Polymorph of novel amide-containing substituted aromatic ring derivative

Abstract

The present invention belongs to the field of medicine, specifically relates to a polymorph of a novel amide-containing substituted aromatic ring derivative, and further relates to a pharmaceutical composition containing the polymorph and the medical use thereof.

Description

Crystal form of novel amide-substituted aromatic ring derivative TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a crystal form of a novel amide-substituted aromatic ring derivative, and also relates to a pharmaceutical composition comprising the crystal form and medical uses. BACKGROUND

[0002] Janus kinases (JAKs) are a family of intracellular non-receptor tyrosine kinases that mediate signaling of most cytokines in cells, such as interleukins (ILs), interferons (IFNs), erythropoietin (EPO), granulocyte and macrophage colony-stimulating factor (GMCSF), growth hormone (GH), prolactin (PRL), thrombopoietin (TPO), platelet-derived growth factor (PDGF), and epidermal growth factor (EGF). Since JAK family members can mediate multiple cytokine signaling pathways, the currently approved JAK inhibitors (targeting JAK1-3) inevitably bring some side effects, and most of the JAK inhibitors approved by FDA have black box warnings in the label, which largely limits their clinical application range. For example, the early JAK inhibitor Tofacitinib has good efficacy, but it has high inhibitory activity against JAK-1, JAK-2 and JAK-3, and has large side effects, and the instructions for tofacitinib are required by FDA to add a black box warning of thrombosis and death risk. Other JAK inhibitors, such as upadacitinib and baricitinib, have "serious infection, malignancy and thrombosis risk". TYK2 is one of the JAK family members and is crucial in the regulatory signaling transduction cascade downstream of IL-12, IL-23 and type I interferon receptors. IL-12 and IL-23 are currently considered to be key cytokines affecting the progression of psoriasis. In addition, TYK2-mediated signaling is also associated with multiple inflammatory and autoimmune diseases such as arthritis, dermatitis, lupus erythematosus, inflammatory bowel disease, etc. Therefore, it is urgent to develop a new TYK2 inhibitor with good efficacy and safety.

[0003] Polymorphs are different crystal forms of the same compound, and polymorphism is of value to the pharmaceutical industry, especially to those industries that design suitable dosage forms for development. Certain polymorphs can exhibit improved thermodynamic stability or can be more readily produced in high purity and in large quantities, and thus are more suitable for use in pharmaceutical formulations. Due to different lattice energies, certain polymorphs can show other advantageous physical properties such as no hygroscopic tendency, improved stability, etc. SUMMARY

[0004] The present application provides a compound of the following formula I:

[0005] The first aspect of the present application relates to a crystalline form A of a compound represented by Formula I,

[0006] In some embodiments of the first aspect of the application, the crystalline form A crystals belong to monoclinic system, space group C2 / c, with unit cell parameters of: Z = 8.

[0007] In some embodiments of the first aspect of the application, the crystalline form A has peaks at 7.75 ± 0.2°, 18.22 ± 0.2°, 22.64 ± 0.2°, 11.27 ± 0.2° and 16.83 ± 0.2° in its X-ray powder diffraction pattern, using Cu-Ka radiation, expressed in terms of the 2 theta angle.

[0008] In some embodiments of the first aspect of the application, the crystalline form A has one or more peaks in its X-ray powder diffraction pattern, using Cu-Ka radiation, expressed in terms of the 2 theta angle, selected from the group consisting of: 23.61 ± 0.2°, 23.44 ± 0.2°, 27.07 ± 0.2°, 19.77 ± 0.2°, 26.58 ± 0.2°, 29.48 ± 0.2°, 22.32 ± 0.2° and 21.43 ± 0.2°.

[0009] In some embodiments of the first aspect of the application, the XRPD pattern of the crystalline form A using Cu-Ka radiation comprises the characteristic peaks expressed in terms of diffraction angles (2 theta) as shown in Table 1 below, wherein the error range for the 2 theta values is ± 0.2°:

[0010] Table 1: XRPD diffraction peak data for crystalline form A

[0011] In some embodiments of the first aspect of the application, the XRPD pattern of the crystalline form A using Cu-Ka radiation comprises peaks at substantially the same diffraction angles (2 theta) as shown in Figure 1-1. In preferred embodiments, the XRPD pattern of the crystalline form A using Cu-Ka radiation is substantially as shown in Figure 1-1 or as shown in Figure 1-1.

[0012] In some embodiments of the first aspect of the application, the DSC pattern of the crystalline form A as determined by differential scanning calorimetry comprises a characteristic peak with a peak temperature of 266 ± 5 °C (preferably 266 ± 2 °C); preferably, the DSC pattern comprises an endothermic peak at 265.6 °C.

[0013] In some embodiments of the first aspect of the application, the crystalline form A is an anhydrous crystalline form.

[0014] In some embodiments of the second aspect of the application, the crystalline Form B has one or more peaks in an X-ray powder diffraction pattern, using Cu-Ka radiation, at 2-theta angles selected from the group consisting of: 19.46±0.2°, 21.33±0.2°, 21.01±0.2°, 24.64±0.2° and 20.20±0.2°.

[0015] In some embodiments of the second aspect of the application, the crystalline Form B has one or more peaks in an X-ray powder diffraction pattern, using Cu-Ka radiation, at 2-theta angles selected from the group consisting of: 19.46±0.2°, 21.33±0.2°, 21.01±0.2°, 24.64±0.2° and 20.20±0.2°.

[0016] In some embodiments of the second aspect of the application, the crystalline Form B has an XRPD pattern using Cu-Ka radiation comprising characteristic peaks at diffraction angles (2-theta) as set out in Table 2 below, wherein the error range in the 2-theta values is ±0.2°:

[0017] Table 2: XRPD diffraction peak data for Form B

[0018] In some embodiments of the second aspect of the application, the crystalline Form B has an XRPD pattern using Cu-Ka radiation comprising peaks at substantially the same diffraction angles (2-theta) as shown in Figure 2. In preferred embodiments, the crystalline Form B has an XRPD pattern using Cu-Ka radiation substantially as shown in Figure 2 or as shown in Figure 2.

[0019] In some embodiments of the second aspect of the application, the crystalline Form B has a DSC pattern as determined by differential scanning calorimetry comprising a peak temperature at 264±5°C (preferably 264±2°C); preferably, the DSC pattern comprises endothermic peaks at 247.8°C, 263.9°C and an exothermic peak at 248.9°C.

[0020] In some embodiments of the second aspect of the application, the crystalline Form B is an anhydrous crystalline form.

[0021] In some embodiments of the third aspect of the application, the crystalline Form C has one or more peaks in an X-ray powder diffraction pattern, using Cu-Ka radiation, at 2-theta angles selected from the group consisting of: 12.60±0.2°, 6.84±0.2°, 12.87±0.2°, 14.84±0.2° and 19.09±0.2°.

[0022] In some embodiments of the third aspect of the application, the Form C has one or more additional peaks in its X-ray powder diffraction pattern, using Cu-Kalphai radiation, at positions 2-theta expressed in degrees, selected from the group consisting of: 16.37±0.2°, 13.64±0.2°, 21.64±0.2°, 18.16±0.2°, 20.14±0.2°, 24.62±0.2°, 24.15±0.2°, 22.26±0.2° and 16.90±0.2°.

[0023] In some embodiments of the third aspect of the application, the Form C has an XRPD pattern using Cu-Kalphai radiation comprising characteristic peaks at diffraction angles (2theta) expressed in degrees as shown in Table 3 below, wherein the error range for the 2theta values is ±0.2°:

[0024] Table 3: XRPD diffraction peak data for Form C

[0025] In some embodiments of the third aspect of the application, the Form C has an XRPD pattern using Cu-Kalphai radiation comprising peaks at substantially the same diffraction angles (2theta) as shown in Figure 3. In preferred embodiments, the Form C has an XRPD pattern using Cu-Kalphai radiation substantially as shown in Figure 3 or as shown in Figure 3.

[0026] In some embodiments of the third aspect of the application, the Form C has a DSC pattern as determined by differential scanning calorimetry comprising a peak temperature at 265±5°C (preferably 265±2°C); preferably, the DSC pattern comprises an endothermic peak at 234.3°C, 264.7°C, and an exothermic peak at 234.9°C.

[0027] In some embodiments of the third aspect of the application, the Form C is an anhydrous form.

[0028] A fourth aspect of the application relates to a crystalline Form D of the compound of Formula I, wherein the Form D has peaks in its X-ray powder diffraction pattern, using Cu-Kalphai radiation, at positions 2-theta expressed in degrees, selected from the group consisting of: 17.07±0.2°, 10.90±0.2°, 9.40±0.2°, 15.20±0.2° and 10.11±0.2°.

[0029] In some embodiments of the fourth aspect of the application, the Form D has one or more additional peaks in its X-ray powder diffraction pattern, using Cu-Kalphai radiation, at positions 2-theta expressed in degrees, selected from the group consisting of: 23.32±0.2°, 21.34±0.2°, 15.05±0.2°, 18.39±0.2°, 20.72±0.2°, 24.45±0.2°, 26.60±0.2°, 26.79±0.2°, 14.66±0.2° and 13.45±0.2°.

[0030] In some embodiments of the fourth aspect of the application, the Form D has an XRPD pattern using Cu-Ka radiation comprising characteristic peaks at diffraction angles (2 theta) essentially the same as shown in Table 4 below, wherein the error range for the 2 theta values is ±0.2°:

[0031] Table 4: XRPD diffraction peak data for Form D

[0032] In some embodiments of the fourth aspect of the application, the Form D has an XRPD pattern using Cu-Ka radiation comprising characteristic peaks at diffraction angles (2 theta) essentially the same as shown in Table 4 below, wherein the error range for the 2 theta values is ±0.2°:

[0033] In some embodiments of the fourth aspect of the application, the Form D has a DSC pattern as determined by differential scanning calorimetry comprising a peak at a temperature of 261 ± 5 °C (preferably 261 ± 2 °C); preferably, the DSC pattern comprises an endothermic peak at 248.3 °C, 260.6 °C, and an exothermic peak at 249.4 °C.

[0034] In some embodiments of the fourth aspect of the application, the Form D is an anhydrous form.

[0035] A fifth aspect of the application relates to a crystalline Form E of the compound of Formula I, wherein the Form E has peaks at 4.94 ± 0.2°, 8.23 ± 0.2°, 18.02 ± 0.2°, 9.85 ± 0.2° and 14.04 ± 0.2° in an X-ray powder diffraction pattern, using Cu-Ka radiation, expressed in terms of 2 theta angles.

[0036] In some embodiments of the fifth aspect of the application, the Form E has one or more peaks selected from the group consisting of peaks at 19.32 ± 0.2°, 21.89 ± 0.2°, 21.07 ± 0.2°, 24.75 ± 0.2°, 20.88 ± 0.2°, 26.09 ± 0.2°, 10.17 ± 0.2° and 6.00 ± 0.2° in an X-ray powder diffraction pattern, using Cu-Ka radiation, expressed in terms of 2 theta angles.

[0037] In some embodiments of the fifth aspect of the application, the Form E has an XRPD pattern using Cu-Ka radiation comprising characteristic peaks at diffraction angles (2 theta) essentially the same as shown in Table 5 below, wherein the error range for the 2 theta values is ±0.2°:

[0038] Table 5: XRPD diffraction peak data for Form E

[0039] In some embodiments of the fifth aspect of the application, the XRPD pattern of Form E using Cu-Ka radiation includes peaks at substantially the same diffraction angles (2 theta) as those set forth in FIG. 5. In preferred embodiments, the XRPD pattern of Form E using Cu-Ka radiation is substantially as depicted in FIG. 5 or as depicted in FIG. 5.

[0040] In some embodiments of the fifth aspect of the application, Form E is an anhydrous form.

[0041] The sixth aspect of the application relates to a process for preparing Form A of the compound of Formula I, comprising the steps of: mixing the compound of Formula I with tetrahydrofuran and ethyl acetate under heating or without heating, stirring, cooling to obtain Form A, the weight volume ratio of the compound of Formula I: tetrahydrofuran: ethyl acetate being 1:(0.5-1.5):(15-25); preferably, the weight volume ratio of the compound of Formula I: tetrahydrofuran: ethyl acetate is 1:(0.5-1):(20-25); further preferably, the weight volume ratio of the compound of Formula I: tetrahydrofuran: ethyl acetate is 1:1:20.

[0042] In some embodiments of the sixth aspect of the application, the stirring is performed at 40-60°C for 5-10h after mixing; preferably, the stirring is performed at 50-60°C for 6-10h after mixing; further preferably, the stirring is performed at 50°C for 6h after mixing.

[0043] In some embodiments of the sixth aspect of the application, the compound of Formula I is added into a mixed solution of 1-fold (weight: volume) tetrahydrofuran and 20-fold (weight: volume) ethyl acetate, stirred at 50°C for 6h, naturally cooled to room temperature while continuing stirring to obtain Form A.

[0044] The seventh aspect of the application relates to a process for preparing Form B of the compound of Formula I, comprising the steps of: dissolving the compound of Formula I in benzyl alcohol under heating or without heating, then adding isopropyl acetate, stirring, cooling to obtain Form B, the weight volume ratio of the compound of Formula I: benzyl alcohol: isopropyl acetate being 1:(10-50):(100-500); preferably, the weight volume ratio of the compound of Formula I: benzyl alcohol: isopropyl acetate is 1:(25-50):(250-500); further preferably, the weight volume ratio of the compound of Formula I: benzyl alcohol: isopropyl acetate is 1:25:250.

[0045] In some embodiments of the seventh aspect of the application, the compound of Formula I is added into 25-fold (weight: volume) benzyl alcohol under heating or without heating, stirred until clear, then 250-fold (weight: volume) isopropyl acetate is added, stirred, naturally cooled to room temperature while continuing stirring to obtain Form B.

[0046] The eighth aspect of the present application relates to a method for preparing the crystalline form C of the compound of formula I, comprising the following steps: dissolving the compound of formula I in dimethyl sulfoxide under heating or without heating, then adding isopropyl alcohol, stirring, and cooling to obtain the crystalline form C, wherein the weight-volume ratio of the compound of formula I: dimethyl sulfoxide: isopropyl alcohol is 1:(10-50):(100-500); preferably, the weight-volume ratio of the compound of formula I: dimethyl sulfoxide: isopropyl alcohol is 1:(25-50):(250-500); further preferably, the weight-volume ratio of the compound of formula I: dimethyl sulfoxide: isopropyl alcohol is 1:25:250.

[0047] In some embodiments of the eighth aspect of the present application, the compound of formula I is added into 25 times (weight: volume) of dimethyl sulfoxide under heating or without heating, stirred until clear, then 250 times (weight: volume) of isopropyl alcohol is added, stirred and naturally cooled to room temperature, and the stirring is continued to obtain the crystalline form C.

[0048] The ninth aspect of the present application relates to a method for preparing the crystalline form D of the compound of formula I, comprising the following steps: dissolving the compound of formula I in DMF under heating or without heating, then adding water, stirring, and cooling to obtain the crystalline form D, wherein the weight-volume ratio of the compound of formula I: DMF: water is 1:(10-50):(100-500); preferably, the weight-volume ratio of the compound of formula I: DMF: water is 1:(25-50):(250-500); further preferably, the weight-volume ratio of the compound of formula I: DMF: water is 1:25:250.

[0049] In some embodiments of the ninth aspect of the present application, the compound of formula I is added into 25 times (weight: volume) of DMF under heating or without heating, stirred until clear, then 250 times (weight: volume) of water is added, stirred and naturally cooled to room temperature, and the stirring is continued to obtain the crystalline form D.

[0050] The tenth aspect of the present application relates to a method for preparing the crystalline form E of the compound of formula I, comprising the following steps: mixing the compound of formula I with 1,4-dioxane and isopropyl ether under heating or without heating, stirring, and cooling to obtain the crystalline form E, wherein the weight-volume ratio of the compound of formula I: 1,4-dioxane: isopropyl ether is 1:(10-30):(10-30); preferably, the weight-volume ratio of the compound of formula I: 1,4-dioxane: isopropyl ether is 1:(10-20):(10-20); further preferably, the weight-volume ratio of the compound of formula I: 1,4-dioxane: isopropyl ether is 1:10:10.

[0051] In some embodiments of the tenth aspect of the present application, the mixture is stirred at 40-60°C for 0.5-3h after mixing; preferably, the mixture is stirred at 50-60°C for 1-3h after mixing; further preferably, the mixture is stirred at 50°C for 1h after mixing.

[0052] In some embodiments of the tenth aspect of the present application, the compound of formula I is added to a mixed solution of 10 times (weight: volume) of 1,4-dioxane and 10 times (weight: volume) of isopropyl ether, stirred at 50°C for 1 h, stirred to cool to room temperature naturally, and continue to stir to obtain the crystal form E.

[0053] The eleventh aspect of the present application relates to a pharmaceutical composition comprising at least one of the crystal form A according to the first aspect of the present application, the crystal form B according to the second aspect of the present application, the crystal form C according to the third aspect of the present application, the crystal form D according to the fourth aspect of the present application, the crystal form E according to the fifth aspect of the present application, the crystal form A prepared by the preparation method according to the sixth aspect of the present application, the crystal form B prepared by the preparation method according to the seventh aspect of the present application, the crystal form C prepared by the preparation method according to the eighth aspect of the present application, the crystal form D prepared by the preparation method according to the ninth aspect of the present application, and the crystal form E prepared by the preparation method according to the tenth aspect of the present application, and optionally a pharmaceutically acceptable carrier and / or excipient.

[0054] The pharmaceutical composition of the present application can be optionally administered in combination with other ingredients which have at least some effect in treating various diseases.

[0055] The crystal form or the pharmaceutical composition of the present application can be administered by parenteral, topical, intravenous, oral, subcutaneous, intra-arterial, intradermal, transdermal, rectal, intracranial, intraperitoneal, intranasal, intramuscular routes. The crystal form or the pharmaceutical composition of the present application can be prepared into various suitable dosage forms according to the administration route.

[0056] The dosage form of the pharmaceutical composition is selected from the group consisting of a cream, an ointment, a gel, a transdermal patch, an intradermal injection, an eye drop, an intraocular injection, an ocular implant, a nasal spray, an inhalation powder spray, an inhalation aerosol, an inhalation spray, an inhalation liquid preparation, a vaginal suppository, a vaginal tablet, a vaginal gel, a preparation for oral or rectal administration.

[0057] The twelfth aspect of the present application relates to the use of the crystal form A according to the first aspect of the present application, the crystal form B according to the second aspect of the present application, the crystal form C according to the third aspect of the present application, the crystal form D according to the fourth aspect of the present application, the crystal form E according to the fifth aspect of the present application, the crystal form A prepared by the preparation method according to the sixth aspect of the present application, the crystal form B prepared by the preparation method according to the seventh aspect of the present application, the crystal form C prepared by the preparation method according to the eighth aspect of the present application, the crystal form D prepared by the preparation method according to the ninth aspect of the present application, the crystal form E prepared by the preparation method according to the tenth aspect of the present application, or the pharmaceutical composition according to the eleventh aspect of the present application in the preparation of a medicament for treating a disease related to TYK2 mediation.

[0058] The twelfth aspect of the present application relates to a method for treating a TYK2-mediated related disease, comprising administering to a subject in need thereof an effective amount of the crystalline form A according to the first aspect of the present application, the crystalline form B according to the second aspect of the present application, the crystalline form C according to the third aspect of the present application, the crystalline form D according to the fourth aspect of the present application, the crystalline form E according to the fifth aspect of the present application, the crystalline form A prepared by the preparation method according to the sixth aspect of the present application, the crystalline form B prepared by the preparation method according to the seventh aspect of the present application, the crystalline form C prepared by the preparation method according to the eighth aspect of the present application, the crystalline form D prepared by the preparation method according to the ninth aspect of the present application, the crystalline form E prepared by the preparation method according to the tenth aspect of the present application, or the pharmaceutical composition according to the eleventh aspect of the present application.

[0059] In some embodiments of the eleventh aspect to the twelfth aspect of the present application, the disease is selected from an inflammatory or autoimmune disease.

[0060] Further preferably, the disease is selected from alopecia areata, psoriasis, psoriatic arthritis, dermatitis, lupus erythematosus, inflammatory bowel disease, hidradenitis suppurativa, rheumatoid arthritis, or uveitis.

[0061] The thirteenth aspect of the present application relates to the use of a compound of formula I for the preparation of a medicament for the treatment of an alopecia areata disease.

[0062] In the present application, the positions of the absorption peaks in the XRPD patterns of the various crystalline forms can be within ±0.2° of the specific values recited in the above application, for example within ±0.1° of the specific values.

[0063] In the present application, the positions of the characteristic peaks in the DSC patterns of the various crystalline forms can be within ±5°C of the specific temperature values recited in the above application, or within ±3°C of the specific temperature, or within ±2°C of the specific temperature, or within ±1°C of the specific temperature values.

[0064] It should be understood that different types of equipment or different test conditions can give slightly different XRPD patterns and characteristic peaks or different DSC patterns and characteristic peaks. The specific values provided should not be taken as absolute.

[0065] Definitions of terms

[0066] The term "about" or "approximately," when used in connection with a numerical value, generally refers to the numerical value and all values within experimental error (e.g., within a 95% confidence interval for a mean) or within ±10% of the specified value, or within a broader range.

[0067] The terms "comprising" or "comprise," when used in this specification including the claims, shall not be construed as necessarily limiting the elements, steps or components to the precise procedures described herein. The term "consisting essentially of when used in this specification including the claims, shall not be construed as excluding particles, steps or components not specified. The term "consisting essentially of means that the range is limited to the specified elements, steps or components plus optional elements, steps or components that do not materially affect the basic and novel characteristics of the claimed subject matter. It is understood that the term "comprising" encompasses the terms "consisting essentially of and "consisting of."

[0068] The term "optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, that the description includes instances where the event or circumstance occurs, and instances where it does not.

[0069] The term "crystal form" or "crystal" refers to any solid material that exhibits three-dimensional ordering, as opposed to amorphous solid material, which produces a characteristic XRPD pattern with well-defined peaks.

[0070] The term "X-ray powder diffraction pattern (XRPD pattern)" refers to the experimentally observed diffraction pattern or parameters, data or values derived therefrom. An XRPD pattern is typically characterized by peak positions (abscissa) and / or peak intensities (ordinate).

[0071] The term "2 theta" refers to the peak position in degrees (°) set based on the X-ray diffraction experiment and is typically the unit of abscissa in the diffraction pattern. The experimental setup requires recording the reflected beam at a 2 theta angle if the reflection is diffracted when the incident beam forms a theta angle with a certain lattice plane. It is understood that the specific 2 theta values mentioned herein for a specific crystal form are intended to represent the 2 theta values (in degrees) measured using the X-ray diffraction experimental conditions described herein. For example, Cu-Ka is used as the radiation source as described herein. The XRPD patterns herein are preferably collected on an X'Pert3 X-ray powder diffractometer in transmission mode at room temperature. The instrument employs Cu-Ka illumination. The scan range is 3.5° to 40° in the 2 theta interval.

[0072] The terms "substantially the same" or "substantially as depicted in Figure X" in reference to X-ray diffraction peaks means that representative peak positions and intensity variations are taken into account. For example, one skilled in the art will appreciate that peak positions (2 theta) can show some variation, typically up to 0.1-0.2 degrees, and that the instrument used to measure the diffraction can also cause some variation. Additionally, one skilled in the art will appreciate that relative peak intensities can vary from instrument to instrument and also due to the degree of crystallinity, preferred orientation, surface of the sample prepared, and other factors known to one skilled in the art.

[0073] For the "peak temperature" of a DSC pattern of a crystalline form, it means the Peak value of the endothermic peak curve of the DSC pattern. The peak temperature of a DSC test can vary due to the purity, weight, particle size, test heating rate, and instrument system error of the test sample, and the provided values cannot be used as absolute values (Reference: Guo Yonghui, Yang Ning, Lv Yang. Application of Differential Scanning Calorimetry in the Study of Crystalline Drugs [C]. China Crystalline Drug Research and Development Technology Seminar. 2010.).

[0074] It should be understood that different types of equipment or different test conditions can give slightly different DSC patterns. For example, a Mettler Toledo DSC1 or TA DSC2500 differential scanning calorimeter can be used to determine the DSC pattern. As used herein, the term "substantially the same" for a DSC pattern takes into account the representative characteristic peak position. For example, a person skilled in the art will understand that the characteristic peak position will show some changes, usually up to 5°C. For a solid sample with multiple polymorphs, the heating rate of the DSC test has a greater effect on the DSC pattern. At a faster heating rate, the instrument thermal hysteresis effect is obvious, and the high melting point solid crystalline form does not have time to recrystallize, so the DSC pattern often only shows a melting endothermic peak of the low melting point crystalline form. At a moderate heating rate, the DSC pattern shows two peaks: a low melting point crystalline form melting endothermic peak and a high melting point crystalline form melting endothermic peak; and only at a lower heating rate, the instrument thermal hysteresis effect is weak, and three peaks appear: a low melting point crystalline form melting peak-re-crystallization exothermic peak-high melting point crystalline form melting endothermic peak. A person skilled in the art will understand that the determination of the heating rate range corresponding to the above different DSC patterns will vary depending on the weight, morphology, particle size and distribution of the test sample (Reference: Giron D. Thermal analysis and calorimetric methods in the characterisation of polymorphs and solvates [J]. Thermochimica Acta, 1995, 248: 1-59.).

[0075] "Thermogravimetric analysis (TGA)" is a common method for determining the thermal stability of a compound. The TGA pattern can be measured, for example, on a Mettler Toledo TGA1 instrument. The error of TGA can be within about ±0.5 mass%. The term "substantially the same" means that such error changes are taken into account. Exemplary test conditions are a temperature range of 35°C to 500°C, a heating rate of 10 K / min, and a purge gas of nitrogen (99.99%).

[0076] The term "treatment" is intended to refer to the alleviation or eradication of the disease state or condition being addressed. It is also understood that the treatment of the disease state or condition as described includes a complete treatment and also includes a treatment that achieves some, but not necessarily complete, alleviation of the disease state or condition.

[0077] The term "room temperature" refers to 20°C ± 5°C.

[0078] Unless otherwise indicated, the technical and scientific terms used herein have the same meaning as those generally understood by one of ordinary skill in the art to which the application pertains. In case of conflict between the definitions provided in this application and those of the art, the definitions provided in this application shall control. When referring to a range, a preferred range, or a preferred upper value and a preferred lower value, it is understood that the disclosure specifically contemplates any and every combination of the range limits with each other, and with the range limits of any other ranges, even if not expressly stated. The numerical ranges listed herein are intended to include the endpoints and all the integers and fractions within that range, unless otherwise indicated.

[0079] The present application achieves at least one of the following beneficial effects:

[0080] (1) The compound of formula I has good TYK2 inhibitory effect, can reduce the side effects of inhibiting other JAK family members, has low off-target risk and is safer. It has good therapeutic effect in animal models of psoriasis, asthma and alopecia areata, has good drug property, high stability and good safety.

[0081] (2) The polymorphs of the compound of formula I have excellent physicochemical properties, such as mechanical stability, high humidity resistance, thermal stability, low or no hygroscopicity, etc.; the flowability of crystal forms A / B / C / D is significantly better than that of crystal form E, and the polymorphs prepared into oral preparations have good stability.

[0082] (3) The preparation method of the polymorphs of the compound of formula I is simple and suitable for popularization and application. BRIEF DESCRIPTION OF DRAWINGS

[0083] Figure 1-1 is the XRPD pattern of crystal form A.

[0084] Figure 1-2 is the calculated XRPD of the single crystal structure model of crystal form A, the XRPD of the single crystal sample, and the superimposed XRPD of crystal form A.

[0085] Figure 1-3 is the polarizing microscope (PLM) photo of the single crystal obtained by culture and the micrograph of the single crystal selected and loaded on a loop for testing (a: PLM photo of the single crystal obtained by culture; b: micrograph of the single crystal selected and loaded on a loop for testing).

[0086] Figure 1-4 are the schematic diagram of asymmetric unit of the single crystal structure of Form A (Note: the second conformation position of disordered group in the structure is highlighted in green semi-transparent in the figure).

[0087] Figure 2 is the XRPD pattern of Form B.

[0088] Figure 3 is the XRPD pattern of Form C.

[0089] Figure 4 is the XRPD pattern of Form D.

[0090] Figure 5 is the XRPD pattern of Form E.

[0091] Figure 6 is the DVS pattern of Form A.

[0092] Figure 7 is the XRPD pattern of Form A before and after DVS test.

[0093] Figure 8 is the DVS pattern of Form B.

[0094] Figure 9 is the XRPD pattern of Form B before and after DVS test.

[0095] Figure 10 is the DVS pattern of Form C.

[0096] Figure 11 is the XRPD pattern of Form C before and after DVS test.

[0097] Figure 12 is the DVS pattern of Form D.

[0098] Figure 13 is the XRPD pattern of Form D before and after DVS test.

[0099] Figure 14 is the XRPD overlay of stability experiment of Form A.

[0100] Figure 15 is the XRPD overlay of stability experiment of Form B.

[0101] Figure 16 is the XRPD overlay of stability experiment of Form C.

[0102] Figure 17 is the XRPD overlay of stability experiment of Form D. DETAILED DESCRIPTION

[0103] The embodiments of the present application will be apparent from the following description of the embodiments, taken in conjunction with the accompanying drawings, illustrating by way of example the principles of the present application. It is to be understood that the description and specific examples are intended for the sole purpose of illustration of the embodiments of the present application, and are not intended to limit the present application as it is to be understood that variations and modifications can be made by those skilled in the art without departing from the scope of the present application. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of the present application.

[0104] Example 1 Preparation of the compound of Formula I

[0105] Step 1

[0106] To 2-methoxy-3-nitrobenzoic acid (1.0 g, 5.1 mmol) was added thionyl chloride (15.0 g) and the reaction was heated to reflux until the reaction was complete by TLC. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 5 mL of acetone and added dropwise to a solution of m-toluidine (0.49 g, 4.6 mmol) and triethylamine (1.0 g, 10.2 mmol) in 10 mL of acetone. The temperature was maintained below 10 °C during the addition. After the addition was complete, the reaction was allowed to warm to room temperature and stirred until the reaction was complete by TLC. The reaction was diluted with 50 mL of water and filtered. The solid was dried at room temperature for 24 h to give 2-methoxy-3-nitro-N-(m-tolyl)benzamide (1.2 g, 92.3 %).

[0107] MS m / z (ESI): 287.09 [M+H]+.

[0108] Step 2

[0109] To 2-methoxy-3-nitro-N-(m-tolyl)benzamide (1.0 g, 3.5 mmol) was added 10% palladium on carbon (0.15 g) in 30 mL of methanol. The reaction was stirred under a hydrogen atmosphere at 25 °C until the reaction was complete by TLC. The reaction was filtered and the filtrate was evaporated to give 2-methoxy-3-amino-N-(m-tolyl)benzamide (0.8 g, 89.4 %).

[0110] MS m / z (ESI): 257.13 [M+H] + .

[0111] Step 3

[0112] To a solution of 2-methoxy-3-amino-N-(m-tolyl)benzamide (0.8 g, 3.1 mmol) and 4,6-dichloro-N-(methyl-d3)pyridazine-3-carboxamide (0.65 g, 3.1 mmol) in tetrahydrofuran (5 mL) was added dropwise a solution of lithium bis(trimethylsilyl)amide (1 M, 9.9 mL, 9.9 mmol) in tetrahydrofuran at room temperature. The reaction was stirred at room temperature until the reaction was complete by TLC. The reaction was diluted with dichloromethane and washed with saturated aqueous sodium chloride. The organic phase was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Column chromatography gave 6-chloro-4-((2-methoxy-3-(m-tolylbenzamido)-N-(methyl-d3)pyridazine-3-carboxamide (0.85 g, 63.4 %).

[0113] MS m / z (ESI): 429.09 [M+H]+ .

[0114] Step 4

[0115] To a solution of 6-chloro-4-((2-methoxy-3-(m-tolylbenzamido)-N-(methyl-d3) pyridazine-3-carboxamide (0.8 g, 1.9 mmol), cyclopropanecarboxamide (0.16 g, 1.9 mmol), tris(dibenzylideneacetone)dipalladium (0.1 g, 0.1 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.12 g, 0.2 mmol), cesium carbonate (1.27 g, 3.9 mmol) in 1,4-dioxane (10 ml) was heated to reflux until the reaction was complete by TLC. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate: dichloromethane = 1:1) to give 6-(cyclopropanecarboxamide)-4-(2-methoxy-3-(m-tolylbenzamido)-N-(methyl-d3) pyridazine-3-carboxamide (0.43 g, 48.3%) as colorless oil.

[0116] MS m / z (ESI): 478.22 [M+H] + .

[0117] The instrument information and methods used in the preparation and testing of the crystalline forms are listed below

[0118] 1. X-ray powder diffraction (XRPD)

[0119] XRPD patterns were collected on a PANalytical X'Pert Pro X-ray powder diffractometer. The scan parameters are shown in Table 6 below.

[0120] Table 6: XRPD test parameters

[0121] 2. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

[0122] TGA and DSC patterns were collected on a TA 5500 thermogravimetric analyzer and a TA 2500 differential scanning calorimeter, respectively. The test parameters are shown in Table 7 below.

[0123] Table 7: DSC and TGA test parameters

[0124] Table 7: DSC and TGA test parameters

[0125] 3. Dynamic vapor sorption (DVS)

[0126] Dynamic vapor sorption (DVS) curves were collected on a DVS Intrinsic Plus from Surface Measurement Systems. The relative humidity at 25 °C was corrected using the deliquescence points of LiCl, Mg(NO3)2, and KCl. The test parameters are shown in Table 8 below.

[0127] Table 8: DVS test parameters

[0128] 4. Single crystal X-ray diffraction (SC-XRD)

[0129] The polarized light microscope (PLM) photographs of the single crystal sample were taken using an OLYMPUS SZX-7 stereomicroscope. The single crystal X-ray diffraction data were collected using a Rigaku XtaLAB Synergy R single crystal X-ray diffractometer. The test parameters are shown in Table 9 below.

[0130] Table 9: X-ray single crystal diffractometer parameters

[0131] Preparation of Example 2 Form A and its single crystal

[0132] Preparation of Example 2-1 Form A

[0133] Example 1 obtained 0.5 g of the compound of formula I, 1 times the amount (weight: volume) of tetrahydrofuran and 20 times the amount (weight: volume) of ethyl acetate were added, 50 °C stirring for 6 h, stirring and cooling to room temperature, continue to stir for 6 h, filter, 50 °C drying for 10 h, to obtain 0.46 g of Form A sample, white powder solid, yield 92%.

[0134] (1) X-ray powder diffraction: The XRPD pattern of Form A of the compound of formula I was determined by X-ray powder diffraction analyzer, as shown in Figure 1-1.

[0135] The XRPD pattern of Form A includes the characteristic peaks at the diffraction angles (2θ) in Table 10, wherein the error range of the 2θ value is ±0.2°.

[0136] Table 10: XRPD diffraction peak data of Form A

[0137] (2) Differential scanning calorimetry (DSC) analysis: The DSC of Form A includes an endothermic peak at 265.6 °C.

[0138] (3) Thermogravimetric analysis (TGA): The sample has no significant weight loss by thermogravimetric analyzer, and Form A is determined to be an anhydrous crystal form.

[0139] Preparation of single crystal of Form A of Example 2-2

[0140] A 3 mL glass vial was charged with 21.3 mg of Form A of the compound of Formula I (Example 2-1) and 0.5 mL of DMF. The sample was completely dissolved by ultrasonic oscillation. The resulting solution was filtered into a clean 3 mL glass vial. The 3 mL glass vial was then placed into a 20 mL glass vial containing 4 mL of methyl tert-butyl ether (MTBE) and the 20 mL glass vial was sealed with a cap. The 20 mL glass vial was then left to stand at room temperature. The system was observed daily and after 7 days, single crystals were observed to have formed in the system, as shown in Figures 1-3(a).

[0141] (1) X-ray powder diffraction: XRPD characterization results showed that the XRPD of the single crystal sample was consistent with the XRPD of Form A of the compound of Formula I (Example 2-1), as shown in Figure 1-2, indicating that the single crystal sample obtained by cultivation was Form A of the compound of Formula I.

[0142] (2) Single crystal X-ray diffraction: A single crystal (0.17 x 0.13 x 0.12 mm3) with suitable size and diffraction quality was selected from the single crystal sample obtained by cultivation above, as shown in Figure 1-3(b), and was subjected to room temperature single crystal X-ray diffraction characterization. Single crystal diffraction characterization results showed that the single crystal belonged to a monoclinic system, with a C2 / c space group, and a unit cell parameter of: Z = 8.

[0143] (3) Figure 1-4 shows the schematic diagram of the asymmetric unit of the single crystal structure of Form A of the compound of Formula I obtained by analysis. The asymmetric unit of the crystal structure contains only one molecule of the compound of Formula I, and does not contain any crystallization water molecules or other solvent molecules. The structure of the compound of Formula I was further confirmed by nuclear magnetic resonance and single crystal detection, and it was further confirmed that Form A of the compound of Formula I was an anhydrous crystal form.

[0144] Preparation of Form B of Example 3

[0145] Example 1 was taken as the starting material, 0.5 g of the compound of Formula I was added, 25 times the amount (weight: volume) of benzyl alcohol was added, and stirring was performed at 50°C for 1 h. Then 250 times the amount (weight: volume) of isopropyl acetate was added, and stirring was performed until the temperature cooled to room temperature. Stirring was continued for 6 h, and then filtration was performed. The filter cake was washed once with 50 times the amount (weight: volume) of isopropyl acetate, and then was dried at 50°C for 10 h. This resulted in 0.35 g of Form B of the compound of Formula I, which was a white powdery solid, and the yield was 70%.

[0146] (1) X-ray powder diffraction: The XRPD pattern of Form B of the compound of Formula I was determined using an X-ray powder diffraction analyzer, as shown in Figure 2.

[0147] The XRPD pattern of Form B includes characteristic peaks at diffraction angles (2Θ) in Table 11, wherein the error range of the 2Θ values is ±0.2°.

[0148] Table 11: XRPD diffraction peak data of Form B

[0149] (2) Differential scanning calorimetry (DSC) analysis: The DSC of Form B includes endothermic peaks at 247.8, 263.9 °C, and an exothermic peak at 248.9 °C.

[0150] (3) Thermogravimetric analysis (TGA): The sample has no significant weight loss determined by the thermogravimetric analyzer, and Form B is determined to be an anhydrous crystal form.

[0151] Preparation of Form C in Example 4

[0152] Take 0.5 g of the compound of Formula I prepared in Example 1, add 25 times the amount (weight: volume) of dimethyl sulfoxide, stir at 50 °C for 1 h, then add 250 times the amount (weight: volume) of isopropyl alcohol, stir and cool to room temperature naturally, continue to stir for 6 h, filter, rinse once with 50 times the amount (weight: volume) of isopropyl alcohol, and dry at 50 °C for 10 h to obtain a sample of Form C 0.38 g, white powder solid, with a yield of 76%.

[0153] (1) X-ray powder diffraction: The XRPD pattern of Form C of the compound of Formula I was determined by an X-ray powder diffraction analyzer, as shown in Figure 3.

[0154] The XRPD pattern of Form C includes characteristic peaks at diffraction angles (2Θ) in Table 12, wherein the error range of the 2Θ values is ±0.2°.

[0155] Table 12: XRPD diffraction peak data of Form C

[0156] (2) Differential scanning calorimetry (DSC) analysis: The DSC of Form C includes endothermic peaks at 234.3, 264.7 °C, and an exothermic peak at 234.9 °C.

[0157] (3) Thermogravimetric analysis (TGA): The sample has no significant weight loss determined by the thermogravimetric analyzer, and Form C is determined to be an anhydrous crystal form.

[0158] Preparation of Form D in Example 5

[0159] To the compound of formula I prepared in Example 1, 0.5 g, was added 25 times the amount (weight: volume) of DMF, stirred at 50°C for 1 h, then 250 times the amount (weight: volume) of water was added, stirred and allowed to cool to room temperature naturally, and stirred for another 6 h, filtered, washed once with 50 times the amount (weight: volume) of water, and dried at 50°C for 10 h to obtain a sample of crystal form D, 0.36 g, as a white powdery solid, in a yield of 72%.

[0160] (1) X-ray powder diffraction: The XRPD pattern of crystal form D of the compound of formula I was determined using an X-ray powder diffraction analyzer, as shown in FIG. 4.

[0161] The XRPD pattern of crystal form D includes characteristic peaks at diffraction angles (2θ) in Table 13, wherein the error range of the 2θ values is ±0.2°.

[0162] Table 13: XRPD diffraction peak data of crystal form D

[0163] (2) Differential scanning calorimetry (DSC) analysis: The DSC of crystal form D includes an endothermic peak at 248.3, 260.6°C and an exothermic peak at 249.4°C.

[0164] (3) Thermogravimetric analysis (TGA): The sample had no significant weight loss determined by a thermogravimetric analyzer, indicating that crystal form D is an anhydrous crystal form.

[0165] Preparation of crystal form E

[0166] To the compound of formula I prepared in Example 1, 0.5 g, was added 10 times the amount (weight: volume) of a mixed solution of 1,4-dioxane and isopropyl ether, stirred at 50°C for 1 h, stirred and allowed to cool to room temperature naturally, and stirred for another 6 h, filtered, and dried at 50°C for 10 h to obtain a sample of crystal form E, 0.31 g, as a white powdery solid, in a yield of 62%.

[0167] (1) X-ray powder diffraction: The XRPD pattern of crystal form E of the compound of formula I was determined using an X-ray powder diffraction analyzer, as shown in FIG. 5.

[0168] The XRPD pattern of crystal form E includes characteristic peaks at diffraction angles (2θ) in Table 14, wherein the error range of the 2θ values is ±0.2°.

[0169] Table 14: XRPD diffraction peak data of crystal form E

[0170] (2) Thermogravimetric analysis (TGA): The sample had no significant weight loss determined by a thermogravimetric analyzer, indicating that crystal form E is an anhydrous crystal form.

[0171] Biological test evaluation

[0172] The present application is further described and explained with the following test examples, which are not meant to limit the scope of the present application.

[0173] Test Example 1: Ex vivo - cell inhibition model

[0174] The inhibitory effect of the compound of Formula I (Example 1) on the cell TYK2 signaling pathway can be determined by the following method:

[0175] Experimental instrument: SpectraMax Paradigm plate reader.

[0176] Experimental method: The Ba / F3 cell line expressing TYK2 is used in this experiment, and the effect of the compound of Formula I (Example 1) on cell proliferation inhibition is evaluated by determining the effect of the compound of Formula I (Example 1) on the activity of cells in vitro.

[0177] Experimental operation: The CellTiter-Glo buffer is thawed and placed at room temperature, the CellTiter-Glo substrate is placed at room temperature and added to the buffer to dissolve the substrate, and the CellTiter-Glo working solution is prepared. Add 98 μL of cell culture solution to a flat-bottomed 96-well transparent drug plate, add 2 μL of gradient-diluted compound of Formula I (Example 1) solution, stand for 10 minutes, then add 50 μL of CellTiter-Glo working solution, shake on the orbital shaker for 2 minutes, then stand at room temperature for 10 minutes, and finally detect the luminescence signal on the SpectraMax Paradigm plate reader.

[0178] Experimental data processing method: The cell proliferation inhibition rate (Inhibition Rate) data is processed using the following formula:

[0179] Inhibition Rate (Inh%) = 100 - (RLUDrug-RLUMin) / (RLUMax-RLUMin)*100%. The inhibition rate corresponding to different concentrations of the compound of Formula I (Example 1) is calculated in EXCEL, then the inhibition rate curve is drawn and the relevant parameters are calculated using GraphPad Prism software, including the maximum and minimum inhibition rates of the cells, IC 50 values.

[0180] The inhibitory activity of the compound of Formula I (Example 1) obtained by the above scheme is as follows:

[0181] Table 15: Inhibitory activity

[0182] Note: A represents 1 nM < IC 50 <5 nM.

[0183] Experimental conclusion: the compound of formula I (example 1) has good inhibitory effect on cell TYK2 signal pathway.

[0184] The oily substance (example 1), anhydrous crystal form (example 2 crystal form A, example 3 crystal form B, example 4 crystal form C, example 5 crystal form D, example 6 crystal form E) of the compound of formula I in the experiment are completely dissolved, and have consistent inhibitory effect on cell TYK2 signal pathway.

[0185] Test example 2: hERG channel inhibitory activity experiment

[0186] Experimental instrument: HEKAEPC 10 patch clamp amplifier.

[0187] Experimental method: In this experiment, the manual patch clamp technique is used to detect the blocking effect of the compound of formula I (example 1) on the current of HEK-293 cell line stably expressing hERG channel, and the risk of inhibitory effect of the compound of formula I (example 1) on cardiac hERG potassium channel is evaluated by fitting concentration effect relationship.

[0188] Experimental instrument: SpectraMax Paradigm plate reader.

[0189] Experimental method: In this experiment, an expression cell line is used to evaluate the toxic effect of the compound of formula I (example 1) on the heart by measuring the inhibition of the compound of formula I (example 1) on the activity of in vitro cells.

[0190] Experimental operation: The patch clamp operation first uses a microelectrode puller to draw a capillary glass tube into a recording electrode, and then the electrode filled with intracellular fluid is installed in a microelectrode holder, and the microelectrode manipulator is operated under an inverted microscope to make the electrode immersed in extracellular fluid and record the electrode resistance (Rpip). Then the electrode is slowly contacted to the cell surface, and negative pressure suction is given to form a GΩ seal. At this time, fast capacitance compensation is performed, and negative pressure suction is continued to break the cell membrane to form a whole cell recording mode. Finally, slow capacitance compensation is performed and experimental parameters such as series resistance (Rs) are recorded. No leakage compensation is given. When the hERG current of the whole cell recording is stable, the drug administration is started, and after about 5 min (or the current is stable) of each drug concentration, the next concentration is detected. The cover glass with cells is placed in the recording bath under the inverted microscope, and the blank control extracellular fluid and the working solution of the compound of formula I (example 1) to be tested are sequentially flowed through the recording bath from low concentration to high concentration by using the method of gravity perfusion to act on the cells, and the peristaltic pump is used for liquid exchange in the recording. The current detected in the extracellular fluid without the compound of formula I (example 1) is used as the control group of each cell. Each concentration is independently detected twice. All electrophysiological experiments are carried out at room temperature.

[0191] Experimental data processing method: first, the tail current (Peak tail current compound ) and blank control tail current (Peak tail current control ) after each drug concentration effect is normalized, then the inhibition rate corresponding to each drug concentration is calculated , and the average (Mean), standard deviation (SD) and standard error (SE) of each concentration inhibition rate are calculated, and the data are expressed as Mean±SE. The IC 50 value of the compound of formula I (Example 1) and the dose-effect curve fitting are completed by using GraphPad Prism software.

[0192] Experimental conclusion: the hERG detection of the compound of formula I (Example 1) of the present application is negative, that is, it has no cardiotoxicity, and has good safety.

[0193] The oily substance (Example 1), anhydrous crystal form (Example 2 crystal form A, Example 3 crystal form B, Example 4 crystal form C, Example 5 crystal form D, Example 6 crystal form E) of the compound of formula I of the present application are all in completely dissolved state in the experiment, and the hERG channel inhibition activity is consistent.

[0194] Test example 3: kinase selectivity experiment

[0195] 3.1 JAK1-3 / TYK2 JH1 in vitro enzyme binding experiment

[0196] Experimental instrument: Envision enzyme marker (PerkinElmer), ECHO550 (LABCYTE)

[0197] Experimental method: the method of fluorescence resonance energy transfer (TR-FRET) is used to test the inhibition effect of the compound of formula I (Example 1) on JAK1-3 / TYK2 JH1 kinase.

[0198] Experimental procedure: Compound of Formula I (Example 1) was prepared as 10 mM stock solution in DMSO, further diluted in DMSO to different concentration gradient of Compound of Formula I (Example 1) dilutions, 100x Compound of Formula I (Example 1) dilutions were transferred to 384 experimental plates using Echo instrument. Assay buffer, SEB, TK-Substrate-biotin, detection reagent, etc. in the experiment were all from TK kit (Cisbio, Cat#62TK0PEC). Three 1x experimental working solutions were prepared as follows: 1. for JAK1 JH1 experimental working solution: Assay buffer final concentration 1x, MgCl2 final concentration 5 mM, EGTA final concentration 0.625 mM, SEB final concentration 60 nM, Brij-35 final concentration 0.01%, DTT final concentration 1 mM, 2. for JAK2-3 JH1 experimental working solution: Assay buffer final concentration 1x, MgCl2 final concentration 5 mM, DTT final concentration 1 mM, 3. for TYK2 JH1 experimental working solution: Assay buffer final concentration 1x, MgCl2 final concentration 5 mM, MnCl2 final concentration 1 mM, SEB final concentration 12.5 nM, DTT final concentration 1 mM. JAK1-3 / TYK2 JH1 kinases were prepared at 2x final concentration with respective 1x working solutions, 2x TK-Substrate-biotin substrate. 5 uL of JAK1-3 / TYK2 JH1 kinases were added to 384 well experimental plates, centrifuged at 1000 rpm for 30 seconds, incubated at room temperature for 15 minutes. 5 uL of TK-Substrate-biotin substrate were added to 384 well experimental plates, centrifuged at 1000 rpm for 30 seconds, JAK1-2 JH1 incubated at room temperature for 45 minutes, JAK3 / TYK2 JH1 incubated at room temperature for 60 minutes. 2x detection reagent was prepared, 10 uL were added to 384 well experimental plates, centrifuged at 1000 rpm for 30 seconds, JAK1-2 JH1 incubated at room temperature for 60 minutes, JAK3 / TYK2 JH1 incubated at room temperature for 120 minutes, then all incubated at 4 °C overnight. Finally, the 665 nm / 615 nm fluorescence signal ratio was read on Envision plate reader (PerkinElmer).

[0199] Experimental data processing method: The experimental data were processed and analyzed by using XLfit software written by IDBS company, which was integrated in Microsoft Excel environment. First, the average values of the high signal control wells and the low signal control wells were calculated, respectively. Then the inhibition rate of the wells of the compound of Formula I (Example 1) was calculated according to the formula “% inhibition of single well = 100% - (average value of high signal control group - signal value of single well) / (average value of high signal control group - average value of low signal control group) * 100%”. Finally, the concentration and corresponding inhibition rate data were imported into the XLfit software, and the IC50value of the compound of Formula I (Example 1) was calculated by using the Dose Response One Site 205 model in the software and the four-parameter inhibition rate-concentration curve fitting. 50

[0200] 3.2 JAK1 JH2 in vitro enzyme binding experiment

[0201] Experimental instrument: Envision microplate reader (PerkinElmer), Echo (LABCYTE)

[0202] Experimental method: In this experiment, the fluorescence resonance energy transfer (TR-FRET) method was used to test the inhibitory effect of the compound of Formula I (Example 1) on JAK1 JH2 pseudokinase.

[0203] Experimental operation: The compound of Formula I (Example 1) was prepared into a 10 mM stock solution using DMSO, and further diluted into different concentration gradients of the compound of Formula I (Example 1) diluent in DMSO. The 200x final concentration of the compound of Formula I (Example 1) diluent was transferred to a 384 experimental plate using an Echo instrument. Prepare the experimental 1x working solution as shown in the following table: HEPES pH 7.5 final concentration 20 mM, MgCl2 final concentration 10 mM, Brij-35 final concentration 0.015%, DTT final concentration 2 mM, BSA final concentration 50 ug / mL. Prepare 3x final concentration of JAK1 JH2 pseudokinase, Tb antibody, Tracer with 1x working solution, respectively. Add 5 uL of JAK1 JH2 pseudokinase to the 384-well experimental plate and centrifuge at 1000 rpm for 30 seconds. Add 5 uL of Tb antibody to the 384-well experimental plate and centrifuge at 1000 rpm for 30 seconds. Add 5 uL of Tracer to the 384-well experimental plate and centrifuge at 1000 rpm for 30 seconds. Incubate at room temperature for 60 minutes and then incubate overnight at 4°C. Finally, read the 495 nm / 520 nm fluorescence signal ratio on the Envision microplate reader (PerkinElmer).

[0204] ​Experimental data processing method: The experimental data were processed and analyzed by using XLfit software written by IDBS company, which was integrated in Microsoft Excel environment. First, the average values of the high signal control wells and the low signal control wells were calculated, respectively. Then the inhibition rate of each well of the compound of Formula I (Example 1) was calculated according to the formula: “Inhibition rate of single well % = 100% - (average value of high signal control group - signal value of single well) / (average value of high signal control group - average value of low signal control group) * 100%”. Finally, the concentration and corresponding inhibition rate data were imported into the XLfit software, and the IC50value of the compound of Formula I (Example 1) was calculated by using the Dose Response One Site 205 model in the software and the four-parameter inhibition rate-concentration curve fitting. 50

[0205] 3.3 TYK2 JH2 in vitro enzyme binding experiment

[0206] Experimental instrument: Envision microplate reader (PerkinElmer), ECHO550 (LABCYTE).

[0207] Experimental method: The fluorescence resonance energy transfer (TR-FRET) method was used to test the inhibition of the compound of Formula I (Example 1) on TYK2 JH2 pseudokinase.

[0208] Experimental operation: The compound of Formula I (Example 1) was dissolved in DMSO to a concentration of 10 mM stock solution, and further diluted in DMSO to different concentration gradients of the compound of Formula I (Example 1) diluent (200x). The Echo instrument was used to transfer the compound of Formula I (Example 1) diluent (200x) to a 384 experimental plate. The dilution buffer (20 mM HEPES pH 7.5, 10 mM MgCl2, 0.015% Brij-35, 2 mM DTT and 50 ug / mL BSA) was prepared. The TYK2 JH2 pseudokinase, Tb antibody and Tracer working solutions were prepared with the dilution buffer. 5 uL of TYK2 JH2 pseudokinase working solution (final concentration 0.5 nM) was added to the 384-well experimental plate and centrifuged at 1000 rpm for 30 seconds. 5 uL of Tb antibody (final concentration 1x) was added to the 384-well experimental plate and centrifuged at 1000 rpm for 30 seconds. 5 uL of Tracer (final concentration 0.5 nM) was added to the 384-well experimental plate and centrifuged at 1000 rpm for 30 seconds. After incubation at room temperature for 60 minutes and overnight incubation at 4°C, the 495 nm / 520 nm fluorescence signal ratio was finally read on the Envision microplate reader (PerkinElmer).

[0209] ​Experimental data processing method: the test data processing and analysis were performed using XLfit software written by IDBS company and integrated in Microsoft Excel environment. First, the average signal values of the high signal control group and the low signal control group were calculated respectively, and then the reaction inhibition rate of the hole of the compound of formula I (Example 1) was calculated according to the formula "inhibition rate of single hole % = 100%-(high signal control group average value-single hole signal value) / (high signal control group average value-low signal control group average value)*100%". Then the concentration and corresponding inhibition rate data were imported into the XLfit software, the Dose Response One Site 205 model in the software was used, the four parameter method inhibition rate-concentration curve was used for fitting, and the IC 50 value of the compound of formula I (Example 1) was calculated.

[0210] Table 16: TYK2 JH2 in vitro enzyme binding experiment results

[0211] Experimental conclusion: the JH1 target point selectivity level of the compound of formula I (Example 1) of the application to JAK 1-3 / TYK2 is equivalent to that of BMS-986165, and neither has inhibitory activity. In addition, the inhibitory activity to JAK1 JH2 is also significantly reduced compared with BMS-986165. The compound of formula I (Example 1) of the application has good TYK2 JH2 selectivity and low off-target risk.

[0212] The oil (Example 1), anhydrous crystal form (Example 2 crystal form A, Example 3 crystal form B, Example 4 crystal form C, Example 5 crystal form D, Example 6 crystal form E) of the compound of formula I of the application are all in a completely dissolved state in the experiment, and the target point selectivity is consistent.

[0213] Test example 4: in vivo-mouse psoriasis model

[0214] The efficacy of the compound of formula I (Example 1) in the imiquimod-induced mouse psoriasis model can be determined by the following method:

[0215] Sample preparation: reference to commercially available binidolol cream preparation. Mix the compound of formula I (Example 1), oil phase (hexadecanol, vaseline, liquid paraffin and glycerol monodouble stearate), water phase (propylene glycol, Tween 80 and water) and suitable additives (ethylparaben), and prepare 1% cream sample by vacuum emulsification.

[0216] Model preparation: the test site on the back of the animal is shaved, the area is 2*3 square centimeters, and the imiquimod ointment is continuously applied to the skin on the back of the mouse for 6 days to construct a mouse psoriasis model. According to the experimental scheme, the animals in each group are administered on Day 1-6, and the experimental design of the imiquimod-induced mouse psoriasis model is shown in the following table:

[0217] Table 17: Experimental design

[0218] The severity of skin inflammation was evaluated on the seventh day using a five-point scale (0-4) for scoring (PASI score):

[0219] Skin thickness: 0: smooth skin without wrinkles; 1: slight wrinkles on the skin at the edge of the treated area; 2: slight wrinkles on the skin throughout the treated area; 3: further deepening of the wrinkles in the treated area; 4: on the basis of a score of 3, the mouse shows weight loss or poor condition, etc.

[0220] Crusting: 0: smooth skin without scales; 1: slight scales on the skin in the treated area; 2: the skin in the treated area is completely covered with scales; 3: further deepening of the scales in the treated area; 4: on the basis of a score of 3, the mouse shows weight loss or poor condition, etc.

[0221] Erythema: 0: smooth skin; 1: slight redness on the skin in the treated area; 2: the skin in the treated area is completely red; 3: further deepening of the redness in the treated area; 4: on the basis of a score of 3, the mouse shows weight loss or poor condition, etc.

[0222] The comparison results of the PASI scores of the compound of formula I (Example 1) in the mouse psoriasis-like model induced by imiquimod are shown in the following table:

[0223] Table 18: PASI score results

[0224] Note: compared with the model group, *** P < 0.001, ** P < 0.01, the data represent the average PASI score within the group.

[0225] Experimental conclusion: the compound of formula I (Example 1) can effectively improve the symptoms of psoriasis in the mouse psoriasis-like model induced by imiquimod, and has a significant difference compared with the model group (P < 0.001), which is better than binimetinib cream and BMS-986165.

[0226] In the mouse psoriasis-like model induced by imiquimod, the compound of formula I (Example 1) can effectively improve the symptoms of psoriasis by oral administration (gavage, 10 mg / kg, 25 mg / kg, 50 mg / kg, bid), and has a significant difference compared with the model group.

[0227] It is found through pharmacodynamic experiments that there is a certain difference in the anti-psoriasis efficacy when the same compound is administered by different routes of administration, such as oral administration or external use.

[0228] In an in vivo mouse psoriasis-like model (oral and topical), the anti-psoriasis efficacy of crystal form A (Example 2), crystal form B (Example 3), crystal form C (Example 4), and crystal form D (Example 5) of the compound of formula I was comparable to that of the oily form of compound I (Example 1); the anti-psoriasis efficacy of crystal form E (Example 6) of compound I was slightly inferior to that of the oily form of compound I (Example 1).

[0229] Test Example 5: In vivo mouse asthma model

[0230] The efficacy of compound I of this invention (Example 1) in an OVA-induced mouse asthma model can be determined by the following method:

[0231] Preparation method of test sample: Test sample: Accurately weigh the compound of formula I (Example 1) and prepare a solution with a concentration of 2 mg / ml using DMSO for later use.

[0232] Model preparation: Thirty female BALB / c mice were acclimatized and fed for one week. Ten mice were randomly selected as blank control group and 20 mice were selected to prepare asthma model. The model mice were sensitized by intraperitoneal injection of 0.2 mL of sensitizing solution on days 0, 7 and 14. The sensitizing solution contained 50 μg OVA and 2 mg Al(OH)3. The normal group was injected with an equal volume of physiological saline.

[0233] On days 21-23 after sensitization, mice in the model group and the drug administration group were challenged by nebulization with 5% OVA for 30 minutes, while mice in the normal group were nebulized with physiological saline for the same duration.

[0234] Grouping and administration: Mice in the blank control group and model group were administered 50 μl of physiological saline intratracheally 30 min before daily challenge, while mice in the compound I (Example 1) group were administered 50 μl of the corresponding drug intratracheally, once daily for 3 consecutive days. The animal grouping and dosage table are as follows:

[0235] Table 19: Animal Grouping and Dosage

[0236] Airway hyperresponsiveness assay: Airway hyperresponsiveness was measured in each group of mice after the last challenge. Mice were placed in a body plethysmography chamber, and the baseline expiratory interval (Penh) value was recorded. Mice were then challenged with nebulized methacholine at concentrations ranging from 0, 6.25, and 12.5 mg / mL, with a nebulization dose of 100 μL per 60 s. After each concentration was nebulized, the presence of hypoxic symptoms such as shortness of breath, head scratching, and agitation was observed. Penh values ​​were recorded for 3 minutes, and the average values ​​were used to compare the airway hyperresponsiveness of each group of mice.

[0237] Inflammatory cell detection in lavage fluid: After the detection of airway hyperresponsiveness in mice, the mice were sacrificed by cervical dislocation, fixed on a dissection plate, the abdomen and chest of the mice were opened, the skin and excess tissue of the neck were cut off, the trachea of the mice was exposed, a small opening was cut on the horizontal axis of the trachea with small scissors, a needle of a 1ml syringe was inserted, and the needle was fixed with surgical thread. 0.5ml of pre-cooled phosphate buffer solution (PBS) was drawn into the 1ml syringe and slowly injected into the lungs of the mice, and then slowly sucked. Each mouse was lavaged once, and about 0.4ml of lavage fluid was collected. The number of inflammatory cells in the lung lavage fluid was detected by a cell counter.

[0238] Statistical analysis: SPSS software was used for statistical analysis. The measurement data was expressed as mean ± standard deviation. One-Way ANOVA was used for comparison between groups, LSD test was used for variance equality, Dunnett-t test was used for variance inequality, and P<0.05 was considered statistically significant.

[0239] Experimental results:

[0240] The results of the influence on the Penh value of mice are as follows:

[0241] Table 20: Influence on the Penh value of mice

[0242] Note: compared with the blank control group, # P<0.05, ### P<0.001; compared with the model group, * P<0.05.

[0243] After 3 weeks of sensitization of mice with OVA+Al(OH)3, continuous stimulation with 5% OVA for 3 days, and stimulation with 6.25mg / ml and 12.5mg / ml acetylcholine, the Penh value was significantly higher than that of the blank control group (P<0.001 or P<0.05), which proved that the mouse asthma model was successfully established.

[0244] Compared with the model group, the Penh value of mice was significantly reduced after administration of the compound of formula I (Example 1) before stimulation (P<0.05).

[0245] Influence on inflammatory cells in the lung lavage fluid of mice: the results are as follows:

[0246] Table 21: Influence on inflammatory cells in the lung lavage fluid of mice

[0247] Note: compared with the blank control group, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05.

[0248] The number of inflammatory cells in the bronchoalveolar lavage fluid of the mice significantly increased (P<0.001 or P<0.01) after the mice were sensitized with OVA+Al(OH)3 for 3 weeks and then continuously challenged with 5% OVA for 3 days. Compared with the model group, the number of inflammatory cells (white blood cells, lymphocytes) significantly decreased after the mice were administered the compound of formula (I) (Example 1) before the challenge.

[0249] The oil (Example 1), anhydrous crystal forms (crystal form A of Example 2, crystal form B of Example 3, crystal form C of Example 4, crystal form D of Example 5, and crystal form E of Example 6) of the compound of formula I of the present application were all completely dissolved in the experiment, and the anti-asthma efficacy was consistent.

[0250] Test Example 6: In vivo-mouse alopecia areata model

[0251] The efficacy of the compound of formula I (Example 1) in the imiquimod-induced C3H / HeJ mouse alopecia areata model can be determined as follows:

[0252] Sample preparation: A commercially available brodalumab cream was used as a reference. The compound of formula I (Example 1), the oil phase (cetyl alcohol, vaseline, liquid paraffin, and glycerol monostearate), the water phase (propylene glycol, Tween 80, and water), and the appropriate additives (ethylparaben) were mixed to prepare a 1% cream sample by vacuum emulsification.

[0253] Positive control: After the litrexin was ground, a 3 mg / ml suspension was prepared using a 0.5% CMC-Na solution.

[0254] Model preparation: C3H / HeJ mice were used in this experiment, with 8 mice in each group. Cotton swabs were used to dip imiquimod ointment and apply it evenly to the skin of the neck, with an application area of about 1.5 cm x 1.5 cm, once a day, for 7 consecutive days. After 1 hour of imiquimod application, the animals in each group were administered the corresponding drug, the blank control group and the model group were applied with the corresponding blank matrix on the back, and the test drug group (compound of formula I (Example 1)) was applied with 100 mg of the test drug (1%). The positive drug control group was administered 30 mg / kg of litrexin by gavage, once a day, for 7 consecutive days.

[0255] Detection index: On the 8th day of administration, the condition of the neck hair was observed, and the score was determined based on the range of hair loss, the degree of hair thinning, and the color of the skin. After the scoring was completed, the mice were sacrificed and the skin was taken, the level of IFN-γ in the skin was detected by ELISA kit, and the histopathological section was observed.

[0256] Scoring criteria:

[0257] Table 22: Alopecia areata scoring criteria

[0258] Results of the experiment:

[0259] Table 23: Score of hair loss

[0260] Note: compared with the control group, ### P<0.001; compared with the model group, *** P<0.001, ** P<0.01.

[0261] Results of the experiment: after the mice were coated with imiquimod on the back for 7 days, obvious alopecia phenomenon appeared (P<0.001); after the model animals were given the test drug (the compound of formula I (Example 1)), the alopecia was obviously relieved (P<0.001); after the model animals were given the positive control drug ruxolitinib, the alopecia was slightly relieved (P<0.01).

[0262] Table 24: IFN-γ level in the skin

[0263] Note: compared with the control group, ### P<0.001; compared with the model group, *** P<0.001.

[0264] Results of the experiment: after the mice were coated with imiquimod on the back for 7 days, the IFN-γ content in the skin was obviously increased (P<0.001); after the model animals were given the test drug (the compound of formula I (Example 1)), the IFN-γ content was significantly reduced (P<0.001); after the model animals were given the positive control drug ruxolitinib, the IFN-γ content was also significantly reduced (P<0.001).

[0265] Conclusion of the experiment: the compound of formula I (Example 1) can effectively improve the symptoms of alopecia areata in the imiquimod-induced mouse alopecia areata model, which is manifested as the alleviation of hair loss and the reduction of IFN-γ content in the skin, and has a significant difference (P<0.001) compared with the model group, and is superior to the positive control drug ruxolitinib. In addition, according to the histopathological section, it is shown that the hair follicle damage in the compound of formula I (Example 1) group is obviously relieved.

[0266] In the in vivo-mouse alopecia areata model, the compounds of formula I in the form of crystal A (Example 2), crystal B (Example 3), crystal C (Example 4), crystal D (Example 5) and the compound of formula I in the form of oil (Example 1) have equivalent anti-alopecia areata efficacy; compared with the compound of formula I in the form of oil (Example 1), the compound of formula I in the form of crystal E (Example 6) has slightly worse anti-alopecia areata efficacy.

[0267] Test Example 7: toxicity study of repeated administration for 4 weeks in rats

[0268] Experimental materials:

[0269] Animals: SD rats, SPF level, half male and half female, 200 ± 20 g.

[0270] Test product: cream of compound of formula I (Example 1) (2%), prepared with reference to commercially available binimetinobe cream.

[0271] Positive control: 10 mg of BMS-986165 was dissolved in 4 ml of PEG400, and after complete dissolution, normal saline was added to prepare a solution with a final concentration of 1 mg / ml.

[0272] Reagents: urethane, reagents for blood cell analyzer, reagents for automatic biochemical analyzer, reagents for automatic coagulation analyzer, reagents for electrolyte analyzer.

[0273] Instruments: electronic balance, blood cell analyzer, automatic biochemical analyzer, automatic coagulation analyzer, electrolyte analyzer.

[0274] Grouping and administration:

[0275] Grouping: the animals were randomly divided into 3 groups according to body weight, namely blank control group, compound of formula I (Example 1) group and BMS-986165 group, 10 animals in each group, half male and half female.

[0276] Dosing regimen: the compound of formula I (Example 1) group was administered twice a day by skin application, the BMS-986165 group was administered twice a day by gavage, for 4 weeks (28 days) in succession, the specific dosing regimen is shown in the table below:

[0277] Table 25: Dosing regimen

[0278] Index detection:

[0279] Daily clinical observation, body weight detection;

[0280] After the last administration, the rats were fasted for more than 16 h, the next day they were anesthetized, blood was collected from the abdominal aorta, and hematological (EDTA-2K anticoagulation), blood biochemistry (serum) and coagulation function (sodium citrate anticoagulation to separate plasma) tests were performed;

[0281] After the animals were sacrificed, the tissues and organs were dissected, gross observation was performed, and the organ weights and coefficients (heart, liver, spleen, kidney, thymus) were measured;

[0282] The skin at the administration site and lung tissue were taken for HE staining to observe the changes in epidermal thickness and inflammatory cells in the dermis layer, as well as lung lesions.

[0283] Experimental results:

[0284] Table 26: Test results

[0285] Note: Compared with the blank control group: * P < 0.05, ** P < 0.01.

[0286] WBC: white blood cell; Lymph: lymphocyte; Mono: monocyte; Gran: granulocyte; PLT: platelet; APTT: activated partial thromboplastin time; TB: total bilirubin.

[0287] Hematological results showed that BMS-986165 could significantly reduce the number of white blood cells (P < 0.01) and platelets (P < 0.05) in rats, and the compound of formula I (Example 1) had no significant effect on the number of white blood cells (P > 0.05);

[0288] Coagulation results showed that BMS-986165 could significantly prolong APTT (P < 0.01), and the compound of formula I (Example 1) had no significant effect on APTT (P > 0.05);

[0289] Biochemical results showed that BMS-986165 could significantly increase the total bilirubin (TB) level in serum (P < 0.05), and the compound of formula I (Example 1) had no significant effect on total bilirubin (P > 0.05).

[0290] In addition, the compound of formula I (Example 1) had no significant effect on the general state, body weight, organ coefficient, skin and lung pathology, and other hematological, coagulation and biochemical indicators of rats, showing good safety.

[0291] In addition, the compound of formula I (Example 1) showed good safety in skin irritation experiment, skin allergy experiment, and oral acute toxicity experiment.

[0292] The compound of formula I (Example 2), the compound of formula I (Example 3), the compound of formula I (Example 4), the compound of formula I (Example 5), and the compound of formula I (Example 6) have similar safety compared with the compound of formula I (Example 1).

[0293] Hygroscopicity evaluation

[0294] The hygroscopicity of the crystal forms A / B / C / D was evaluated using DVS.

[0295] The DVS test results and the XRPD results of the samples after DVS test are shown in Figures 6 to 13.

[0296] The results show that the moisture adsorption of Form A at 25°C / 80% RH is 0.0806%, almost no hygroscopicity, and the crystal form does not change after the test. The moisture adsorption of Form B at 25°C / 80% RH is 0.0698%, almost no hygroscopicity, and the crystal form does not change after the test. The moisture adsorption of Form C at 25°C / 80% RH is 0.0702%, almost no hygroscopicity, and the crystal form does not change after the test. The moisture adsorption of Form D at 25°C / 80% RH is 0.4062%, slightly hygroscopic, and the crystal form does not change after the test. The hygroscopicity evaluation of Form E under the same conditions is slightly hygroscopic, and there is no crystal form change after the test.

[0297] Solid state stability evaluation

[0298] The Form A / B / C / D samples were placed at 60°C / closed / 1 day, 25°C / 60% RH / open / 1 week and 40°C / 75% RH / open / 1 week, respectively, and their physical and chemical stability was evaluated by XRPD and HPLC. The XRPD results are listed in Figures 14 to 17.

[0299] Procedure: weigh the solid into the HPLC vial, and store the sample under the corresponding experimental condition. The samples under 25°C / 60% RH and 40°C / 75% RH conditions were sealed with a sealing film, and 10 pinholes were additionally punched on the film.

[0300] Table 27: Summary of stability test results

[0301] *: compared with the starting sample = HPLC purity / starting sample HPLC purity * 100%

[0302] The results show that Form A, Form B, Form C, Form D have no significant change in HPLC purity after being placed at 60°C / closed / 1 day, 25°C / 60% RH / open / 7 days, 40°C / 75% RH / open / 7 days, and the crystal form does not change. Form E was also evaluated for solid state stability under the same conditions, and there was no significant change in HPLC purity, and the crystal form did not change.

[0303] Flowability test

[0304] When the particles slide on the free slope of the powder accumulation layer, they are in a state of rest when the gravity and the inter-particle friction reach a balance, and at this time the angle between the slope of the powder accumulation layer and the horizontal plane is the angle of repose (θ). The angle of repose is the simplest method to test the flowability of the powder. The angle of repose can be directly measured, and can be calculated according to the height of the powder layer and the radius of the disc.

[0305] Table 28: Angle of repose and flowability evaluation

[0306] Compressibility is also a simple way to predict the flowability. It is obtained by measuring the apparent volume (V0) and the tap volume (V F ) of the bulk powder, and then calculating. It can be calculated according to the following formula: Compressibility = 100 x [(V0-V F ) / V0]

[0307] Table 29: Compressibility and flowability evaluation

[0308] To simulate the production process, the raw materials of different crystal forms were crushed by a small universal crusher at a speed of about 20000 rpm, and then passed through a 60 mesh sieve. The excipients were mixed with the specific crystal form raw materials to form a powder. The rest angle of the mixed material was directly detected by BT-1000 powder comprehensive performance tester. The apparent volume and tap volume were determined according to the bulk density and tap density determination method (0993) in Chinese Pharmacopoeia 2020 edition.

[0309] Table 30: Flowability evaluation of polymorphs

[0310] The results show that the flowability of crystal forms A / B / C / D is significantly better than that of crystal form E, which will greatly facilitate the production control of tablets or capsules, simplify the process, reduce the production cost, and improve the qualified rate of products. Especially for the process of powder mixing and direct filling of capsules, or powder direct compression process, crystal forms A / B / C / D have greater advantages compared with E.

[0311] Stability of oral capsule preparation

[0312] (1) Prescription

[0313] An oral capsule preparation is prepared from different crystal forms of the compound of formula I.

[0314] Table 31: Prescription of capsule preparation

[0315] (2) Preparation

[0316] 1) Active ingredient treatment: The raw materials of different crystal forms were crushed by a small universal crusher at a speed of about 20000 rpm, and then passed through an 80 mesh sieve. The prescription amount of active ingredient was weighed for standby.

[0317] 2) The prescription amount of povidone K30 was prepared into a 10% aqueous solution as a binder. The microcrystalline cellulose, active ingredient, corn starch and cross-linked povidone were put into a small wet granulator, and the povidone K30 aqueous solution was added, and granulation was carried out for 5 min. The wet granules were taken out and placed in an oven at 60°C for drying, and then passed through a 24 mesh sieve.

[0318] 3) Mixing

[0319] The dry granules and magnesium stearate were added to a small mixer in sequence and mixed at 15 rpm for 10 minutes.

[0320] 4) Capsule filling

[0321] The mixed drug granules were filled into 3# gelatin capsules by hand, and the content control weight range was 105-115 mg.

[0322] 5) Packaging

[0323] The capsules were packed into 60 ml high-density polyethylene bottles, and a desiccant was added, and the bottles were sealed using foil-type induction sealing gaskets and child-resistant closures.

[0324] (4) Stability observation of the capsules

[0325] The capsule samples prepared from different crystal forms were placed under the conditions of 60°C / open / 10 days, 25°C / 92.5% RH / open / 10 days, 40°C / 75% RH / bottled / 3 months, and 25°C / 60% RH / bottled / 3 months, respectively, and their physical and chemical stability was evaluated by various methods such as Raman spectroscopy and HPLC.

[0326] Table 32: Summary of the stability evaluation of the formulations

[0327] *: Compared with the starting sample = HPLC purity / starting sample HPLC purity * 100%

[0328] The results showed that the HPLC purity of the samples did not change significantly under the storage conditions, and no crystal form change was observed. The HPLC purity of the E crystal formulation changed significantly under certain conditions, but no crystal form change was observed.

[0329] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application, and are not limited thereto; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can still modify the technical solutions described in the foregoing examples, or make equivalent substitutions for part or all of the technical features; and these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. The crystal form A of the compound shown in Formula I, wherein, The crystal of crystal form A belongs to the monoclinic crystal system, space group C2 / c, and its unit cell parameters are: { α=90°, β=102.8680(10)°, γ=90°, }, Z = 8.

2. Crystal form A of the compound shown in Formula I, wherein, The crystal form A, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, has peaks at the following positions: 7.75±0.2°, 18.22±0.2°, 22.64±0.2°, 11.27±0.2° and 16.83±0.2°. Preferably, the crystal form A, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, further has one or more peaks selected from the following positions: 23.61±0.2°, 23.44±0.2°, 27.07±0.2°, 19.77±0.2°, 26.58±0.2°, 29.48±0.2°, 22.32±0.2°, and 21.43±0.2°; More preferably, the XRPD pattern of crystal form A is basically as shown in Figure 1-1.

3. Crystal form B of the compound shown in Formula I, wherein, The crystal form B, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, has peaks at the following positions: 6.21±0.2°, 12.42±0.2°, 13.55±0.2°, 8.17±0.2° and 16.37±0.2°. Preferably, the crystal form B, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, also has one or more peaks selected from the following positions: 19.46±0.2°, 21.33±0.2°, 21.01±0.2°, 24.64±0.2° and 20.20±0.2°; More preferably, the XRPD pattern of the crystal form B is essentially as shown in Figure 2.

4. The crystal form C of the compound shown in Formula I, wherein, The crystal form C, when irradiated with Cu-Kα, has peaks at the following positions in the X-ray powder diffraction pattern expressed in 2θ angles: 12.60±0.2°, 6.84±0.2°, 12.87±0.2°, 14.84±0.2° and 19.09±0.2°. Preferably, the crystal form C, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, further has one or more peaks selected from the following positions: 16.37±0.2°, 13.64±0.2°, 21.64±0.2°, 18.16±0.2°, 20.14±0.2°, 24.62±0.2°, 24.15±0.2°, 22.26±0.2°, and 16.90±0.2°; More preferably, the XRPD pattern of crystal form C is shown in Figure 3.

5. The crystal form D of the compound shown in Formula I, wherein, The crystal form D, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, has peaks at the following positions: 17.07±0.2°, 10.90±0.2°, 9.40±0.2°, 15.20±0.2°, and 10.11±0.2°. Preferably, the crystal form D, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, further has one or more peaks selected from the following positions: 23.32±0.2°, 21.34±0.2°, 15.05±0.2°, 18.39±0.2°, 20.72±0.2°, 24.45±0.2°, 26.60±0.2°, 26.79±0.2°, 14.66±0.2°, and 13.45±0.2°; More preferably, the XRPD pattern of crystal form D is shown in Figure 4.

6. The crystal form E of the compound shown in Formula I, wherein, The crystal form E, when irradiated with Cu-Kα, has peaks at the following positions in the X-ray powder diffraction pattern expressed in 2θ angles: 4.94±0.2°, 8.23±0.2°, 18.02±0.2°, 9.85±0.2°, and 14.04±0.2°. Preferably, the crystal form E, in the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation, further has one or more peaks selected from the following positions: 19.32±0.2°, 21.89±0.2°, 21.07±0.2°, 24.75±0.2°, 20.88±0.2°, 26.09±0.2°, 10.17±0.2°, and 6.00±0.2°; More preferably, the XRPD pattern of crystal form E is shown in Figure 5.

7. A pharmaceutical composition comprising at least one selected from crystal form A of claim 1 or 2, crystal form B of claim 3, crystal form C of claim 4, crystal form D of claim 5, and crystal form E of claim 6, and optionally a pharmaceutically acceptable carrier and / or excipient.

8. The pharmaceutical composition according to claim 7, characterized in that, The dosage form of the pharmaceutical composition is selected from creams, ointments, gels, transdermal patches, intradermal injections, eye drops, intraocular injections, ophthalmic implants, nasal sprays, inhaled powders, inhaled aerosols, inhaled sprays, inhaled liquid preparations, vaginal suppositories, vaginal tablets, vaginal gels, and preparations administered orally or rectally.

9. Use of the crystal form A of claim 1 or 2, the crystal form B of claim 3, the crystal form C of claim 4, the crystal form D of claim 5, the crystal form E of claim 6, or the pharmaceutical composition of claim 7 or 8 in the preparation of a medicament for treating TYK2-mediated diseases; Preferably, the disease is selected from inflammatory or autoimmune diseases; More preferably, the disease is selected from alopecia areata, psoriasis, psoriatic arthritis, dermatitis, lupus erythematosus, inflammatory bowel disease, hidradenitis suppurativa, rheumatoid arthritis, or uveitis.

10. Use of the compound shown in Formula I in the preparation of a drug for treating alopecia areata.

Images