Salts and / or crystalline forms of compounds as casein kinase inhibitors

Abstract

The present disclosure provides salts and / or crystalline forms of compounds, particularly compounds that are casein kinase inhibitors.

Description

Technical Field

[0001] The present application relates to salts and / or crystal forms of compounds that are tyrosine kinase inhibitors, pharmaceutical compositions comprising the salts and / or crystal forms, uses of the salts and / or crystal forms and pharmaceutical compositions, and methods for preparing the salts and / or crystal forms. Background Art

[0002] Compound HY-A is 2-(4-fluorophenyl)-3-(2-methylpyridin-4-yl)-5-(methylsulfonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine, which is a potent inhibitor of casein kinase. However, its crystalline form has not been reported.

[0003] The present application includes the unexpected discovery of novel solid forms of HY-A. The novel salts and / or crystalline forms of HY-A disclosed herein have surprising and useful properties. Summary of the invention

[0004] The present disclosure provides salts and / or crystalline forms of compounds that are casein kinase inhibitors.

[0005] The present disclosure also provides a method for preparing the salt and / or crystal form of the compound as a casein kinase inhibitor.

[0006] The present disclosure also provides a pharmaceutical composition comprising a salt and / or a crystal form of the compound as a tyrosine kinase inhibitor and a pharmaceutically acceptable carrier or excipient.

[0007] The present disclosure also provides a method of inhibiting casein kinase, comprising the step of administering an effective amount of a salt and / or a crystalline form of a compound that is a casein kinase inhibitor to a subject in need thereof.

[0008] Additional aspects and advantages of the present disclosure will become apparent to those skilled in the art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be appreciated, the present disclosure is capable of other and different embodiments, and its several details are capable of modification in various obvious respects, all without departing from the present disclosure. Therefore, the drawings and description are to be regarded as illustrative in nature, and not restrictive.

[0009] Incorporated by Reference

[0010] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The novel features of the present invention are particularly set forth in the appended claims. The features and advantages of the present invention will be better understood by reference to the following detailed description and accompanying drawings (also referred to herein as "figures") which set forth illustrative embodiments in which the principles of the invention are employed, wherein:

[0012] Figure 1-1 An interconversion diagram of HY-A free base polymorphs is shown;

[0013] Figure 2-1 The XRPD pattern of the free base Form A (823121-01-A) is shown;

[0014] Figure 2-2 The TGA / DSC graph of the free base Form A (823121-01-A) is shown;

[0015] Figure 2-3 The 1H NMR spectrum of the free base Form A (823121-01-A) is shown;

[0016] Figure 2-4 HPLC chromatogram of free base Form A (823121-01-A) is shown;

[0017] Figure 2-5 VT-XRPD overlays of free base Form A (823121-01-A) are shown;

[0018] Figure 2-6 The XRPD pattern of free base Form C (823121-16-A) is shown;

[0019] Figure 2-7 The TGA / DSC curve of free base Form C (823121-16-A) is shown;

[0020] Figure 2-8 The 1H NMR spectrum of the free base Form C (823121-16-A) is shown;

[0021] Figure 2-9 UPLC chromatogram of free base Form C (823121-16-A) is shown;

[0022] Figure 2-10 Shown is an XRPD overlay of the residual solids after equilibrium solubility testing;

[0023] Figure 3-1 Shown is an XRPD overlay of HY-A free base (I / IV);

[0024] Figure 3-2 Shown is an XRPD overlay of HY-A free base (II / IV);

[0025] Figure 3-3Shown is an XRPD overlay of HY-A free base (III / IV);

[0026] Figure 3-4 Shown is an XRPD overlay of HY-A free base (IV / IV);

[0027] Figure 3-5 The XRPD pattern of the free base P form (823121-36-A2-TO 150C) is shown;

[0028] Figure 3-6 The TGA / DSC curve of the free base P form (823121-36-A2-TO 150C) is shown;

[0029] Figure 3-7 The 1H NMR spectrum of the free base P form (823121-36-A2-TO 150C) is shown;

[0030] Figure 3-8 VT-XRPD overlays of the free base P form (823121-36-A2-TO 150C) are shown;

[0031] Figure 3-9 The XRPD pattern of the free base Form N (823121-20-A10-DRY) is shown;

[0032] Figure 3-10 The TGA / DSC curve of the free base N form (823121-20-A10-DRY) is shown;

[0033] Figure 3-11 The 1H NMR spectrum of the free base N-form (823121-20-A10-DRY) is shown;

[0034] Figure 3-12 An XRPD overlay of a re-prepared sample of the free base Form N is shown;

[0035] Figure 3-13 The XRPD pattern of the free base Form Q (823121-45-A1_AFT 90C) is shown;

[0036] Figure 3-14 The TGA / DSC curve of the free base Q form (823121-45-A1_AFT 90C) is shown;

[0037] Figure 3-15 The 1H NMR spectrum of the free base Q form (823121-45-A1_AFT 90C) is shown;

[0038] Figure 3-16 An XRPD overlay of a free base Form B sample is shown;

[0039] Figure 3-17 The TGA / DSC curve of free base Form B (823121-43-A2) is shown;

[0040] Figure 3-18 The 1H NMR spectrum of the free base Form B (823121-43-A2) is shown;

[0041] Figure 3-19 VT-XRPD overlays of free base Form B (823121-43-A2) (I / II) are shown;

[0042] Figure 3-20 VT-XRPD overlays of free base Form B (823121-43-A2) (II / II) are shown;

[0043] Figure 3-21 An XRPD overlay of a free base Form I sample is shown;

[0044] Figure 3-22 The TGA / DSC curve of free base Form I (823121-45-A1) is shown;

[0045] Figure 3-23 The 1H NMR spectrum of the free base Form I (823121-45-A1) is shown;

[0046] Figure 3-24 Shown is an XRPD overlay of free base Form I (823121-45-A1) before and after heating;

[0047] Figure 3-25 VT-XRPD overlays of free base Form I (823121-45-A1) are shown;

[0048] Figure 3-26 Shown is an XRPD overlay of a free base Form L sample;

[0049] Figure 3-27 The TGA / DSC curve of the free base L-form (823121-36-A3) is shown;

[0050] Figure 3-28 The 1H NMR spectrum of the free base L-form (823121-36-A3) is shown;

[0051] Figure 3-29 VT-XRPD overlays of the free base L-form (823121-36-A3) are shown;

[0052] Figure 3-30 The XRPD pattern of the free base D-form (823121-20-A6-DRY) is shown;

[0053] Figure 3-31The TGA / DSC curve of the free base D-form (823121-20-A6-DRY) is shown;

[0054] Figure 3-32 The 1H NMR spectrum of the free base D-form (823121-20-A6-DRY) is shown;

[0055] Figure 3-33 Shown is an XRPD overlay of the free base Form D (823121-20-A6-DRY) before and after heating;

[0056] Figure 3-34 An XRPD overlay of a free base Form F sample is shown;

[0057] Figure 3-35 The TGA / DSC curve of free base Form F (823121-17-A8-DRY) is shown;

[0058] Figure 3-36 The 1H NMR spectrum of the free base Form F (823121-17-A8-DRY) is shown;

[0059] Figure 3-37 Shown is an XRPD overlay of free base Form F (823121-17-A8-DRY) before and after heating;

[0060] Figure 3-38 The TGA curve of free base Form F (823121-39-A1-1210) is shown;

[0061] Figure 3-39 The 1H NMR spectrum of the free base Form F (823121-39-A1-1210) is shown;

[0062] Figure 3-40 The XRPD pattern of the free base Form G (823121-18-A8-DRY) is shown;

[0063] Figure 3-41 The TGA / DSC curve of the free base Form G (823121-18-A8-DRY) is shown;

[0064] Figure 3-42 The 1H NMR spectrum of the free base Form G (823121-18-A8-DRY) is shown;

[0065] Figure 3-43 Shown are XRPD overlays of the free base Form G (823121-18-A8-DRY) before and after heating;

[0066] Figure 3-44 An XRPD overlay of a free base Form H sample is shown;

[0067] Figure 3-45 The TGA / DSC curve of the free base H form (823121-37-A3) is shown;

[0068] Figure 3-46 The 1H NMR spectrum of the free base H form (823121-37-A3) is shown;

[0069] Figure 3-47 Shown is an XRPD overlay of the free base Form H (823121-20-A16-DRY) before and after heating;

[0070] Figure 3-48 The XRPD pattern of the free base Form J (823121-19-A5-DRY) is shown;

[0071] Figure 3-49 The TGA / DSC curve of the free base Form J (823121-19-A5-DRY) is shown;

[0072] Figure 3-50 The 1H NMR spectrum of the free base Form J (823121-19-A5-DRY) is shown;

[0073] Figure 3-51 Shown is an XRPD overlay of the free base Form J (823121-19-A5-DRY) before and after heating;

[0074] Figure 3-52 The XRPD pattern of the free base Form K (823121-19-A8-DRY) is shown;

[0075] Figure 3-53 The TGA / DSC curve of the free base K form (823121-19-A8-DRY) is shown;

[0076] Figure 3-54 The 1H NMR spectrum of the free base K form (823121-19-A8-DRY) is shown;

[0077] Figure 3-55 Shown is an XRPD overlay of the free base Form K (823121-19-A8-DRY) before and after heating;

[0078] Figure 3-56 The XRPD pattern of the free base Form M (823121-23-A4-DRY) is shown;

[0079] Figure 3-57 The TGA / DSC curve of the free base M form (823121-23-A4-DRY) is shown;

[0080] Figure 3-58The 1H NMR spectrum of the free base M form (823121-23-A4-DRY) is shown;

[0081] Figure 3-59 Shown are XRPD overlays of the free base Form M (823121-23-A4-DRY) before and after heating;

[0082] Figure 3-60 The XRPD pattern of the free base O form (823121-18-A9-DRY) is shown;

[0083] Figure 3-61 The TGA / DSC curve of the free base O form (823121-18-A9-DRY) is shown;

[0084] Figure 3-62 The 1H NMR spectrum of the free base O form (823121-18-A9-DRY) is shown;

[0085] Figure 3-63 Shown is an XRPD overlay of the free base O-form (823121-18-A9-DRY) before and after heating;

[0086] Figure 3-64 Shown is an XRPD overlay of the free base Form C (823121-16-A) before / after soaking in MeOH and after drying;

[0087] Figure 3-65 Shown is an XRPD overlay of the free base Form N (823121-45-A1 -check) before / after soaking in MeOH and after drying;

[0088] Figure 3-66 The XRPD pattern of the free base Form E (823121-23-A4) is shown;

[0089] Figure 3-67 The XRPD pattern of the free base R form (823121-43-A2-N2_30.0°C) is shown;

[0090] Figure 4-1 Shown is an XRPD overlay of solids from competitive slurrying for 3 days;

[0091] Figure 4-2 Shown is an XRPD overlay of solids from competitive slurrying for 5 days;

[0092] Figure 4-3 Shown is an XRPD overlay of solids from competitive slurries in Forms C, B, I and L (I / II);

[0093] Figure 4-4Shown is an XRPD overlay of solids from competitive slurries in Forms C, B, I and L (II / II);

[0094] Figure 5-1 The XRPD pattern of HCl Salt Form A (823121-28-A1) is shown;

[0095] Figure 5-2 The TGA / DSC curve of HCl salt Form A (823121-28-A1) is shown;

[0096] Figure 5-3 The 1H NMR spectrum of the HCl salt Form A (823121-28-A1) is shown;

[0097] Figure 5-4 The TGA / DSC curve of citrate salt Form A (823121-28-A3) is shown;

[0098] Figure 5-5 The TGA / DSC curve of citrate salt Form A (823121-28-A3) is shown;

[0099] Figure 5-6 The 1H NMR spectrum of the citrate salt Form A (823121-28-A3) is shown;

[0100] Figure 5-7 The XRPD pattern of Phosphate Form A (823121-32-A) is shown;

[0101] Figure 5-8 The TGA / DSC curve of phosphate type A (823121-32-A) is shown;

[0102] Figure 5-9 The 1H NMR spectrum of phosphate form A (823121-32-A) is shown;

[0103] Figure 6-1 A graph showing kinetic solubility;

[0104] Figure 6-2 Shown is an XRPD overlay of the residual solid from solubility testing of HCl salt Form A in FaSSIF;

[0105] Figure 6-3 Shown is an XRPD overlay of the residual solid from solubility testing of HCl salt Form A in FeSSIF;

[0106] Figure 6-4 Shown is an XRPD overlay of residual solids from solubility testing of citrate Form A in FaSSIF;

[0107] Figure 6-5Shown is an XRPD overlay of residual solids from solubility testing of citrate Form A in FeSSIF;

[0108] Figure 6-6 Shown is an XRPD overlay of residual solids from solubility testing of phosphate Form A in FeSSIF;

[0109] Figure 6-7 Shown is an XRPD overlay of the residual solid from solubility testing of free base Form A in H2O;

[0110] Figure 6-8 Shown is an XRPD overlay of the residual solid from solubility testing of free base Form A in FaSSIF;

[0111] Figure 6-9 Shown is an XRPD overlay of residual solids from solubility testing of free base Form A in FeSSIF;

[0112] Figure 6-10 The DVS pattern of the free base Form C (823121-16-A) is shown;

[0113] Figure 6-11 Shown is an XRPD overlay of free base Form C (823121-16-A) before and after DVS;

[0114] Figure 6-12 The DVS diagram of HCl salt Form A (823121-28-A1) is shown;

[0115] Figure 6-13 Shown is an XRPD overlay of HCl salt Form A (823121-28-A1) before and after DVS;

[0116] Figure 6-14 The DVS diagram of citrate salt Form A (823121-28-A3) is shown;

[0117] Figure 6-15 Shown is an XRPD overlay of citrate salt Form A (823121-28-A3) before and after DVS;

[0118] Figure 6-16 The DVS diagram of phosphate form A (823121-32-A) is shown;

[0119] Figure 6-17 Shown is an XRPD overlay of phosphate Form A (823121-32-A) before and after DVS;

[0120] Figure 6-18 Shown is an XRPD overlay of the free base Form A (823121-01-A) before and after stability evaluation;

[0121] Figure 6-19 Shown is an XRPD overlay of the free base Form C (823121-16-A) before and after stability evaluation;

[0122] Figure 6-20 Shown is an XRPD overlay of HCl salt Form A (823121-28-A1) before and after stability evaluation;

[0123] Figure 6-21 Shown is an XRPD overlay of Citrate Form A (823121-28-A3) before and after stability evaluation;

[0124] Figure 6-22 Shown is an XRPD overlay of Phosphate Form A (823121-32-A) before and after stability evaluation;

[0125] Figure 6-23 UPLC chromatogram of free base Form A (823121-01-A) is shown after stability evaluation at 25°C / 60%RH for one week;

[0126] Figure 6-24 UPLC chromatogram of free base Form A (823121-01-A) is shown after stability evaluation at 40°C / 75%RH for one week;

[0127] Figure 6-25 UPLC chromatogram of free base Form C (823121-16-A) is shown after stability assessment at 25°C / 60%RH for one week;

[0128] Figure 6-26 UPLC chromatogram of free base Form C (823121-16-A) is shown after stability evaluation at 40°C / 75%RH for one week;

[0129] Figure 6-27 UPLC chromatogram of HCl salt Form A (823121-28-A1) is shown after stability evaluation at 25°C / 60%RH for one week;

[0130] Figure 6-28 UPLC chromatogram of HCl salt Form A (823121-28-A1) is shown after stability evaluation at 40°C / 75%RH for one week;

[0131] Figure 6-29 UPLC chromatogram of citrate salt Form A (823121-28-A3) is shown after stability evaluation at 25°C / 60%RH for one week;

[0132] Figure 6-30UPLC chromatogram showing citrate Form A (823121-28-A3) after stability assessment at 40°C / 75%RH for one week;

[0133] Figure 6-31 UPLC chromatogram showing the phosphate form A (823121-32-A) after stability evaluation at 25°C / 60%RH for one week;

[0134] Figure 6-32 UPLC chromatogram showing the phosphate salt Form A (823121-32-A) after stability evaluation at 40°C / 75%RH for one week;

[0135] Figure 7-1 The XRPD pattern of the free base Form T (823121-63-A) is shown;

[0136] Figure 7-2 XRPD overlays of Form T (823121-63-A) before and after heating are shown;

[0137] Figure 7-3 The 1H NMR spectrum of the free base T form (823121-63-A) after heating is shown;

[0138] Figure 7-4 Shown is an XRPD overlay of HCl Salt Form A / B (823121-06-D1 / D2);

[0139] Figure 7-5 The TGA / DSC curve of HCl salt Form A (823121-06-D1) is shown;

[0140] Figure 7-6 The 1H NMR spectrum of the HCl salt Form A (823121-06-D1) is shown;

[0141] Figure 7-7 VT-XRPD overlays showing HCl Salt Form A (823121-06-D1);

[0142] Figure 7-8 The TGA / DSC curve of HCl salt Form B (823121-06-D2) is shown;

[0143] Figure 7-9 The 1H NMR spectrum of the HCl salt Form B (823121-06-D2) is shown;

[0144] Figure 7-10 The XRPD pattern of sulfate salt Form A (823121-06-C3) is shown;

[0145] Figure 7-11The TGA / DSC curve of sulfate form A (823121-06-C3) is shown;

[0146] Figure 7-12 The 1H NMR spectrum of sulfate salt Form A (823121-06-C3) is shown;

[0147] Figure 7-13 Shown is an XRPD overlay of maleate salt Form A / B / C (823121-06-A4 / D4 / C4);

[0148] Figure 7-14 The TGA / DSC curve of maleate Form A (823121-06-A4) is shown;

[0149] Figure 7-15 The 1H NMR spectrum of the maleate salt Form A (823121-06-A4) is shown;

[0150] Figure 7-16 The TGA / DSC curve of maleate Form B (823121-06-D4) is shown;

[0151] Figure 7-17 The 1H NMR spectrum of maleate Form B (823121-06-D4) is shown;

[0152] Figure 7-18 The TGA / DSC curve of maleate Form C (823121-06-C4) is shown;

[0153] Figure 7-19 The 1H NMR spectrum of the maleate salt Form C (823121-06-C4) is shown;

[0154] Figure 7-20 The XRPD pattern of Phosphate Form A (823121-06-A6) is shown;

[0155] Figure 7-21 The TGA / DSC curve of phosphate type A (823121-06-A6) is shown;

[0156] Figure 7-22 The 1H NMR spectrum of phosphate form A (823121-06-A6) is shown;

[0157] Figure 7-23 Shown is an XRPD overlay of L-tartrate salt Form A / B (823121-06-A7 / C7);

[0158] Figure 7-24 The TGA / DSC curve of L-tartrate Form A (823121-06-A7) is shown;

[0159] Figure 7-25 The 1H NMR spectrum of L-tartrate salt Form A (823121-06-A7) is shown;

[0160] Figure 7-26 The TGA / DSC curve of L-tartrate Form B (823121-06-C7) is shown;

[0161] Figure 7-27 The 1H NMR spectrum of L-tartrate Form B (823121-06-C7) is shown;

[0162] Figure 7-28 The XRPD pattern of the fumarate salt Form A (823121-06-A8) is shown;

[0163] Figure 7-29 The TGA / DSC curve of fumarate form A (823121-06-A8) is shown;

[0164] Figure 7-30 The 1H NMR spectrum of the fumarate salt Form A (823121-06-A8) is shown;

[0165] Figure 7-31 The TGA / DSC curve of citrate salt Form A (823121-06-C9) is shown;

[0166] Figure 7-32 The TGA / DSC curve of citrate salt Form A (823121-06-C9) is shown;

[0167] Figure 7-33 The 1H NMR spectrum of the citrate salt Form A (823121-06-C9) is shown;

[0168] Figure 7-34 The XRPD pattern of the tosylate salt Form A (823121-06-A10) is shown;

[0169] Figure 7-35 The TGA / DSC curve of the tosylate salt Form A (823121-06-A10) is shown;

[0170] Figure 7-36 The 1H NMR spectrum of the tosylate salt Form A (823121-06-A10) is shown;

[0171] Figure 7-37 Shown is an XRPD overlay of Tosylate Form A / B (823121-06-C11 / D11);

[0172] Figure 7-38 The TGA / DSC curve of the tosylate salt Form B (823121-06-D11) is shown;

[0173] Figure 7-39 The 1H NMR spectrum of the tosylate salt Form B (823121-06-D11) is shown;

[0174] Figure 7-40 Shown is an XRPD overlay of besylate salt Form A / B / C (823121-06-A12 / B12 / C12);

[0175] Figure 7-41 The TGA / DSC curve of benzenesulfonate salt Form A (823121-06-A12) is shown;

[0176] Figure 7-42 The 1H NMR spectrum of benzenesulfonate salt Form A (823121-06-A12) is shown;

[0177] Figure 7-43 The TGA / DSC curve of benzenesulfonate Form B (823121-06-B12) is shown;

[0178] Figure 7-44 The 1H NMR spectrum of benzenesulfonate salt Form B (823121-06-B12) is shown;

[0179] Figure 7-45 The TGA / DSC curve of benzenesulfonate Form C (823121-06-C12) is shown;

[0180] Figure 7-46 The 1H NMR spectrum of benzenesulfonate salt Form C (823121-06-C12) is shown;

[0181] Figure 7-47 The XRPD pattern of Gentisate Form A (823121-06-C13) is shown;

[0182] Figure 7-48 The TGA / DSC curve of gentisate Form A (823121-06-C13) is shown;

[0183] Figure 7-49 The 1H NMR spectrum of gentisate Form A (823121-06-C13) is shown;

[0184] Figure 7-50 Shown is a UPLC overlay of supernatants from equilibrium solubility experiments in water and pH buffer;

[0185] Figure 7-51 LC-MS results of supernatants from equilibrium solubility experiments in pH 6.0 buffer are shown;

[0186] Figure 7-52LC-MS results of supernatants from equilibrium solubility experiments in pH 8.0 buffer are shown;

[0187] Figure 7-53 LC-MS results of supernatants from equilibrium solubility experiments in water are shown;

[0188] Figure 7-54 Shown is a UPLC overlay of the supernatant from a kinetic solubility experiment in water for 1 hour;

[0189] Figure 7-55 Shown is a UPLC overlay of supernatants from a kinetic solubility experiment in FaSSIF lasting 1 hour;

[0190] Figure 7-56 XRPD overlays showing residual solids after slurrying in water for 24 hours or 6 days;

[0191] Figure 7-57 Shown is a UPLC overlay of the supernatant after slurrying in water for 24 hours or 6 days;

[0192] Figure 7-58 UPLC chromatogram showing the residual solids after slurrying the free base Form C in water for 24 hours;

[0193] Figure 7-59 Shown is a UPLC chromatogram of the residual solids after slurrying the free base Form C in water for 6 days. DETAILED DESCRIPTION

[0194] Although various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Those skilled in the art may conceive of a variety of variations, changes, and substitutions without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0195] As used herein, unless otherwise indicated, the term "treat" generally refers to reversing, slowing the progression of, or preventing the disease or condition to which such term applies, or one or more symptoms of such disease or condition. Unless otherwise indicated, the term "treat" as used herein generally refers to the act of treating, as "treat" is defined above. The term "treat" may also include adjuvant therapy and neoadjuvant therapy of a subject.

[0196] As used herein, unless otherwise indicated, the term "prevent" generally means to exclude, avoid, eliminate, forestall, stop or hinder something from happening, especially by preemptive action. It is understood that where reduce, inhibit or prevent is used herein, the use of the other two words may also be explicitly disclosed unless otherwise expressly indicated.

[0197] The term "pharmaceutically acceptable salt" as used herein generally refers to a salt that is pharmaceutically acceptable and that may have the desired pharmacological activity of the parent compound. Such salts may include: acid addition salts formed with inorganic acids or with organic acids, or base addition salts formed with the conjugate base of any inorganic acid, wherein the conjugate base contains a cationic component.

[0198] As used herein, the term "pharmaceutically acceptable carrier" generally refers to aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders that are reconstituted into sterile injectable solutions or dispersions before use. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or excipients may include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Appropriate fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the desired particle size in the case of dispersions, and by using surfactants. These compositions may also include adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, etc. It may also be necessary to include isotonic agents such as sugars, sodium chloride, etc. The extended absorption of injectable drug forms may be caused by including agents that delay absorption, such as aluminum monostearate and gelatin. Injectable depot forms can be made by forming a microcapsule matrix of the drug in a biodegradable polymer such as polylactic acid-polyglycolic acid, poly(orthoesters) and poly(anhydrides). According to the ratio of the drug to the polymer and the properties of the specific polymer used, the rate of drug release can be controlled. Depot injectable formulations can also be prepared by encapsulating the drug in a liposome or microemulsion compatible with body tissues. Injectable formulations can be, for example, sterilized by filtering through a bacteria-retaining filter or by adding a sterilizing agent in the form of a sterile solid composition, which can be dissolved or dispersed in sterile water or other sterile injectable media before use. Suitable inert carriers can include sugars such as lactose. Ideally, at least 95% of the active ingredient particles by weight can have an effective particle size in the range of 0.01 micron to 10 microns.

[0199] As used herein, the term "prodrug" generally refers to a compound that is metabolized, e.g., hydrolyzed or oxidized, in a host to form a compound of the invention. Typical examples of prodrugs include compounds having biologically unstable protecting groups on the functional moieties of the active compound. Prodrugs may include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrated, alkylated, dealkylated, acylated, deacylated, phosphorylated, dephosphorylated to produce an active compound.

[0200] As used herein, the term "casein kinase" generally refers to a protein having the activity of catalyzing serine / threonine selective phosphorylation of a protein. This activity may be referred to as "casein kinase activity". The gene ID of a gene encoding a casein kinase may be 1453 or 1454.

[0201] As used herein, the term "subject" generally refers to an animal, which may include, but is not limited to, cattle, pigs, sheep, chickens, turkeys, buffaloes, camels, ostriches, dogs, cats, and humans, and the subject may be a human. It is contemplated that in the method for treating a subject of the sixth embodiment, any compound may be used alone or in combination with another compound of the present invention.

[0202] As used herein, the term "effective amount" generally refers to the amount of an agent or compound used to treat a disease or condition or some or all of the symptoms. The result can be a reduction and / or alleviation of the signs, symptoms or causes of a disease or condition, or any other desired change in a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein that is required to provide a clinically significant reduction in the symptoms of a disease or condition without undue adverse side effects.

[0203] As used herein, the term "administering" generally means that the compound can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously or topically in liquid or solid form.

[0204] As used herein, the term "molecular formula" may be referred to hereinafter as "compound of the present invention". These terms are also defined to include all forms of the compound of the molecular formula, including hydrates, solvates, isomers, crystalline and amorphous forms, isomorphs, polymorphs, and metabolites thereof. For example, the compound of the molecular formula or a pharmaceutically acceptable salt thereof may exist in unsolvated and solvated forms. When the solvent or water is tightly bound, the complex may have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, such as in channel solvates and hygroscopic compounds, the water / solvent content may depend on humidity and drying conditions. In such cases, non-stoichiometry will become the norm.

[0205] Compounds of the "formula" may have asymmetric carbon atoms. The carbon-carbon bonds of the formula compounds may be depicted herein using solid lines, solid wedges, or dashed wedges. The use of solid lines to depict bonds to asymmetric carbon atoms may mean that all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) on the carbon atom are included. The use of solid lines or dashed wedges to depict bonds to asymmetric carbon atoms may mean that only the stereoisomers shown are included. Possibly, the compounds of the present application may contain more than one asymmetric carbon atom. In these compounds, the use of solid lines to depict bonds to asymmetric carbon atoms may mean that all possible stereoisomers are included. For example, unless otherwise indicated, it may mean that the formula compounds may exist as enantiomers and diastereomers or as racemates and mixtures thereof. The use of solid lines to depict bonds to one or more asymmetric carbon atoms in the formula compounds and the use of solid lines or dashed wedges to depict bonds to other asymmetric carbon atoms in the same compound may mean that there is a mixture of diastereomers.

[0206] The compounds of the present application (e.g., compounds of formula) can exist as inclusion compounds or other complexes. Included within the scope of the present invention are complexes, such as clathrates, drug-host inclusion complexes, wherein, in contrast to the above-mentioned solvates, the drug and the host can be present in stoichiometric or non-stoichiometric amounts. Complexes of chemical formulas comprising two or more organic and / or inorganic components can also be included, and these components can be in stoichiometric or non-stoichiometric amounts. The resulting complex can be ionized, partially ionized or non-ionized. For a review of such complexes, see Haleblian's J. Pharm. Sci., 64 (8), 1269-1288 (August 1975).

[0207] Molecular formula stereoisomers may include cis and trans isomers, optical isomers such as R and S enantiomers, diastereomers, geometric isomers, rotational isomers, conformational isomers and tautomers of the molecular formula compound, including compounds exhibiting more than one type of isomerism; and mixtures thereof (such as racemates and diastereoisomer pairs). Also included are acid addition salts or base addition salts in which the counterion is optically active, for example, D-lactate or L-lysine, or racemic, for example, DL-tartrate or DL-arginine.

[0208] When any racemate is crystallized, two different types of crystals may be formed. The first type is the racemic compound mentioned above (true racemate), in which a homogeneous form of crystals containing two enantiomers in equimolar amounts is produced. The second type is a racemic mixture or aggregate, in which two crystalline forms are produced in equimolar amounts, each crystalline form comprising a single enantiomer.

[0209] Compounds of the formula may exhibit tautomerism and structural isomerism. For example, compounds of the formula may exist in several tautomeric forms, including enol and imine forms, as well as ketone and enamine forms, and their geometric isomers and mixtures. All such tautomeric forms may be included within the scope of compounds of the formula. Tautomers may exist in solution as mixtures of tautomeric sets. In solid form, one tautomer is usually dominant. Although one tautomer may be described, the present invention includes all tautomers of compounds of the formula.

[0210] The present invention also includes isotopically labeled compounds, which are identical to those recited in the above formula, but for the fact that one or more atoms may be replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that may be incorporated into compounds of the formula include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as, but not limited to, 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl. Certain isotopically labeled compounds, for example radioactive isotopes such as 3 H and 14 Those having C incorporated therein may be used in drug and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e., 14 C) isotopes can be particularly useful due to their ease of preparation and detectability. In addition, heavier isotopes such as deuterium (i.e., 2 H) substitution may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore, may be used in some circumstances. Isotopically labeled compounds of the formula can generally be prepared by carrying out the procedures disclosed in the schemes and / or in the examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0211] The compound of the present application can be used in the form of a salt derived from an inorganic acid or an organic acid. According to the specific compound, the salt of the compound can have advantages due to the physical properties of one or more salts, such as the drug stability enhanced under different temperatures and humidity or the solubility desired in water or oil. In some cases, the salt of the compound can also be used as an adjuvant for separation, purification and / or separation of the compound.

[0212] In one aspect, the present application provides a crystalline form of compound HY-A, 2-(4-fluorophenyl)-3-(2-methylpyridin-4-yl)-5-(methylsulfonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-θ degrees, wherein the characteristic peaks are selected from the group consisting of:

[0213] (1) 11.3±0.3 degrees, 22.7±0.3 degrees, 23.2±0.3 degrees;

[0214] (2) 14.4±0.3 degrees, 19.2±0.3 degrees, 21.6±0.3 degrees;

[0215] (3) 12.8±0.3 degrees, 16.7±0.3 degrees, 19.2±0.3 degrees;

[0216] (4) 7.5±0.3 degrees, 7.8±0.3 degrees, 19.7±0.3 degrees;

[0217] (5) 10.1±0.3 degrees, 12.5±0.3 degrees, 19.7±0.3 degrees;

[0218] (6) 11.5±0.3 degrees, 17.5±0.3 degrees, 24.5±0.3 degrees;

[0219] (7) 13.9±0.3 degrees, 20.6±0.3 degrees, 23.8±0.3 degrees;

[0220] (8) 7.0±0.3 degrees, 12.6±0.3 degrees, 19.5±0.3 degrees;

[0221] (9) 7.1±0.3 degrees, 10.9±0.3 degrees, 12.3±0.3 degrees;

[0222] (10) 6.4±0.3 degrees, 15.6±0.3 degrees, 20.0±0.3 degrees;

[0223] (11) 8.6±0.3 degrees, 13.2±0.3 degrees, 20.7±0.3 degrees;

[0224] (12) 5.6±0.3 degrees, 17.1±0.3 degrees, 21.4±0.3 degrees;

[0225] (13) 5.6±0.3 degrees, 17.1±0.3 degrees, 22.5±0.3 degrees;

[0226] (14) 18.9±0.3 degrees, 19.7±0.3 degrees, 25.0±0.3 degrees;

[0227] (15) 5.3±0.3 degrees, 8.7±0.3 degrees, 17.2±0.3 degrees;

[0228] (16) 7.9±0.3 degrees, 18.2±0.3 degrees, 18.7±0.3 degrees;

[0229] (17) 5.5±0.3 degrees, 12.3±0.3 degrees, 17.2±0.3 degrees;

[0230] (18) 5.2 ± 0.3 degrees, 10.4 ± 0.3 degrees, 26.1 ± 0.3 degrees; and

[0231] (19) 6.5±0.3 degrees, 18.7±0.3 degrees, 22.2±0.3 degrees;

[0232] For example, wherein the crystalline form further exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of:

[0233] (1) 5.6±0.3 degrees, 19.0±0.3 degrees, 19.4±0.3 degrees;

[0234] (2) 13.6±0.3 degrees, 17.9±0.3 degrees, 27.4±0.3 degrees;

[0235] (3) 14.7±0.3 degrees, 15.1±0.3 degrees, 26.3±0.3 degrees;

[0236] (4) 6.6±0.3 degrees, 12.5±0.3 degrees, 23.4±0.3 degrees;

[0237] (5) 8.8±0.3 degrees, 17.5±0.3 degrees, 23.5±0.3 degrees;

[0238] (6) 6.3±0.3 degrees, 17.8±0.3 degrees, 21.0±0.3 degrees;

[0239] (7) 8.9±0.3 degrees, 17.3±0.3 degrees, 27.0±0.3 degrees;

[0240] (8) 13.9±0.3 degrees, 18.4±0.3 degrees, 21.0±0.3 degrees;

[0241] (9) 18.9±0.3 degrees, 20.6±0.3 degrees, 27.0±0.3 degrees;

[0242] (10) 10.6±0.3 degrees, 13.1±0.3 degrees, 16.8±0.3 degrees;

[0243] (11) 5.3±0.3 degrees, 12.7±0.3 degrees, 17.5±0.3 degrees;

[0244] (12) 8.7±0.3 degrees, 12.6±0.3 degrees, 24.7±0.3 degrees;

[0245] (13)12.6±0.3 degrees;

[0246] (14) 11.5±0.3 degrees, 17.5±0.3 degrees, 21.7±0.3 degrees;

[0247] (15) 9.2±0.3 degrees, 12.3±0.3 degrees, 21.4±0.3 degrees;

[0248] (16) 12.7±0.3 degrees, 20.7±0.3 degrees, 23.4±0.3 degrees;

[0249] (17) 9.1±0.3 degrees, 18.5±0.3 degrees, 25.1±0.3 degrees;

[0250] (18) 15.6 ± 0.3 degrees, 18.8 ± 0.3 degrees, 20.8 ± 0.3 degrees; and

[0251] (19) 17.7±0.3 degrees, 21.9±0.3 degrees, 24.0±0.3 degrees;

[0252] For example, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of:

[0253] (1) 13.9±0.3 degrees, 18.6±0.3 degrees, 25.2±0.3 degrees;

[0254] (2) 9.9±0.3 degrees, 10.4±0.3 degrees, 23.2±0.3 degrees;

[0255] (3) 19.7±0.3 degrees, 20.6±0.3 degrees, 21.0±0.3 degrees;

[0256] (4) 11.7±0.3 degrees, 17.5±0.3 degrees, 21.9±0.3 degrees;

[0257] (5) 6.3±0.3 degrees, 21.8±0.3 degrees, 22.1±0.3 degrees;

[0258] (6) 19.4±0.3 degrees, 19.8±0.3 degrees, 25.2±0.3 degrees;

[0259] (7) 15.2±0.3 degrees, 17.9±0.3 degrees, 24.6±0.3 degrees;

[0260] (8) 9.2±0.3 degrees, 16.0±0.3 degrees, 21.7±0.3 degrees;

[0261] (9) 14.2±0.3 degrees, 17.9±0.3 degrees, 25.3±0.3 degrees;

[0262] (10) 9.5±0.3 degrees, 16.2±0.3 degrees, 19.6±0.3 degrees;

[0263] (11) 16.6±0.3 degrees, 20.3±0.3 degrees, 21.4±0.3 degrees;

[0264] (12) 19.3±0.3 degrees, 26.5±0.3 degrees;

[0265] (14) 12.0±0.3 degrees, 16.3±0.3 degrees, 22.3±0.3 degrees;

[0266] (15) 17.5±0.3 degrees, 19.3±0.3 degrees, 26.8±0.3 degrees;

[0267] (16) 8.5±0.3 degrees, 21.9±0.3 degrees, 23.8±0.3 degrees;

[0268] (18) 28.1 ± 0.3 degrees; and

[0269] (19) 15.6±0.3 degrees, 19.2±0.3 degrees, 26.3±0.3 degrees;

[0270] For example, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of:

[0271] (1) 11.3±0.3 degrees, 22.7±0.3 degrees, 23.2±0.3 degrees, 5.6±0.3 degrees, 19.0±0.3 degrees, 19.4±0.3 degrees, 13.9±0.3 degrees, 18.6±0.3 degrees, 25.2±0.3 degrees;

[0272] (2) 14.4±0.3 degrees, 19.2±0.3 degrees, 21.6±0.3 degrees, 13.6±0.3 degrees, 17.9±0.3 degrees, 27.4±0.3 degrees, 9.9±0.3 degrees, 10.4±0.3 degrees, 23.2±0.3 degrees;

[0273] (3) 12.8±0.3 degrees, 16.7±0.3 degrees, 19.2±0.3 degrees, 14.7±0.3 degrees, 15.1±0.3 degrees, 26.3±0.3 degrees, 19.7±0.3 degrees, 20.6±0.3 degrees, 21.0±0.3 degrees;

[0274] (4) 7.5±0.3 degrees, 7.8±0.3 degrees, 19.7±0.3 degrees, 6.6±0.3 degrees, 12.5±0.3 degrees, 23.4±0.3 degrees, 11.7±0.3 degrees, 17.5±0.3 degrees, 21.9±0.3 degrees;

[0275] (5) 10.1±0.3 degrees, 12.5±0.3 degrees, 19.7±0.3 degrees, 8.8±0.3 degrees, 17.5±0.3 degrees, 23.5±0.3 degrees, 6.3±0.3 degrees, 21.8±0.3 degrees, 22.1±0.3 degrees;

[0276] (6) 11.5±0.3 degrees, 17.5±0.3 degrees, 24.5±0.3 degrees, 6.3±0.3 degrees, 17.8±0.3 degrees, 21.0±0.3 degrees, 19.4±0.3 degrees, 19.8±0.3 degrees, 25.2±0.3 degrees;

[0277] (7) 13.9±0.3 degrees, 20.6±0.3 degrees, 23.8±0.3 degrees, 8.9±0.3 degrees, 17.3±0.3 degrees, 27.0±0.3 degrees, 15.2±0.3 degrees, 17.9±0.3 degrees, 24.6±0.3 degrees;

[0278] (8) 7.0±0.3 degrees, 12.6±0.3 degrees, 19.5±0.3 degrees, 13.9±0.3 degrees, 18.4±0.3 degrees, 21.0±0.3 degrees, 9.2±0.3 degrees, 16.0±0.3 degrees, 21.7±0.3 degrees;

[0279] (9) 7.1±0.3 degrees, 10.9±0.3 degrees, 12.3±0.3 degrees, 18.9±0.3 degrees, 20.6±0.3 degrees, 27.0±0.3 degrees, 14.2±0.3 degrees, 17.9±0.3 degrees, 25.3±0.3 degrees;

[0280] (10) 6.4±0.3 degrees, 15.6±0.3 degrees, 20.0±0.3 degrees, 10.6±0.3 degrees, 13.1±0.3 degrees, 16.8±0.3 degrees, 9.5±0.3 degrees, 16.2±0.3 degrees, 19.6±0.3 degrees;

[0281] (11) 8.6±0.3 degrees, 13.2±0.3 degrees, 20.7±0.3 degrees, 5.3±0.3 degrees, 12.7±0.3 degrees, 17.5±0.3 degrees, 16.6±0.3 degrees, 20.3±0.3 degrees, 21.4±0.3 degrees;

[0282] (12) 5.6±0.3 degrees, 17.1±0.3 degrees, 21.4±0.3 degrees, 8.7±0.3 degrees, 12.6±0.3 degrees, 24.7±0.3 degrees, 19.3±0.3 degrees, 26.5±0.3 degrees;

[0283] (13) 5.6±0.3 degrees, 17.1±0.3 degrees, 22.5±0.3 degrees, 12.6±0.3 degrees;

[0284] (14) 18.9±0.3 degrees, 19.7±0.3 degrees, 25.0±0.3 degrees, 11.5±0.3 degrees, 17.5±0.3 degrees, 21.7±0.3 degrees, 12.0±0.3 degrees, 16.3±0.3 degrees, 22.3±0.3 degrees;

[0285] (15) 5.3±0.3 degrees, 8.7±0.3 degrees, 17.2±0.3 degrees, 9.2±0.3 degrees, 12.3±0.3 degrees, 21.4±0.3 degrees, 17.5±0.3 degrees, 19.3±0.3 degrees, 26.8±0.3 degrees;

[0286] (16) 7.9±0.3 degrees, 18.2±0.3 degrees, 18.7±0.3 degrees, 12.7±0.3 degrees, 20.7±0.3 degrees, 23.4±0.3 degrees, 8.5±0.3 degrees, 21.9±0.3 degrees, 23.8±0.3 degrees;

[0287] (17) 5.5±0.3 degrees, 12.3±0.3 degrees, 17.2±0.3 degrees, 9.1±0.3 degrees, 18.5±0.3 degrees, 25.1±0.3 degrees;

[0288] (18) 5.2 ± 0.3 degrees, 10.4 ± 0.3 degrees, 26.1 ± 0.3 degrees, 15.6 ± 0.3 degrees, 18.8 ± 0.3 degrees, 20.8 ± 0.3 degrees, 28.1 ± 0.3 degrees; and

[0289] (19) 6.5 ± 0.3 degrees, 18.7 ± 0.3 degrees, 22.2 ± 0.3 degrees, 17.7 ± 0.3 degrees, 21.9 ± 0.3 degrees, 24.0 ± 0.3 degrees, 15.6 ± 0.3 degrees, 19.2 ± 0.3 degrees, 26.3 ± 0.3 degrees. As used herein, when used to refer to a 2-theta value, it is meant that value ± 0.3 degrees 2-theta. As used herein, when used to refer to a 2-theta value, it is meant that value ± 0.2 degrees 2-theta. As used herein, when used to refer to a 2-theta value, it is meant that value ± 0.1 degrees 2-theta.

[0290] For example, wherein the crystalline form exhibits a mass loss in TGA (thermogravimetric) analysis selected from the group consisting of:

[0291] (1) 3.2% mass loss up to 120°C;

[0292] (2) 2% mass loss up to 120°C;

[0293] (3) 0.9% mass loss up to 120°C;

[0294] (4) 2.2% mass loss up to 150°C;

[0295] (5) 0.6% mass loss up to 120°C;

[0296] (6) 7% mass loss up to 120°C;

[0297] (7) 3.5% mass loss up to 120°C;

[0298] (8) 1.1% mass loss up to 120°C;

[0299] (9) up to 13.6% mass loss at 120°C and / or up to 4.3% mass loss at 120°C for crystalline forms with water of crystallization;

[0300] (10) 7.6% mass loss up to 120 °C;

[0301] (11) 13.9% mass loss up to 120 °C;

[0302] (12) 8.9% mass loss up to 120 °C;

[0303] (13) 24.5% mass loss up to 150°C and / or 9.8% mass loss from 90°C to 120°C;

[0304] (14) 10.7% mass loss up to 120 °C;

[0305] (15) 11.2% mass loss up to 120 °C;

[0306] (16) 13.9% mass loss up to 120 °C;

[0307] (17) 7.1% mass loss up to 150 °C;

[0308] (18) 2.5% mass loss up to 61°C and / or 8.8% mass loss from 61°C to 90°C; and

[0309] (19) Up to 11.1% mass loss at 120 °C.

[0310] For example, wherein the crystalline form exhibits an endothermic peak and / or an exothermic peak selected from the group consisting of:

[0311] (1) 77.7°C (endothermic), 231.4°C (endothermic);

[0312] (2) 164.2°C (endothermic);

[0313] (3) 214.7°C (endothermic);

[0314] (4) 175.6°C (endothermic), 121.5°C (exothermic);

[0315] (5) 176.2°C (endothermic);

[0316] (6) 176.1°C (endothermic);

[0317] (7) 170.4°C (endothermic), 176.1°C (endothermic);

[0318] (8) 115.9°C (endothermic), 170.3°C (endothermic), 176.1°C (endothermic), 120.0°C (exothermic);

[0319] (9) 91.5°C (endothermic), 175.8°C (endothermic);

[0320] (10) 89.0°C (endothermic), 116.2°C (endothermic), 170.5°C (endothermic), 176.9°C (endothermic), 122.5°C (exothermic);

[0321] (11) 66.2°C (endothermic), 70.8°C (endothermic), 79.1°C (endothermic), 176.0°C (endothermic);

[0322] (12) 99.9°C (endothermic), 175.7°C (endothermic);

[0323] (13) 126.3°C (endothermic), 175.3°C (endothermic);

[0324] (14) 72.5°C (endothermic), 176.0°C (endothermic);

[0325] (15) 116.4°C (endothermic), 175.9°C (endothermic);

[0326] (16) 102.1°C (endothermic), 176.3°C (endothermic);

[0327] (17) 115.9°C (endothermic), 176.1°C (endothermic);

[0328] (18) 83.6°C (endothermic), 176.0°C (endothermic); and

[0329] (19) 118.4°C (endothermic), 175.2°C (endothermic).

[0330] For example, wherein the crystalline form exhibits a salt stoichiometry (molar ratio of acid to free base) selected from the group consisting of:

[0331] (1) 1:1 (molar ratio of hydrochloric acid to free base);

[0332] (2) 1:1 (molar ratio of citric acid to free base); and

[0333] (3) 2.1:1 (molar ratio of phosphoric acid to free base).

[0334] For example, the crystalline form comprises no residual solvent or comprises a residual solvent selected from the group consisting of: 0.2 wt % EtOAc; 6.3 wt % 2-MeTHF; 9.3 wt % 1,4-dioxane; 7.2 wt % CHCl3; 9.0 wt % toluene; 0.7 wt % 2-MeTHF; 11.9 wt % anisole; 6.6 wt % toluene; 5.3 wt % n-hexane; and 12.3 wt % DMSO.

[0335] For example, wherein the crystalline form shows a purity rate selected from the group consisting of: (1) 99.8 area %; (2) 99.7 area %; (3) 100.0 area %; (4) 99.80 area %; and (5) 99.7 area % in LC-MS (liquid chromatography-mass spectrometry) analysis.

[0336] For example, wherein the crystalline form has a solubility selected from the group consisting of:

[0337] (1) 4.8 mg / mL for H2O, 4.9 mg / mL for SGF, 1.4 mg / mL for FaSSIF, and 1.3 mg / mL for FeSSIF;

[0338] (2) 3.9 mg / mL for H2O, 3.7 mg / mL for SGF, 1.9 mg / mL for FaSSIF, and 1.4 mg / mL for FeSSIF;

[0339] (3) 5.0 mg / mL for H2O, 4.3 mg / mL for SGF, 4.2 mg / mL for FaSSIF, and 1.6 mg / mL for FeSSIF; and

[0340] (4) 0.036 mg / mL for H2O, 5.1 mg / mL for SGF, 0.064 mg / mL for FaSSIF, and 0.96 mg / mL for FeSSIF.

[0341] For example, wherein the crystalline form exhibits a hygroscopicity selected from the following in DVS (dynamic vapor sorption) analysis at 25° C. and 80% RH (room humidity): 4.12%, 0.38%, 0.97% and 0.50%.

[0342] In one aspect, the present application provides an acid addition salt of the crystalline form of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or a solvate or hydrate of any of the foregoing substances.

[0343] In one aspect, the present application provides a method for preparing the crystalline form of the present application, the method being selected from the group consisting of:

[0344] (1) Compound HY-A was added to EtOAc, HCl (1.05 molar ratio) was slowly added to the solution under magnetic stirring, and the solution was dried by stirring to obtain a crystalline form;

[0345] (2) adding compound HY-A and citric acid, adding EtOH, and drying by stirring to obtain a crystalline form;

[0346] (3) Compound HY-A and 10 mL of EtOH were added, and 95.4 μL of 85% (2.05 molar ratio) H3PO4 was slowly added to the solution under magnetic stirring. After stirring, the solid was separated by centrifugation and continued to be slurried in EtOAc. After slurrying for 2 hours, the solid was centrifuged and dried under vacuum at room temperature to obtain a crystalline form;

[0347] (4) Compound HY-A was purged with N2 for 20 min;

[0348] (5) Compound HY-A was heated to 130°C and cooled to 30°C under N2 atmosphere;

[0349] (6) Compound HY-A was heated to 120°C under N2 atmosphere and then cooled to room temperature;

[0350] (7) Compound HY-A was added to IPA / MIBK (1:1, v / v) at 50 °C for 5 days and then dried under vacuum;

[0351] (8) Compound HY-A was heated to 90°C under N2 atmosphere and then cooled to room temperature;

[0352] (9) Slowly evaporating the solution of compound HY-A together with polymer mixture B;

[0353] (10) Compound HY-A was added to MeOH for 5 days and then dried at room temperature;

[0354] (11) Compound HY-A was added to ACN / H2O (v / v, 1:4) for 5 days and then dried at room temperature;

[0355] (12) Compound HY-A was added to 2-MeTHF at 50 °C for 5 days, then slurried at 5 °C for 1 day and dried at room temperature;

[0356] (13) adding the anti-solvent MTBE to the 1,4-dioxane solution of compound HY-A, and then drying at room temperature;

[0357] (14) Compound HY-A was exposed to a CHCl3 atmosphere for one day and then dried at room temperature;

[0358] (15) Compound HY-A was added to 2-MeTHF / toluene (v / v, 1:1) at 50 °C for 5 days and then dried at room temperature;

[0359] (16) Compound HY-A was added to anisole at room temperature for 5 days and then dried at room temperature;

[0360] (17) Compound HY-A was added to toluene at room temperature for 5 days and then dried at room temperature;

[0361] (18) drying compound HY-A at room temperature (temperature range: 20-22° C.; humidity range: 44-77% RH); and

[0362] (19) Compound HY-A was exposed to a DMSO atmosphere for 7 days and then dried at room temperature.

[0363]

[0364]

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373] On the one hand, the present application provides a pharmaceutical composition, comprising: the crystalline form of the present application and / or the acid addition salt of the crystalline form of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate of the foregoing, and a pharmaceutically acceptable carrier.

[0374] On the one hand, the present application provides a kit, comprising: the crystalline form of the present application, the acid addition salt of the crystalline form of the present application and / or the pharmaceutical composition of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate of the foregoing, and a pharmaceutically acceptable carrier.

[0375] On the one hand, the present application provides: a method for inhibiting the activity of casein kinase (CK), the method comprising administering an effective amount of the crystalline form of the present application, the acid addition salt of the crystalline form of the present application and / or the pharmaceutical composition of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite of the pharmaceutical composition, or any of the aforementioned solvates or hydrates and / or the kit of the present application to a subject in need thereof. For example, the casein kinase (CK) can be selected from the group consisting of: casein kinase Iα (CK1α), casein kinase Iδ (CK1δ) and casein kinase Iε (CK1ε). For example, the method can be selected from the group consisting of: an in vitro method, an ex vivo method, and an in vivo method.

[0376] In another embodiment, the present application provides a crystalline form of the present application, an acid addition salt of the crystalline form of the present application and / or a pharmaceutical composition of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite of the pharmaceutical composition, or any of the aforementioned solvates or hydrates and / or the use of a kit of the present application in the preparation of a drug and / or kit for inhibiting the activity of casein kinase (CK). For example, the casein kinase (CK) can be selected from the group consisting of casein kinase Iα (CK1α), casein kinase Iδ (CK1δ) and casein kinase Iε (CK1ε). For example, the method can be selected from the group consisting of: an in vitro method, an ex vivo method, and an in vivo method.

[0377] In another embodiment, the present application provides a crystalline form of the present application, an acid addition salt of the crystalline form of the present application and / or a pharmaceutical composition of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite of the pharmaceutical composition, or any of the aforementioned solvates or hydrates and / or a kit of the present application for inhibiting casein kinase (CK) activity. For example, the casein kinase (CK) can be selected from the group consisting of casein kinase Iα (CK1α), casein kinase Iδ (CK1δ) and casein kinase Iε (CK1ε). For example, the method can be selected from the group consisting of: an in vitro method, an ex vivo method, and an in vivo method.

[0378] On the other hand, the present application provides a method for preventing and / or treating a disease or condition, the method comprising administering an effective amount of a compound of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any of the aforementioned solvates or hydrates to a subject in need thereof. For example, the disease or condition may be selected from the group consisting of a neurological disease and a mental illness. For example, the disease or condition may be selected from the group consisting of a mood disorder, a sleep disorder, and a circadian rhythm disorder. For example, the disease or condition may be selected from the group consisting of depression and bipolar disorder.

[0379] In another embodiment, the present application provides the use of the crystalline form of the present application, the acid addition salt of the crystalline form of the present application and / or the pharmaceutical composition of the present application, or the pharmaceutically acceptable salt, prodrug or metabolite of the pharmaceutical composition, or any of the aforementioned solvates or hydrates and / or the kit of the present application in the preparation of a drug and / or kit for preventing and / or treating a disease or condition. For example, the disease or condition can be selected from the group consisting of a neurological disease and a mental illness. For example, the disease or condition can be selected from the group consisting of a mood disorder, a sleep disorder and a circadian rhythm disorder. For example, the disease or condition can be selected from the group consisting of depression and bipolar disorder.

[0380] In another embodiment, the present application provides a crystalline form of the present application, an acid addition salt of the crystalline form of the present application and / or a pharmaceutical composition of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite of the pharmaceutical composition, or any of the aforementioned solvates or hydrates and / or a kit of the present application for preventing and / or treating a disease or condition. For example, the disease or condition can be selected from the group consisting of a neurological disease and a mental illness. For example, the disease or condition can be selected from the group consisting of a mood disorder, a sleep disorder, and a circadian rhythm disorder. For example, the disease or condition can be selected from the group consisting of depression and bipolar disorder.

[0381] In another embodiment, the present application provides a composition comprising: a compound of the present application or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate thereof, and optionally a pharmaceutically acceptable carrier.

[0382] The compounds of the present application can be administered orally. Oral administration may involve swallowing, so that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be used, whereby the compound enters the bloodstream directly from the mouth.

[0383] In some cases, the compound of the present application can also be directly administered to the blood, muscle, or internal organs. Suitable means for parenteral administration may include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular and subcutaneous. Applicable devices for parenteral administration may include needle (including microneedle) syringes, needle-free syringes and infusion techniques.

[0384] The compounds of the present application can also be topically administered to the skin or mucous membrane, i.e., percutaneously or transdermally. In some cases, the compounds of the present application can also be administered intranasally or by inhalation. In some cases, the compounds of the present application can be administered rectally or vaginally. In another embodiment, the compounds of the present application can also be directly administered to the eyes or ears.

[0385] The dosage regimen of the compound and / or the composition comprising the compound is based on a variety of factors, including the type, age, weight, sex and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the specific compound employed. Thus, the dosage regimen may vary widely. A dosage level of about 0.01 mg to about 100 mg per kilogram of body weight per day may be effective in treating the above-mentioned conditions.

[0386] Suitable subjects according to the present invention include mammalian subjects. Mammals according to the present invention may include, but are not limited to, canines, felines, bovines, caprines, equines, ovines, porcines, rodents, lagomorphs, primates, and the like, and include mammals in utero. In one embodiment, humans are suitable subjects. Human subjects may be of any gender and may be at any stage of development.

[0387] In another embodiment, the present application provides the use of one or more compounds of the present application in the preparation of a medicament for treating the conditions listed herein.

[0388] For the treatment of the above mentioned conditions, the compounds of the present application may be administered as the compounds themselves. Alternatively, pharmaceutically acceptable salts may be suitable for medical use due to their greater water solubility relative to the parent compound.

[0389] In another embodiment, the present application provides a composition. Such a composition may include a compound of the present application with a pharmaceutically acceptable carrier. The carrier may be a solid product, a liquid, or both, and may be formulated with the compound into a unit dose composition, for example, a tablet, which may contain from 0.05% to 95% by weight of the active compound. The compound of the present application may be coupled with a suitable polymer as a targetable drug carrier. Other pharmacologically active substances may also be present.

[0390] The compounds of the present invention can be administered by any suitable route, in the form of a pharmaceutical composition suitable for the route, and in a dose effective for the intended treatment. The active compounds and compositions, for example, can be administered orally, rectally, parenterally or topically.

[0391] The compounds of the present application can be used alone or in combination with other therapeutic agents to treat various disorders or disease states. The compounds of the present application and other therapeutic agents can be administered simultaneously (in the same dosage form or in separate dosage forms) or sequentially.

[0392] Administration of two or more compounds "in combination" may mean that the two compounds are administered sufficiently closely in time so that the presence of one compound alters the biological effect of the other. The two or more compounds may be administered simultaneously, concurrently or sequentially. In addition, simultaneous administration may be performed by mixing the compounds prior to administration or by administering the compounds at the same time point but at different anatomical sites or using different routes of administration.

[0393] The phrases "concurrent administration," "co-administration," "simultaneous administration," and "administered simultaneously" may refer to administration of the compounds in combination.

[0394] Example

[0395] The following examples are presented so as to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the invention and are not intended to limit the scope of what the inventors regard as their invention nor to represent that the experiments below are all or the only experiments performed. Every effort has been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. Standard abbreviations may be used, e.g., bp, base pairs; kb, kilobase; p1, picoliter; s or sec, seconds; min, minutes; h or hr, hours; aa, amino acid; nt, nucleotide; im, intramuscular (IM); ip, intraperitoneal (IP); sc, subcutaneous (IP); etc.

[0396] Example 1 Preparation of Compounds

[0397] Preparation of compounds

[0398]

[0399] Synthesis route of compound HY-A

[0400]

[0401] Step 1: 2-(4-Fluorophenyl)-3-(2-methylpyridin-4-yl)-5-(methylsulfonyl)-4,5,6,7-tetrahydropyridine Oxalo[1,5-a]pyrazine

[0402] To a mixture of 2-(4-fluorophenyl)-3-(2-methylpyridin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine (85 mg, 0.28 mmol) in DCM (10 mL) was added DIPEA (108.5 mg, 0.84 mmol) at 25°C, and then methanesulfonyl chloride (0.027 ml, 0.34 mmol) in DCM (2 mL) was added dropwise to the mixture at -20°C. The reaction was stirred at -20°C for 0.5 h. LCMS showed that the desired MS was detected as the main peak. The mixture was quenched with 2 drops of MeOH and extracted with DCM (10 mL*2) and H2O (20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated, and then purified by column chromatography (DCM / MeOH=20 / 1) to give a white solid (10.1 mg, yield: 9.3%). Chemical formula: calculated as (M+H + )C 19 H 19 FN4O2S:386.12, found:387. 1H NMR(400MHz, CDCl3)δ8.46(d,J=5.2Hz,1H),7.40–7.35(m,2H),7.04–6.98(m,2H),6.97–6 .88(m,2H),4.57(s,2H),4.43–4.38(m,2H),3.90–3.85(m,2H),2.96(s,3H),2.56(s,3H).

[0403] Biological activity assay

[0404] CK1δ kinase assay was performed with a buffer (40 μL, pH 7.5) containing 50 mM Tris, 10 mM MgCl2, 1 mM dithiothreitol, 100 μg / mL BSA with 10 μM ATP, 2 nM wild-type CK1δ, and 42 μM peptide substrate PLSRTLpSVASLPGL (Flotow et al., 1990) in the presence of 1 μL of CK1δ inhibitor (e.g., compounds of the present application) or 4% DMSO (e.g., as a control). The reaction mixture was incubated at 25°C for 85 min; detection was performed as described for the Kinase-Glo assay (Promega). Luminescent output was measured on a Perkin Elmer Envision microplate reader (PerkinElmer, Waltham, MA).

[0405] Bmal1-dLuc or Per2-dLuc U2OS cells were suspended in culture medium (DMEM supplemented with 10% fetal bovine serum, 0.29 mg / mL L-glutamine, 100 units / mL penicillin and 100 mg / mL streptomycin) and seeded in 96-well white solid bottom plates at 200 μL (10,000 cells) per well. After 2 days, 100 μL of explant culture medium (DMEM supplemented with 2% B27, 10 mM HEPES, 0.38 mg / mL sodium bicarbonate, 0.29 mg / mL L-glutamine, 100 units / mL penicillin, 100 mg / mL streptomycin, 0.1 mg / mL gentamicin and 1 mM luciferin, pH 7.2) was dispensed into each well, and then 1 μL of the compound of the present application (dissolved in DMSO; the final concentration in DMSO is 0.7%) was applied. The plate was covered with an optically transparent film and set to a microplate reader (Infinite M200, Tecan). Luminescence was recorded every 1 hour for 3-4 days. The cycle parameters were obtained from the luminescence rhythm by curve fitting programs CellulaRhythm or MultiCycle (Actimetrics), both of which produced similar results.

[0406] The CK1δ inhibition result (IC50) of HY-A is 369.4 nM. The CK1δ inhibition result (EC50) of HY-A is 2.2 μM.

[0407] The present disclosure provides HY-A as a potent inhibitor of casein kinase.

[0408] Drug transport trials

[0409] 1. Preparation of Caco-2 Cells

[0410] 1) 50 μL and 25 mL of cell culture medium were added to each well of the Transwell insert and reservoir, respectively. The HTS transwell plates were then incubated at 37° C., 5% CO 2 for 1 hour before cell seeding.

[0411] 2) Dilute Caco-2 cells to 6.86 x 10 5 cells / mL, and dispense 50 μL of the cell suspension into the filter wells of a 96-well HTS transwell plate. The cells were cultured in a cell culture incubator at 37°C, 5% CO2, and 95% relative humidity for 14-18 days. The cell culture medium was changed every other day, starting at the latest within 24 hours after the initial seeding.

[0412] 2. Preparation of Stock Solutions

[0413] Prepare 10 mM stock solutions of test compounds in DMSO. Prepare 10 mM stock solutions of positive controls in DMSO. Digoxin and propranolol were used as control compounds in this assay.

[0414] 3. Assessment of Cell Monolayer Integrity

[0415] 1) Remove the culture medium from the reservoir and each Transwell insert and replace with pre-warmed fresh culture medium.

[0416] 2) The transepithelial electrical resistance (TEER) across the monolayer was measured using the Millicell Epithelial Volt-Ohm measurement system (Millipore, USA).

[0417] 3) After the measurement is completed, return the plate to the incubator.

[0418] Calculate the TEER value according to the following equation:

[0419] TEER measurement (ohms)*membrane area (cm 2 ) = TEER value (ohm cm 2 )

[0420] TEER value should be greater than 230ohm·cm2 , which indicates a well-formed Caco-2 monolayer.

[0421] 4. Assay Procedure

[0422] 1) Remove the Caco-2 plate from the incubator and wash twice with pre-warmed HBSS (10 mM HEPES, pH 7.4), then incubate at 37°C for 30 minutes.

[0423] 2) The stock solutions of the control compound and the test compound were diluted in DMSO to obtain 1 mM solutions, and then diluted with HBSS (10 mM HEPES, pH 7.4) to obtain 5 μM working solutions. The final concentration of DMSO in the incubation system was 0.5%.

[0424] 3) Determine the drug transport rate in the apical to basolateral direction. 125 μL of 5 μM control compound and working solution of test compound are added to the Transwell insert (apical compartment), and 50 μL sample (D0 sample) is immediately transferred from the apical compartment to a new 96-well plate. The wells in the receiving plate (basolateral compartment) are filled with 235 μL of HBSS (10 mM HEPES, pH 7.4).

[0425] 4) Determine the drug transport rate in the basolateral to apical direction. 285 μL of 5 μM control compound and working solution of test compound are added to the receiving plate well (basolateral compartment), and 50 μL sample (D0 sample) is immediately transferred from the basolateral compartment to a new 96-well plate. The holes in the Transwell insert (apical compartment) are filled with 75 μL of HBSS (10 mM HEPES, pH 7.4). The determination is repeated.

[0426] 5) Incubate the plate at 37°C for 2 hours.

[0427] 6) At the end of the incubation, 50 μL samples from the donor side (apical compartment for Ap→Bl flux, and basolateral compartment for Bl→Ap) and the acceptor side (basolateral compartment for Ap→Bl flux, and apical compartment for Bl→Ap) were transferred to wells of a new 96-well plate, followed by the addition of 4 volumes of cold methanol containing the appropriate internal standard (IS). The samples were vortexed for 5 minutes and then centrifuged at 3,220 g for 40 minutes. Prior to LC-MS / MS analysis, a 100 μL aliquot of the supernatant was mixed with an appropriate volume of ultrapure water.

[0428] 7) To determine the leakage of fluorescein after a 2-hour transport period, a stock solution of fluorescein was prepared in water and diluted with HBSS (10mM HEPES, pH 7.4) to a final concentration of 100 μM. 100 μL of fluorescein solution was added to each Transwell insert (top compartment), and then the holes in the receiving plate (basolateral compartment) were filled with 300 μL of HBSS (10mM HEPES, pH 7.4). The plate was incubated at 37°C for 30 minutes. 80 μL of sample was taken directly from the top hole and the basolateral hole (using the basolateral access hole) and transferred to the hole of a new 96-well plate. Measure the fluorescein fluorescence (to monitor the integrity of the monolayer) signal at 485 nM excitation and 530 nM emission in a fluorescent plate reader.

[0429] 5. Data Analysis

[0430] The apparent permeability coefficient (P) for Caco-2 drug transport assay can be calculated using the following equation: app ), in cm / s:

[0431] P app =(V A ×[Drug] 受体 ) / (area×time×[drug] 初始，供体 )

[0432] Where V A is the volume in the receptor pore (in mL), and area is the surface area of ​​the membrane (0.143 cm for Transwell-96 permeable scaffolds). 2 ), and time is the total transit time in seconds.

[0433] The outflow ratio will be determined using the following equation:

[0434] Outflow ratio = P app(B-A) / P app(A -B)

[0435] Where P app(B-A) represents the apparent permeability in the basolateral to apical direction, P app(A-B) Represents the apparent permeability in the apical to basolateral direction.

[0436] Recycling can be determined using the following equation:

[0437] Recovery rate % = (V A ×[Drug] 受体 +V D ×[Drug] 供体 ) / (V D ×[Drug] 初始，供体 )

[0438] Where V A is the volume in mL in the acceptor pore (0.235 mL for Ap→Bl flux and 0.075 mL for Bl→Ap), V D is the volume in the donor well (in mL) (0.075 mL for Ap→Bl flux and 0.235 mL for Bl→Ap)

[0439] The leakage of fluorescent yellow, in percent (%), can be calculated using the following equation:

[0440] %LY leakage = 100 × [LY] 受体 / ([LY] 供体 +[LY] 受体 )

[0441] Less than 1% LY leakage is acceptable to indicate that the Caco-2 monolayer is of good quality.

[0442] HY-A's P app(B-A) , P app(A-B) and outflow ratios are summarized as follows.

[0443]

[0444] Intrinsic clearance assay

[0445] 1. Prepare the master solution as follows.

[0446] Reagents Reserve concentration volume Final concentration Phosphate buffered saline 200mM 200μL 100mM <![CDATA[Ultra-pure H2O]]> - 106μL - <![CDATA[MgCl2 solution]]> 50mM 40μL 5mM

[0447] 2. Three separate experiments were performed as follows: a) With NADPH: 10 μL of 20 mg / mL liver microsomes and 40 μL of 10 mM NADPH were added to the incubation. The final concentrations of microsomes and NADPH were 0.5 mg / mL and 1 mM, respectively. b) Without NADPH: 10 μL of 20 mg / mL liver microsomes and 40 μL of ultrapure water were added to the incubation. The final concentration of microsomes was 0.5 mg / mL. c) Heat-inactivated microsomes without NADPH: 10 μL of 20 mg / mL heat-inactivated liver microsomes and 40 μL of ultrapure H2O were added to the incubation. The final concentration of microsomes was 0.5 mg / mL.

[0448] 3. The reaction was started by adding 4 μL of 200 μM test compound solution or control compound solution (final concentration 2 μM) and carried out at 37°C.

[0449] 4. Take 50 μL aliquots from the reaction solution at 0 min, 15 min, 30 min, 45 min and 60 min. Terminate the reaction by adding 4 volumes of cold acetonitrile with IS (100 nM alprazolam, 200 nM labetalol, 200 nM caffeine and 2 μM ketoprofen). Centrifuge the samples at 3,220 g for 40 minutes. Mix 100 μL aliquots of the supernatant with 100 μL of ultrapure H2O and use for LC-MS / MS analysis.

[0450] 5. Data Analysis

[0451] All calculations were performed using Microsoft Excel.

[0452] Peak areas were determined from the extracted ion chromatograms. The slope value, k, was determined by linear regression of the natural logarithm of the percentage of parent drug remaining versus the incubation time curve.

[0453] The in vitro half-life (in vitro t 1 / 2 ):

[0454] In vitro 1 / 2 =-(0.693 / k)

[0455] The in vitro t was calculated using the following equation (average of two determinations): 1 / 2 (min) In vitro internal clearance (in vitro CL int , in μL / min / mg protein):

[0456] In vitro CL int =(0.693*incubation volume (μl)) / (in vitro t 1 / 2 *Amount of protein (mg)

[0457] The in vitro t was calculated using the following equation (average of two determinations): 1 / 2 (min) to amplify the intrinsic clearance of unbound int , in mL / min / kg):

[0458] The scaling factors used for the prediction of intrinsic clearance in liver microsomes are summarized below.

[0459]

[0460]

[0461] a. Iwatosubo et al., Davies and Morris, 1993, 10(7) pp. 1093-1095.

[0462] b. Barter et al., 2007, Curr Drug Metab, 8(1), pp. 33-45; Iwatsubo et al., 1997, JPET, 283, pp. 462-469.

[0463] Papp(BA), Papp(AB) and outflow ratio are summarized below.

[0464]

[0465] HY-A has desirable intrinsic scavenging properties.

[0466] The purpose of this application is to perform salt screening and polymorph screening of selected salts or free base of compound HY-A to identify solid forms for toxicology studies and further development.

[0467] For the free base form A, equilibrium solubility studies were performed in pH 2.0, 4.0, 6.0, 8.0 buffers and pure water at room temperature (room temperature: 20-22°C). The solubility in pH 2.0 and 4.0 was found to be 2.36 mg / mL and 1.47 mg / mL, respectively, which is relatively high compared to the solubility in other media (0.023-0.046 mg / mL).

[0468] Using free base Form C as the material, 100 polymorph screening experiments were performed using the following methods: antisolvent addition, solid vapor diffusion, slurrying at room temperature (RT) and 50°C, slow evaporation, slow cooling, liquid vapor diffusion, polymer induced crystallization, and grinding. A total of 20 polymorphs were obtained from the screening and further experiments, designated as free base Form AT, which were characterized using X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and proton nuclear magnetic resonance (NMR) spectrometry. 1 HNMR). Based on the characterization results, A, C, P, N and Q types are anhydrates; B, I and L types are hydrates; D, F, G, H, J, K, M, O, S and T types are solvates; E type (converted to M type after drying at 20-22°C / 44-77%RH) and R type (can be converted to B type after exposure to air for 10min) were not identified. The characterization results are summarized in Table 1-1.

[0469] The interconversion relationships between the anhydrates (free base A, C, P, N and Q forms) were studied via competitive slurrying in EtOH or IPAc at room temperature or 50°C. The results showed that free base form C was obtained in all experiments, indicating that free base form C was the most stable anhydrate from room temperature to 50°C. Competitive slurrying experiments of free base anhydrate form C, hydrate form B, hydrate form I and hydrate form L in EtOH / H2O solvent system at different water activities (aw The results showed that when a w ≤0.6, the free base type C is obtained; when a w When the free base L form is obtained, w The free base B form is obtained in ~1). Figure 1-1 The conversion methods are shown in Tables 1-2. Based on the interconversion results, the free base Form C shows: good performance in the solid state; most stable at room temperature and 50°C in the anhydrate; stable in a relatively wide range of water activities (0-0.6).

[0470] Using free base Form C as material, salt screening was performed under 52 different conditions using 12 acids (applied HCl at different molar ratios) in 4 solvent systems. A total of 11 crystalline salt hits (19 crystalline forms) were obtained during the screening, which were characterized by XRPD, TGA and DSC. Salt stoichiometry was used 1 H NMR or ultra-performance liquid chromatography (UPLC) coupled with ion chromatography (IC) was used for determination. Based on the characterization results (small TGA weight loss, pure endothermic signal on the DSC curve, safety level of the acid, and simple polymorphism), HCl salt Form A, citrate Form A, and phosphate Form A were selected for re-preparation, and the results showed that HCl salt Form A, citrate Form A, and phosphate Form A were successfully obtained.

[0471] The three reconstituted salt forms and the free base were evaluated for kinetic solubility, hygroscopicity (DVS), and solid stability. The salt evaluation results are summarized in Tables 1-3. The evaluation results are summarized as follows.

[0472] Kinetic solubility results: All three salts showed higher solubility in water than the free base. Both salts and free base were completely dissolved in SGF. In FaSSIF, phosphate salt Form A showed higher solubility than other salts and free base, and HCl salt Form A, citrate salt Form A, and free base Form A transformed into other crystalline forms of free base after 1 hour. In FeSSIF, the solubility of these three salts was higher than that of free base, while the solubility of salts and free base became comparable after 24 hours; these three salts disproportionated into other crystalline forms of free base after 1 hour.

[0473] Hygroscopicity results: At 25°C / 80%RH, the water absorption of free base Form C, citrate Form A, and phosphate Form A were 0.50%, 0.38%, and 0.97% (slightly hygroscopic), respectively. HCl salt Form A showed 2.95% water absorption at 25°C / 30%RH, and 4.07% water absorption at 25°C / 80%RH.

[0474] Solid Stability Assessment: Free base Form A converted to Form C after storage at 40°C / 75%RH for one week. No form change was observed for the other samples. No significant purity decrease was observed for all samples after storage at 25°C / 60%RH or 40°C / 75%RH for one week.

[0475] Based on the interconversion results, the free base Form C may be suitable for further development and toxicology studies. If considering developing a salt as an alternative to the free base Form C, the salt candidate may be selected based on kinetic solubility assessments and PK results.

[0476] Table 1-1 Summary of characterization results of polymorphs of compound HY010101

[0477]

[0478]

[0479] *: Exotherm (peak); **: TGA weight loss (%, up to 150°C); TGA weight loss (%, up to 90°C)

[0480] Table 1-2 Summary of transformation methods of HY010101 free base polymorphs

[0481]

[0482]

[0483] Table 1-3 Summary of evaluation of salts and free bases

[0484]

[0485]

[0486] *: Free base Form A (200754-075-P3; 823121-01-A1) was used in this evaluation study; -: No acid; N / A: 1 No detectable solvent residues were found in H NMR.

[0487] Example 2

[0488] Characterization of materials

[0489] This application received two batches of materials: the first batch (200754-075-P3, used for approximate solubility testing, salt screening experiments, equilibrium solubility, kinetic solubility and solid stability evaluation); the second batch (200754-075-P5, used for free base polymorph screening experiments, hygroscopicity and solid stability evaluation). These two batches were tested by XRPD, TGA, DSC, 1The samples were characterized by H NMR and purity test.

[0490] Free base type A

[0491] Test material (200754-075-P3; 823121-01-A). XRPD results are shown in Figure 2-1 It is named as free base type A. The TGA / DSC curve is shown in Figure 2-2 The TGA results showed a weight loss of 2.2% up to 150°C. The DSC results showed an endotherm (peak) at 175.6°C and an exotherm (peak) at 121.5°C. 1 H NMR results showed Figure 2-3 Due to the small TGA weight loss, the free base Form A was presumed to be anhydrous. The HPLC results showed Figure 2-4 and Table 2-1, which shows that the purity of this free base Form A material is 99.8 area %. Free base Form A was subjected to variable temperature XRPD (VT-XRPD). Results ( Figure 2-5 ) shows no form change after N2 purging for 20 min. After heating the sample to 130°C under N2 atmosphere, it is converted to free base Form C (see the embodiment for characterization data of free base Form C). After cooling to 30°C under N2 atmosphere and exposure to air for 10 min, no further form change is observed.

[0492] Table 2-1 Summary of impurities in free base type A (823121-01-A)

[0493]

[0494] Free base type C

[0495] Test material (200754-075-P5; 823121-16-A). XRPD results are shown in Figure 2-6 It is named as free base type C. The TGA / DSC curve is shown in Figure 2-7 The TGA results showed a weight loss of 0.6% up to 120°C. The DSC results showed an endotherm (peak) at 176.2°C. 1 H NMR results showed Figure 2-8 Due to the small TGA weight loss, the free base Form A was assumed to be anhydrous. The UPLC results showed Figure 2-9 and Table 2-2, which shows that the purity of this free base Type C material is 99.70 area %.

[0496] Table 2-2 Summary of impurities in free base type C (823121-16-A)

[0497]

[0498] Equilibrium solubility of free base

[0499] The equilibrium solubility of the free base Type A material (823121-01-A) was measured in pH 2.0, 4.0, 6.0, 8.0 buffers (preparation of buffer media in reference example) and ultrapure water. The material (solid loading: 5 mg / mL) was added to the corresponding media and then slurried (1000 rpm) for 24 hours at room temperature. After centrifugation and filtration (0.45 μm PTFE filter), the supernatant was tested for solubility and pH, and residual solids were detected by XRPD. The results are summarized in Figure 2-10 and Table 2-3. After slurrying the free base type A material (823121-01-A) in buffers of pH 2.0, 4.0, 6.0, and 8.0 for 24 hours, a new form was obtained, named free base type B. The solubility in pH 2.0 and 4.0 was 2.36 mg / mL and 1.47 mg / mL, respectively, which was relatively high compared to the solubility in other media (0.023-0.046 mg / mL).

[0500] Table 2-3 Summary of equilibrium solubility test of free base

[0501]

[0502] *: Separation of API signals was observed by using the HPLC method (Table 8-20). By using the optimized UPLC method (Table 8 21), impurity signals were observed, which are summarized in the application.

[0503] Example 3

[0504] Based on the XRPD results of polymorph screening and further experiments, a total of 20 polymorphs (free base AT form) were obtained, including anhydrates A, C, P, N and Q forms, hydrates B, I and L forms, solvates D, F, G, H, J, K, M, O, S and T forms, and other forms E form (converted to M form after drying at 0-22°C / 44-77%RH, so it was not identified) and R form (can be converted to B form after exposure to air, so it was not identified). The characterization results of each form are summarized in Table 1-1 and are from Figure 3-1 to Figure 3-4 The XRPD results are shown in .

[0505] Anhydrous

[0506] Free base type A

[0507] Refer to the characterization results of the free base Form A of the Examples.

[0508] Free base type C

[0509] Refer to the characterization results of the free base type C in the examples. The method for obtaining the free base type C can refer to the polymorph screening experiment in the examples.

[0510] Free base P type

[0511] The free base P form (823121-36-A2-TO 150C) was obtained by heating the free base K form (823121-36-A2) to 120°C under N2 atmosphere and then cooling to room temperature in air. The XRPD pattern is shown in Figure 3-5 The TGA / DSC curve is shown in Figure 3-6 The TGA results showed a weight loss of 7.0% up to 120°C. The DSC results showed an endothermic peak at 176.1°C. 1 H NMR results ( Figure 3-7 ) showed no detectable solvent residues. VT-XRPD was performed on the free base P form. Results ( Figure 3-8 ) showed no form change after N2 purging for 20 min, heating to 130°C under N2 atmosphere (assuming that the peak shift at 130°C is due to lattice expansion), cooling to 30°C under N2 atmosphere, or exposure to air. Therefore, the free base P form is presumed to be an anhydrate.

[0512] Free base N type

[0513] The free base N form (823121-20-A10-DRY) was obtained by slurrying the free base C form material (823121-16-A) in IPA / MIBK (1:1, v / v) at 50°C for 5 days and then vacuum drying. The XRPD pattern is shown in Figure 3-9 The TGA / DSC curve is shown in Figure 3-10 The TGA results showed a weight loss of 3.5% up to 120°C. The DSC results showed two endothermic peaks at 170.4°C and 176.1°C. 1 H NMR results ( Figure 3-11 ) showed no detectable IPA or MIBK solvent residues. Since free base Form N was also obtained from the VT-XRPD experiment of free base Form I, a mixture of free base Form N and Form C was obtained by heating the sample to 130°C under N2 atmosphere (Example). Therefore, it is speculated that free base Form N is anhydrous. An attempt was made to re-prepare free base Form N by using free base Form A (823121-01-A) as a material, and the results are summarized in Table 3-1. The XRPD results are shown in Table 3-1. Figure 3-12 In the figure, all re-prepared samples were shown to be the free base C form.

[0514] Table 3-1 Summary of the re-preparation of free base type N

[0515]

[0516] Free base Q type

[0517] Free base Form Q (823121-45-A1_AFT 90C) was obtained by heating free base Form I (823121-45-A1) to 90°C under N2 atmosphere and then cooling to room temperature (Example). The XRPD pattern is shown in Figure 3-13 The TGA / DSC curve is shown in Figure 3-14 The TGA results showed a weight loss of 1.1% up to 120°C. The DSC results showed three endotherms (peaks) at 115.9°C, 170.3°C and 176.1°C and one exotherm (peak) at 120.0°C. 1 H NMR results ( Figure 3-15 ) showed that no detectable solvent residue was found. Since the free base Q form was obtained under heating conditions and N2 atmosphere, combined with its small TGA weight loss, it was speculated that the free base Q form was anhydrous.

[0518] Hydrate

[0519] Free base type B

[0520] The free base Form B (823121-24-A5-DRY) obtained in the polymorph screening experiment was obtained by slowly evaporating a solution of the material (823121-16-A) together with polymer mixture B at room temperature. The re-prepared free base Form B was obtained by slurrying the free base Form C material (823121-16-A) in H2O for one day and then drying at room temperature. The XRPD pattern is shown at Figure 3-16 The TGA / DSC curve is shown in Figure 3-17 The TGA results showed a weight loss of 13.6% up to 120°C (the weight loss of the monohydrate was about 4.3%). The DSC results showed two endotherms (peaks) at 91.5°C and 175.8°C. 1 H NMR results showed Figure 3-18 In. Figure 3-19 and Figure 3-20 The VT-XRPD results of free base type B (823121-43-A2) shown in Figure 1 show peak shifts after N2 purge for 20 min, and the form after N2 purge is named free base type R. After being exposed to air for 10 min, free base type R is converted back to free base type B. Under N2 atmosphere, free base type C is obtained after free base type B is heated to 100°C. After cooling to 30°C and exposed to air for 10 min under N2 atmosphere, no further form changes are observed. Combined with the form change after heating and the large TGA weight loss, it is speculated that free base type B is a hydrate.

[0521] Free base type I

[0522] Free base Form I (823121-19-A1-DRY) was obtained by slurrying the free base Form C material (823121-16-A) in MeOH for 5 days and then drying at room temperature. A re-prepared free base Form I sample (823121-45-A1) was obtained under the same procedure. The XRPD pattern is shown at Figure 3-21 The TGA / DSC curve is shown in Figure 3-22 The TGA results showed a weight loss of 7.6% up to 120°C. The DSC results showed four endotherms (peaks) at 89.0°C, 116.2°C, 170.5°C and 176.9°C and one exotherm (peak) at 122.5°C. 1 H NMR ( Figure 3-23 ) The results showed that no detectable MeOH residue was found. XRPD results ( Figure 3-24 ) shows that free base Form T, after heating to 90°C and then cooling to room temperature, transforms into a new form called free base Form Q (see Example). Combined with the endotherm at 89.0°C and the large TGA weight loss, it is speculated that free base Form I is a hydrate. After heating to 120°C and then cooling to room temperature, a mixture of free base Forms C and N is obtained. Figure 3-25 The VT-XRPD results of the free base Form I shown in show that after N2 purging for 20 min, the form is converted to the free base Form Q. After heating to 130°C under N2 atmosphere, a mixture of Form N and Form C is obtained again, and no form change is observed after cooling to 30°C under N2 atmosphere and exposure to air for 10 min.

[0523] Free base L-form

[0524] The free base L-form (823121-19-A1-DRY) was obtained by slurrying the free base C-form material (823121-16-A) in ACN / H2O (v / v, 1:4) for 5 days and then drying at room temperature. The re-prepared free base L-form sample (823121-36-A3) was obtained under the same procedure. The XRPD pattern is shown in Figure 3-26 The TGA / DSC curve is shown in Figure 3-27 The TGA results showed a weight loss of 13.9% up to 120°C. The DSC results showed four endotherms (peaks) at 66.2°C, 70.8°C, 79.1°C and 176.0°C. 1 H NMR ( Figure 3-28 ) The results showed that no detectable ACN residues were found. Figure 3-29The VT-XRPD results of the free base L-type (823121-36-A3) shown in Figure 1 show that after N2 purging for 20 min, the form is converted to the free base A-type. After heating to 120°C under N2 atmosphere, cooling to 30°C under N2 atmosphere, and exposure to air for 10 min, no form change was observed. Combined with the endothermic signal and the large TGA weight loss, it is speculated that the free base L-type is a hydrate.

[0525] Solvate

[0526] Free base D-type

[0527] The free base Form D (823121-20-A6-DRY) was obtained by slurrying the free base Form C material (823121-16-A) in 2-MeTHF at 50°C for 5 days, then at 5°C for 1 day, and drying at room temperature. The XRPD pattern is shown at Figure 3-30 The TGA / DSC curve is shown in Figure 3-31 The TGA results showed a weight loss of 8.9% up to 120°C. The DSC results showed two endotherms (peaks) at 99.9°C and 175.7°C. 1 H NMR ( Figure 3-32 ) The results showed that the residual 2-MeTHF / API molar ratio was 0.3 (6.3 wt %). XRPD results ( Figure 3-33 ) shows that the free base D form is converted to the free base P form after heating to 120°C and then cooling to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base Form D was a 2-MeTHF solvate.

[0528] Free base type F

[0529] The free base F form (823121-17-A8-DRY) was obtained by adding the anti-solvent of MTBE to a 1,4-dioxane solution of the material (823121-16-A) and then drying at room temperature. The XRPD pattern is shown in Figure 3-34 The TGA / DSC curve is shown in Figure 3-35 The TGA results showed a weight loss of 24.5% up to 150°C. The DSC results showed two endotherms (peaks) at 126.3°C and 175.3°C. 1 H NMR ( Figure 3-36 ) The results showed that the residual 1,4-dioxane / API molar ratio was 0.5 (9.3 wt%), and no detectable MTBE residue was found. XRPD results ( Figure 3-37 ) shows that the free base F form is converted to the free base P form after heating to 150°C and then cooling to room temperature. 1The amount of 1,4-dioxane shown in the H NMR was significantly lower than the TGA weight loss, so the free base Form F was re-prepared and re-characterized. TGA results for the re-prepared free base Form F (823121-39-A1-1210) ( Figure 3-38 ) shows a step-wise weight loss of 10.4% up to 150 °C. 1 HNMR Figure 3-39 ) The results showed that the residual 1,4-dioxane / API molar ratio was 0.5 (9.3 wt%), which is consistent with the TGA weight loss. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base Form F was a 1,4-dioxane solvate.

[0530] Free base G type

[0531] The free base G form (823121-18-A8-DRY) was obtained by exposing the free base C form material (823121-16-A) to an atmosphere of CHCl3 for one day and then drying at room temperature. The XRPD pattern is shown at Figure 3-40 The TGA / DSC curve is shown in Figure 3-41 The TGA results showed a weight loss of 10.7% up to 120°C. The DSC results showed two endotherms (peaks) at 72.5°C and 176.0°C. 1 H NMR ( Figure 3-42 ) The results showed that the molar ratio of residual CHCl3 / API was 0.25 (7.2 wt%). XRPD results ( Figure 3-43 ) shows that the free base form G is converted to the free base form C after being heated to 120°C and then cooled to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base Form G was a CHCl3 solvate.

[0532] Free base H type

[0533] The free base H form (823121-20-A16-DRY) was obtained by slurrying the free base C form material (823121-16-A) in 2-MeTHF / toluene (v / v, 1:1) at 50°C for 5 days and then drying at room temperature. The re-prepared free base H form (823121-37-A3) was obtained by the same method. The XRPD pattern is shown in Figure 3-44 The TGA / DSC curve is shown in Figure 3-45 The TGA results showed a weight loss of 11.2% up to 120°C. The DSC results showed two endotherms (peaks) at 116.4°C and 175.9°C. 1 H NMR ( Figure 3-46 ) The results showed that the residual toluene / API molar ratio was 0.4 (9.0 wt%) and the residual 2-MeTHF / API molar ratio was 0.03 (0.7 wt%). XRPD results ( Figure 3-47 ) shows that the free base H form (823121-20-A16-DRY) is converted to the free base P form after heating to 130°C and then cooling to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base H form was a toluene solvate.

[0534] Free base J type

[0535] Free base Form J (823121-19-A5-DRY) was obtained by slurrying the free base Form C material (823121-16-A) in anisole at room temperature for 5 days and then drying at room temperature. The XRPD pattern is shown at Figure 3-48 The TGA / DSC curve is shown in Figure 3-49 The TGA results showed a weight loss of 13.9% up to 120°C. The DSC results showed two endotherms (peaks) at 102.1°C and 176.3°C. 1 H NMR ( Figure 3-50 ) The results showed that the residual anisole / API molar ratio was 0.5 (11.9 wt %). XRPD results ( Figure 3-51 ) shows that free base form J (823121-19-A5-DRY) transforms into free base form C after heating to 120°C and then cooling to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base Form J was an anisole solvate.

[0536] Free base K type

[0537] The free base Form K (823121-19-A8-DRY) was obtained by slurrying the free base Form C material (823121-16-A) in toluene at room temperature for 5 days and then drying at room temperature. The XRPD pattern is shown at Figure 3-52 The TGA / DSC curve is shown in Figure 3-53 The TGA results showed a weight loss of 7.1% up to 150°C. The DSC results showed two endotherms (peaks) at 115.9°C and 176.1°C. 1 H NMR ( Figure 3-54 ) The results showed that the residual toluene / API molar ratio was 0.3 (6.6 wt %). XRPD results ( Figure 3-55) shows that the free base K form (823121-19-A8-DRY) is converted to the free base P form after heating to 150°C and then cooling to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base Form K was a toluene solvate.

[0538] Free base M type

[0539] The free base M form (823121-23-A4-DRY) was obtained by drying the free base E form (823121-23-A4) at room temperature (temperature range: 20-22°C; humidity range: 44-77% RH). The XRPD pattern is shown in Figure 3-56 The TGA / DSC curve is shown in Figure 3-57 The TGA results showed a weight loss of 2.5% up to 61°C and a weight loss of 8.8% from 61°C to 90°C. The DSC results showed two endotherms (peaks) at 83.6°C and 176.0°C. 1 H NMR ( Figure 3-58 ) The results showed that the residual n-hexane / API molar ratio was 0.3 (5.3 wt %). XRPD results ( Figure 3-59 ) shows that the free base M form (823121-23-A4-DRY) is converted to the free base P form after heating to 120°C and then cooling to room temperature. 1 The residual solvent in H NMR and TGA weight loss indicated that the free base M form was a hexane solvate.

[0540] Free base O form

[0541] The free base O form (823121-18-A9-DRY) was obtained by exposing the free base C form material (823121-16-A) to a DMSO atmosphere for 7 days and then drying at room temperature. The XRPD pattern is shown at Figure 3-60 The TGA / DSC curve is shown in Figure 3-61 The TGA results showed a weight loss of 11.1% up to 120°C. The DSC results showed two endotherms (peaks) at 118.4°C and 175.2°C. 1 H NMR ( Figure 3-62 ) The results showed that the residual DMSO / API molar ratio was 0.7 (12.3 wt %). XRPD results ( Figure 3-63 ) shows that the free base O form (823121-18-A9-DRY) is converted to the free base C form after heating to 130°C and then cooling to room temperature. 1The residual solvent in HNMR and TGA weight loss indicated that the free base O-form was a DMSO solvate.

[0542] Free base S type

[0543] It is noteworthy that all free base Form I samples were obtained from experimental systems involving methanol. Therefore, further experiments with the compounds in MeOH were investigated. Figure 3-64 and Figure 3-65 As shown, after soaking the free base form C (823121-16-A) and form N (823121-45-A1-check) in MeOH for 2 hours, XRPD characterization was performed. The results showed that both experiments gave a new form, named free base form S (823121-73-A1 and 823121-73-A2, respectively). After drying the free base form S (823121-73-A1 and 823121-73-A2) at room temperature for 2 hours (temperature: 17°C; humidity range: 23-26% RH), free base form Q was obtained. Therefore, TGA, DSC and 1 H NMR characterization showed that MeOH was involved in the formation of its crystal lattice. Therefore, it was speculated that the free base S-type was a MeOH solvate.

[0544] other

[0545] Free base E type

[0546] The free base E form (823121-23-A4) was obtained by liquid vapor diffusion by exposing a THF solution of the material (823121-16-A) to an atmosphere of n-hexane. The XRPD pattern is shown in Figure 3-66 After drying at room temperature (temperature range: 20-22°C; humidity range: 44-77% RH), this form was converted to the free base M form (see Example). Therefore, TGA, DSC and 1 H NMR characterization.

[0547] Free base R type

[0548] The free base R form (823121-43-A2-N2_30.0°C) was obtained by purging N2 for 20 min on the free base B form (823121-43-A2) (see Example). The XRPD pattern is shown in Figure 3-67 After exposure to air for 10 min, this form converted back to the free base B form. Therefore, TGA, DSC and 1 H NMR characterization.

[0549] Example 4

[0550] Competitive slurrying experiment of anhydrous substances

[0551] In this application, five anhydrous products (free base type A, type C, type P, type N and type Q) were obtained. In order to study the mutual conversion relationship between the anhydrous products, competitive slurrying was carried out.

[0552] About 2 mg of free base Form A, Form C, Form P, Form N, and Form Q were added to 1 mL of a saturated solution of free base Form A material (823121-01-A) in EtOH and IPAc (slurried for 2 hours at the desired temperature and then filtered). After slurrying for 3 or 5 days at room temperature or 50°C, the solids were tested by XRPD. The results are summarized in Table 4-1, and the XRPD results are shown in Table 4-1. Figure 4-1 and Figure 4-1 The free base form C was obtained in all experiments, indicating that form C is the most stable anhydrous form from room temperature to 50°C.

[0553] Table 4-1 Summary of competitive slurrying between anhydrous substances

[0554]

[0555] Competitive slurry experiment of anhydrate and hydrate

[0556] About 2 mg of free base Form C, Form B, Form I, and Form L were added to 1 mL of a saturated solution of free base Form A material (823121-01-A) in EtOH / H2O with different volume ratios (slurried at room temperature for 4 hours and then filtered). After slurrying at room temperature for 3 days, the solid was tested by XRPD. The results are summarized in Table 4-2, and the XRPD results are shown in Table 4-2. Figure 4-3 and Figure 4-4 When a w ≤0.6, the free base type C is obtained. w When the concentration of free alkali is 0.8, the free alkali type L is obtained, while the free alkali type B is obtained in pure water (a w -1). Based on the above results, at room temperature, the anhydrous C type can be converted into a w The hydrate L-form can be converted into a hydrate with an a of 0.8-1.0 w Hydrate type B.

[0557] Table 4-2 Summary of competitive slurrying between anhydrate and hydrate

[0558]

[0559]

[0560] Example 5

[0561] Free base salt screening

[0562] Salt Screening Summary

[0563] Using free base type A (823121-01-A) as material, a total of 100 salt screening experiments were performed using 25 acids in 4 solvent systems. About 20 mg of material and equimolar corresponding acid were added to 0.5 mL solvent and then slurried at room temperature for 2 days. The resulting suspension was centrifuged to recover the solid for XRPD testing, and the results are summarized in Table 5-1. A total of 11 crystalline salt hits (19 crystalline forms) were obtained from the salt screening, which were characterized by XRPD, TGA, DSC and NMR or UPLC / IC. The detailed results of the salt hit characterization are listed in the examples.

[0564] Table 5-1 Summary of salt screening

[0565]

[0566]

[0567] a : Slurry at room temperature → anti-solvent addition (MTBE) → slurry at 5°C for one day → evaporate at room temperature. b :Slurry at room temperature → Anti-solvent addition (MTBE) → Slurry at 5°C for one day c : After 8 days of storage (temperature range: 20-23°C; humidity range: 63-87% RH), mesylate Form A converted to a mixture of Form A and Form B. No further characterization was performed. c : After 8 days of storage (temperature range: 20-23°C; humidity range: 63-87% RH), benzenesulfonate form D converted to form B. No further characterization was performed.

[0568] Re-preparation of candidate salts

[0569] Based on the characterization results of the salt hits from the salt screen (small TGA weight loss, pure DSC endotherm), simple polymorphism, frequency of use and safety level of the salt formers, HCl Salt Form A, Citrate Form A and Phosphate Form A were selected for de novo preparation.

[0570] All forms were re-prepared by slurrying at a 300 mg scale, and the results showed that the HCl salt Form A, citrate salt Form A, and phosphate salt Form A were successfully obtained. XRPD, TGA, DSC, 1 The re-prepared samples were characterized by HNMR or UPLC / IC and are summarized in Table 5-2.

[0571] Table 5-2 Summary of characterization of newly prepared salts

[0572]

[0573] *: No detectable solvent residues were found

[0574] HCl salt type A

[0575] HCl salt type A (823121-28-A1) preparation process: 300.8 mg of free base type A material (823121-01-A) and 10 mL of EtOAc were added to a 20-mL glass vial. 68 μL of 12M HCl (1.05 molar ratio) was slowly added to the solution under magnetic stirring. After stirring for 3 days, a solid (275.0 mg, yield -83.8%) was obtained by centrifugation and vacuum drying at room temperature. XRPD results showed Figure 5-1 In, it is shown that HCl salt type A was successfully obtained. The TGA / DSC curve is shown in Figure 5-2 It shows a weight loss of 3.2% up to 120°C and two endotherms (peaks) at 77.7°C and 231.4°C. Figure 5-3 In 1 H NMR results showed that the molar ratio of EtOAc / API was 0.01 (0.2 wt%). UPLC / IC results showed that the molar ratio of acid / FB was 1.0.

[0576] Citrate Type A

[0577] Citrate Form A (823121-28-A3) Preparation: 299.8 mg of free base Form A material (823121-01-A) and 162.9 mg of citric acid were added to a 20-mL glass vial. Then, 10 mL of EtOH was added. After stirring at room temperature (1000 rpm) for 3 days, a solid (320 mg, yield -69.1%) was obtained by centrifugation and vacuum drying at room temperature. The XRPD pattern was shown in Figure 5-4 In the figure, it is shown that citrate type A was successfully obtained. The TGA / DSC curve of citrate type A is shown in Figure 5-5 It shows a weight loss of 2.0% up to 120°C and one endotherm at 164.2°C. Figure 5-6 In 1 HNMR results showed that the molar ratio of acid / FB was 1.0 and no detectable EtOH residue was found.

[0578] Phosphate Type A

[0579] Preparation of Phosphate Type A (823121-32-A): 300.0 mg of free base Type A material (823121-01-A) and 10 mL of EtOH were added to a 20-mL glass vial. 95.4 μL of 85% (2.05 molar ratio) H3PO4 was slowly added to the solution under magnetic stirring. After stirring for 3 days, the solid was separated by centrifugation and continued to be slurried in EtOAc. After slurrying for 2 hours, the solid was dried under vacuum at room temperature after centrifugation to obtain a solid (297.0 mg, yield -66.3%). XRPD results show Figure 5-7 In the figure, it is shown that phosphate type A was successfully obtained. The TGA / DSC curve is shown in Figure 5-8 It shows a weight loss of 0.9% up to 120°C and two endotherms (peaks) at 214.7°C. Figure 5-9 In 1 H NMR results showed no detectable EtOH or EtOAc solvent residues were found. UPLC / IC results showed that the molar ratio of acid / FB was 2.1.

[0580] Example 6

[0581] Salt and Free Base Evaluation

[0582] Three re-prepared salt samples: HCl salt Form A (823121-28-A1), citrate salt Form A (823121-28-A3), and phosphate salt Form A (823121-32-A) were used for evaluations to compare with the free base, including kinetic solubility, hygroscopicity, and solid stability. Free base Form A (200754-075-P3; 823121-01-A) was used in the kinetic solubility evaluation. Free base Form C (200754-075-P5; 823121-16-A) was used in the hygroscopicity evaluation. Two free base batches were used in the solid stability evaluation.

[0583] Kinetic solubility

[0584] The kinetic solubility of the reconstituted HCl salt Form A (823121-28-A1), citrate salt Form A (823121-28-A3), phosphate salt Form A (823121-32-A), and free base Form A (823121-01-A) in water and three biologically relevant media was measured.

[0585] The materials were added to H2O, SGF, FaSSIF or FeSSIF (solid loading: 5 mg / mL based on free base) and then rolled at 25 rpm at 37°C for 1 hour, 2 hours, 4 hours and 24 hours. For each time point, centrifugation and filtration (0.45 μm PTFE filter) were performed. The supernatant was tested for solubility and pH by UPLC. The solids were tested by XRPD. The results are summarized in Table 6-1. The solubility curve is shown in Figure 6-1 The XRPD results are shown in Figure 6-2 to Figure 6-9 Based on the results, all three salts showed higher solubility in water compared to the free base. Both salts and free base were completely dissolved in SGF. In FaSSIF, phosphate salt form A showed higher solubility than other salts and free base, and HCl salt form A, citrate salt form A and free base form A were converted into other crystalline forms of free base after rolling for 1 hour. In FeSSIF, the solubility of these three salts was higher than that of free base, while the solubility of salts and free base became comparable after 24 hours; these three salts were disproportionated into other crystalline forms of free base after rolling for 1 hour.

[0586] Table 6-1 Summary of kinetic solubility tests

[0587]

[0588]

[0589] S: solubility based on free base (mg / mL); FC: form change; -: sample was clear, therefore no solids were detected by XRPD; a : The form changes to type B; b : The form changes to a mixture of type B + type C; c : The form changes to C type; c : The form changes to N type; e : The form changes to a mixture of type N + type B.

[0590] Hygroscopicity

[0591] To evaluate hygroscopicity, the free base Form C, HCl Salt Form A, Citrate Salt Form A, and Phosphate Salt Form A were characterized by DVS. DVS isotherms were collected between 0% RH and 95% RH at 25°C. Since the free base Form C was presumed to be an anhydrous form, the initial RH% of the free base Form C started at 0 RH%, while the initial RH% of the salts started at 50 RH% (room humidity). After the DVS test, the samples were subjected to XRPD characterization. The DVS graphs and XRPD results are shown in Figure 6-10 to Figure 6-17Based on the results, at 25°C / 80%RH, the water absorption of free base Form C, citrate Form A, and phosphate Form A were 0.50%, 0.38%, and 0.97%, respectively, which indicated that they were slightly hygroscopic. HCl salt Form A showed 2.95% water absorption at 25°C / 30%RH, and 4.07% water absorption at 25°C / 80%RH. After DVS testing, no form change was observed for all samples.

[0592] Solid stability

[0593] The free base A / C forms, the re-prepared HCl salt A forms, the citrate salt A forms and the phosphate salt A forms were placed under conditions of 25°C / 60%RH and 40°C / 75%RH for one week for solid stability evaluation. Physical and chemical stability were evaluated by XRPD and purity, respectively. The results are summarized in Table 6-2 and the XRPD results are shown in Figure 6-18 to Figure 6-22 , which showed that after one week of storage at 40°C / 75% RH, the free base Form A converted to Form C, and no form conversion was observed for the other samples. The UPLC results and impurity summary are as follows Figure 6-23 to Figure 6-32 As shown in Tables 6-3 to 6-12, no significant decrease in purity was observed for all samples after one week of storage under both conditions.

[0594] Table 6-2 Summary of solid stability evaluation

[0595]

[0596]

[0597] Table 6-3 Summary of impurities of free base Form A (823121-01-A) after stability evaluation at 25°C / 60%RH for one week

[0598]

[0599] Table 6-4 Summary of impurities of free base Form A (823121-01-A) after stability evaluation at 25°C / 75%RH for one week

[0600]

[0601] Table 6-5 Summary of impurities of free base Form C (823121-16-A) after stability evaluation at 25°C / 60%RH for one week

[0602]

[0603] Table 6-6 Summary of impurities of free base Form C (823121-16-A) after stability evaluation at 40°C / 75%RH for one week

[0604]

[0605] Table 6-7 Summary of impurities of HCl salt Form A (823121-28-A1) after stability evaluation at 25°C / 60%RH for one week

[0606]

[0607] Table 6-8 Summary of impurities of HCl salt Form A (823121-28-A1) after stability evaluation at 40°C / 75%RH for one week

[0608]

[0609] Table 6-9 Summary of impurities of citrate form A (823121-28-A3) after stability evaluation at 25°C / 60%RH for one week

[0610]

[0611]

[0612] Table 6-10 Summary of impurities of citrate form A (823121-28-A3) after stability evaluation at 40°C / 75%RH for one week

[0613]

[0614] Table 6-11 Summary of impurities of phosphate type A (823121-32-A) after stability evaluation at 25°C / 60%RH for one week

[0615]

[0616] Table 6-12 Summary of impurities of phosphate type A (823121-32-A) after stability evaluation at 40°C / 75%RH for one week

[0617]

[0618] Example 7

[0619] Test two batches of material free base A type (200754-075-P3; 823121-01-A) and free base C type (200754-075-P5; 823121-16-A). For free base A type, equilibrium solubility studies were carried out at room temperature (room temperature: 20-22°C) in pH 2.0, 4.0, 6.0, 8.0 buffers and pure water. It was found that the solubility in pH 2.0 and 4.0 was 2.36 mg / mL and 1.47 mg / mL, respectively, which was relatively high compared to the solubility in other media (0.023-0.046 mg / mL).

[0620] Using free base C as the material, 100 polymorph screening experiments were performed. A total of 20 polymorphs were obtained from the screening and further experiments, named free base AT, which were characterized using XRPD, TGA, DSC and 1 H NMR was used for characterization. Based on the characterization results, A, C, P, N and Q forms are anhydrates; B, I and L forms are hydrates; D, F, G, H, J, K, M, O, S and T forms are solvates; E form (converted to M form after drying at 20-22°C / 44-77%RH) and R form (which can be converted to B form after exposure to air) were not tested.

[0621] The interconversion relationships between the anhydrates (free base A, C, P, N and Q forms) were studied via competitive slurrying in EtOH or IPAc at room temperature or 50°C. The results showed that free base form C was obtained in all experiments, indicating that form C was the most stable anhydrate from room temperature to 50°C. Competitive slurrying experiments of free base anhydrate form C, hydrate form B, hydrate form I and hydrate form L in EtOH / H2O solvent system at different water activities (a w The results showed that when a w ≤0.6, the free base type C is obtained; when a w ~0.8 to obtain the free base L-type, and at the same time in pure water (a w Based on the interconversion results, free base Form C showed: good performance in the solid state; more stable than other anhydrates at room temperature and 50°C; stable in a relatively wide water activity range (0-0.6). Therefore, free base Form C is suitable for further development and toxicology studies.

[0622] Using free base Form C as material, salt screening was performed under 52 different conditions using 12 acids (applying different molar ratios of HCl) in 4 solvent systems. During the screening, a total of 11 crystalline salt hits (19 crystalline forms) were obtained, which were characterized by XRPD, TGA, and DSC. UPLC, combined IC or1 H NMR confirmed the salt stoichiometry. Based on the characterization results (small TGA weight loss, pure endothermic signal on the DSC curve, safety level and simple polymorphism), HCl Salt Form A, Citrate Form A and Phosphate Form A were selected for re-preparation, and the results showed that HCl Salt Form A, Citrate Form A and Phosphate Form A were successfully obtained. The three re-prepared salt forms and the free base were evaluated, including kinetic solubility, hygroscopicity and solid stability.

[0623] The kinetic solubility results show that all three salts show higher solubility in water than the free base. Both the salt and the free base are completely dissolved in SGF. In FaSSIF, the phosphate type A shows a higher solubility than other salts and the free base, and the HCl salt type A, the citrate type A and the free base type A are converted into other crystalline forms of the free base after 1 hour. In FeSSIF, the solubility of these three salts is higher than that of the free base, and the solubility of the salt and the free base becomes comparable after 24 hours; these three salts are disproportionated into other crystalline forms of the free base after 1 hour. The hygroscopicity results show that at 25°C / 80%RH, the water absorption rates of the free base type C, the citrate type A and the phosphate type A are 0.50%, 0.38% and 0.97% (slightly hygroscopic), respectively. The HCl salt type A shows a water absorption rate of 2.95% at 25°C / 30%RH, and a water absorption rate of 4.07% at 25°C / 80%RH. The solid stability evaluation results showed that the free base Form A was converted to Form C after storage at 40°C / 75%RH for one week. No form change was observed for the other samples. After storage at 25°C / 60%RH or 40°C / 75%RH for one week, no significant purity decrease was observed for all samples.

[0624] Based on the interconversion results, the free base Form C is suitable for further development and toxicology studies. If considering developing a salt as a backup to the free base Form C, the salt candidate can be selected based on the kinetic solubility assessment and PK results.

[0625] Example 8

[0626] Approximate solubility of free base Type A material (823121-01-A)

[0627] At room temperature, the solubility of the free base A-type material (823121-01-A) in 20 solvents was estimated. About 2 mg of solid was added to a 3-mL glass vial. Then the solvent was added to the vial stepwise (each step 50-50-100-100-200-500-500-500 μL) until the solid dissolved or reached a total volume of 2 mL. The results summarized in Table 8-1 are used to guide solvent selection in polymorph screening.

[0628] Table 8-1 Approximate solubility of free base type A material (823121-01-A) at room temperature

[0629]

[0630]

[0631] *: Abbreviation of the solvent used that dissolved after storage at 50°C overnight

[0632] The solvent abbreviations are listed in Table 8-2.

[0633] Table 8-2 List of solvent abbreviations

[0634]

[0635] Free base polymorph screening

[0636] A total of 100 polymorph screening experiments were performed using different crystallization or solid transformation methods. The methods used and the results are summarized in Table 8-3.

[0637] Table 8-3 Summary of polymorph screening experiments

[0638]

[0639]

[0640] Antisolvent addition

[0641] A total of 12 antisolvent addition experiments were performed. About 20 mg of the material (823121-16-A) was dissolved in the corresponding solvent to obtain a clear solution, and the solution was magnetically stirred (~1000 rpm), and then the antisolvent was added until the precipitant appeared or the total amount of the antisolvent reached 10 mL. The solid was isolated for XRPD analysis. The results in Table 8-4 show that free base C, F, L, B type + additional peaks and C type + additional peaks were obtained.

[0642] Table 8-4 Summary of Antisolvent Addition Experiment

[0643]

[0644] *: the sample was obtained by slurrying at 5°C; **: the solution was clear at 5°C and the sample was obtained after evaporation at room temperature;

[0645] Solid Vapor Diffusion

[0646] Use 9 kinds of different solvents to carry out solid vapor diffusion experiment.About 20mg of material (823121-16-A) is weighed in a 3-mL bottle, and this bottle is put into a 20-mL bottle of the volatile solvent of 4mL. Seal the 20-mL bottle with a lid and remain at room temperature, allowing solvent vapor to interact with the sample. Test solid by XRPD and the result of summarizing in table 8-5 shows that free alkali C / G / I / O type is obtained.

[0647] Table 8-5 Summary of solid vapor diffusion experiments

[0648]

[0649] Slurry at room temperature

[0650] A total of 25 slurry experiments were performed at room temperature using different solvents. Approximately 20 mg of material (823121-16-A) was suspended in the solvent of 0.5 mL in a HPLC glass vial. After 5 days of magnetic stirring (1000 rpm) of the suspension at room temperature, the remaining solid was centrifuged for XRPD analysis. The results summarized in Table 8-6 show that free alkali C / D / F / I / J / K / L type is generated.

[0651] Table 8-6 Summary of slurry conversion experiment at room temperature

[0652]

[0653]

[0654] *: A clear solution was obtained at room temperature and then transferred to 5°C. # Since all free base Form I samples were obtained from experimental systems involving MeOH, further experiments with compound HY010101 in MeOH were investigated, see Examples.

[0655] Slurry at 50°C

[0656] A total of 18 slurry experiments were performed at 50°C using different solvents. Approximately 20 mg of material (823121-16-A) was suspended in 0.5 mL of solvent in a HPLC glass vial. After the suspension was stirred (1000 rpm) at 50°C for approximately 5 days, the remaining solid was centrifuged for XRPD analysis. The results summarized in Table 8-7 show that free base C / D / K / H / I / J / N forms were generated.

[0657] Table 8-7 Summary of slurry conversion experiments at 50°C

[0658]

[0659]

[0660] *: A clear solution was obtained at 50°C, then transferred and stirred at 5°C for one day. # Since all free base Form I samples were obtained from experimental systems involving MeOH, further experiments with compound HY010101 in MeOH were investigated, see Examples.

[0661] Slow evaporation

[0662] Slow evaporation experiments were performed under 7 conditions. About 20 mg of the material (823121-16-A) was dissolved in the corresponding solvent in a 3-mL glass vial. The resulting solution was heated at room temperature with The vial was sealed and punctured with several pinholes for slow evaporation. The solid was isolated for XRPD analysis and the results summarized in Table 8-8 indicated that the free base C / G / F form was obtained.

[0663] Table 8-8 Summary of slow evaporation experiment

[0664]

[0665] Slow cooling

[0666] Slow cooling experiments were carried out in 6 solvent systems. At room temperature, about 20 mg of material (823121-16-A) was suspended in 1.0 mL of solvent in a 3-mL glass vial. The suspension was then heated to 50 ° C, balanced for about 1 h and filtered to a new vial (0.45 μm PTFE membrane). The filtrate was slowly cooled to 5 ° C at a rate of 0.1 ° C / min. The solid obtained was kept isothermal at 5 ° C and then tested by XRPD solid. The results summarized in Table 8-9 show free alkali H / G / I / C / C+J type.

[0667] Table 8-9 Summary of slow cooling experiment

[0668]

[0669] Liquid vapor diffusion

[0670] A total of 12 liquid vapor diffusion experiments were carried out. About 20 mg of material (823121-16-A) was dissolved in a suitable solvent to obtain a clear solution in a 3-mL vial. The solution was then placed in a 20-mL vial containing 4 mL of a volatile solvent. The 20-mL vial was sealed with a lid and maintained at room temperature, allowing enough time for organic vapor to interact with the solution. Separation precipitation agents were used for XRPD analysis. The results summarized in Table 8-10 showed that free alkali O / C / E / F / C+N type was observed.

[0671] Table 8-10 Summary of Liquid Vapor Diffusion Experiment

[0672]

[0673] *: Antisolvent was added to give a clear solution, which was then transferred to room temperature for evaporation. **: Free base Form E was transferred to Form M after drying at room temperature (temperature range: 20-22°C; humidity range: 44-77% RH).

[0674] Polymer-induced crystallization

[0675] A total of 6 polymer-induced crystallization experiments were performed using two sets of polymer mixtures in different solvent systems. Approximately 20 mg of material (823121-16-A) was dissolved in the corresponding solvent in a 3-mL glass vial. Approximately 2 mg of the polymer mixture was added to the 3-mL glass vial. The resulting solution was heated at room temperature with The sealed vial was evaporated to slow evaporation. The solid was isolated for XRPD analysis. The results summarized in Tables 8-11 show that the free base B / C / H / C form + additional peaks and amorphous form were obtained.

[0676] Table 8-11 Summary of polymer induced crystallization experiments

[0677]

[0678] Polymer mixture A: polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA), polyvinyl chloride (PVC), polyvinyl acetate (PVAC), hydroxypropyl methylcellulose (HPMC), methyl cellulose (MC) (mass ratio is 1:1:1:1:1:1:1).

[0679] Polymer mixture B: polycaprolactone (PCL), polyethylene glycol (PEG), polymethyl methacrylate (PMMA), sodium alginate (SA) and hydroxyethyl cellulose (HEC) (mass ratio is 1:1:1:1:1).

[0680] Grinding

[0681] A total of 5 grinding experiments were performed with or without the addition of solvent. About 15 mg of material (823121-16-A) was weighed into a mortar. 10 μL of solvent was added to the mortar. The solid was ground for 3-5 min. The solid was isolated for XRPD analysis. The results summarized in Tables 8-12 show that only free base Form C was obtained.

[0682] Table 8-12 Grinding experiment summary

[0683]

[0684] Free base T-solvate

[0685] The samples (A09954-038-A4; 823121-63-A) were characterized by XRPD. The XRPD results are shown in Figure 8-1, indicating a new form, named free base T-form. TGA / DSC and 1 HNMR data showed a weight loss of 6.0% up to 120°C and two endotherms (peaks) at 109.2°C and 176.2°C; the residual acetone / API molar ratio was 0.5 (7.0 wt%). XRPD results (Figure 8-2) showed that the free base Form T converted to Form C after heating to 120°C and then cooling to room temperature. 1 H NMR results (Figure 8-3) show that the residual acetone is reduced to 0.003 (acetone / API molar ratio). Combined with the form change after heating, 1 Based on the residual solvent (before and after heating) and TGA weight loss in the H NMR results, the free base T-form was presumed to be an acetone solvate.

[0686] Salt Hits

[0687] A total of 11 salt hits (19 crystalline forms) were obtained from the salt screening and characterized by XRPD, TGA and DSC. 1 H NMR was used to determine the salt stoichiometry. All characterization results are summarized in 8-13.

[0688] Table 8-13 Summary of salt hits characterization

[0689]

[0690]

[0691] *: Exothermic (peak).

[0692] HCl salt

[0693] HCl salt Form A / B (823121-06-D1 / D2) was obtained by dissolving 20 mg of free base Form A (823121-01-A) in 2-MeTHF, then adding HCl (molar ratios of 1:1 and 1:2 acid / FB, respectively) at room temperature for slurrying for 3 days and drying under vacuum. XRPD overlays are shown in Figure 8-4.

[0694] The TGA / DSC curve of HCl Salt Form A (823121-06-D1) is shown in FIG8-5 , which shows a weight loss of 3.7% up to 120° C. and three endotherms (peaks) at 83.9° C., 232.2° C., and 243.7° C. 1 H NMR results (FIG. 8-6) showed that no 2-MeTHF residue was detected. UPLC / IC results showed a molar ratio of 1.0:1.0 (acid / FB). VT-XRPD results (FIG. 8-7) showed a peak shift after N2 purging for 20 min, and a peak shift was also observed after heating to 120°C and cooling to 30°C under N2 atmosphere. After the sample was exposed to air for 10 min, HCl salt Form A was reformed. Therefore, HCl salt Form A is presumed to be anhydrous.

[0695] The TGA / DSC curve of HCl Salt Form B (823121-06-D2) is shown in Figure 8-8, which shows a weight loss of 7.0% up to 120°C and three endotherms (peaks) at 104.4°C, 174.0°C and 224.9°C. 1 H NMR results (Figures 8-9) showed no detectable 2-MeTHF residual. UPLC / IC results showed a molar ratio of 1.1:1.0 (acid / FB).

[0696] Sulfate

[0697] Sulfate salt Form A (823121-06-C3) was obtained by dissolving 20 mg of free base Form A (823121-01-A) in EtOAc, then adding H2SO4 (molar ratio 1:1, acid / FB) for slurrying at room temperature for 3 days, and drying under vacuum. XRPD overlays are shown in Figures 8-10. The TGA / DSC curve of Sulfate Salt Form A (823121-06-C3) is shown in Figures 8-11, which shows a weight loss of 6.5% up to 120°C and three endotherms (peaks) at 92.2°C and 145.0°C. 1 HNMR results (Figures 8-12) showed that no EtOAc residue was detected. UPLC / IC results showed that the molar ratio was 1.0:1.0 (acid / FB).

[0698] Maleate

[0699] Maleate salt A / B / C forms (823121-06-A4 / D4 / C4) were prepared by dissolving 20 mg of free base A form (823121-01-A) and maleic acid (molar ratio 1:1, acid / FB) in EtOH, 2-MeTHF and EtOAc, slurrying at room temperature for 3 days and drying under vacuum. XRPD overlays are shown in Figures 8-13.

[0700] The TGA / DSC curve of maleate salt Form A (823121-06-A4) is shown in FIG8-14, which shows a weight loss of 11.0% up to 120°C and an endotherm (peak) at 120.2°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.1:1.0, and the molar ratio of EtOH / API was 0.4:1.0 (3.2 wt %).

[0701] The TGA / DSC curve of maleate salt Form B (823121-06-D4) is shown in FIG8-16, which shows a weight loss of 2.1% up to 120°C and an endotherm (peak) at 177.4°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable 2-MeTHF residue was found.

[0702] The TGA / DSC curve of maleate salt form C (823121-06-C4) is shown in FIG8-18, which shows a weight loss of 3.7% up to 120°C and an endotherm (peak) at 166.1°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable EtOAc residue was found.

[0703] Phosphate

[0704] Phosphate Form A (823121-06-A6) was obtained by dissolving 20 mg of free base Form A (823121-01-A) in EtOH, then adding H3PO4 (molar ratio 1:1, acid / FB) for slurrying at room temperature for 3 days, and drying under vacuum. The XRPD overlay is shown in Figure 8-20. The TGA / DSC curve of Phosphate Form A (823121-06-A6) is shown in Figure 8-21. The TGA results show a weight loss of 2.6% up to 120°C. The DSC results show an endotherm (peak) at 214.8°C. 1 The H NMR results (Figure 8-22) showed that no EtOH residue was detected. The UPLC / IC results showed that the molar ratio was 2.0:1.0 (acid / FB).

[0705] L-Tartrate

[0706] L-Tartrate A / B Form (823121-06-A7 / C7) was obtained by dissolving 20 mg of free base A Form (823121-01-A) and L-tartaric acid (molar ratio 1:1, acid / FB) in EtOH and EtOAc, slurrying at room temperature for 3 days and drying under vacuum. XRPD overlays are shown in Figure 8-23.

[0707] The TGA / DSC curve of L-tartrate salt form A (823121-06-A7) is shown in FIG8-24, which shows a weight loss of 3.7% up to 120°C and two endotherms (peaks) at 170.0°C and 190.6°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and the molar ratio of EtOH / API was 0.13:1.0 (1.1 wt %).

[0708] The TGA / DSC curve of L-tartrate Form B (823121-06-C7) is shown in FIG8-26, which shows a weight loss of 5.9% up to 120°C and an endotherm (peak) at 198.1°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and the molar ratio of EtOAc / API was 0.13:1.0 (0.2 wt %).

[0709] Fumarate

[0710] Fumarate Form A (823121-06-A8) was obtained by dissolving 20 mg of free base Form A (823121-01-A) and fumaric acid (molar ratio 1:1, acid / FB) in EtOH for slurrying at room temperature for 3 days and drying under vacuum. The XRPD overlay is shown in Figure 8-28. The TGA / DSC curve of Fumarate Form A (823121-06-A8) is shown in Figure 8-29, which shows a weight loss of 4.8% up to 120°C and endotherms (peaks) at 120.7°C and 201.5°C. The XRPD overlay in Figure 8-30 shows a weight loss of 4.8% up to 120°C and endotherms (peaks) at 120.7°C and 201.5°C. 1 H NMR results showed that the molar ratio of acid / FB was 0.9:1.0 and the molar ratio of EtOH / API was 0.03:1 (0.3 wt %).

[0711] Citrate

[0712] Citrate Form A (823121-06-C9) was obtained by dissolving 20 mg of free base Form A (823121-01-A) and citric acid (molar ratio 1:1, acid / FB) in EtOAc for slurrying at room temperature for 3 days and drying under vacuum. The XRPD overlay is shown in Figure 8-31. The TGA / DSC curve of Citrate Form A (823121-06-C9) is shown in Figure 8-32, which shows a weight loss of 2.3% up to 120°C and endotherms (peaks) at 167.4°C and 181.0°C. The XRPD overlay in Figure 8-33 shows a weight loss of 2.3% up to 120°C and endotherms (peaks) at 167.4°C and 181.0°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable EtOAc residue was found.

[0713] Tosylate

[0714] Tosylate salt Form A (823121-06-A10) was obtained by dissolving 20 mg of free base Form A (823121-01-A) and p-toluenesulfonic acid (molar ratio 1:1, acid / FB) in EtOH for slurrying at room temperature for 3 days and drying under vacuum. The XRPD overlay is shown in Figure 8-34. The TGA / DSC curve of Tosylate Salt Form A (823121-06-A10) is shown in Figure 8-35, which shows a weight loss of 2.5% up to 120°C and two endotherms (peaks) at 81.2°C and 242.9°C. The XRPD overlay in Figure 8-36 shows a weight loss of 2.5% up to 120°C and two endotherms (peaks) at 81.2°C and 242.9°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable EtOH residue was found.

[0715] Methanesulfonate

[0716] Methanesulfonate Form A / B (823121-06-C11 / D11) was obtained by slurrying 20 mg of free base Form A (823121-01-A) and methanesulfonic acid (molar ratio 1:1, acid / FB) in EtOH and 2-MeTHF, respectively, at room temperature for 3 days and drying under vacuum. After storage for 8 days (temperature range: 20-23°C; humidity range: 63-87% RH), mesylate Form A (823121-06-C11) was partially converted to mesylate Form B. Therefore, it was not analyzed by TGA / DSC and 1 H NMR was used for further characterization. The XRPD overlay is shown in Figure 8-37. The TGA / DSC curve of the mesylate salt Form B (823121-06-D11) is shown in Figure 8-38, which shows a weight loss of 8.8% up to 120°C and an endotherm (peak) at 111.6°C. 1H NMR results showed that the molar ratio of acid / FB was 1.1:1.0, and no detectable 2-MeTHF residue was found.

[0717] Benzenesulfonate

[0718] Benzenesulfonate salt A / B / C forms (823121-06-A12 / B12 / C12) were prepared by dissolving 20 mg of free base A form (823121-01-A) and benzenesulfonic acid (molar ratio 1:1, acid / FB) in EtOH, acetone / H2O (19:1, v / v) and EtOAc, respectively, for slurrying at room temperature for 3 days and drying under vacuum. XRPD overlays are shown in Figures 8-40.

[0719] The TGA / DSC curve of benzenesulfonate salt form A (823121-06-A12) is shown in FIG8-41, which shows a weight loss of 6.7% up to 120°C and three endotherms (peaks) at 75.6°C, 177.3°C and 192.8°C. 1 HNMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable EtOH residue was found.

[0720] The TGA / DSC curve of benzenesulfonate salt Form B (823121-06-B12) is shown in FIG8-43, which shows a weight loss of 2.1% up to 120°C and three endotherms (peaks) at 78.9°C, 176.6°C and 192.4°C. 1 HNMR results showed that the molar ratio of acid / FB was 1.0:1.0, and the molar ratio of acetone / API was 0.07:1.0 (0.8 wt %).

[0721] The TGA / DSC curve of benzenesulfonate salt form C (823121-06-C12) is shown in FIG8-45, which shows a weight loss of 2.0% up to 120°C and an endotherm (peak) at 191.9°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and the molar ratio of EtOAc / API was 0.02:1.0 (0.3 wt %).

[0722] Gentisate

[0723] Gentisate Form A (823121-06-C13) was obtained by dissolving 20 mg of free base Form A (823121-01-A) and gentisic acid (molar ratio 1:1, acid / FB) in EtOAc for slurrying at room temperature for 3 days and drying under vacuum. The XRPD overlay is shown in Figure 8-47. The TGA / DSC curve of Gentisate Form A (823121-06-C13) is shown in Figure 8-48, which shows a weight loss of 2.3% up to 120°C and an endotherm (peak) at 214.6°C. The XRPD overlay in Figure 8-49 shows a weight loss of 2.3% up to 120°C and an endotherm (peak) at 214.6°C. 1 H NMR results showed that the molar ratio of acid / FB was 1.0:1.0, and no detectable EtOAc residue was found.

[0724] Impurities Summary

[0725] Since an impurity signal at RRT = 1.10 (UPLC) was observed in the supernatant in both equilibrium and kinetic solubility experiments of free base Form A (823121-01-A), those samples were characterized by LC-MS (Liquid Chromatography-Mass Spectrometry).

[0726] UPLC results for samples from equilibrium solubility experiments in water, pH 6.0 buffer, and pH 8.0 buffer are shown in Figures 8-50.

[0727] LC-MS results of samples from equilibrium solubility experiments in water, pH 6.0 buffer, and pH 8.0 buffer are shown in Figures 8-51 to 8-53.

[0728] Results from water, pH 6.0 buffer and pH MS are shown in Figures 8-54 to 8-55.

[0729] Slurry of free base Form C in water.

[0730] To investigate whether free base type C can produce the same impurity (RRT = 1.10), slurry experiments in ultrapure water were performed using free base type C material (823121-16-A). The material (solid loading: 10 mg / mL) was added to H2O and then slurried (1000 rpm) at room temperature for 24 hours (823121-43-B) or 6 days (823121-43-A). After centrifugation and filtration (0.45 μm PTFE filter), the solubility and pH of the supernatant were tested, and the residual solids were tested by XRPD. The results are summarized in Table 8-14. The XRPD results show that after slurrying in H2O for 24 hours or 6 days, the form becomes free base type B. The UPLC results (Figure 8-57) show that the impurity (RRT = 1.10) is observed again in the corresponding filtrate. The residual solids tested by UPLC, shown in Figure 8-58, Figure 8-59, Table 8-15 and Table 8-16, showed that no impurities were detected (RRT = 1.10).

[0731] Table 8-14 Summary of equilibrium solubility tests for free base type C

[0732]

[0733]

[0734] Table 8-15 Summary of impurities in the residual solids after slurrying the free base Form C in water for 24 hours

[0735]

[0736] Table 8-16 Summary of impurities in the residual solids after slurrying the free base Form C in water for 6 days

[0737]

[0738] Preparation of biologically relevant culture media

[0739] Simulated gastric fluid (SGF)

[0740] 200 mg of NaCl and 100 mg of Triton X-100 were weighed into a 100-mL volumetric flask. Purified water was added to dissolve the solid. 136 μL of HCl (12 M) was added after the solid was completely dissolved, and then the pH was adjusted to 1.8. Purified water was added to dissolve the solid.

[0741] Fasting state simulated intestinal fluid (FaSSIF)

[0742] 0.17 g of anhydrous NaH2PO4, 0.021 g of NaOH and 0.31 g of NaCl were weighed into a 50-mL volumetric flask. 48 mL of purified water was added to dissolve the solid. 1 M HCl or 1 M NaOH was used to adjust the pH to 6.5. Purified water was then added to the volume. 22 mg of SIF powder was added to a 10-mL volumetric flask, and the prepared buffer was then added to the volume.

[0743] Fed State Simulated Intestinal Fluid (FeSSIF)

[0744] 0.41 mL of acetic acid, 202 mg of NaOH and 594 mg of NaCl are weighed into a 50-mL volumetric flask. 48 mL of purified water is added to dissolve the solid. 1 M HCl or 1 M NaOH is used to adjust the pH to 5.0. Purified water is then added to the volume. 0.56 g of SIF powder is added into a 10-mL volumetric flask, and the prepared buffer is then added to the volume.

[0745] pH Buffers

[0746] pH 2.0 buffer: Add 149.1 mg of KCl and 0.5 mol of 1M HCl solution to a 50-mL volumetric flask. Then add purified water to the volume. The pH is 2.04.

[0747] pH 4.0 buffer: Add 283.4 mg of anhydrous citric acid and 301.5 mg of trisodium citrate dihydrate to a 50-mL volumetric flask. Then add purified water to the volume. The pH is 4.00.

[0748] pH 6.0 buffer: Add 55.2 mg of anhydrous citric acid and 650.7 mg of trisodium citrate dihydrate to a 50-mL volumetric flask. Then add purified water to the volume. The pH is 5.95.

[0749] pH 8.0 buffer: Add 336.1 mg Na2HPO4 and 15.9 mg NaH2PO4 to a 50-mL volumetric flask. Then add purified water to the volume. The pH is 8.01.

[0750] Instruments and methods

[0751] XRPD

[0752] For XRPD analysis, PANalytical Empyrean and X'Pert3 X-ray powder diffractometers were used. The XRPD parameters used are listed in Tables 8-17.

[0753] Table 8-17 Parameters of XRPD test

[0754]

[0755] TGA and DSC

[0756] TGA data were collected using a TA Q5000 / Discovery 5500 TGA from TA Instruments. The detailed parameters used for DSC using a TA Discovery 2500 DSC from TA Instruments are listed in Tables 8-18.

[0757] Table 8-18 Parameters of TGA and DSC tests

[0758]

[0759] DVS

[0760] DVS was measured by SMS (Surface Measurement Systems) DVS Intrinsic. Relative humidity at 25°C was calibrated based on the deliquescent points of LiCl, Mg(NO3)2 and KCl. The parameters of the DVS test are listed in Tables 8-19.

[0761] Table 8-19 DVS test parameters

[0762]

[0763] Solution NMR

[0764] Solution NMR was collected on a Bruker 400M NMR spectrometer using DMSO-d6.

[0765] HPLC / UPLC / IC / LC-MS

[0766] In table 8-20, list the HPLC method for the purity test of free alkali A-type (823121-01-A) and the equilibrium solubility experiment in pH 2.0 buffer and pH 4.0 buffer. Remaining solubility and purity test are carried out by the UPLC method listed in table 8-21.Use UPLC (table 8-21) and IC (table 8-22) to determine the salt stoichiometry.LC-MS method is listed in table 8-23.

[0767] Table 8-20 HPLC method

[0768]

[0769] Table 8-21 UPLC method

[0770]

[0771]

[0772] Table 8-22 IC parameters for chemometric testing

[0773]

[0774] Table 8-23 LC-MS method

[0775]

[0776]

[0777] Although preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. The present invention is not intended to be limited by the specific embodiments provided in this specification. Although the present invention has been described with reference to the foregoing description, the description and illustration of the embodiments herein are not meant to be interpreted in a limiting sense. Without departing from the present invention, those skilled in the art will appreciate many variations, changes and substitutions. In addition, it should be understood that all aspects of the present invention are not limited to the specific depictions, configurations or relative proportions depending on various conditions and variables set forth herein. It should be understood that various alternatives to the embodiments of the present invention described herein can be adopted when practicing the present invention. Therefore, it is contemplated that the present invention should also encompass any such substitutions, modifications, variations or equivalents. The following claims are intended to define the scope of the present invention, and thus encompass methods and structures within the scope of these claims and their equivalents.

Claims

1. Compound HY-A, a crystalline form of 2-(4-fluorophenyl)-3-(2-methylpyridin-4-yl)-5-(methylsulfonyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine, wherein: The crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of: (1) 11.3±0.3 degrees, 22.7±0.3 degrees, 23.2±0.3 degrees; (2) 14.4±0.3 degrees, 19.2±0.3 degrees, 21.6±0.3 degrees; (3) 12.8±0.3 degrees, 16.7±0.3 degrees, 19.2±0.3 degrees; (4) 7.5±0.3 degrees, 7.8±0.3 degrees, 19.7±0.3 degrees; (5) 10.1±0.3 degrees, 12.5±0.3 degrees, 19.7±0.3 degrees; (6) 11.5±0.3 degrees, 17.5±0.3 degrees, 24.5±0.3 degrees; (7) 13.9±0.3 degrees, 20.6±0.3 degrees, 23.8±0.3 degrees; (8) 7.0±0.3 degrees, 12.6±0.3 degrees, 19.5±0.3 degrees; (9) 7.1±0.3 degrees, 10.9±0.3 degrees, 12.3±0.3 degrees; (10) 6.4±0.3 degrees, 15.6±0.3 degrees, 20.0±0.3 degrees; (11) 8.6±0.3 degrees, 13.2±0.3 degrees, 20.7±0.3 degrees; (12) 5.6±0.3 degrees, 17.1±0.3 degrees, 21.4±0.3 degrees; (13) 5.6±0.3 degrees, 17.1±0.3 degrees, 22.5±0.3 degrees; (14) 18.9±0.3 degrees, 19.7±0.3 degrees, 25.0±0.3 degrees; (15) 5.3±0.3 degrees, 8.7±0.3 degrees, 17.2±0.3 degrees; (16) 7.9±0.3 degrees, 18.2±0.3 degrees, 18.7±0.3 degrees; (17) 5.5±0.3 degrees, 12.3±0.3 degrees, 17.2±0.3 degrees; (18) 5.2 ± 0.3 degrees, 10.4 ± 0.3 degrees, 26.1 ± 0.3 degrees; and (19) 6.5±0.3 degrees, 18.7±0.3 degrees, 22.2±0.3 degrees.

2. The crystalline form according to claim 1, wherein The crystalline form further exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks selected from the group consisting of: (1) 5.6±0.3 degrees, 19.0±0.3 degrees, 19.4±0.3 degrees; (2) 13.6±0.3 degrees, 17.9±0.3 degrees, 27.4±0.3 degrees; (3) 14.7±0.3 degrees, 15.1±0.3 degrees, 26.3±0.3 degrees; (4) 6.6±0.3 degrees, 12.5±0.3 degrees, 23.4±0.3 degrees; (5) 8.8±0.3 degrees, 17.5±0.3 degrees, 23.5±0.3 degrees; (6) 6.3±0.3 degrees, 17.8±0.3 degrees, 21.0±0.3 degrees; (7) 8.9±0.3 degrees, 17.3±0.3 degrees, 27.0±0.3 degrees; (8) 13.9±0.3 degrees, 18.4±0.3 degrees, 21.0±0.3 degrees; (9) 18.9±0.3 degrees, 20.6±0.3 degrees, 27.0±0.3 degrees; (10) 10.6±0.3 degrees, 13.1±0.3 degrees, 16.8±0.3 degrees; (11) 5.3±0.3 degrees, 12.7±0.3 degrees, 17.5±0.3 degrees; (12) 8.7±0.3 degrees, 12.6±0.3 degrees, 24.7±0.3 degrees; (13)12.6±0.3 degrees; (14) 11.5±0.3 degrees, 17.5±0.3 degrees, 21.7±0.3 degrees; (15) 9.2±0.3 degrees, 12.3±0.3 degrees, 21.4±0.3 degrees; (16) 12.7±0.3 degrees, 20.7±0.3 degrees, 23.4±0.3 degrees; (17) 9.1±0.3 degrees, 18.5±0.3 degrees, 25.1±0.3 degrees; (18) 15.6 ± 0.3 degrees, 18.8 ± 0.3 degrees, 20.8 ± 0.3 degrees; and (19) 17.7±0.3 degrees, 21.9±0.3 degrees, 24.0±0.3 degrees.

3. The crystalline form according to any one of claims 1 to 2, wherein The crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of: (1) 13.9±0.3 degrees, 18.6±0.3 degrees, 25.2±0.3 degrees; (2) 9.9±0.3 degrees, 10.4±0.3 degrees, 23.2±0.3 degrees; (3) 19.7±0.3 degrees, 20.6±0.3 degrees, 21.0±0.3 degrees; (4) 11.7±0.3 degrees, 17.5±0.3 degrees, 21.9±0.3 degrees; (5) 6.3±0.3 degrees, 21.8±0.3 degrees, 22.1±0.3 degrees; (6) 19.4±0.3 degrees, 19.8±0.3 degrees, 25.2±0.3 degrees; (7) 15.2±0.3 degrees, 17.9±0.3 degrees, 24.6±0.3 degrees; (8) 9.2±0.3 degrees, 16.0±0.3 degrees, 21.7±0.3 degrees; (9) 14.2±0.3 degrees, 17.9±0.3 degrees, 25.3±0.3 degrees; (10) 9.5±0.3 degrees, 16.2±0.3 degrees, 19.6±0.3 degrees; (11) 16.6±0.3 degrees, 20.3±0.3 degrees, 21.4±0.3 degrees; (12) 19.3±0.3 degrees, 26.5±0.3 degrees; (14) 12.0±0.3 degrees, 16.3±0.3 degrees, 22.3±0.3 degrees; (15) 17.5±0.3 degrees, 19.3±0.3 degrees, 26.8±0.3 degrees; (16) 8.5±0.3 degrees, 21.9±0.3 degrees, 23.8±0.3 degrees; (18) 28.1 ± 0.3 degrees; and (19) 15.6±0.3 degrees, 19.2±0.3 degrees, 26.3±0.3 degrees.

4. The crystalline form according to any one of claims 1 to 3, wherein The crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2-theta degrees, the characteristic peaks being selected from the group consisting of: (1) 11.3±0.3 degrees, 22.7±0.3 degrees, 23.2±0.3 degrees, 5.6±0.3 degrees, 19.0±0.3 degrees, 19.4±0.3 degrees, 13.9±0.3 degrees, 18.6±0.3 degrees, 25.2±0.3 degrees; (2) 14.4±0.3 degrees, 19.2±0.3 degrees, 21.6±0.3 degrees, 13.6±0.3 degrees, 17.9±0.3 degrees, 27.4±0.3 degrees, 9.9±0.3 degrees, 10.4±0.3 degrees, 23.2±0.3 degrees; (3) 12.8±0.3 degrees, 16.7±0.3 degrees, 19.2±0.3 degrees, 14.7±0.3 degrees, 15.1±0.3 degrees, 26.3±0.3 degrees, 19.7±0.3 degrees, 20.6±0.3 degrees, 21.0±0.3 degrees; (4) 7.5±0.3 degrees, 7.8±0.3 degrees, 19.7±0.3 degrees, 6.6±0.3 degrees, 12.5±0.3 degrees, 23.4±0.3 degrees, 11.7±0.3 degrees, 17.5±0.3 degrees, 21.9±0.3 degrees; (5) 10.1±0.3 degrees, 12.5±0.3 degrees, 19.7±0.3 degrees, 8.8±0.3 degrees, 17.5±0.3 degrees, 23.5±0.3 degrees, 6.3±0.3 degrees, 21.8±0.3 degrees, 22.1±0.3 degrees; (6) 11.5±0.3 degrees, 17.5±0.3 degrees, 24.5±0.3 degrees, 6.3±0.3 degrees, 17.8±0.3 degrees, 21.0±0.3 degrees, 19.4±0.3 degrees, 19.8±0.3 degrees, 25.2±0.3 degrees; (7) 13.9±0.3 degrees, 20.6±0.3 degrees, 23.8±0.3 degrees, 8.9±0.3 degrees, 17.3±0.3 degrees, 27.0±0.3 degrees, 15.2±0.3 degrees, 17.9±0.3 degrees, 24.6±0.3 degrees; (8) 7.0±0.3 degrees, 12.6±0.3 degrees, 19.5±0.3 degrees, 13.9±0.3 degrees, 18.4±0.3 degrees, 21.0±0.3 degrees, 9.2±0.3 degrees, 16.0±0.3 degrees, 21.7±0.3 degrees; (9) 7.1±0.3 degrees, 10.9±0.3 degrees, 12.3±0.3 degrees, 18.9±0.3 degrees, 20.6±0.3 degrees, 27.0±0.3 degrees, 14.2±0.3 degrees, 17.9±0.3 degrees, 25.3±0.3 degrees; (10) 6.4±0.3 degrees, 15.6±0.3 degrees, 20.0±0.3 degrees, 10.6±0.3 degrees, 13.1±0.3 degrees, 16.8±0.3 degrees, 9.5±0.3 degrees, 16.2±0.3 degrees, 19.6±0.3 degrees; (11) 8.6±0.3 degrees, 13.2±0.3 degrees, 20.7±0.3 degrees, 5.3±0.3 degrees, 12.7±0.3 degrees, 17.5±0.3 degrees, 16.6±0.3 degrees, 20.3±0.3 degrees, 21.4±0.3 degrees; (12) 5.6±0.3 degrees, 17.1±0.3 degrees, 21.4±0.3 degrees, 8.7±0.3 degrees, 12.6±0.3 degrees, 24.7±0.3 degrees, 19.3±0.3 degrees, 26.5±0.3 degrees; (13) 5.6±0.3 degrees, 17.1±0.3 degrees, 22.5±0.3 degrees, 12.6±0.3 degrees; (14) 18.9±0.3 degrees, 19.7±0.3 degrees, 25.0±0.3 degrees, 11.5±0.3 degrees, 17.5±0.3 degrees, 21.7±0.3 degrees, 12.0±0.3 degrees, 16.3±0.3 degrees, 22.3±0.3 degrees; (15) 5.3±0.3 degrees, 8.7±0.3 degrees, 17.2±0.3 degrees, 9.2±0.3 degrees, 12.3±0.3 degrees, 21.4±0.3 degrees, 17.5±0.3 degrees, 19.3±0.3 degrees, 26.8±0.3 degrees; (16) 7.9±0.3 degrees, 18.2±0.3 degrees, 18.7±0.3 degrees, 12.7±0.3 degrees, 20.7±0.3 degrees, 23.4±0.3 degrees, 8.5±0.3 degrees, 21.9±0.3 degrees, 23.8±0.3 degrees; (17) 5.5±0.3 degrees, 12.3±0.3 degrees, 17.2±0.3 degrees, 9.1±0.3 degrees, 18.5±0.3 degrees, 25.1±0.3 degrees; (18) 5.2 ± 0.3 degrees, 10.4 ± 0.3 degrees, 26.1 ± 0.3 degrees, 15.6 ± 0.3 degrees, 18.8 ± 0.3 degrees, 20.8 ± 0.3 degrees, 28.1 ± 0.3 degrees; and (19) 6.5±0.3 degrees, 18.7±0.3 degrees, 22.2±0.3 degrees, 17.7±0.3 degrees, 21.9±0.3 degrees, 24.0±0.3 degrees, 15.6±0.3 degrees, 19.2±0.3 degrees, 26.3±0.3 degrees.

5. The crystalline form according to any one of claims 1 to 4, wherein The crystalline form exhibits a mass loss selected from the following in TGA (thermogravimetric) analysis: (1) 3.2% mass loss up to 120°C; (2) 2% mass loss up to 120°C; (3) 0.9% mass loss up to 120°C; (4) 2.2% mass loss up to 150°C; (5) 0.6% mass loss up to 120°C; (6) 7% mass loss up to 120°C; (7) 3.5% mass loss up to 120°C; (8) 1.1% mass loss up to 120°C; (9) up to 13.6% mass loss at 120°C and / or up to 4.3% mass loss at 120°C for crystalline forms with water of crystallization; (10) 7.6% mass loss up to 120 °C; (11) 13.9% mass loss up to 120 °C; (12) 8.9% mass loss up to 120 °C; (13) 24.5% mass loss up to 150°C and / or 9.8% mass loss from 90°C to 120°C; (14) 10.7% mass loss up to 120 °C; (15) 11.2% mass loss up to 120 °C; (16) 13.9% mass loss up to 120 °C; (17) 7.1% mass loss up to 150 °C; (18) 2.5% mass loss up to 61°C and / or 8.8% mass loss from 61°C to 90°C; and (19) Up to 11.1% mass loss at 120 °C.

6. The crystalline form according to any one of claims 1 to 5, wherein The crystalline form exhibits endothermic peaks and / or exothermic peaks selected from the group consisting of: (1) 77.7°C (endothermic), 231.4°C (endothermic); (2) 164.2°C (endothermic); (3) 214.7°C (endothermic); (4) 175.6°C (endothermic), 121.5°C (exothermic); (5) 176.2°C (endothermic); (6) 176.1°C (endothermic); (7) 170.4°C (endothermic), 176.1°C (endothermic); (8) 115.9°C (endothermic), 170.3°C (endothermic), 176.1°C (endothermic), 120.0°C (exothermic); (9) 91.5°C (endothermic), 175.8°C (endothermic); (10) 89.0°C (endothermic), 116.2°C (endothermic), 170.5°C (endothermic), 176.9°C (endothermic), 122.5°C (exothermic); (11) 66.2°C (endothermic), 70.8°C (endothermic), 79.1°C (endothermic), 176.0°C (endothermic); (12) 99.9°C (endothermic), 175.7°C (endothermic); (13) 126.3°C (endothermic), 175.3°C (endothermic); (14) 72.5°C (endothermic), 176.0°C (endothermic); (15) 116.4°C (endothermic), 175.9°C (endothermic); (16) 102.1°C (endothermic), 176.3°C (endothermic); (17) 115.9°C (endothermic), 176.1°C (endothermic); (18) 83.6°C (endothermic), 176.0°C (endothermic); and (19) 118.4°C (endothermic), 175.2°C (endothermic).

7. The crystalline form according to any one of claims 1 to 6, wherein The crystalline form exhibits a salt stoichiometry (molar ratio of acid to free base) selected from the group consisting of: (1) 1:1 (molar ratio of hydrochloric acid to free base); (2) 1:1 (molar ratio of citric acid to free base); and (3) 2.1:1 (molar ratio of phosphoric acid to free base).

8. The crystalline form according to any one of claims 1 to 7, wherein The crystalline form does not include residual solvent or includes a residual solvent selected from the group consisting of: 0.2 wt % EtOAc; 6.3 wt % 2-MeTHF; 9.3 wt % 1,4-dioxane; 7.2 wt % CHCl3; 9.0 wt% toluene; 0.7 wt% 2-MeTHF; 11.9 wt% anisole; 6.6 wt% toluene; 5.3 wt% n-hexane; and 12.3 wt% DMSO.

9. The crystalline form according to any one of claims 1 to 8, wherein The crystalline form shows a purity rate selected from the group consisting of: (1) 99.8 area %; (2) 99.7 area %; (3) 100.0 area %; (4) 99.80 area %; and (5) 99.7 area % in LC-MS (liquid chromatography-mass spectrometry) analysis.

10. The crystalline form according to any one of claims 1 to 9, wherein The crystalline form has a solubility selected from the group consisting of: (1) 4.8 mg / mL for H2O, 4.9 mg / mL for SGF, 1.4 mg / mL for FaSSIF, and 1.3 mg / mL for FeSSIF; (2) 3.9 mg / mL for H2O, 3.7 mg / mL for SGF, 1.9 mg / mL for FaSSIF, and 1.4 mg / mL for FeSSIF; (3) 5.0 mg / mL for H2O, 4.3 mg / mL for SGF, 4.2 mg / mL for FaSSIF, and 1.6 mg / mL for FeSSIF; and (4) 0.036 mg / mL for H2O, 5.1 mg / mL for SGF, 0.064 mg / mL for FaSSIF, and 0.96 mg / mL for FeSSIF.

11. The crystalline form according to any one of claims 1 to 10, wherein The crystalline form exhibits a hygroscopicity selected from the following in DVS (Dynamic Vapor Sorption) analysis at 25° C. and 80% RH (room humidity): 4.12%, 0.38%, 0.97% and 0.50%.

12. An acid addition salt of a crystalline form according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or a solvate or hydrate of any of the foregoing.

13. A method for preparing the crystalline form according to any one of claims 1 to 11, wherein the method is selected from the group consisting of: (1) Compound HY-A was added to EtOAc, HCl (1.05 molar ratio) was slowly added to the solution under magnetic stirring, and the solution was dried by stirring to obtain the crystalline form; (2) adding compound HY-A and citric acid, adding EtOH, and obtaining the crystalline form by stirring and drying; (3) Compound HY-A and 10 mL of EtOH were added, and 95.4 μL of 85% (2.05 molar ratio) H3PO4 was slowly added to the solution under magnetic stirring. After stirring, the solid was separated by centrifugation and continued to be slurried in EtOAc. After slurrying for 2 hours, the solid was centrifuged and vacuum dried at room temperature to obtain the crystalline form; (4) Compound HY-A was purged with N2 for 20 min; (5) Compound HY-A was heated to 130°C and cooled to 30°C under N2 atmosphere; (6) Compound HY-A was heated to 120°C under N2 atmosphere and then cooled to room temperature; (7) Compound HY-A was added to IPA / MIBK (1:1, v / v) at 50 °C for 5 days and then dried under vacuum; (8) Compound HY-A was heated to 90°C under N2 atmosphere and then cooled to room temperature; (9) Slowly evaporating the solution of compound HY-A together with polymer mixture B; (10) Compound HY-A was added to MeOH for 5 days and then dried at room temperature; (11) Compound HY-A was added to ACN / H2O (v / v, 1:4) for 5 days and then dried at room temperature; (12) Compound HY-A was added to 2-MeTHF at 50 °C for 5 days, then slurried at 5 °C for 1 day and dried at room temperature; (13) adding the anti-solvent MTBE to the 1,4-dioxane solution of compound HY-A, and then drying at room temperature; (14) Compound HY-A was exposed to a CHCl3 atmosphere for one day and then dried at room temperature; (15) Compound HY-A was added to 2-MeTHF / toluene (v / v, 1:1) at 50 °C for 5 days and then dried at room temperature; (16) Compound HY-A was added to anisole at room temperature for 5 days and then dried at room temperature; (17) Compound HY-A was added to toluene at room temperature for 5 days and then dried at room temperature; (18) Drying compound HY-A at room temperature (temperature range: 20-22°C; humidity range: 44-77% RH); and (19) Compound HY-A was exposed to a DMSO atmosphere for 7 days and then dried at room temperature.

14. A pharmaceutical composition comprising: the crystalline form according to any one of claims 1 to 11 and / or the acid addition salt of the crystalline form according to claim 12, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate thereof, and a pharmaceutically acceptable carrier.

15. A kit comprising: the crystalline form according to any one of claims 1 to 11, the acid addition salt of the crystalline form according to claim 12 and / or the pharmaceutical composition according to claim 14, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate of the foregoing, and a pharmaceutically acceptable carrier.

16. A method for inhibiting casein kinase (CK) activity, the method comprising providing: the crystalline form of any one of claims 1 to 11, the acid addition salt of the crystalline form of claim 12 and / or the pharmaceutical composition of claim 14, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate of the foregoing and / or the kit of claim 15.

17. The method of claim 16, wherein: The casein kinase (CK) is selected from the group consisting of casein kinase Iα (CK1α), casein kinase Iδ (CK1δ) and casein kinase Iε (CK1ε).

18. The method according to any one of claims 16 to 17, wherein: The method is selected from the group consisting of an in vitro method, an ex vivo method, and an in vivo method.

19. A method for preventing and / or treating a disease or condition, the method comprising administering to a subject in need thereof an effective amount of: the crystalline form of any one of claims 1 to 11, the acid addition salt of the crystalline form of claim 12, and / or the pharmaceutical composition of claim 14, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any solvate or hydrate of the foregoing and / or the kit of claim 15.

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