, 35.256±0.2°, 35.818±0.2°, 36.759±0.2°, 39.259±0.2° and 39.494±0.2°.

[0015] In some embodiments of the present invention, the XRPD pattern of Form 1 is substantially as shown in FIG2 .

[0016] In some embodiments of the present invention, the crystalline form 1 loses less than 0.40% of weight during the temperature range from 29.20°C to 150°C;

[0017] And / or, the differential scanning calorimetry curve of the crystalline form 1 has an endothermic peak at 183.47±5°C;

[0018] and / or, the crystalline form 1 has a moisture absorption weight gain of less than 0.2% at 80% RH;

[0019] And / or, the X-ray powder diffraction pattern is measured using Cu-Kα radiation spectrum.

[0020] In some embodiments of the present invention, the crystalline form 1 loses about 0.2653% of weight at 29.2°C to 150°C; preferably, the thermogravimetric analysis curve of the crystalline form 1 is substantially as shown in FIG4 ;

[0021] And / or, the differential scanning calorimetry spectrum of the crystalline form 1 is substantially as shown in FIG6 ;

[0022] And / or, the dynamic moisture adsorption spectrum of the crystal form 1 is substantially as shown in FIG8 .

[0023] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 1 expressed at 2θ angles also has diffraction peaks at the diffraction angles shown in Table 1:

[0024] Table 1

[0025] In some embodiments of the present invention, the diffraction peaks, d values ​​and peak height percentages of the X-ray powder diffraction pattern of Form 1 expressed in 2θ angles may be shown in Table 2:

[0026] Table 2

[0027] In the present invention, the target used in the X-ray powder diffraction is a Cu target.

[0028] The present invention also provides a method for preparing the aforementioned crystalline form 1 of the compound as shown in Formula I, which is Scheme 1, Scheme 2 or Scheme 3;

[0029] The scheme 1 comprises the following steps: slurrying and crystallizing the compound represented by Formula I in ethyl acetate, collecting the solid to obtain the crystalline form 1 of the compound represented by Formula I, wherein the volume mass ratio of the ethyl acetate to the compound represented by Formula I is 5 to 50 mL / g;

[0030] The scheme 2 comprises the following steps: cooling the solution of the compound represented by Formula I to precipitate a solid, collecting the solid to obtain the crystalline form 1 of the compound represented by Formula I, wherein the solvent of the solution is a mixed solvent of acetone, methanol and water; the volume ratio of the acetone, methanol and water is 10:(2-4):(1-2); the volume mass ratio of the mixed solvent to the compound represented by Formula I is 10-100 mL / g;

[0031] The scheme 3 comprises the following steps: suspending the crystalline form 2 of the compound represented by formula I in a solvent for crystallization to obtain the crystalline form 1 of the compound represented by formula I;

[0032] The solvent is one, two or three of water, acetone and ethanol;

[0033] The X-ray powder diffraction pattern of the crystalline form 2 of the compound represented by formula I expressed in 2θ angles is 9.743 ± 0.2°, 12.042 ± 0.2°, 15.062 ± 0.2°, 15.285 ± 0.2°, 17.908 ± 0.2°, 18.146 ± 0.2°, 19.531 ± 0.2°, 20.029 ± 0.2°, 21.307 ± 0.2°, 23.251 ± 0.2° and 25.254 ± 0.2°. There are diffraction peaks (for example, in the X-ray powder diffraction pattern of the crystalline form 2 expressed in 2θ angles, the diffraction peaks, d values ​​and peak height percentages are shown in Table 3 below). In some embodiments of the present invention, the schemes 1, 2 and 3 further include drying; preferably, the drying temperature is 55 ± 5°C.

[0034] In some embodiments of the present invention, in the scheme 1, the volume mass ratio of the mixed solvent to the compound represented by formula I is 10 to 20 mL / g; for example, 9.1 mL / g;

[0035] In some embodiments of the present invention, in Scheme 2, the volume mass ratio of the mixed solvent to the compound represented by Formula I may be 15.1 mL / g.

[0036] In some embodiments of the present invention, in Scheme 3, crystallization is carried out under the seed crystals of Form 1 of the compound shown in Formula I.

[0037] In some embodiments of the present invention, in Scheme 3, the temperature for crystallization is room temperature, for example, 10-30°C.

[0038] The present invention provides a crystalline form 2 of a compound as represented by formula I, having an X-ray powder diffraction pattern expressed in 2θ angles at 9.743±0.2°, 12.042±0.2°, 15.062±0.2°, 15.285±0.2°, 17.908±0.2°, 18.146±0.2°, 19.531±0.2°, 20.029±0.2°, 21.307±0.2°, 23.251±0.2°, and 25.254±0.2°;

[0039] In some embodiments of the present invention, preferably, the X-ray powder diffraction pattern of the crystalline form 2 expressed in 2θ angles further has diffraction peaks at one or more of 6.071±0.2°, 13.465±0.2°, 14.503±0.2°, 15.686±0.2°, 20.747±0.2°, 21.727±0.2°, 22.390±0.2°, 23.828±0.2° and 25.112±0.2°.

[0040] In some embodiments of the present invention, more preferably, the X-ray powder diffraction pattern of the crystalline form 2 expressed in 2θ angle is 8.991±0.2°, 14.125±0.2°, 15.867±0.2°, 16.870±0.2°, 18.786±0.2°, 24.087±0.2°, 26.211±0.2°, 26.511±0.2°, 27.051±0.2°, 27.8 There are diffraction peaks at one or more of 79±0.2°, 28.247±0.2°, 28.431±0.2°, 28.895±0.2°, 29.909±0.2°, 30.517±0.2°, 30.811±0.2°, 31.653±0.2°, 31.928±0.2°, 32.170±0.2°, 33.355±0.2° and 36.674±0.2°.

[0041] In some embodiments of the present invention, preferably, the diffraction peaks, d values ​​and peak height percentages of the X-ray powder diffraction pattern of the crystalline form 2 expressed in 2θ angles can also be shown in Table 3:

[0042] Table 3

[0043] In some embodiments of the present invention, the X-ray powder diffraction pattern of the crystalline form 2 is substantially as shown in FIG3 .

[0044] In some embodiments of the present invention, the weight loss of the crystalline form 2 during the temperature range from 31.30°C to 150°C is ≤1.4%; preferably, the weight loss of the crystalline form 1 during the temperature range from 31.30°C to 150°C is about 1.394%.

[0045] In some embodiments of the present invention, the thermogravimetric analysis curve of the crystal form 2 is substantially as shown in FIG5 .

[0046] In some embodiments of the present invention, the differential scanning calorimetry diagram of the crystalline form 2 has an endothermic peak at 181.95±5°C.

[0047] In some embodiments of the present invention, the differential scanning calorimetry spectrum of the crystalline form 2 is substantially as shown in FIG7 .

[0048] In some embodiments of the present invention, the X-ray powder diffraction pattern is measured using Cu-Kα radiation line.

[0049] The present invention also provides a method for preparing the aforementioned crystalline form 2 of the compound as shown in Formula I, comprising the following steps: the amorphous sample of the compound as shown in Formula I is left open for 8 days under conditions of 25°C-60% RH, and the solid is collected to obtain the crystalline form 2 of the compound as shown in Formula I.

[0050] The present invention also provides a method for preparing the aforementioned crystalline form 2 of the compound as shown in Formula I, comprising the following steps: the amorphous sample of the compound as shown in Formula I is left open for 8 days under RT-12% RH or RT-58% RH conditions, and the solid is collected to obtain the crystalline form 2 of the compound as shown in Formula I.

[0051] The present invention provides an amorphous compound as shown in Formula I, whose X-ray powder diffraction pattern has no obvious sharp diffraction peaks.

[0052] Preferably, the amorphous substance has an XRPD pattern as shown in FIG1 .

[0053] The present invention also provides a method for preparing the amorphous form of the aforementioned compound as shown in Formula I, comprising the following steps: suspending the compound as shown in Formula I in a mixed solvent of acetone and tetrahydrofuran, heating to dissolve, and then filtering, concentrating the filtrate to dryness under reduced pressure at 40°C-60°C, and collecting the solid to obtain the amorphous sample of the compound as shown in Formula I.

[0054] The volume ratio of acetone to tetrahydrofuran is preferably (5:1 to 4:1). The volume mass ratio of the mixed solvent to the compound of formula I is preferably 20 to 100 mL / g; for example, 60 mL / g.

[0055] The present invention also provides a pharmaceutical composition comprising a substance X and at least one pharmaceutical excipient; the substance X is the aforementioned crystalline form 1 of the compound represented by formula I or its amorphous form.

[0056] The selection of the pharmaceutical excipients varies depending on the route of administration and the characteristics of the action, and can generally be fillers, diluents, binders, wetting agents, disintegrants, lubricants, emulsifiers, suspending agents, etc. conventional in the art.

[0057] The pharmaceutical composition can be administered orally, by injection (intravenous, intramuscular, subcutaneous and intracoronary), sublingually, buccally, rectally, urethrally, vaginally, nasally, by inhalation or topically, with oral administration being preferred.

[0058] The present invention provides a use of the aforementioned crystalline form 1, amorphous form or the aforementioned pharmaceutical composition as shown in Formula I in the preparation of a drug, wherein the drug is used to prevent or treat diseases related to the RORγt protein receptor.

[0059] In some embodiments of the present invention, the disease associated with RORγt protein receptor is selected from one or more of psoriasis, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, ankylosing spondylitis, systemic lupus erythematosus, Behcet's disease and chronic obstructive pulmonary disease.

[0060] The present invention provides a pharmaceutical solid dispersion, which comprises the compound represented by formula I and a pharmaceutical carrier.

[0061] Preferably, the compound I is an amorphous form of the compound represented by formula I.

[0062] Preferably, the mass ratio of the crystalline form 1 of the compound shown in Formula I to the drug carrier is 1:(2-4), for example 1:3.

[0063] Preferably, the drug carrier is hypromellose acetate succinate.

[0064] Preferably, the particle size of the pharmaceutical solid dispersion is ≤100 mesh.

[0065] The present invention provides a solid form of a crystalline form 1 of a compound as shown in Formula I, comprising Compound I and hypromellose acetate succinate;

[0066] In some embodiments of the present invention, in the solid phase of the crystalline form 1 of Compound I, the mass ratio of Compound I to Hydroxypropyl Methylcellulose Acetate Succinate is 1:(2-4), for example, 1:3.

[0067] The present invention also provides a method for preparing a pharmaceutical solid dispersion, which comprises the following steps: extruding the compound represented by Formula I and a pharmaceutical carrier by a melt method to obtain the pharmaceutical solid dispersion.

[0068] Preferably, the compound of formula I is preferably a crystalline form or amorphous form of the compound of formula I. The crystalline form of the compound of formula I is preferably the above-mentioned crystalline form 1 of formula I.

[0069] Preferably, the drug carrier is hypromellose acetate succinate.

[0070] Preferably, the temperature of the melting process is 180-200°C, such as 190°C.

[0071] The present invention also provides a method for preparing the solid form 1 of the aforementioned compound as shown in Formula I, which comprises the following steps: extruding the aforementioned compound I and hydroxypropyl methylcellulose acetate succinate by a melt method to obtain the solid form 1 of the compound as shown in Formula I.

[0072] Preferably, the temperature of the melting process is 180-200°C, such as 190°C.

[0073] In some embodiments of the present invention, the method for preparing the solid form of the compound represented by Formula I comprises the following steps:

[0074] The crystalline form 1 of compound I and hydroxypropyl methylcellulose acetate succinate (HPMCAS) were weighed in a mass ratio of 1:3 and mixed evenly to obtain a mixed powder; the temperature of the main heating section of the hot melt extruder was set to 190° C., the heating was turned on, and when the equipment reached the set temperature and stabilized, the mixed powder was added to the feeding bin; the automatic feeder speed and the screw speed were gradually increased. When the feeder speed and the screw speed both reached a feeder speed of 25 rpm and a screw speed of 300 rpm, the hot melt extrudate was collected and pulverized using a multifunctional pulverizer. After pulverization, the sample was passed through a 100-mesh sieve to obtain a solid body of the crystalline form 1 of compound I.

[0075] In some embodiments of the present invention, the method for preparing the solid form 1 of the compound of Formula I comprises the following steps:

[0076] The crystalline form 1 of compound I and hydroxypropyl methylcellulose acetate succinate (HPMCAS) were weighed in a mass ratio of 1:3 and mixed evenly to obtain a mixed powder for hot melt extrusion; the temperature of the main heating section of the hot melt extruder was set to 190°C, the heating was turned on, and when the equipment reached the set temperature and stabilized, the mixed powder was added to the feeding bin; the automatic feeder speed and the screw speed were gradually increased, and attention was paid to coordinated adjustment to ensure that the material did not accumulate in the twin screw; when the feeder speed and the screw speed both reached the target value (feeder speed: 25 rpm, screw speed: 300 rpm), the hot melt extrudate was collected, and the extrudate was transparent; the extrudate was crushed using a multi-functional crusher, and the crushed sample was passed through a 100-mesh sieve to obtain a solid body of the crystalline form 1 of compound I.

[0077] The present invention also provides a pharmaceutical solid dispersion prepared by the above-mentioned pharmaceutical solid dispersion preparation method.

[0078] The present invention also provides a pharmaceutical composition comprising the above-mentioned pharmaceutical solid dispersion and at least one pharmaceutical excipient.

[0079] The present invention also provides a pharmaceutical composition comprising a substance X and at least one pharmaceutical excipient; the substance X is the solid phase of the crystalline form 1 of the compound shown in Formula I.

[0080] The selection of the pharmaceutical excipients varies depending on the route of administration and the characteristics of the action, and can generally be fillers, diluents, binders, wetting agents, disintegrants, lubricants, emulsifiers, suspending agents, etc. conventional in the art.

[0081] The pharmaceutical composition can be administered orally, by injection (intravenous, intramuscular, subcutaneous and intracoronary), sublingually, buccally, rectally, urethrally, vaginally, nasally, by inhalation or topically, with oral administration being preferred.

[0082] The present invention provides a use of the aforementioned solid phase represented by Formula I or the aforementioned pharmaceutical composition in the preparation of a drug, wherein the drug is used to prevent or treat diseases associated with the RORγt protein receptor.

[0083] In some embodiments of the present invention, the disease associated with RORγt protein receptor is selected from one or more of psoriasis, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, ankylosing spondylitis, systemic lupus erythematosus, Behcet's disease and chronic obstructive pulmonary disease.

[0084] In the present invention, "solid fraction" and "solid dispersion" have the same meaning.

[0085] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0086] The reagents and raw materials used in the present invention are commercially available.

[0087] The positive progress of the present invention is that: the crystal form 1 of the present invention has high stability and low hygroscopicity, and can be used to treat TNBS-induced colitis;

[0088] The crystal form 1, amorphous form and drug solid form of the present invention have excellent pharmacokinetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0089] FIG1 is an X-ray powder diffraction pattern of the amorphous sample obtained in Example 1;

[0090] FIG2 is an X-ray powder diffraction pattern of Form 1 obtained in Example 2;

[0091] FIG3 is an X-ray powder diffraction pattern of Form 2 obtained in Example 4;

[0092] FIG4 is a thermogravimetric analysis diagram of Form 1 obtained in Example 2;

[0093] FIG5 is a thermogravimetric analysis diagram of Form 2 obtained in Example 4;

[0094] FIG6 is a differential scanning calorimetry diagram of Form 1 obtained in Example 2;

[0095] FIG7 is a differential scanning calorimetry diagram of Form 2 obtained in Example 4;

[0096] FIG8 is a dynamic water adsorption diagram of Form 1 obtained in Example 2;

[0097] FIG9 is a dynamic water adsorption diagram of Form 2 obtained in Example 4;

[0098] FIG10 is an X-ray powder diffraction pattern of the mixed sample of the crystal transformation experiment in Effect Example 1;

[0099] FIG11 is an X-ray powder diffraction pattern of the crystal form transformation experiment in water in Example 1;

[0100] FIG12 is an X-ray powder diffraction pattern of the crystal form conversion experiment in ethanol in Example 1;

[0101] FIG13 is an X-ray powder diffraction pattern of the crystal form conversion experiment in acetone in Example 1;

[0102] FIG14 is a comparison diagram of X-ray powder diffraction of the stability experiment in Example 2;

[0103] FIG15 is a differential scanning calorimetry comparison diagram of the stability experiment in Effect Example 2.

[0104] FIG16 is a comparison diagram of differential scanning calorimetry of the stability experiment in Example 5 DETAILED DESCRIPTION

[0105] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0106] Instruments and methods used to collect data:

[0107] The X-ray powder diffraction patterns described in this application were collected on a Bruker D8 Advance Diffractometer X-ray powder diffractometer.

[0108] The technical specifications of the instrument are as follows:

[0109] The wavelength of copper target is Kα radiation (40kV, 40mA), θ-2θ goniometer, nickel filter, Lynxeye detector.

[0110] Acquisition software: Diffrac Plus XRD Commander

[0111] Calibration material: corundum (Al2O3)

[0112] Analysis software: MDI Jade

[0113] The attachments are as follows:

[0114] The parameters are as follows:

[0115] The differential scanning calorimetry (DSC) data described in this application were collected using a TA Instruments Q200 differential scanning calorimeter, controlled by Thermal Advantage and analyzed using Universal Analysis software. Typically, 0.5-5 mg of sample was placed in an aluminum crucible (covered but not perforated). The temperature was raised from room temperature to 220°C or 250°C at a rate of 10°C / min under a flow of 50 mL / min of dry nitrogen. The TA software simultaneously recorded the heat gain during the heating process. In this application, melting points are reported based on the onset temperature.

[0116] Thermogravimetric analysis (TGA) data described in this application were collected on a TA Instruments Q500 thermogravimetric analyzer, using Thermal Advantage software for instrument control and Universal Analysis software for analysis. Typically, a 1-10 mg sample was placed in a platinum crucible. Using segmented high-resolution detection, the sample was heated from room temperature to 350°C or 400°C at a rate of 10°C / min under a flow of 40 mL / min of dry nitrogen. The TA software simultaneously recorded the sample weight change during the heating process.

[0117] The dynamic moisture sorption graphs described herein were collected on a TA Instruments Q5000 dynamic moisture sorption instrument, using Thermal Advantage software for instrument control and Universal Analysis software for analysis. The dynamic moisture sorption test parameters described herein are as follows:

[0118] Temperature: 25℃

[0119] Protective gas and flow rate: N2, 10 ml / min

[0120] Minimum dm / dt balancing time: 15 minutes

[0121] Maximum equilibration time: 90 minutes

[0122] Relative humidity range: 0%RH-80%RH-0%RH

[0123] Relative humidity gradient: 10% (0% RH-80% RH-0% RH)

[0124] Example 1 Preparation of amorphous sample of Compound I

[0125] A sample of compound I (100 mg) was weighed, acetone (5 mL) and tetrahydrofuran (1 mL) were added, the mixture was heated to dissolve, and then filtered. The filtrate was concentrated to dryness under reduced pressure at 60° C. to obtain an amorphous sample.

[0126] Example 2 Preparation of Compound I Crystalline Form 1

[0127] A sample of Compound I (33 g) was dispersed in ethyl acetate (300 mL), heated to reflux under stirring for 18 hours, then slowly cooled to room temperature under stirring. The mixture was filtered and the filter cake was dried to obtain a solid sample, which was Compound I Form 1.

[0128] Example 3 Preparation of Compound I Crystalline Form 1

[0129] A sample of compound I (10.21 g) was dispersed in a mixed solvent of acetone (100 mL), methanol (34 mL) and water (20 mL), and the temperature was raised to reflux under stirring until the system was clear. The temperature was slowly lowered to room temperature under stirring to precipitate a solid, which was filtered and dried to obtain a solid sample, namely, compound I Form 1.

[0130] After testing, the X-ray powder diffraction data of the solids obtained in Example 2 and Example 3 are shown in Table 6, the X-ray powder diffraction pattern is shown in Figure 2, the TGA pattern is shown in Figure 4, and the DSC pattern is shown in Figure 6. The results show that the obtained solid product is the crystalline form 1 described in the present application. The TGA data show that the crystalline form sample loses approximately 0.2653% of its weight from 29.20°C to 150°C, and there is a single melting endothermic peak at 183.47°C (peak temperature) in the DSC.

[0131] Table 6

[0132] Hygroscopicity test of Form 1

[0133] Approximately 9.698 mg of Form 1 was collected and tested for hygroscopicity using a dynamic moisture sorption (DVS) instrument. The experimental results are shown in Table 7. The DVS graph of the hygroscopicity experiment is shown in Figure 8.

[0134] Table 7 Hygroscopicity test of Form 1

[0135] Regarding the description of hygroscopic characteristics and the definition of hygroscopic weight gain (Appendix XIX J of the Chinese Pharmacopoeia 2010 edition, Guiding Principles for Hygroscopicity Test of Drugs, experimental conditions: 25℃±1℃, 80% relative humidity):

[0136] Deliquescent: Absorbs enough water to form a liquid

[0137] Highly hygroscopic: weight gain due to moisture absorption is not less than 15%

[0138] Hygroscopic: Weight gain due to moisture absorption is less than 15% but not less than 2%

[0139] Slightly hygroscopic: weight gain due to moisture absorption is less than 2% but not less than 0.2%

[0140] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0141] Example 4 Preparation of Compound I Crystal Form 2

[0142] 30 mg of the amorphous sample was sampled and left to stand in an open state at 25°C-60% RH for 8 days. The solid collected was Form 2 of Compound I.

[0143] After testing, the X-ray powder diffraction data of the solid obtained in Example 4 are shown in Table 8, its X-ray powder diffraction pattern is shown in Figure 3, its TGA pattern is shown in Figure 5, and its DSC pattern is shown in Figure 7. The results show that the obtained solid product is the crystalline form 2 described in this application. The TGA data show that the crystalline form sample loses approximately 1.394% of its weight from 31.30°C to 150°C, and there is a single melting endothermic peak at 181.95°C (peak temperature) in the DSC.

[0144] Table 8

[0145] Hygroscopicity test of Form 2

[0146] Approximately 2.3020 mg of Form 2 was collected and tested for hygroscopicity using a dynamic moisture sorption (DVS) instrument. The experimental results are shown in Table 9. The DVS graph of the hygroscopicity experiment is shown in Figure 9.

[0147] Table 9 Hygroscopicity test of Form 2

[0148] Example 5 Preparation of Solid Dispersion of Compound I

[0149] The crystal form 1 of compound I and hydroxypropyl methylcellulose acetate succinate (HPMCAS) were weighed in a mass ratio of 1: 3, and mixed evenly to obtain a mixed powder for hot melt extrusion. The temperature of the main heating section of the hot melt extruder was set to 190 ° C, and the heating was turned on. When the equipment reached the set temperature and was stable, the mixed powder was added to the feeding bin. The automatic feeder speed and the screw speed were gradually increased, and attention was paid to coordinated adjustment to ensure that the material did not accumulate in the twin screw. When the feeder speed and the screw speed both reached the target value (feeder speed: 25 rpm, screw speed: 300 rpm), the hot melt extrudate was collected, and the extrudate was transparent. The extrudate was pulverized using a multifunctional pulverizer, and the sample was sieved through a 100 mesh sieve to obtain the solid body of compound I after pulverization.

[0150] The obtained solid was subjected to X-ray powder diffraction, as shown in FIG16 , and the form of compound I in the solid phase of compound I was amorphous.

[0151] Effect Example 1: Crystal form conversion experiment

[0152] The X-ray powder diffraction pattern of the mixed sample in the crystal transformation experiment is shown in FIG10 .

[0153] As shown in FIG11 , after the mixture of Form 1 and Form 2 was slurried in water at room temperature for 4 days, Form 2 transformed into Form 1.

[0154] As shown in FIG12 , after the mixture of Form 1 and Form 2 was slurried in ethanol at room temperature for 4 days, Form 2 transformed into Form 1.

[0155] As shown in FIG13 , after the mixture of Form 1 and Form 2 was slurried in acetone at room temperature for 4 days, Form 2 transformed into Form 1.

[0156] Conclusion: Crystal transformation experiments were conducted on Form 1 and Form 2. The experimental results showed that at room temperature, Form 1 was the most stable crystal form in water, acetone and ethanol.

[0157] Effect Example 2: Stability Study of Crystal Form 1

[0158] in conclusion:

[0159] Under the four conditions of long-term, accelerated, high humidity and high temperature, as shown in FIG14 , the X-ray powder diffraction patterns of the samples were compared, and Form 1 did not change and no crystal transformation occurred.

[0160] Under the four conditions of long-term, accelerated, high humidity and high temperature, as shown in FIG15 , the melting point of Form 1 did not change by comparing the differential scanning calorimetry graphs of the samples.

[0161] Effect Example 3: Absorption of Compound I Crystal Form 1 and Compound I Solid Dispersion in Rats after Administration

[0162] SD rats (provided by Sino-British SIPPR Lab Animal Ltd, Shanghai) were divided into groups of three and administered Compound I Form 1 (see Table 9) by intravenous injection or oral gavage. Blood samples (0.4 mL) were collected from the retinal venous plexus of the rats before and at 0.033 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after administration in the intravenous group. Blood samples (0.4 mL) were collected from the retinal venous plexus of the rats before and at 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after administration in the oral gavage group. Blood samples were centrifuged at 8000 rpm for 5 min to separate the upper plasma layer. 40 μL of plasma sample was added with 400 μL of methanol containing internal standard to precipitate protein, vortexed for 1 min, and centrifuged at 18000 g at 4°C for 10 min. 400 μL of the supernatant was injected into a 96-well plate. The injection volume of 1 μL was used for LC / MS / MS detection to obtain the plasma drug concentration, and then the corresponding pharmacokinetic parameters were calculated, as shown in Table 10.

[0163] Table 9 Animal grouping and drug administration Note: Solutol is polyethylene glycol stearate 15, Saline is normal saline, DI water is ultrapure water

[0164] Table 10 Pharmacokinetic parameters of Form 1 after injection / oral administration

[0165] Conclusion: In this experiment, the crystal form 1 of compound I was administered intravenously and the solid form of compound I was administered orally. The results showed that the half-life of the crystal form 1 of compound I administered intravenously was 0.19h, and the bioavailability of the solid form of compound I administered orally was 1.24%.

[0166] The crystalline form 1 in Table 9 was obtained according to the preparation method of Example 2, and the solid form of Compound I was obtained according to the preparation method of Example 5.

[0167] Effect Example 4: Tissue distribution of compound I in rats after administration

[0168] SD rats (provided by Sino-British SIPPR Lab Animal Ltd, Shanghai), 3 rats / time point, were gavage-administered with the solid fraction of Compound I (see Table 11). Plasma, stomach, duodenum, jejunum, colon, rectum, apical blood, portal vein plasma, liver, and other tissues and organs were collected at 0.25h, 1h, 2h, and 4h for the gavage group. Blood samples were centrifuged at 6800g for 6min, and the upper plasma layer was separated. 30μL of plasma sample was added to 300μL of methanol containing internal standard to precipitate protein, vortexed for 1min, and centrifuged at 18000g at 4°C for 10min. 400μL of the supernatant was injected into a 96-well plate. A 1μL injection volume was used for LC / MS / MS analysis to obtain plasma drug concentrations. The corresponding pharmacokinetic parameters were then calculated as shown in Table 13.

[0169] Tissue samples were homogenized with 50% MeOH (1:5), and 400 μL of methanol containing internal standard was added to 40 μL of homogenate to precipitate protein. The mixture was vortexed for 1 min and centrifuged at 18,000 g at 4°C for 10 min. 400 μL of the supernatant was injected into a 96-well plate. The injection volume was 1 μL and the tissue drug concentration was detected by LC / MS / MS, as shown in Table 12.

[0170] Table 11 Animal grouping and drug administration Note: DI water is ultrapure water, and the solid phase of compound I was prepared according to the description in Example 3.

[0171] Table 12 Plasma and tissue drug concentrations of Compound I after oral administration of solid fraction

[0172] Conclusion: In this experiment, the solid fraction of Compound I was administered orally. The results showed that the solid fraction of Compound I was widely distributed in the gastrointestinal tract after oral administration, and the amount entering the blood was very small.

[0173] Effect Example 5: Solid dispersion of Compound I significantly improves TNBS-induced colitis model in rats

[0174] Fifty male SD rats, 6 weeks old, weighing about 200 g, were randomly divided into 5 groups (Table 13), with 10 rats in each group.

[0175] Rats in the sham group and the model group were gavaged with blank excipient (hydroxypropyl methylcellulose acetate succinate, HPMCAS) twice a day with an interval of 8 hours, starting from Day -1 to Day 5.

[0176] The positive drug Mesalamine group and the solid dispersion of Compound I group were administered from Day-1 to Day 5, and Day 6 was the end point of the experiment.

[0177] Model rats: On experimental day 0, SD rats weighing approximately 200 g were anesthetized with 3.2 ml of 1.25% avertin (Easycheck, M2910). Rats in the model group and each dose group were rectal instilled with 1 ml of 1% TNBS solution (final concentration 50% ethanol).

[0178] The rats in the sham group were rectally instilled with an equal volume of 50% ethanol. During the experiment, the body weight of the animals was measured daily, and the fecal characteristics and occult blood scores of the rats were evaluated to calculate the disease activity index (DAI) (Table 14).

[0179] At the end of the experiment, the animals were euthanized by CO2, and blood was quickly collected from the heart to prepare serum and stored at -80°C. The colon (from the anus to the cecum) was collected, photographed, and the length of the colon was measured. The fat and connective tissue around the colon were removed, and the contents and cecum were removed and weighed. The rat colon was then cut open longitudinally, and half of the colon tissue was made into a Swiss roll shape (such as the rectal segment, the part of each animal must be unified), fixed with neutral paraformaldehyde fixative, and used for subsequent histopathological H&E staining. The pathologist performed a specific pathological score on the staining results (Table 15). The other half of the colon tissue was used for intestinal mucosa collection, tissue homogenization, and protein concentration testing.

[0180] Table 13. Animal grouping and dosing regimen Note: The positive drug is Mesalamine; PO is oral, BID is twice a day, QD is once a day;

[0181] The solid dispersion of Compound I (the solid dispersion of Compound I in Example 5) was dissolved in ultrapure water for administration.

[0182] Table 14. DAI scoring criteria

[0183] Table 15. Colon histopathology scoring criteria

[0184] Table 16 Body weight changes Note: *P<0.05, **P<0.01, ***p<0.001, ****P<0.0001 vs. model group, two-way ANOVA analysis and Dunnett's test were used for comparison between groups.

[0185] Table 17. Disease activity score Note: *P<0.05, **P<0.01, ***p<0.001, ****P<0.0001 vs. model group, two-way ANOVA analysis and Dunnett's test were used for comparison between groups.

[0186] Table 18. Colon parameters Note: *P<0.05, ****P<0.0001 vs. model group, one-way ANOVA analysis and Dunnett's test were used for comparison between groups.

[0187] Table 19. Pathological scores of rat colon tissue Note: ****P<0.0001 vs. model group, one-way ANOVA analysis and Dunnett's test were used for comparison between groups.

[0188] Results: Combined with the data from the in-life experiment and pathological analysis, the solid dispersion of compound I of this patent: oral administration of 5 mg / kg and 5 mg / kg (twice a day) can significantly alleviate TNBS-induced colitis and show good dose-dependence such as body weight (Table 16) and disease severity (Table 17).

[0189] The results showed that the weight loss of rats with enteritis was slowed down, diarrhea and bloody stools were improved, the length-to-weight ratio of the colon of rats with enteritis was reversed (Table 18), and the loss of crypt structure and inflammatory cell infiltration were alleviated (Table 19).

[0190] Conclusion: The solid dispersion of Compound I significantly alleviated the symptoms of TNBS-induced colitis in rats at a dose as low as 5 mg / kg (twice a day) and has the potential to treat inflammatory bowel disease.

Claims

1. Polymorph 1 of a compound represented by Formula I, characterized in that, Its X-ray powder diffraction pattern expressed in terms of 2θ has diffraction peaks at 9.862 ± 0.2°, 15.446 ± 0.2°, 19.406 ± 0.2° and 23.809 ± 0.2°; 2. The crystalline form 1 of the compound represented by formula I as described in claim 1, characterized in that, Its X-ray powder diffraction pattern expressed in terms of 2θ has diffraction peaks at 9.862 ± 0.2°, 15.446 ± 0.2°, 16.802 ± 0.2°, 18.167 ± 0.2°, 19.406 ± 0.2°, 19.705 ± 0.2°, 23.809 ± 0.2°, 27.070 ± 0.2° and 27.629 ± 0.2°.

3. The crystalline form 1 of the compound represented by formula I as described in claim 2, characterized in that, The X-ray powder diffraction pattern expressed in terms of 2θ of the described crystalline form 1 further has diffraction peaks at one or more of 9.091 ± 0.2°, 11.502 ± 0.2°, 15.285 ± 0.2° and 17.729 ± 0.2°.

4. The crystalline form 1 of the compound shown in Formula I as described in claim 3, characterized in that, The X-ray powder diffraction pattern expressed in terms of 2θ of the described crystalline form 1 further has diffraction peaks at one or more of 15.884 ± 0.2°, 18.773 ± 0.2°, 20.414 ± 0.2°, 20.922 ± 0.2°, 22.881 ± 0.2°, 23.202 ± 0.2°, 24.050 ± 0.2°, 24.568 ± 0.2°, 25.490 ± 0.2°, 25.769 ± 0.2°, 26.491 ± 0.2°, 27.852 ± 0.2°, 30.331 ± 0.2°, 30.477 ± 0.2°, 33.450 ± 0.2°, 34.311 ± 0.2°, 34.813 ± 0.2°, 37.472 ± 0.2° and 38.460 ± 0.2°.

5. The crystalline form 1 of the compound shown in formula I as described in claim 4, characterized in that, The X-ray powder diffraction pattern expressed in terms of 2θ of the described crystalline form 1 further has diffraction peaks at one or more of 4.718 ± 0.2°, 10.719 ± 0.2°, 15.064 ± 0.2°, 17.148 ± 0.2°, 18.595 ± 0.2°, 22.180 ± 0.2°, 22.399 ± 0.2°, 22.682 ± 0.2°, 23.526 ± 0.2°, 24.310 ± 0.2°, 25.232 ± 0.2°, 28.218 ± 0.2°, 28.650 ± 0.2°, 29.689 ± 0.2°, 30.731 ± 0.2°, 31.170 ± 0.2°, 31.649 ± 0.2°, 31.975 ± 0.2°, 32.612 ± 0.2°, 32.975 ± 0.2°, 33.954 ± 0.2°, 34.592 ± 0.2°, 35.256 ± 0.2°, 35.818 ± 0.2°, 36.759 ± 0.2°, 39.259 ± 0.2° and 39.494 ± 0.2°.

6. The crystalline form 1 of the compound shown in Formula I as described in any one of claims 1-5, characterized in that, The X-ray powder diffraction pattern of the described polymorph 1 also has diffraction peaks at the diffraction angles shown below:

7. The crystalline form 1 of the compound represented by formula I as described in any one of claims 1-6, characterized in that It satisfies one or more of the following conditions: (1) The described crystalline form 1 has a weight loss of < 0.40% during the process from 29.2 °C to 150 °C; (2) The differential scanning calorimetry curve of the described crystalline form 1 has an endothermic peak at 183.47 ± 5 °C; (3) The described crystalline form 1 has a moisture absorption weight gain of < 0.2% at 80% RH; (4) The X-ray powder diffraction pattern is measured using Cu-Kα radiation spectrum; In the X-ray powder diffraction pattern of the described polymorph 1 expressed in 2θ angles, its diffraction peaks, d values, and peak height percentages are as follows:

8. The crystalline form 1 of the compound represented by formula I as described in any one of claims 1-7, characterized in that, It satisfies one or more of the following conditions: (1) The crystalline form 1 described above has a weight loss of about 0.2653% from 29.20 °C to 150 °C; preferably, the thermogravimetric analysis curve pattern of the crystalline form 1 is substantially as shown in Figure 4; (2) The differential scanning calorimetry pattern of the crystalline form 1 is substantially as shown in Figure 6; (3) The dynamic moisture sorption pattern of the crystalline form 1 is substantially as shown in Figure 8; (4) The XRPD pattern of the crystalline form 1 is substantially as shown in Figure 2.

9. A method for preparing polymorph 1 of a compound represented by formula I as described in any one of claims 1-8, characterized in that, It is Scheme 1, Scheme 2 or Scheme 3; The said Scheme 1 includes the following steps: The compound shown in Formula I is slurried and crystallized in ethyl acetate, and the solid is collected to obtain the crystalline form 1 of the compound shown in Formula I. The volume-mass ratio of the ethyl acetate to the compound shown in Formula I is 5-50 mL / g; The said Scheme 2 includes the following steps: The solution of the compound shown in Formula I is cooled to precipitate a solid, and the solid is collected to obtain the crystalline form 1 of the compound shown in Formula I. The solvent of the solution is a mixed solvent of acetone, methanol and water; the volume ratio of the acetone, methanol and water is 10:(2-4):(1-2); the volume-mass ratio of the mixed solvent to the compound shown in Formula I is 10-100 mL / g; The said Scheme 3 includes the following steps: The crystalline form 2 of the compound shown in Formula I is suspended in a solvent for polymorph conversion to obtain the crystalline form 1 of the compound shown in Formula I; The said solvent is 1, 2 or 3 of water, acetone and ethanol; The X-ray powder diffraction pattern of the crystalline form 2 of the compound shown in Formula I has diffraction peaks at 9.743±0.2°, 12.042±0.2°, 15.062±0.2°, 15.285±0.2°, 17.908±0.2°, 18.146±0.2°, 19.531±0.2°, 20.029±0.2°, 21.307±0.2°, 23.251±0.2° and 25.254±0.2° in terms of 2θ angle.

10. The preparation method of polymorph 1 of the compound represented by Formula I as described in claim 9, characterized in that, It satisfies one or more of the following conditions: (1) The said Scheme 1 and Scheme 2 also include drying; preferably, the drying temperature is 55±5 °C; (2) In the said Scheme 1, the volume-mass ratio of the mixed solvent to the compound shown in Formula I is 10-20 mL / g; for example, 9.1 mL / g; (3) In the said Scheme 2, the volume-mass ratio of the mixed solvent to the compound shown in Formula I is 15.1 mL / g; (4) In the said Scheme 3, the polymorph conversion is carried out under the crystal seed of the crystalline form 1 of the compound shown in Formula I; (5) In the said Scheme 3, the polymorph conversion temperature is 10-30 °C.

11. An amorphous form of a compound represented by Formula I, characterized in that, Its X-ray powder diffraction pattern has no obvious sharp diffraction peaks; preferably, the amorphous substance has an XRPD pattern as shown in Figure 1; 12. A pharmaceutical solid dispersion, characterized in that, It comprises the compound shown by formula I as described above and a pharmaceutical carrier; 13. The pharmaceutical solid dispersion according to claim 12, characterized in that ， It satisfies one or more of the following conditions: (1) The said Compound I is the amorphous form of the compound shown in Formula I; (2) The mass ratio of the crystalline form 1 of the compound shown in Formula I to the pharmaceutical carrier is 1:(2-4), for example, 1:3; (3) The said pharmaceutical carrier is hypromellose acetate succinate; (4) The particle size of the said pharmaceutical solid dispersion ≤ 100 mesh sieve.

14. The pharmaceutical solid dispersion according to claim 13, characterized in that, It includes Compound I and hypromellose acetate succinate; Preferably, in the solid dispersion of polymorph 1 of Compound I, the mass ratio of Compound I to hypromellose acetate succinate is 1:(2 - 4), such as 1:

3.

15. A preparation method of a pharmaceutical solid dispersion, characterized in that, It includes the following steps: extruding the compound shown in Formula I and a pharmaceutical carrier by a melting method to obtain the pharmaceutical solid dispersion; Preferably, the compound represented by Formula I is a polymorph or amorphous form of the compound represented by Formula I; the polymorph of the compound represented by Formula I is preferably polymorph 1 represented by Formula I as described in any one of Claims 1 - 8; Preferably, the pharmaceutical carrier is hypromellose acetate succinate; Preferably, the temperature of the melting method is 180 - 200 °C, such as 190 °C.

16. A pharmaceutical solid dispersion prepared by the method for preparing a pharmaceutical solid dispersion as described in Claim 15.

17. A pharmaceutical composition, characterized in that, It comprises Substance X and at least one pharmaceutical excipient; Substance X is polymorph 1 of the compound represented by Formula I as described in any one of Claims 1 - 8, the amorphous form of the compound of Formula I as described in Claim 11, or the pharmaceutical solid dispersion as described in any one of Claims 12 - 14 and 16.

18. Use of polymorph 1 represented by Formula I as described in any one of Claims 1 - 8, the amorphous form of the compound of Formula I as described in Claim 11, or the pharmaceutical solid dispersion as described in any one of Claims 12 - 14 and 16 in the preparation of a medicament for preventing or treating a disease associated with the RORγt protein receptor; Preferably, the disease associated with the RORγt protein receptor is selected from one or more of psoriasis, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, ankylosing spondylitis, systemic lupus erythematosus, Behcet's disease, and chronic obstructive pulmonary disease.

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