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-B is 2-(8-(3-(4-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)imidazo[1,2-b]pyridazin-2-yl)propan-2-ol, 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-B. The novel salts and / or crystalline forms of HY-B 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 crystalline 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 invention are set forth with particularity in the appended claims. The features and advantages of the invention will be better understood by reference to the following detailed description of illustrative embodiments in which the principles of the invention are employed and the accompanying drawings (also referred to herein as "Figures"), in which:

[0012] Figure 1-1 The structure of HY-B is shown.

[0013] Figure 2-1 The XRPD pattern of the starting material (823129-01-A / B) is shown;

[0014] Figure 2-2 The TGA / DSC curve of the starting material (823129-01-A) is shown;

[0015] Figure 2-3 The 1H NMR spectrum of the starting material (823129-01-A) is shown;

[0016] Figure 2-4 UPLC chromatogram of the starting material (823129-01-A) is shown;

[0017] Figure 2-5 The TGA / DSC curve of the starting material (823129-01-B) is shown;

[0018] Figure 2-6 The 1H NMR spectrum of the starting material (823129-01-B) is shown;

[0019] Figure 2-7 UPLC chromatogram of the starting material (823129-01-B) is shown;

[0020] Figure 3-1 The XRPD pattern of the re-prepared sulfate salt Form A (823129-12-A) is shown;

[0021] Figure 3-2 The TGA / DSC curve of the re-prepared sulfate salt Form A (823129-12-A) is shown;

[0022] Figure 3-3 The 1H NMR spectrum of the re-prepared sulfate salt Form A (823129-12-A) is shown;

[0023] Figure 3-4 UPLC chromatogram showing the re-prepared sulfate salt Form A (823129-12-A);

[0024] Figure 3-5 The XRPD pattern of the re-prepared maleate salt Form B (823129-13-A) is shown;

[0025] Figure 3-6 The TGA / DSC curve of the re-prepared maleate salt Form B (823129-13-A) is shown;

[0026] Figure 3-7 The 1H NMR spectrum of the re-prepared maleate salt Form B (823129-13-A) is shown;

[0027] Figure 3-8 UPLC chromatogram showing the re-prepared maleate salt Form B (823129-13-A);

[0028] Figure 3-9 The XRPD pattern of the re-prepared tosylate salt Form A (823129-14-A) is shown;

[0029] Figure 3-10 The TGA / DSC curve of the re-prepared tosylate salt Form A (823129-14-A) is shown;

[0030] Figure 3-11 The 1H NMR spectrum of the re-prepared tosylate salt Form A (823129-14-A) is shown;

[0031] Figure 3-12 UPLC chromatogram showing the re-prepared tosylate salt Form A (823129-14-A);

[0032] Figure 4-1 The DVS pattern of the free base Form A (823129-01-A) is shown;

[0033] Figure 4-2 Shown is an XRPD overlay of the free base salt Form A (823129-01-A) before and after DVS;

[0034] Figure 4-3 The DVS diagram of sulfate form A (823129-12-A) is shown;

[0035] Figure 4-4 Shown is an XRPD overlay of sulfate Form A (823129-12-A) before and after DVS;

[0036] Figure 4-5 The DVS pattern of maleate salt Form B (823129-13-A) is shown;

[0037] Figure 4-6 Shown is an XRPD overlay of maleate Form B (823129-13-A) before and after DVS;

[0038] Figure 4-7 The DVS pattern of the tosylate salt Form A (823129-14-A) is shown;

[0039] Figure 4-8 Shown is an XRPD overlay of Tosylate Form A (823129-14-A) before and after DVS;

[0040] Figure 4-9 A graph showing kinetic solubility;

[0041] Figure 4-10 Shown is an XRPD overlay of the free base Form A (823129-01-A) after stability evaluation;

[0042] Figure 4-11 Shows an XRPD overlay of sulfate Form A (823129-12-A) after stability evaluation;

[0043] Figure 4-12 Shown is an XRPD overlay of maleate salt Form B (823129-13-A) after stability evaluation;

[0044] Figure 4-13 Shown is an XRPD overlay of Tosylate Salt Form A (823129-14-A) after stability evaluation;

[0045] Figure 5-1 An XRPD overlay of HY-B maleate salt polymorph is shown;

[0046] Figure 5-2 VT-XRPD pattern of maleate salt Form B (823129-13-A) is shown;

[0047] Figure 5-3 The XRPD spectrum of maleate salt Form C (823129-27-A) is shown;

[0048] Figure 5-4 The TGA / DSC curve of maleate Form C (823129-27-A) is shown;

[0049] Figure 5-5 The 1H NMR spectrum of the maleate salt Form C (823129-27-A) is shown;

[0050] Figure 5-6 UPLC chromatogram showing maleate Form C (823129-27-A);

[0051] Figure 5-7 The XRPD pattern of maleate salt Form D (823129-35-A9) is shown;

[0052] Figure 5-8 The TGA / DSC curve of maleate form D (823129-35-A9) is shown;

[0053] Figure 5-9 The 1H NMR spectrum of the maleate salt form D (823129-35-A9) is shown;

[0054] Figure 6-1 Shown is an XRPD overlay of solids from competitive slurrying for 3 days (I / II);

[0055] Figure 6-2 Shown is an XRPD overlay of solids from competitive slurrying for 3 days (II / II);

[0056] Figure 6-3 Shown is an XRPD overlay of solids from competitive slurry for an additional day (I / II);

[0057] Figure 6-4 Shown is an XRPD overlay of solids from competitive slurry for an additional day (I / II);

[0058] Figure 7-1 XRPD overlay showing maleate Form B (823129-13-A) after stability evaluation Figure 9-1 The XRPD pattern of HCl Salt Form A / B / C (823129-03-B1 / D1 / A2) is shown;

[0059] Figure 9-2 The TGA / DSC curve of HCl salt Form A (823129-03-B1) is shown;

[0060] Figure 9-3 The 1H NMR spectrum of the HCl salt Form A (823129-03-B1) is shown;

[0061] Figure 9-4 The TGA / DSC curve of HCl salt Form B (823129-03-D1) is shown;

[0062] Figure 9-5 The 1H NMR spectrum of HCl salt Form B (823129-03-D1) is shown;

[0063] Figure 9-6 The TGA / DSC curve of HCl salt Form C (823129-03-A2) is shown;

[0064] Figure 9-7 The 1H NMR spectrum of the HCl salt Form C (823129-03-A2) is shown;

[0065] Figure 9-8 The XRPD pattern of sulfate salt Form A (823129-03-B3) is shown;

[0066] Figure 9-9The TGA / DSC curve of sulfate form A (823129-03-B3) is shown;

[0067] Figure 9-10 The 1H NMR spectrum of sulfate salt Form A (823129-03-B3) is shown;

[0068] Figure 9-11 The XRPD pattern of the maleate salt Form A / B (823129-03-A4 / B4) is shown;

[0069] Figure 9-12 The TGA / DSC curve of maleate Form A (823129-03-A4) is shown;

[0070] Figure 9-13 The 1H NMR spectrum of the maleate salt Form A (823129-03-A4) is shown;

[0071] Figure 9-14 The TGA / DSC curve of maleate Form B (823129-03-B4) is shown;

[0072] Figure 9-15 The 1H NMR spectrum of maleate Form B (823129-03-B4) is shown;

[0073] Figure 9-16 Shown is the XRPD pattern of the tosylate salt Form A (823129-03-B9);

[0074] Figure 9-17 The TGA / DSC curve of the tosylate salt Form A (823129-03-B9) is shown;

[0075] Figure 9-18 The 1H NMR spectrum of the tosylate salt Form A (823129-03-B9) is shown;

[0076] Figure 9-19 The XRPD pattern of the mesylate salt Form A (823129-03-A11) is shown;

[0077] Figure 9-20 The TGA / DSC curve of the mesylate salt Form A (823129-03-A11) is shown;

[0078] Figure 9-21 The 1H NMR spectrum of the mesylate salt Form A (823129-03-A11) is shown;

[0079] Figure 9-22 The XRPD pattern of the oxalate salt Form A / B (823129-03-B12 / C12) is shown;

[0080] Figure 9-23The TGA / DSC curve of oxalate Form A (823129-03-B12) is shown;

[0081] Figure 9-24 The 1H NMR spectrum of oxalate Form A (823129-03-B12) is shown;

[0082] Figure 9-25 The TGA / DSC curve of oxalate Form B (823129-03-C12) is shown;

[0083] Figure 9-26 The 1H NMR spectrum of oxalate Form B (823129-03-C12) is shown;

[0084] Figure 9-27 The XRPD pattern of HBr salt Form A / B (823129-03-A13 / B13) is shown;

[0085] Figure 9-28 The TGA / DSC curve of HBr salt Form A (823129-03-A12) is shown;

[0086] Figure 9-29 The 1H NMR spectrum of HBr salt Form A (823129-03-A13) is shown;

[0087] Figure 9-30 The TGA / DSC curve of HBr salt Form B (823129-03-B13) is shown;

[0088] Figure 9-31 The 1H NMR spectrum of HBr salt Form B (823129-03-B13) is shown;

[0089] Figure 9-32 Shown is an XRPD overlay of residual solids from solubility testing of free base Form A in SGF;

[0090] Figure 9-33 Shown is an XRPD overlay of the residual solid from solubility testing of free base Form A in FaSSIF;

[0091] Figure 9-34 Shown is an XRPD overlay of residual solids from solubility testing of free base Form A in FeSSIF;

[0092] Figure 9-35 Shown is an XRPD overlay of the residual solid from solubility testing of the free base Form A in water;

[0093] Figure 9-36 Shown is an XRPD overlay of residual solids from solubility testing of sulfate Form A in SGF;

[0094] Figure 9-37 Shown is an XRPD overlay of residual solids from solubility testing of sulfate Form A in FaSSIF;

[0095] Figure 9-38 Shown is an XRPD overlay of residual solids from solubility testing of sulfate Form A in FeSSIF;

[0096] Figure 9-39 Shown is an XRPD overlay of residual solids from solubility testing of the sulfate salt Form A in water;

[0097] Figure 9-40 Shown is an XRPD overlay of the residual solids from solubility testing of the maleate salt Form B in SGF;

[0098] Figure 9-41 Shown is an XRPD overlay of the residual solid from solubility testing of maleate Form B in FaSSIF;

[0099] Figure 9-42 Shown is an XRPD overlay of the residual solid from solubility testing of maleate Form B in FeSSIF;

[0100] Figure 9-43 Shown is an XRPD overlay of the residual solid from solubility testing of the maleate salt Form B in water;

[0101] Figure 9-44 Shown is an XRPD overlay of the residual solids from solubility testing of the tosylate salt Form A in SGF;

[0102] Figure 9-45 Shown is an XRPD overlay of the residual solid from solubility testing of the tosylate salt Form A in FaSSIF;

[0103] Figure 9-46 Shown is an XRPD overlay of the residual solid from solubility testing of maleate Form B in FeSSIF;

[0104] Figure 9-47 Shown is an XRPD overlay of the residual solids from solubility testing of the tosylate salt Form A in water;

[0105] Figure 9-48 The XRPD pattern of the re-prepared maleate salt Form B (823129-24-A) is shown;

[0106] Figure 9-49 The TGA / DSC curve of the re-prepared maleate salt Form B (823129-24-A) is shown;

[0107] Figure 9-50The 1H NMR spectrum of the re-prepared maleate salt Form B (823129-24-A) is shown;

[0108] Figure 9-51 UPLC chromatogram showing the re-prepared maleate salt Form B (823129-24-A);

[0109] Figure 9-52 The XRPD pattern of the re-prepared tosylate salt Form A (823129-25-A) is shown;

[0110] Figure 9-53 The TGA / DSC curve of the re-prepared tosylate salt Form A (823129-25-A) is shown;

[0111] Figure 9-54 The 1H NMR spectrum of the re-prepared tosylate salt Form A (823129-25-A) is shown;

[0112] Figure 9-55 Shown is the UPLC chromatogram of the re-prepared tosylate salt Form A (823129-25-A). DETAILED DESCRIPTION

[0113] 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 multiple 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.

[0114] 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.

[0115] 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.

[0116] 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.

[0117] 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.

[0118] 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.

[0119] 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.

[0120] 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.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] 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.

[0125] 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).

[0126] 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.

[0127] 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.

[0128] 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.

[0129] 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. Some isotopically labeled compounds of formula, for example radioactive isotopes such as 3H and 14C are incorporated therein, can be used for drug and / or matrix tissue distribution assays. 3H) and carbon-14 (i.e., 14C) isotopes can be used particularly because they are easy to prepare and detectability. In addition, substitution with heavier isotopes such as deuterium (i.e., 2H) can provide certain therapeutic advantages produced by higher metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore, can be used in some cases. Isotopically labeled compounds of formula can be prepared generally by carrying out the procedures disclosed in the following schemes and / or embodiments, by replacing non-isotopically labeled reagents with isotopically labeled reagents.

[0130] 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.

[0131] In one aspect, the present application provides a crystalline form of the compound HY-B 2-(8-(3-(4-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)imidazo[1,2-b]pyridazin-2-yl)propan-2-ol, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2θ degrees, wherein the characteristic peaks are selected from the group consisting of:

[0132] (1) 18.8±0.3 degrees, 19.3±0.3 degrees, 21.8±0.3 degrees;

[0133] (2) 8.2±0.3 degrees, 18.3±0.3 degrees, 23.4±0.3 degrees;

[0134] (3) 11.3±0.3 degrees, 18.5±0.3 degrees, 26.1±0.3 degrees;

[0135] (4) 12.6±0.3 degrees, 18.3±0.3 degrees, 18.9±0.3 degrees;

[0136] (5) 10.5±0.3 degrees, 18.4±0.3 degrees, 28.8±0.3 degrees;

[0137] (6) 9.5±0.3 degrees, 10.8±0.3 degrees, 18.3±0.3 degrees;

[0138] (7) 9.2±0.3 degrees, 18.4±0.3 degrees, 24.7±0.3 degrees;

[0139] (8) 13.3±0.3 degrees, 18.0±0.3 degrees, 20.1±0.3 degrees;

[0140] (9) 6.5±0.3 degrees, 9.7±0.3 degrees, 19.2±0.3 degrees;

[0141] (10) 9.5±0.3 degrees, 19.1±0.3 degrees, 24.5±0.3 degrees;

[0142] (11) 10.4±0.3 degrees, 11.9±0.3 degrees, 16.9±0.3 degrees;

[0143] (12) 14.5±0.3 degrees, 21.2±0.3 degrees, 23.0±0.3 degrees;

[0144] (13) 8.0 ± 0.3 degrees, 10.3 ± 0.3 degrees, 18.9 ± 0.3 degrees; and

[0145] (14) 11.9±0.3 degrees, 18.3±0.3 degrees, 20.3±0.3 degrees.

[0146] For example, wherein the crystalline form further exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2θ degrees, the characteristic peaks being selected from the group consisting of:

[0147] (1) 10.1±0.3 degrees, 12.5±0.3 degrees, 24.0±0.3 degrees;

[0148] (2) 12.0±0.3 degrees, 15.4±0.3 degrees, 16.5±0.3 degrees;

[0149] (3) 8.5±0.3 degrees, 8.9±0.3 degrees, 25.6±0.3 degrees;

[0150] (4) 8.2±0.3 degrees, 17.0±0.3 degrees, 21.8±0.3 degrees;

[0151] (5) 20.3±0.3 degrees, 24.8±0.3 degrees, 27.4±0.3 degrees;

[0152] (6) 14.3±0.3 degrees, 20.7±0.3 degrees, 25.2±0.3 degrees;

[0153] (7) 10.3±0.3 degrees, 18.0±0.3 degrees, 24.1±0.3 degrees;

[0154] (8) 13.8±0.3 degrees, 20.4±0.3 degrees, 25.3±0.3 degrees;

[0155] (9) 18.6±0.3 degrees, 19.5±0.3 degrees, 23.5±0.3 degrees;

[0156] (10) 17.2±0.3 degrees, 18.4±0.3 degrees, 24.9±0.3 degrees;

[0157] (11) 15.2±0.3 degrees, 19.5±0.3 degrees, 20.7±0.3 degrees;

[0158] (12) 9.8±0.3 degrees, 17.5±0.3 degrees, 20.4±0.3 degrees;

[0159] (13) 14.0 ± 0.3 degrees, 16.1 ± 0.3 degrees, 24.3 ± 0.3 degrees; and

[0160] (14) 24.7±0.3 degrees, 27.3±0.3 degrees, 28.6±0.3 degrees.

[0161] For example, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2θ degrees, the characteristic peaks being selected from the group consisting of:

[0162] (1) 10.7±0.3 degrees, 20.3±0.3 degrees, 25.2±0.3 degrees;

[0163] (2) 9.5±0.3 degrees, 16.8±0.3 degrees, 21.7±0.3 degrees;

[0164] (3) 20.8±0.3 degrees, 22.7±0.3 degrees, 23.7±0.3 degrees;

[0165] (4) 9.4±0.3 degrees, 12.0±0.3 degrees, 25.3±0.3 degrees;

[0166] (5) 11.9±0.3 degrees, 12.9±0.3 degrees, 28.2±0.3 degrees;

[0167] (6) 10.1±0.3 degrees, 17.3±0.3 degrees, 23.9±0.3 degrees;

[0168] (7) 14.3±0.3 degrees, 19.0±0.3 degrees, 22.1±0.3 degrees;

[0169] (8) 19.7±0.3 degrees, 23.0±0.3 degrees, 24.0±0.3 degrees;

[0170] (9) 11.2±0.3 degrees, 25.0±0.3 degrees, 27.7±0.3 degrees;

[0171] (10) 21.4±0.3 degrees, 23.7±0.3 degrees, 25.6±0.3 degrees;

[0172] (11) 7.6±0.3 degrees, 24.7±0.3 degrees, 26.5±0.3 degrees;

[0173] (12) 10.1±0.3 degrees, 13.7±0.3 degrees, 22.6±0.3 degrees;

[0174] (13) 15.4 ± 0.3 degrees, 18.5 ± 0.3 degrees, 22.9 ± 0.3 degrees; and

[0175] (14) 14.6±0.3 degrees, 20.8±0.3 degrees, 23.3±0.3 degrees.

[0176] For example, wherein the crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2θ degrees, the characteristic peaks being selected from the group consisting of:

[0177] (1) 18.8±0.3 degrees, 19.3±0.3 degrees, 21.8±0.3 degrees, 10.1±0.3 degrees, 12.5±0.3 degrees, 24.0±0.3 degrees, 10.7±0.3 degrees, 20.3±0.3 degrees, 25.2±0.3 degrees;

[0178] (2) 8.2±0.3 degrees, 18.3±0.3 degrees, 23.4±0.3 degrees, 12.0±0.3 degrees, 15.4±0.3 degrees, 16.5±0.3 degrees, 9.5±0.3 degrees, 16.8±0.3 degrees, 21.7±0.3 degrees;

[0179] (3) 11.3±0.3 degrees, 18.5±0.3 degrees, 26.1±0.3 degrees, 8.5±0.3 degrees, 8.9±0.3 degrees, 25.6±0.3 degrees, 20.8±0.3 degrees, 22.7±0.3 degrees, 23.7±0.3 degrees;

[0180] (4) 12.6±0.3 degrees, 18.3±0.3 degrees, 18.9±0.3 degrees, 8.2±0.3 degrees, 17.0±0.3 degrees, 21.8±0.3 degrees, 9.4±0.3 degrees, 12.0±0.3 degrees, 25.3±0.3 degrees;

[0181] (5) 10.5±0.3 degrees, 18.4±0.3 degrees, 28.8±0.3 degrees, 20.3±0.3 degrees, 24.8±0.3 degrees, 27.4±0.3 degrees, 11.9±0.3 degrees, 12.9±0.3 degrees, 28.2±0.3 degrees;

[0182] (6) 9.5±0.3 degrees, 10.8±0.3 degrees, 18.3±0.3 degrees, 14.3±0.3 degrees, 20.7±0.3 degrees, 25.2±0.3 degrees, 10.1±0.3 degrees, 17.3±0.3 degrees, 23.9±0.3 degrees;

[0183] (7) 9.2±0.3 degrees, 18.4±0.3 degrees, 24.7±0.3 degrees, 10.3±0.3 degrees, 18.0±0.3 degrees, 24.1±0.3 degrees, 14.3±0.3 degrees, 19.0±0.3 degrees, 22.1±0.3 degrees;

[0184] (8) 13.3±0.3 degrees, 18.0±0.3 degrees, 20.1±0.3 degrees, 13.8±0.3 degrees, 20.4±0.3 degrees, 25.3±0.3 degrees, 19.7±0.3 degrees, 23.0±0.3 degrees, 24.0±0.3 degrees;

[0185] (9) 6.5±0.3 degrees, 9.7±0.3 degrees, 19.2±0.3 degrees, 18.6±0.3 degrees, 19.5±0.3 degrees, 23.5±0.3 degrees, 11.2±0.3 degrees, 25.0±0.3 degrees, 27.7±0.3 degrees;

[0186] (10) 9.5±0.3 degrees, 19.1±0.3 degrees, 24.5±0.3 degrees, 17.2±0.3 degrees, 18.4±0.3 degrees, 24.9±0.3 degrees, 21.4±0.3 degrees, 23.7±0.3 degrees, 25.6±0.3 degrees;

[0187] (11) 10.4±0.3 degrees, 11.9±0.3 degrees, 16.9±0.3 degrees, 15.2±0.3 degrees, 19.5±0.3 degrees, 20.7±0.3 degrees, 7.6±0.3 degrees, 24.7±0.3 degrees, 26.5±0.3 degrees;

[0188] (12) 14.5±0.3 degrees, 21.2±0.3 degrees, 23.0±0.3 degrees, 9.8±0.3 degrees, 17.5±0.3 degrees, 20.4±0.3 degrees, 10.1±0.3 degrees, 13.7±0.3 degrees, 22.6±0.3 degrees;

[0189] (13) 8.0 ± 0.3 degrees, 10.3 ± 0.3 degrees, 18.9 ± 0.3 degrees, 14.0 ± 0.3 degrees, 16.1 ± 0.3 degrees, 24.3 ± 0.3 degrees, 15.4 ± 0.3 degrees, 18.5 ± 0.3 degrees, 22.9 ± 0.3 degrees; and

[0190] (14) 11.9 ± 0.3 degrees, 18.3 ± 0.3 degrees, 20.3 ± 0.3 degrees, 24.7 ± 0.3 degrees, 27.3 ± 0.3 degrees, 28.6 ± 0.3 degrees, 14.6 ± 0.3 degrees, 20.8 ± 0.3 degrees, 23.3 ± 0.3 degrees, ± 0.3 degrees. As used herein, when referring to a 2θ degree value, it is meant that value ± 0.3 degrees 2θ. As used herein, when referring to a 2θ degree value, it is meant that value ± 0.2 degrees 2θ. As used herein, when referring to a 2θ degree value, it is meant that value ± 0.1 degrees 2θ.

[0191] For example, wherein the crystalline form exhibits a mass loss in TGA (thermogravimetric) analysis selected from the group consisting of:

[0192] (1) Up to 1.3% mass loss at 120°C;

[0193] (2) up to 0.4% mass loss at 120°C;

[0194] (3) up to 0.7% mass loss at 120°C;

[0195] (4) up to 5.7% mass loss at 120°C;

[0196] (5) Up to 9.7% mass loss at 120°C;

[0197] (6) Up to 16.7% mass loss at 120°C;

[0198] (7) Up to 14.2% mass loss at 120°C;

[0199] (8) Up to 8.7% mass loss at 120°C;

[0200] (9) Up to 1.8% mass loss at 120°C;

[0201] (10) up to 6.8% mass loss at 120 °C;

[0202] (11) Up to 4.1% mass loss at 120°C;

[0203] (12) up to 6.8% mass loss at 120 °C;

[0204] (13) 0.6% mass loss up to 120°C; and

[0205] (14) Up to 8.3% mass loss at 120 °C.

[0206] For example, wherein the crystalline form exhibits an endothermic peak and / or an exothermic peak selected from the group consisting of:

[0207] (1) 135.3°C (endothermic);

[0208] (2) 134.6°C (endothermic);

[0209] (3) 136.5°C (endothermic);

[0210] (4) 127.7°C (endothermic);

[0211] (5) 115.3°C (endothermic);

[0212] (6) 99.4°C (endothermic);

[0213] (7) 112.3°C (endothermic);

[0214] (8) 121.9°C (endothermic);

[0215] (9) 151.8°C (endothermic);

[0216] (10) 91.8°C (endothermic), 122.4°C (endothermic);

[0217] (11) 113.8°C (endothermic);

[0218] (12) 102.1°C (endothermic), 118.4°C (endothermic);

[0219] (13) 155.7°C (endothermic); and

[0220] (14) 125.5℃ (endothermic).

[0221] For example, wherein the crystalline form exhibits a salt stoichiometry (molar ratio of acid to free base) selected from the group consisting of:

[0222] (1) 0.5 (molar ratio of maleic acid to free base);

[0223] (2) 1 (molar ratio of maleic acid to free base);

[0224] (3) 1 (molar ratio of maleic acid to free base);

[0225] (4) 1.5 (molar ratio of maleic acid to free base);

[0226] (5) 1 (molar ratio of hydrochloric acid to free base);

[0227] (6) 1 (molar ratio of hydrochloric acid to free base);

[0228] (7) 1.2 (molar ratio of hydrochloric acid to free base);

[0229] (8) 1.1 (molar ratio of sulfuric acid to free alkali);

[0230] (9) 1 (moles of p-toluenesulfonic acid and free base);

[0231] (10) 1 (molar ratio of methanesulfonic acid to free base);

[0232] (11) 1.1 (molar ratio of oxalic acid to free base);

[0233] (12) 1.1 (molar ratio of oxalic acid to free base);

[0234] (13) 1 (molar ratio of hydrobromic acid to free base); and

[0235] (14) 1 (molar ratio of hydrobromic acid to free base).

[0236] For example, wherein the crystalline form comprises no residual solvent or comprises a residual solvent (wt %) selected from the group consisting of: 2.9 wt % ACN; and 0.5 wt % IPA.

[0237] For example, wherein the crystalline form shows a purity rate selected from the group consisting of: 99.50 area %; and 98.37 area % in LC-MS (liquid chromatography-mass spectrometry) analysis.

[0238] For example, the crystalline form has a solubility selected from the group consisting of: 2.8 mg / mL for SGF, 0.17 mg / mL for FaSSIF, 0.11 mg / mL for FeSSIF, and 0.77 mg / mL for H2O; 5.0 mg / mL for SGF, 0.68 mg / mL for FaSSIF, 0.12 mg / mL for FeSSIF, and 3.1 mg / mL for H2O; 2.4 mg / mL for SGF, 0.21 mg / mL for FaSSIF, 0.12 mg / mL for FeSSIF, and 1.3 mg / mL for H2O.

[0239] 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): 0.03%, 9.1% and 1.2%.

[0240] 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.

[0241] 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:

[0242] (1) Compound HY-A was mixed with an equal molar amount of maleic acid in IPA and then slurried at room temperature for 3 days;

[0243] (2) Compound HY-A was mixed with maleic acid in 5 mL IPAc and then slurried at room temperature for 3 days;

[0244] (3) Grind compound HY-A for 3-5 min;

[0245] (4) Using compound HY-A, solution vapor diffusion was performed in CH2Br2 / n-hexane at room temperature for 4 days;

[0246] (5) Compound HY-A was mixed with an equimolar amount of HCl in IPAc and then slurried at room temperature for 3 days;

[0247] (6) Compound HY-A was mixed with equimolar HCl in ACN / H2O (19:1, v / v) and then slurried at room temperature for 3 days;

[0248] (7) Compound HY-A was mixed with HCl (molar ratio 1:2, FB / acid) in IPA and slurried at room temperature for 3 days, and then evaporated at room temperature.

[0249] (8) Compound HY-A was mixed with an equimolar amount of H2SO4 in IPAc and then slurried at room temperature for 3 days;

[0250] (9) Compound HY-A was mixed with an equimolar amount of p-toluenesulfonic acid in IPAc and then slurried at room temperature for 3 days;

[0251] (10) Compound HY-A was mixed with methanesulfonic acid (molar ratio 1:2, FB / acid) in IPA and then slurried at room temperature for 3 days;

[0252] (11) Compound HY-A was mixed with an equimolar amount of oxalic acid in IPAc and then slurried at room temperature for 3 days;

[0253] (12) Compound HY-A was mixed with an equimolar amount of oxalic acid in MIBK and then slurried at room temperature for 3 days;

[0254] (13) Compound HY-A was mixed with an equimolar amount of HBr in IPA and then slurried at room temperature for 3 days; and

[0255] (14) Compound HY-A was mixed with an equimolar amount of HBr in IPAc and then slurried at room temperature for 3 days.

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264] 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.

[0265] 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.

[0266] On the one hand, the present application provides a method for inhibiting the activity of casein kinase (CK), the method comprising administering to a subject in need thereof 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 thereof, or any of the aforementioned solvates or hydrates and / or the kit of the present application. 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.

[0267] 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 thereof, or any of the aforementioned solvates or hydrates and / or a kit of the present application for 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.

[0268] 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 thereof, 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.

[0269] On the other hand, the present application provides 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: a compound of the present application, or a pharmaceutically acceptable salt, prodrug or metabolite thereof, or any of the aforementioned solvates or hydrates. 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 a depressive disorder and a bipolar disorder.

[0270] 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 its pharmaceutically acceptable salt, prodrug or metabolite, 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 disorder. 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 a depressive disorder and a bipolar disorder.

[0271] 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 thereof, 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, a disease or condition may be selected from a group consisting of a neurological disease and a mental illness. For example, a disease or condition may be selected from a group consisting of a mood disorder, a sleep disorder, and a circadian rhythm disorder. For example, a disease or condition may be selected from a group consisting of a depressive disorder and a bipolar disorder.

[0272] 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 of the foregoing, and optionally a pharmaceutically acceptable carrier.

[0273] 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.

[0274] In some cases, the compound of the present application can also be directly administered into 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.

[0275] 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.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] 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 applications due to their greater water solubility relative to the parent compound.

[0280] In another embodiment, the application provides a composition. Such a composition may include a compound of the 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% of the active compound by weight. The compound of the application may be coupled with a suitable polymer as a targetable drug carrier. Other pharmacologically active substances may also be present.

[0281] 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.

[0282] 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.

[0283] 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.

[0284] The phrases "concurrent administration," "co-administration," "simultaneous administration," and "administered simultaneously" may refer to administration of the compounds in combination.

[0285] Example

[0286] 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.

[0287] Example 1 Preparation of Compounds

[0288] Preparation of compounds

[0289]

[0290] Synthesis scheme of HY-B

[0291]

[0292]

[0293] Step 1: 8-Bromo-6-chloroimidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester

[0294] A mixture of 4-bromo-6-chloropyridazine-3-amine (5 g, 24 mmol, 1.0 equivalent) and methyl 3-bromo-2-oxopropanoate (17 g, 96 mmol, 4.0 equivalent) in DME (50 mL) was stirred at room temperature under an air atmosphere. The reaction was stirred at 90 ° C for 16 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (3: 1) to give 8-bromo-6-chloroimidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester (5.7 g, 81%) as an off-white solid. 1 H NMR(300MHz, CDCl3)δ8.50(s,1H),7.47(s,1H),4.00(s,3H).LC / MS(ESI,m / z):[(M+1)] + =290,292.

[0295] Step 2: 6-Chloro-8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazine-2-carboxylate

[0296] To a stirred mixture of 8-bromo-6-chloroimidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester (1 g, 3.4 mmol, 1.0 eq) and 3-(4-fluorophenyl)-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (1 g, 3.4 mmol, 1.0 eq) in toluene (15 mL) was added CsCO (2.3 g, 6.9 mmol, 2.0 eq) and Pd(dppf)ClCHCl (280 mg, 0.34 mmol, 0.1 eq) under nitrogen atmosphere at room temperature. The resulting mixture was stirred at 100° C. for 2 hours under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with PE / EA (1:2) to give methyl 6-chloro-8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazine-2-carboxylate (390 mg, 29%) as a light brown solid. 1 H NMR (400MHz, CDCl3) δ 8.87 (s, 1H), 8.43 (d, J = 0.8Hz, 1H), 7.51 (dd, J = 8.6, 5.6Hz, 2H), 7.16 (t, J = 8.6Hz, 2H), 6.79 (s, 1H), 4.05 (s, 3H), 4.01 (s, 3H). LC / MS(ESI,m / z):[(M+1)] + =386.

[0297] Step 3:

[0298] To a solution of 6-chloro-8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester (390 mg, 1.0 mmol, 1.0 equiv) in EA (10 mL) was added Pd / C (97 mg, 10%) under a nitrogen atmosphere. The mixture was hydrogenated at 50 ° C for 16 hours under a hydrogen atmosphere using a hydrogen tire. The resulting mixture was filtered and the filter cake was washed with MeOH (3×10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC using the following conditions (column: XSelect CSH Prep C18 OBD column, 19*250 mm, 5 um; mobile phase A: water (0.05% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 30B to 55B in 7 minutes, 254 / 220 nm; RT1: 6.47 minutes; injection volume: 0.4 ml; number of runs: 8) to give 8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester (29.5 mg, 8%) as an off-white solid. 1 H NMR (400MHz, CDCl3) δ8.84 (s, 1H), 8.52 (s, 1H), 8.12 (d, J = 4.9Hz, 1H), 7.52-7. 47(m,2H),7.15-7.09(m,2H),6.77(d,J=5.0Hz,1H),4.04(s,3H),4.01(s,3H). LC / MS(ESI,m / z):[(M+1)] + =352.

[0299] Step 4:

[0300] To a stirred mixture of 8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazine-2-carboxylic acid methyl ester (100 mg, 0.285 mmol, 1.00 equiv) in THF (10.00 mL) at 0°C under air atmosphere was added MeMgBr (0.60 mL, 1.800 mmol, 6.32 equiv) dropwise. The reaction was allowed to react at room temperature for 1 hour. The reaction was monitored by LCMS and then quenched with aqueous NHCl4 and concentrated under reduced pressure. The residue was purified by reverse flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN in 0.1% FA aqueous solution, gradient from 35% to 55% in 15 minutes; detector, UV 254 nm. 2-[8-[3-(4-fluorophenyl)-1-methylpyrazol-4-yl]imidazo[1,2-b]pyridazin-2-yl]propan-2-ol (29.5 mg, 29.50%) was obtained as a light yellow solid. 1H NMR(300MHz,Chloroform-d)δ8.80(s,1H),8.08(s,1H),7.89(s,1H),7.53(dd,J =8.6,5.4Hz,2H),7.14(t,J=8.7Hz,2H),6.76(s,1H),4.08(s,3H),1.74(s,6H). LC / MS(ESI,m / z):[(M+1)] + =352.2.

[0301] Biological activity assay

[0302] 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).

[0303] 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.

[0304] The CK1δ inhibition result (EC50) of HY-B was 0.4 μM. The present disclosure provides HY-B as a potent inhibitor of casein kinase.

[0305] Drug transport trials

[0306] 1. Preparation of Caco-2 Cells

[0307] 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% CO2 for 1 hour before cell seeding.

[0308] 2) Dilute Caco-2 cells to 6.86x105 cells / mL with culture medium and dispense 50 μL of the cell suspension into the filter wells of a 96-well HTS transwell plate. Culture the cells in a cell culture incubator at 37°C, 5% CO2, and 95% relative humidity for 14-18 days. Change the cell culture medium every other day, starting within 24 hours after the initial inoculation at the latest.

[0309] 2. Preparation of Stock Solutions

[0310] 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.

[0311] 3. Assessment of Cell Monolayer Integrity

[0312] 1) Remove the culture medium from the reservoir and each Transwell insert and replace with pre-warmed fresh culture medium.

[0313] 2) The transepithelial electrical resistance (TEER) across the monolayer was measured using the Millicell Epithelial Volt-Ohm measurement system (Millipore, USA).

[0314] 3) After the measurement is completed, return the plate to the incubator.

[0315] Calculate the TEER value according to the following equation:

[0316] TEER measurement (ohms)*membrane area (cm 2 ) = TEER value (ohm cm 2 )

[0317] TEER value should be greater than 230 ohm cm 2 , indicating a well-formed Caco-2 monolayer.

[0318] 4. Assay Procedure

[0319] 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.

[0320] 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%.

[0321] 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).

[0322] 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.

[0323] 5) Incubate the plate at 37°C for 2 hours.

[0324] 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.

[0325] 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.

[0326] 5. Data Analysis

[0327] The apparent permeability coefficient (P) for Caco-2 drug transport assay can be calculated using the following equation: app ), in cm / s:

[0328] P app =(V A ×[Drug] 受体 ) / (area × time × [drug] 初始，供体 )

[0329] 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.

[0330] The outflow ratio will be determined using the following equation:

[0331] Outflow ratio = P app(B-A) / P app(A-B)

[0332] 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.

[0333] The recovery rate can be determined using the following equation:

[0334] Recovery rate % = (V A ×[Drug] 受体 +V D ×[Drug] 供体 ) / (V D ×[Drug] 初始，供体 )

[0335] 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)

[0336] The leakage of fluorescent yellow, in percent (%), can be calculated using the following equation:

[0337] %LY leakage = 100 × [LY] 受体 / ([LY] 供体 +[LY] 受体 )

[0338] Less than 1% LY leakage is acceptable to indicate that the Caco-2 monolayer is of good quality.

[0339] HY-B's P app(B-A) , P app(A-B) and outflow ratios are summarized as follows.

[0340]

[0341] Intrinsic clearance assay

[0342] 1. Prepare the master solution as follows.

[0343] Reagents Reserve concentration volume Final concentration Phosphate buffered saline 200mM 200μL 100mM <![CDATA[Ultra-pure H2O]]> - 106μL - <![CDATA[MgCl2 solution]]> 50mM 40μL 5mM

[0344] 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 H2O 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.

[0345] 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.

[0346] 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.

[0347] 5. Data Analysis

[0348] All calculations were performed using Microsoft Excel.

[0349] 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.

[0350] The in vitro half-life (in vitro t 1 / 2 ):

[0351] In vitro 1 / 2 =-(0.693 / k)

[0352] 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):

[0353] In vitro CL int =(0.693*incubation volume (μl)) / (in vitro t 1 / 2 *Amount of protein (mg)

[0354] 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):

[0355] The scaling factors used for the prediction of intrinsic clearance in liver microsomes are summarized below.

[0356]

[0357] a. Iwatosubo et al., Davies and Morris, 1993, 10(7) pp. 1093-1095.

[0358] b. Barter et al., 2007, Curr Drug Metab, 8(1), pp. 33-45; Iwatsubo et al., 1997, JPET, 283, pp. 462-469.

[0359] Papp(BA), Papp(AB) and outflow ratio are summarized below.

[0360]

[0361] HY-B has desirable intrinsic scavenging properties.

[0362] The purpose of this application is to perform salt screening on compound HY-B and to perform polymorph screening on the selected salts to determine the form suitable for further development.

[0363] Salt screening was performed using HY-B free base Form A as starting material (201328-029-P1, 823129-01-A) in 4 solvent systems using 11 acids under 52 different conditions (two acids in two molar ratios). A total of 12 crystalline salt hits were obtained during the screening and characterized by X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). Salt stoichiometry was determined using ultra-performance liquid chromatography (UPLC) coupled with ion chromatography (IC) or 1 H solution NMR ( 1 Nuclear magnetic resonance (NMR) 1 H NMR). The approximate solubility of the salt hits in water at room temperature was tested. Based on the approximate solubility and characterization results (small TGA weight loss, pure endothermic signal on the DSC curve), sulfate Form A, maleate Form B, and toluenesulfonate Form A were selected for re-preparation.

[0364] Salt evaluations, including hygroscopicity, kinetic solubility, and solid stability, were performed using the three reconstituted salt forms and the free base Form A. The salt evaluation results are summarized in Table 1-1.

[0365] Dynamic vapor sorption (DVS) was used to evaluate the hygroscopicity, and the results showed that at 25°C / 80%RH, the water absorption rates of free base Form A, sulfate Form A, maleate Form B, and tosylate Form A were 0.02%, 9.1%, 0.03%, and 1.2%, respectively. After DVS testing, no form change was observed for all samples.

[0366] For the kinetic solubility results in FeSSIF, similar solubility was observed in free base A-type and 3 salt hits (24-hour solubility was about 0.12 mg / mL). In SGF, FaSSIF and water, the solubility of 3 salts was higher than that of free base A-type, and sulfate A-type showed the highest solubility (24-hour solubility was 5.0 mg / mL, 0.68 mg / mL and 3.1 mg / mL respectively). Maleate B-type and tosylate A-type showed similar solubility. After the solubility test, all forms were observed to change in form.

[0367] The solid stability evaluation results showed that all samples showed no form change or significant purity loss after storage at 25°C / 60%RH or 40°C / 75%RH for 12 days.

[0368] According to the salt evaluation results, no significant stability differences were observed between the three salt hits and the free base type A. The solubility of sulfate type A is high, and sulfate type A is hygroscopic, which may indicate the risk of further development. The solubility of maleate type B and toluenesulfonate type A is higher than that of free base type A, while the hygroscopicity is lower than that of sulfate type A. Therefore, maleate type B and toluenesulfonate type A were used in PK studies to compare with free base type A. According to the PK results, the bioavailability of both salt hits was higher than that of free base type A, and no significant differences were observed between the two salt hits. Considering the safety level of the conformational isomers (maleic acid is level 1 and p-toluenesulfonic acid is level 2), and the hygroscopicity of maleate type B is lower than that of toluenesulfonate type A, maleate was selected as a suitable salt for further polymorph screening studies.

[0369] HY-B maleate material is used as material (N210824-073-01, 823129-27-A). According to the XRPD results, the sample is crystalline, so it is named as maleate C-type. Using the method of anti-solvent addition, slow evaporation, slow cooling, slurrying (room temperature and 50°C), temperature cycling, solid vapor diffusion, solution vapor diffusion, polymer induced crystallization and grinding, 106 polymorph screening experiments were carried out. According to the XRPD characterization results, a total of four polymorphs were obtained, including hemimaleate A-type (anhydrous), monomaleate B-type and C-type (anhydrous) and sesquimaleate D-type (anhydrous or hydrate). The characterization results are summarized in Table 1-2.

[0370] The interconversion relationship between the monomaleate anhydrate Form B and Form C was studied by competitive slurry in different solvents at room temperature and 50°C. Form B maleate was obtained in all experiments, indicating that Form B is more thermodynamically stable than Form C from room temperature to 50°C, and therefore Form B maleate was selected for polymorph evaluation. Since hygroscopicity, kinetic solubility, and solid stability were performed under conditions of 25°C / 60%RH and 40°C / 75%RH during salt evaluation, solid stability at 80°C for 24 hours was performed in the polymorph evaluation. The results showed that after storage at 80°C for 24 hours, Form B maleate had no form change or significant purity decrease.

[0371] Based on all evaluation results, the monomaleate salt Form B was selected as a suitable solid form for further development.

[0372] Table 1-1 Summary of salt assessment

[0373]

[0374] Table 1-2 Summary of characterization of HY-B maleate polymorphs

[0375]

[0376] *: Only maleate forms B and C are monomaleate salts.

[0377] Example 2

[0378] Characterization of starting materials

[0379] This application received two batches of HY-B materials (201328-029-P1, 823129-01-A; 201328-007, 823129-01-B) for salt screening and salt re-preparation / evaluation. The materials were analyzed by XRPD, TGA, DSC, 1 The purity was characterized by H NMR and UPLC. Figure 2-1 The XRPD results in showed that the two materials were in the same form, designated as free base Form A.

[0380] The TGA / DSC curve of the material (823129-01-A) is shown in Figure 2-2 It shows a weight loss of 0.3% up to 150°C and an endotherm (onset) at 176.5°C. 1 H NMR spectroscopy showed Figure 2-3 UPLC results (Table 2-1 and Figure 2-4 ) shows that the purity of the material (823129-01-A) is 98.32 area %.

[0381] The TGA / DSC curve of the material (823129-01-B) is shown in Figure 2-5 It shows a weight loss of 0.6% up to 150°C and an endotherm (onset) at 176.2°C. 1 H NMR spectroscopy showed Figure 2-6 UPLC results (Table 2-2 and Figure 2-7 ) shows that the purity of the material (823129-01-B) is 96.41 area %.

[0382] The approximate solubility of the material (823129-01-A) was determined in 21 solvents / co-solvents at room temperature. Approximately 2 mg of sample was added to a 3-mL glass vial. The solvents listed in Tables 2-3 were then added to the vial in a stepwise manner until the solids were visibly dissolved or a total volume of 2 mL was reached. The solubility results summarized in Tables 2-3 were used to guide solvent selection in the salt screening experimental design.

[0383] Table 2-1 Summary of impurities in material (823129-01-A)

[0384]

[0385] Table 2-2 Summary of impurities in material (823129-01-B)

[0386]

[0387]

[0388] Table 2-3 Approximate solubility of material (823129-01-A)

[0389]

[0390] Example 3

[0391] Salt screening

[0392] Salt Screening Summary

[0393] A total of 52 salt screening experiments were performed using HY-B free base type A (823129-01-A) as the material and 11 acids (two acids using two feed ratios) in four solvent systems. About 20 mg of free base type A (823129-01-A) and the corresponding acid were added to 0.5 mL of solvent and then slurried at room temperature for 3 days. The clarified sample was transferred to stirring at 5°C. If no solid was obtained, it was transferred to stirring at -20°C. Then, the antisolvent was used to clarify the solution, and if no solid was still obtained, it was evaporated at room temperature. The resulting suspension was centrifuged to recover the solid for XRPD testing, and the results are summarized in Table 3-1. A total of 12 crystalline salt hits were obtained from the salt screening, which were characterized by XRPD, TGA, DSC and NMR or UPLC / IC. The detailed results of the characterization of the salt hits are listed in Example 9.2.

[0394] Table 3-1 Summary of salt screening

[0395]

[0396] a : Transfer the clarified sample to 5 °C and stir for 3 hours;

[0397] b : Transfer the clarified sample to 5°C for 3 hours and -20°C for 6 hours;

[0398] c : The clarified sample was transferred to 5°C and stirred for 3 hours, then -20°C and stirred for 6 hours, and then the anti-solvent (MTBE) was added;

[0399] d : The clarified sample was transferred to 5 °C and stirred for 3 hours, then -20 °C for 6 hours, then anti-solvent (MTBE) was added and evaporated in vacuo at room temperature;

[0400] e : The gel sample was transferred to a stirring oven with temperature cycling (50°C-5°C, 0.01°C / min, 2 cycles).

[0401] Approximate solubility in water

[0402] To preliminarily assess the solubility of each salt hit, approximate solubility tests were performed on the salt hits from the salt screen in water. Approximately 2 mg of sample was added to a glass vial. Solvent was then added to the vial stepwise (100 μL, 500 μL, or 1000 μL per step) until the solid was visibly dissolved or a total volume of 10 mL was reached. The solubility results are summarized in Table 3-2.

[0403] Table 3-2 Summary of approximate solubility of salt hits

[0404]

[0405] *: Precipitation was observed after adjusting the pH to 2.26 using 0.1 M NaOH.

[0406] # : A suspension was observed after magnetization (750 rpm) overnight.

[0407] Re-preparation of candidate salts

[0408] Sulfate Form A, Maleate Form B, and Tosylate Form A were selected for de novo preparation based on approximate solubility in water and characterization results (small TGA weight loss, pure DSC endotherm), safety level and frequency of use of the salt conformers.

[0409] All salt forms were prepared de novo by slurrying on a 350 mg scale (based on free base). 1 The re-prepared samples were characterized by H NMR or UPLC / IC and are summarized in Table 3-3. The re-prepared salt forms were used for salt evaluation.

[0410] Table 3-3 Summary of characterization of the re-prepared salts

[0411]

[0412] Sulfate

[0413] Sulfate salt Form A (823129-12-A) was prepared by adding 349.9 mg of free base Form A (823129-01-A) to 5 mL IPAc, followed by 261.5 μL of 4M H2SO4 (acid / FB=1.05) and slurrying at room temperature for 27 hours (1000 rpm). The solid was obtained by centrifugation and vacuum drying at room temperature. The XRPD pattern is shown in Figure 3-1 The TGA / DSC curve is shown in Figure 3-2 It shows a weight loss of 7.7% up to 120°C and an endotherm at 121.8°C. Figure 3-3 In 1 H NMR results showed that no residual IPAc was detected. UPLC results (Tables 3-4 and Figure 3-4 ) showed a purity of 99.56 area %. UPLC / IC results showed that the molar ratio of acid / FB was 1.1.

[0414] Table 3-4 Summary of impurities in the re-prepared sulfate type A (823129-12-A)

[0415]

[0416] Maleate

[0417] Maleate salt Form B (823129-13-A) was obtained by adding 350.1 mg of free base Form A (823129-01-A) and 115.9 mg of maleic acid to 5 mL of IPAc for 3 days at room temperature (1000 rpm). After centrifugation, the solid was dried under vacuum at room temperature. The XRPD pattern is shown at Figure 3-5 The TGA / DSC curve is shown in Figure 3-6 It shows a weight loss of 0.4% up to 120°C and one endotherm at 134.6°C. Figure 3-7 In 1 H NMR results showed that the molar ratio of acid / FB was 1.0, and no residual IPAc was detected. UPLC results (Tables 3-5 and Figure 3-8 ) showed a purity of 99.51 area %.

[0418] Table 3-5 Summary of impurities of the re-prepared maleate salt type B (823129-13-A)

[0419]

[0420] Tosylate

[0421] Tosylate salt Form A (823129-14-A) was prepared by adding 350.0 mg of free base Form A (823129-01-A) and 189.6 mg of p-toluenesulfonic acid to 5 mL of IPAc and then slurrying at room temperature for 3 days (1000 rpm). After centrifugation, the solid was dried under vacuum at room temperature. The XRPD pattern was shown at Figure 3-9 The TGA / DSC curve is shown in Figure 3-10 It shows a weight loss of 0.4% up to 120°C and one endotherm at 150.0°C. Figure 3-11 In 1 The HNMR results showed that the molar ratio of acid / FB was 1.0 and the molar ratio of IPAc / API was 0.02 (0.4 wt %). UPLC results (Tables 3-6 and Figure 3-12 ) showed a purity of 98.91 area %.

[0422] Table 3-6 Summary of impurities of the re-prepared toluenesulfonate salt type A (823129-14-A)

[0423]

[0424] Example 4

[0425] Salt Assessment

[0426] Three de novo prepared salt samples (sulfate Form A, maleate Form B, and tosylate Form A) were used along with the free base Form A for salt evaluations including hygroscopicity, kinetic solubility, and solid stability.

[0427] Hygroscopicity

[0428] DVS testing was performed on the free base Form A and the three salt hits to evaluate hygroscopicity. For the sulfate Form A, the DVS testing was started at ambient humidity (approximately 40% RH), while for the other samples it was started at 0% RH. The DVS isotherms were collected at 25°C, between 0% RH and 95% RH. XRPD characterization was performed on the samples after DVS testing. The DVS evaluation results are summarized in Table 4-1. The DVS plots and XRPD results are shown in Figure 4-1 to Figure 4-8 According to the results, the water absorption of free base type A, sulfate type A, maleate type B, and toluenesulfonate type A at 25°C / 80%RH was 0.02%, 9.1%, 0.03%, and 1.2%, respectively. After the DVS test, no form change was observed for all samples.

[0429] Table 4-1 DVS evaluation summary

[0430]

[0431] Kinetic solubility

[0432] The kinetic solubility of the free base Form A and three re-formulated salt hits in water and three biologically relevant media was measured.

[0433] The materials were added to H2O, SGF, FaSSIF, or FeSSIF with a solid loading of 5 mg / mL (10 mg / mL in SGF and 12 mg / L for sulfate in water) and then rolled at 25 rpm at 37°C for 1, 2, 4, and 24 hours. For each time point, centrifugation and filtration (0.45 μm PTFE filter) were performed. The solubility of the supernatant was checked by UPLC and pH was tested. The solids were tested by XRPD. The results are summarized in Table 4-2 and the solubility plot is shown in Figure 4-9 middle.

[0434] According to the results, free alkali type A and three kinds of salts show similar solubility in FeSSIF (about 0.12mg / mL at 24 hours). In SGF, FaSSIF and water, the solubility of three kinds of salts is higher than free alkali type A, and sulfate type A shows the highest solubility (24-h solubility is 5.0mg / mL, 0.68mg / mL and 3.1mg / mL respectively). Maleate type B and tosylate type A show similar solubility. After solubility test, all forms are observed to change in form.

[0435] Table 4-2 Summary of kinetic solubility test

[0436]

[0437] S: solubility (mg / mL).

[0438] -: The sample was clear and no XRPD test was performed.

[0439] FC: Change of form. a : Free base type A; b : New form I; c : Free base type A + peak; d : New Form II; e : New form I + free base type A.

[0440] Solid stability

[0441] The free base Form A and the three re-prepared salt hits were stored at 25°C / 60% RH and 40°C / 75% RH for 12 days for solid stability evaluation. Physical stability and chemical stability were evaluated by XRPD and UPLC purity, respectively. The results are summarized in Table 4-3 and the XRPD results are shown in Figure 4-10 to Figure 4-13 The results showed no form change or significant loss of purity after 12 days of storage under both conditions.

[0442] Table 4-3 Summary of solid stability evaluation

[0443]

[0444] Salt selection

[0445] According to the salt evaluation results, no significant difference in stability was observed between the three salt hits and the free base type A. The solubility of sulfate type A is high, however, sulfate type A is hygroscopic, which may indicate a high risk for further development. The solubility of maleate type B and toluenesulfonate type A is higher than that of free base type A, while the hygroscopicity is lower than that of sulfate type A. Therefore, maleate type B and toluenesulfonate type A were used in PK studies to compare with free base type A. According to the PK results, the bioavailability of both salt hits was higher than that of free base type A, and no significant difference was observed between the two salt hits. Considering the safety level of the conformer (maleic acid is level 1 and p-toluenesulfonic acid is level 2), and the hygroscopicity of maleate type B is lower than that of toluenesulfonate type A, maleate was selected as the lead salt for further polymorph screening studies.

[0446] Example 5

[0447] Polymorph Screening of Maleate Salt

[0448] Based on the PK results, the monomaleate salt was selected for polymorph screening. A total of 106 polymorph screening experiments were performed using HY-B maleate Form C as the material, including antisolvent addition, slow evaporation, slow cooling, slurrying (room temperature and 50°C), temperature cycling, solid vapor diffusion, solution vapor diffusion, polymer-induced crystallization, and grinding. Based on the XRPD results of the polymorph screening, a total of four polymorphs were obtained. The characterization is summarized in Table 5-1, and the XRPD overlay is shown in Figure 5-1 The detailed procedure of polymorph screening experiment is shown in Appendix 9.6.

[0449] Table 5-1 Summary of characterization of maleate polymorphs

[0450]

[0451] *: Only maleate forms B and C are monomaleate salts.

[0452] Maleate Type A

[0453] Maleate A (823129-03-A4) was obtained from salt screening. The preparation and characterization results are shown in Appendix 9.2.3. Due to the small TGA weight loss, it is presumed that the maleate A is anhydrous.

[0454] Maleate B

[0455] Maleate salt Form B (823129-13-A) was obtained by mixing 350.1 mg of free base Form A (823129-01-A) with 115.9 mg of maleic acid in 5 mL of IPAc and then slurrying at room temperature for 3 days (1000 rpm). See the Examples for characterization results. VT-XRPD (Variable Temperature XRPD) Results ( Figure 5-2 ) showed that no form change was observed for the maleate salt Form B after N2 purge for 20 min, heating to 100°C under N2 purge (presumably the peak shift was due to lattice swelling), cooling to 30°C under N2 purge, or exposure to air. Due to the small TGA weight loss and the relatively clear VT-XRPD results, the maleate salt Form B was presumed to be an anhydrate.

[0456] Maleate C

[0457] Test maleate C form (N210824-073-01, 23129-27-A). XRPD pattern is shown in Figure 5-3 The TGA / DSC curve is shown in Figure 5-4 It shows a weight loss of 0.7% up to 120°C and one endotherm at 136.5°C. Figure 5-5 In 1The H NMR results showed that the molar ratio of acid / FB was 1.0 and the molar ratio of residual IPAc / API was 0.05 (1.1 wt %). UPLC results (Table 5-2 and Figure 5-6 ) showed a purity of 98.37 area %. Due to the small TGA weight loss, it was inferred that the maleate salt Form C was anhydrous.

[0458] Table 5-2 Summary of impurities of maleate type C (823129-27-A)

[0459]

[0460] Maleate D

[0461] Maleate salt form D (823129-35-A9) was obtained by vapor diffusion of a solution of material (823129-27-A) in CH2Br2 / n-hexane at room temperature for 4 days. The XRPD pattern is shown in Figure 5-7 The TGA / DSC curve is shown in Figure 5-8 It shows a weight loss of 5.7% up to 120°C and an endotherm at 127.7°C. Figure 5-5 In 1 H NMR results showed that the molar ratio of acid / FB was 1.5, and no residual CH2Br2 or n-hexane was detected. Therefore, it was speculated that the maleate D-type was an anhydrate or a hydrate. Due to limited quantity, no further study was performed.

[0462] Example 6

[0463] Research on mutual transformation relationship

[0464] Two anhydrates of the monomaleate salt (maleate B / C forms) were observed in this application. To investigate the interconversion relationship between the anhydrates, competitive slurries were performed at room temperature and 50°C.

[0465] Monomaleate C-type (823129-27-A) is added to IPAc, anisole and MEK. Filtered after slurrying for 2 hours (1000rpm) at room temperature and 50°C. About 6mg of maleate B-type and C-type are added to saturated solution, with slurrying for 3 days (1000rpm) at room temperature and 50°C. Then about 0.5mg of half maleate A-type is added, with slurrying for 1 day (1000rpm) at room temperature and 50°C. Solid is tested by XRPD.

[0466] The results are summarized in Table 6-1. The XRPD results are shown in Figure 6-1 to Figure 6-4 Maleate form B was obtained in all experiments, indicating that form A is more stable than form C from room temperature to 50 °C.

[0467] Table 6-1 Summary of competitive slurrying between anhydrous substances

[0468]

[0469] *: After three days of slurrying, yellow samples were observed.

[0470] # : After 3 days of slurrying, the hemimaleate Form A was added to the suspended sample and the sample was then stirred for another day.

[0471] N / A: A clear yellow solution was obtained after slurrying for 1 day. A yellow gel sample was obtained after vacuum drying at room temperature, so XRPD analysis was not performed.

[0472] Example 7

[0473] Polymorph Assessment

[0474] Monomaleate Form B is a more stable anhydrous form than Form C from room temperature to 50°C and was therefore selected for evaluation. During salt screening, Form B was evaluated for hygroscopicity, kinetic solubility, and solid stability at 25°C / 60%RH and 40°C / 75%RH, so only Form B was evaluated for solid stability at 80°C for 24 hours in this example. The results are summarized in Table 7-1, and the XRPD results are shown in Figure 7-1 The results showed that no change in form or significant purity was observed for the maleate salt Form B after storage at 80°C for 24 hours.

[0475] Table 7-1 Summary of solid stability evaluation

[0476]

[0477] Example 8

[0478] Using HY-B free base Form A as the material, salt screening was performed under 52 different conditions using 11 acids (two acids using two molar ratios) in 4 solvent systems. A total of 12 crystalline salt hits were obtained during the screening. Based on the approximate solubility and characterization results, sulfate Form A, maleate Form B, and tosylate Form A were selected for re-preparation. Salt evaluations were performed using the three re-prepared salt forms and free base Form A, including hygroscopicity, kinetic solubility, and solid stability.

[0479] According to the salt evaluation results, the solubility of sulfate salt form A is high, however, sulfate salt form A is hygroscopic, which may indicate a high risk for further development. Maleate salt form B and tosylate salt form A have higher solubility than free base form A and lower hygroscopicity than sulfate form A. PK results showed that the bioavailability of both salt hits was higher than that of free base form A, while no significant difference was observed between the two salt hits. Considering the safety level of the conformers and the solid-state properties of the three salts, the maleate salt was selected as the lead salt for further polymorph screening studies.

[0480] HY-B maleate type C was used to perform 106 polymorph screening experiments. According to the XRPD characterization results, a total of four polymorphs were obtained, including hemimaleate type A (anhydrate), monomaleate type B and type C (anhydrate) and sesquimaleate type D (anhydrate or hydrate). The mutual conversion relationship between the monoanhydrate anhydrate type B and type C was studied by competitive slurrying in different solvents at room temperature and 50°C. Maleate type B was obtained in all experiments, indicating that type B is more thermodynamically stable than type C from room temperature to 50°C. Stability results show that maleate type B has no form change or obvious purity reduction after storage at 80°C for 24 hours.

[0481] Based on all evaluation results, the monomaleate salt Form B was selected as a suitable solid form for further development.

[0482] Example 9

[0483] Abbreviation of the solvent used

[0484] The solvent abbreviations are listed in Table 9-1.

[0485] Table 9-1 List of solvent abbreviations

[0486]

[0487] Characterization of salt hits

[0488] A total of 12 salt hits 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 Table 9-2.

[0489] Table 9-2 Summary of characterization of salt hits

[0490]

[0491]

[0492]

[0493] HCl salt

[0494] HCl salt Form A / B (823129-03-B1 / D1) was prepared by mixing the free base Form A (823129-01-A) with equimolar HCl in IPAc and ACN / H2O (19:1, v / v), respectively, followed by slurrying at room temperature for 3 days. HCl salt Form C (823129-03-A2) was prepared by mixing the free base Form A (823129-01-A) with HCl (molar ratio 1:2, FB / acid) in IPA, slurrying at room temperature for 3 days, and then evaporating at room temperature. The XRPD pattern is shown in Figure 9-1 middle.

[0495] The TGA / DSC curve of HCl salt type A (823129-03-B1) is shown in Figure 9-2 It shows a weight loss of 9.7% up to 120°C and one endotherm at 115.3°C. 1 HNMR results ( Figure 9-3 ) showed that no residual IPAc was detected. UPLC / IC results showed that the molar ratio was 1.0 (acid / FB).

[0496] The TGA / DSC curve of HCl salt type B (823129-03-D1) is shown in Figure 9-4 It shows a weight loss of 16.7% up to 120°C and an endotherm at 99.4°C. 1 HNMR results ( Figure 9-5 ) showed that the molar ratio of ACN / API was 0.3 (2.9 wt%). UPLC / IC results showed that the molar ratio was 1.0 (acid / FB).

[0497] The TGA / DSC curve of HCl salt type C (823129-03-A2) is shown in Figure 9-6 It shows a weight loss of 14.2% up to 120°C and an endotherm at 112.3°C. 1 H NMR results ( Figure 9-7 ) showed that the molar ratio of IPA / API was 0.04 (0.9 wt%). UPLC / IC results showed that the molar ratio was 1.2 (acid / FB).

[0498] Sulfate

[0499] Sulfate salt Form A (823129-03-B3) was prepared by mixing free base Form A (823129-01-A) with equimolar H2SO4 in IPAc and slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-8The TGA / DSC curve of sulfate type A (823129-03-B3) is shown in Figure 9-9 It shows a weight loss of 8.7% up to 120°C and an endotherm at 121.9°C. 1 H NMR results ( Figure 9-10 ) showed that no residual IPAc was detected. UPLC / IC results showed that the molar ratio was 1.1 (acid / FB).

[0500] Maleate

[0501] Maleate salt Form A / B (823129-03-A4 / B4) was prepared by mixing the free base Form A (823129-01-A) with equimolar maleic acid in IPA and IPAc, respectively, followed by slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-11 middle.

[0502] The TGA / DSC curve of maleate type A (823129-03-A4) is shown in Figure 9-12 It shows a weight loss of 1.3% up to 120°C and one endotherm at 135.3°C. Figure 9-13 In 1 H NMR results showed that the acid / FB molar ratio was 0.5, and no residual IPA was detected.

[0503] The TGA / DSC curve of maleate B (823129-03-B4) is shown in Figure 9-14 It shows a weight loss of 0.9% up to 120°C and one endotherm at 134.4°C. Figure 9-15 In 1 H NMR results showed that the acid / FB molar ratio was 1.1, and no residual IPA was detected.

[0504] Tosylate

[0505] The tosylate salt Form A (823129-03-B9) was prepared by mixing the free base Form A (823129-01-A) with an equimolar amount of p-toluenesulfonic acid in IPAc and then slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-16 The TGA / DSC curve of toluenesulfonate type A (823129-03-B9) is shown in Figure 9-17 It shows a weight loss of 1.8% up to 120°C and one endotherm at 151.8°C. Figure 9-18 In 1 H NMR results showed that the molar ratio of acid / FB was 1.0, and no residual IPAc was detected.

[0506] Methanesulfonate

[0507] The mesylate salt Form A (823129-03-A11) was prepared by mixing the free base Form A (823129-01-A) with methanesulfonic acid (molar ratio 1:2, FB / acid) in IPA and then slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-19 The TGA / DSC curve of the mesylate type A (823129-03-A11) is shown in Figure 9-20 It shows a weight loss of 6.8% up to 120°C and two endotherms (peaks) at 91.8°C and 122.4°C. Figure 9-21 In 1 H NMR results showed that the acid / FB molar ratio was 1.0, and no residual IPA was detected.

[0508] Oxalate

[0509] Oxalate Form A / B (823129-03-B12 / C12) was prepared by mixing the free base Form A (823129-01-A) with equimolar oxalic acid in IPAc and MIBK, respectively, followed by slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-22 middle.

[0510] The TGA / DSC curve of oxalate type A (823129-03-B12) is shown in Figure 9-23 It shows a weight loss of 4.1% up to 120°C and one endotherm at 113.8°C. Figure 9-24 In 1 H NMR results showed that no residual IPAc was detected. UPLC / IC results showed that the molar ratio was 1.1 (acid / FB).

[0511] The TGA / DSC curve of oxalate type A (823129-03-C12) is shown in Figure 9-25 It shows a weight loss of 6.8% up to 120°C and two endotherms (peaks) at 102.1°C and 118.4°C. Figure 9-26 In 1 H NMR results showed that no residual MIBK was detected. UPLC / IC results showed that the molar ratio was 1.1 (acid / FB).

[0512] HBr salt

[0513] HBr salt A / B form (823129-03-A13 / B13) was prepared by mixing free base A form (823129-01-A) with equimolar HBr in IPA and IPAc, respectively, followed by slurrying at room temperature for 3 days. The XRPD pattern is shown in Figure 9-27 middle.

[0514] The TGA / DSC curve of HBr salt type A (823129-03-A13) is shown in Figure 9-28 It shows a weight loss of 0.6% up to 120°C and one endotherm at 155.7°C. Figure 9-29 In 1 H NMR results showed that no residual IPA was detected. UPLC / IC results showed that the molar ratio was 1.0 (acid / FB).

[0515] The TGA / DSC curve of HBr salt type B (823129-03-B13) is shown in Figure 9-30 It shows a weight loss of 8.3% up to 120°C and an endotherm (peak) at 125.5°C. Figure 9-31 In 1 H NMR results showed that no residual IPAc was detected. UPLC / IC results showed that the molar ratio was 1.0 (acid / FB).

[0516] Preparation of biorelevant media

[0517] Simulated gastric fluid (SGF)

[0518] Weigh 200 mg of NaCl and 100 mg of Triton X-100 into a 100-mL volumetric flask. Add purified water to dissolve the solid. After the solid is completely dissolved, add 136 μL of HCl (12 M), and then adjust the pH to 1.8 using 1 M HCl or 1 M NaOH. Add purified water to volume.

[0519] Fasting state simulated intestinal fluid (FaSSIF)

[0520] 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. About 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 volume. 22 mg of SIF powder was added to a 10-mL volumetric flask, and the prepared buffer was then added to volume.

[0521] Fed State Simulated Intestinal Fluid (FeSSIF)

[0522] 0.41 mL of acetic acid, 202 mg of NaOH, and 594 mg of NaCl were weighed into a 50-mL volumetric flask. About 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 5.0. Purified water was then added to constant volume. 0.56 g of SIF powder was added to a 10-mL volumetric flask, and the prepared buffer was then added to constant volume.

[0523] XRPD results of solids from kinetic solubility measurements

[0524] The XRPD results of the residual solid from the kinetic solubility measurements are shown at Figure 9-32 to Figure 9-47 middle.

[0525] Re-preparation of candidate salts

[0526] Maleate

[0527] Maleate salt Form B (823129-24-A) was prepared by adding 370.0 mg of free base Form A (823129-01-A) and 122.5 mg of maleic acid to 6 mL of IPAc, followed by the addition of seeds (823129-13-A) and slurrying at room temperature for 27 hours (1000 rpm). The solid was obtained by centrifugation and vacuum drying at room temperature overnight. The XRPD pattern is shown in Figure 9-48 The TGA / DSC curve is shown in Figure 9-49 It shows a weight loss of 0.4% up to 120°C and one endotherm at 133.6°C. Figure 9-50 In 1 H NMR results showed that the molar ratio of acid / FB was 1.0, and no residual IPAc was observed. UPLC results (Table 9-3 and Figure 9-51 ) showed a purity of 99.46 area %.

[0528] Table 9-3 Summary of impurities in the re-prepared maleate salt type B (823129-24-A)

[0529]

[0530]

[0531] Tosylate

[0532] Tosylate salt Form A (823129-14-A) was prepared by adding 370.1 mg of free base Form A (823129-01-A) and 200.6 mg of p-toluenesulfonic acid to 6 mL of IPAc, then adding seeds (823129-14-A) and slurrying at room temperature for 27 hours (1000 rpm). The solid was obtained by centrifugation and vacuum drying at room temperature overnight. The XRPD pattern is shown in Figure 9-52 The TGA / DSC curve is shown in Figure 9-53 It shows a weight loss of 2.5% up to 120°C and an endotherm at 150.6°C. Figure 9-54 In 1 The H NMR results showed that the molar ratio of acid / FB was 1.0 and the molar ratio of IPAc / API was 0.02 (0.4 wt %). UPLC results (Table 9-4 and Figure 9-55 ) showed a purity of 98.87 area %.

[0533] Table 9-4 Summary of impurities in the re-prepared toluenesulfonate salt type A (823129-25-A)

[0534]

[0535] Polymorph screening

[0536] Based on the approximate solubility of maleate salt form C (823129-27-A) in Table 9-5, a total of 106 polymorph screening experiments were performed on the maleate salt using different crystallization or solid phase transformation methods. The methods used and the results are summarized in Table 9-6.

[0537] Table 9-5 Approximate solubility of maleate type C (823129-27-A)

[0538]

[0539]

[0540] Table 9-6 Summary of polymorph screening experiments

[0541]

[0542] Antisolvent addition

[0543] A total of 20 antisolvent addition experiments were carried out. About 20 mg of maleate C type (823129-27-A) was dissolved in 0.2-2.0 mL solvent to obtain a clear solution, and the solution was magnetically stirred (about 1000 rpm), and then antisolvent was added until precipitation occurred or the total amount of antisolvent reached 10.0 mL. The sample without precipitation was transferred to slurry at 5 ° C. After slurrying, the clear solution was further transferred to slurry at -20 ° C, and then if it was still clear, it was transferred to evaporate at room temperature. Separation of solids was used for XRPD analysis. The results in Table 9-7 show that maleate A / B / C / A+B / A type+free alkali A type / free alkali A type / gel was obtained.

[0544] Table 9-7 Summary of Antisolvent Addition Experiment

[0545]

[0546] *: Solid obtained by slurrying at 5°C;

[0547] # : A solid was obtained by evaporation at room temperature.

[0548] Slow evaporation

[0549] Slow evaporation experiments were performed under 10 conditions. Briefly, approximately 20 mg of maleate salt Form C (823129-27-A) was dissolved in 1.0-3.0 mL of solvent in a 3-mL glass vial. The sample was filtered using a 0.45 μm PTFE filter and the resulting solution was washed with The sealed vial, punctured with 4 pinholes, was subjected to slow evaporation at room temperature. The solid was isolated for XRPD analysis and the results summarized in Tables 9-8 indicated that maleate salt Form A / B / C / A + free base Form A / B + peak / gel were obtained.

[0550] Table 9-8 Summary of slow evaporation experiment

[0551]

[0552] Slow cooling

[0553] Slow cooling experiments were performed in 10 solvent systems. At room temperature, about 20 mg of maleate C type (823129-27-A) was suspended in 1.0-2.0 mL of solvent in a 3-mL glass vial. The suspension was then heated to 50 ° C, balanced for about 30 min and filtered into a new vial. The filtrate was slowly cooled to 5 ° C at a rate of 0.1 ° C / min. The resulting solid was kept at a constant temperature at 5 ° C. If no solid was obtained, the sample was transferred to -20 ° C. If still no solid was obtained, the sample was transferred to room temperature for evaporation. The solid was tested by XRPD. The results summarized in Table 9-9 show that maleate B / C / B + C / B + D type was obtained.

[0554] Table 9-9 Summary of slow cooling experiment

[0555]

[0556] *: Solid obtained by evaporation at room temperature;

[0557] # : A solid was obtained at -20°C.

[0558] Room temperature pulping

[0559] About 20 mg of maleate C-type (823129-27-A) was suspended in 0.5 mL of solvent 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 Tables 9-10 show that maleate A / B / C / A+C / A-type+free alkali A-type has been generated.

[0560] Table 9-10 Summary of slurry conversion experiment at room temperature

[0561]

[0562]

[0563] *: A clear solution was obtained at room temperature, which was transferred for slurrying at 5°C for 16 hours and a solid was obtained.

[0564] Slurry at 50℃

[0565] About 20 mg of maleate Form C (823129-27-A) was suspended in 0.5 mL of solvent in a HPLC glass vial. After the suspension was stirred (1000 rpm) for about 5 days at 50° C., the remaining solid was centrifuged for XRPD analysis. The results summarized in Tables 9-11 indicate that maleate Form A / B / B / free base Form A was generated.

[0566] Table 9-11 Summary of slurry conversion experiment at 50℃

[0567]

[0568] *: After slurrying at 50°C for 6 days a clear solution was obtained which was transferred to room temperature to evaporate and a solid was obtained.

[0569] Temperature Cycle

[0570] About 20 mg of maleate C (823129-27-A) was suspended in 0.5 mL of solvent in a HPLC glass vial. After slurrying (1000 rpm) with temperature cycling (50-5°C, 0.1°C / min, 2 cycles), the remaining solid was centrifuged for XRPD analysis. The results summarized in Tables 9-12 show that maleate B / C / B forms + free base A forms were obtained.

[0571] Table 9-12 Summary of temperature cycle experiment

[0572]

[0573] Solid Vapor Diffusion

[0574] Use 6 kinds of different solvents to carry out solid vapor diffusion experiment.The maleate C type (823129-27-A) of about 20mg is weighed in a 3-mL bottle, and this bottle is put into a 20-mL bottle of the volatile solvent of 4mL being housed.The 20-mL bottle is sealed with a lid and remains at room temperature for 6 days, allowing solvent vapor to interact with the sample.Test solid by XRPD, the result display that is summarized in Table 9-13 has obtained maleate A / B / C type.

[0575] Table 9-13 Summary of solid vapor diffusion experiments

[0576]

[0577] Solution vapor diffusion

[0578] Thirteen solution vapor diffusion experiments were carried out. About 20 mg of maleate C type (823129-27-A) was dissolved in 0.2-1.6 mL of an appropriate solvent to obtain a clear solution in a 3-mL vial. The solution was then placed in a 20-mL vial containing 3 mL of a volatile solvent. The 20-mL vial was sealed with a lid and stored at room temperature, allowing enough time for organic vapor to interact with the solution. If a clear solution is obtained, the sample is transferred to store at 5 ° C. If a solid is still not obtained, the sample is transferred to room temperature for evaporation. Separation of the precipitate was used for XRPD analysis. The results summarized in Tables 9-14 show that maleate A / B / C / D types / free base A types / gels were obtained.

[0579] Table 9-14 Summary of solution vapor diffusion experiment

[0580]

[0581] *: Solid obtained by evaporation at room temperature.

[0582] # : A solid was obtained at 5°C.

[0583] Polymer-induced crystallization

[0584] Polymer-induced crystallization experiments were performed under 10 conditions. Briefly, approximately 20 mg of maleate salt Form C (823129-27-A) was dissolved in 1.0-2.0 mL of solvent in a 3-mL glass vial. The resulting solution was filtered using a 0.45 μm PTFE filter, and then approximately 2 mg of polymer was added. The vial was filled with The mixture was sealed and punctured with 4 pinholes for slow evaporation at room temperature. The solid was isolated for XRPD analysis and the results summarized in Tables 9-15 indicated that maleate salt Form A / B / C / A+B / B+peak / free base Form A was obtained.

[0585] Table 9-15 Summary of polymer induced crystallization experiments

[0586]

[0587]

[0588] 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).

[0589] 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).

[0590] Grinding

[0591] Grinding experiments were performed with or without the addition of solvent. About 20 mg of maleate Form C (823129-27-A) was weighed into a mortar. 10 μL of solvent was added to the mortar. The solid was ground for 3-5 minutes. The solid was isolated for XRPD analysis. The results summarized in Tables 9-16 show that only maleate Form C was obtained.

[0592] Table 9-16 Grinding experiment summary

[0593]

[0594] Instruments and methods

[0595] XRPD

[0596] For XRPD analysis, a PANalytical X'Pert3 X-ray powder diffractometer was used. The XRPD parameters used are listed in Tables 9-17.

[0597] Table 9-17 XRPD test parameters

[0598]

[0599] TGA and DSC

[0600] TGA data were collected using a TA Q5000 / Discovery 5500 TGA from TA Instruments. DSC was performed using a TA Discovery 2500 DSC from TA Instruments. The detailed parameters used are listed in Tables 9-18.

[0601] Table 9-18 TGA and DSC test parameters

[0602]

[0603] DVS

[0604] DVS was measured by SMS (Surface Measurement Systems) DVS Intrinsic. Relative humidity at 25°C was calibrated against the deliquescent points of LiCl, Mg(NO3)2 and KCl. The DVS test parameters are listed in Tables 9-19.

[0605] Table 9-19DVS test parameters

[0606]

[0607] Solution NMR

[0608] Solution NMR was collected on a Bruker 400M NMR spectrometer using DMSO-d6.

[0609] UPLC / IC

[0610] Waters H-Class UPLC was used for purity, solubility and stability tests, and the detailed chromatographic conditions are listed in Tables 9-20 and 9-21. IC parameters are listed in Table 9-22.

[0611] Table 9-20 UPLC methods for solubility testing

[0612]

[0613]

[0614] Table 9-21 UPLC methods for purity testing

[0615]

[0616] Table 9-22 IC parameters for chemometric testing

[0617]

[0618] 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-B, a crystalline form of 2-(8-(3-(4-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)imidazo[1,2-b]pyridazin-2-yl)propan-2-ol, wherein: The crystalline form exhibits an X-ray powder diffraction pattern having one or more characteristic peaks expressed in 2θ degrees, the characteristic peaks being selected from the group consisting of: (1) 18.8±0.3 degrees, 19.3±0.3 degrees, 21.8±0.3 degrees; (2) 8.2±0.3 degrees, 18.3±0.3 degrees, 23.4±0.3 degrees; (3) 11.3±0.3 degrees, 18.5±0.3 degrees, 26.1±0.3 degrees; (4) 12.6±0.3 degrees, 18.3±0.3 degrees, 18.9±0.3 degrees; (5) 10.5±0.3 degrees, 18.4±0.3 degrees, 28.8±0.3 degrees; (6) 9.5±0.3 degrees, 10.8±0.3 degrees, 18.3±0.3 degrees; (7) 9.2±0.3 degrees, 18.4±0.3 degrees, 24.7±0.3 degrees; (8) 13.3±0.3 degrees, 18.0±0.3 degrees, 20.1±0.3 degrees; (9) 6.5±0.3 degrees, 9.7±0.3 degrees, 19.2±0.3 degrees; (10) 9.5±0.3 degrees, 19.1±0.3 degrees, 24.5±0.3 degrees; (11) 10.4±0.3 degrees, 11.9±0.3 degrees, 16.9±0.3 degrees; (12) 14.5±0.3 degrees, 21.2±0.3 degrees, 23.0±0.3 degrees; (13) 8.0 ± 0.3 degrees, 10.3 ± 0.3 degrees, 18.9 ± 0.3 degrees; and (14) 11.9±0.3 degrees, 18.3±0.3 degrees, 20.3±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 2θ degrees, the characteristic peaks selected from the group consisting of: (1) 10.1±0.3 degrees, 12.5±0.3 degrees, 24.0±0.3 degrees; (2) 12.0±0.3 degrees, 15.4±0.3 degrees, 16.5±0.3 degrees; (3) 8.5±0.3 degrees, 8.9±0.3 degrees, 25.6±0.3 degrees; (4) 8.2±0.3 degrees, 17.0±0.3 degrees, 21.8±0.3 degrees; (5) 20.3±0.3 degrees, 24.8±0.3 degrees, 27.4±0.3 degrees; ±0.3 degrees, 20.7±0.3 degrees, 25.2±0.3 degrees; (7) 10.3±0.3 degrees, 18.0±0.3 degrees, 24.1±0.3 degrees; (8) 13.8±0.3 degrees, 20.4±0.3 degrees, 25.3±0.3 degrees; (9) 18.6±0.3 degrees, 19.5±0.3 degrees, 23.5±0.3 degrees; (10) 17.2±0.3 degrees, 18.4±0.3 degrees, 24.9±0.3 degrees; (11) 15.2±0.3 degrees, 19.5±0.3 degrees, 20.7±0.3 degrees; (12) 9.8±0.3 degrees, 17.5±0.3 degrees, 20.4±0.3 degrees; (13) 14.0 ± 0.3 degrees, 16.1 ± 0.3 degrees, 24.3 ± 0.3 degrees; and (14) 24.7±0.3 degrees, 27.3±0.3 degrees, 28.6±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 2θ degrees, the characteristic peaks being selected from the group consisting of: (1) 10.7±0.3 degrees, 20.3±0.3 degrees, 25.2±0.3 degrees; (2) 9.5±0.3 degrees, 16.8±0.3 degrees, 21.7±0.3 degrees; (3) 20.8±0.3 degrees, 22.7±0.3 degrees, 23.7±0.3 degrees; (4) 9.4±0.3 degrees, 12.0±0.3 degrees, 25.3±0.3 degrees; (5) 11.9±0.3 degrees, 12.9±0.3 degrees, 28.2±0.3 degrees; (6) 10.1±0.3 degrees, 17.3±0.3 degrees, 23.9±0.3 degrees; (7) 14.3±0.3 degrees, 19.0±0.3 degrees, 22.1±0.3 degrees; (8) 19.7±0.3 degrees, 23.0±0.3 degrees, 24.0±0.3 degrees; (9) 11.2±0.3 degrees, 25.0±0.3 degrees, 27.7±0.3 degrees; (10) 21.4±0.3 degrees, 23.7±0.3 degrees, 25.6±0.3 degrees; (11) 7.6±0.3 degrees, 24.7±0.3 degrees, 26.5±0.3 degrees; (12) 10.1±0.3 degrees, 13.7±0.3 degrees, 22.6±0.3 degrees; (13) 15.4 ± 0.3 degrees, 18.5 ± 0.3 degrees, 22.9 ± 0.3 degrees; and (14) 14.6±0.3 degrees, 20.8±0.3 degrees, 23.3±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 2θ degrees, the characteristic peaks being selected from the group consisting of: ±0.3 degrees, 19.3±0.3 degrees, 21.8±0.3 degrees, 10.1±0.3 degrees, 12.5±0.3 degrees, 24.0±0.3 degrees, 10.7±0.3 degrees, 20.3±0.3 degrees, 25.2±0.3 degrees; (2) 8.2±0.3 degrees, 18.3±0.3 degrees, 23.4±0.3 degrees, 12.0±0.3 degrees, 15.4±0.3 degrees, 16.5±0.3 degrees, 9.5±0.3 degrees, 16.8±0.3 degrees, 21.7±0.3 degrees; (3) 11.3±0.3 degrees, 18.5±0.3 degrees, 26.1±0.3 degrees, 8.5±0.3 degrees, 8.9±0.3 degrees, 25.6±0.3 degrees, 20.8±0.3 degrees, 22.7±0.3 degrees, 23.7±0.3 degrees; (4) 12.6±0.3 degrees, 18.3±0.3 degrees, 18.9±0.3 degrees, 8.2±0.3 degrees, 17.0±0.3 degrees, 21.8±0.3 degrees, 9.4±0.3 degrees, 12.0±0.3 degrees, 25.3±0.3 degrees; (5) 10.5±0.3 degrees, 18.4±0.3 degrees, 28.8±0.3 degrees, 20.3±0.3 degrees, 24.8±0.3 degrees, 27.4±0.3 degrees, 11.9±0.3 degrees, 12.9±0.3 degrees, 28.2±0.3 degrees; (6) 9.5±0.3 degrees, 10.8±0.3 degrees, 18.3±0.3 degrees, 14.3±0.3 degrees, 20.7±0.3 degrees, 25.2±0.3 degrees, 10.1±0.3 degrees, 17.3±0.3 degrees, 23.9±0.3 degrees; (7) 9.2±0.3 degrees, 18.4±0.3 degrees, 24.7±0.3 degrees, 10.3±0.3 degrees, 18.0±0.3 degrees, 24.1±0.3 degrees, 14.3±0.3 degrees, 19.0±0.3 degrees, 22.1±0.3 degrees; (8) 13.3±0.3 degrees, 18.0±0.3 degrees, 20.1±0.3 degrees, 13.8±0.3 degrees, 20.4±0.3 degrees, 25.3±0.3 degrees, 19.7±0.3 degrees, 23.0±0.3 degrees, 24.0±0.3 degrees; (9) 6.5±0.3 degrees, 9.7±0.3 degrees, 19.2±0.3 degrees, 18.6±0.3 degrees, 19.5±0.3 degrees, 23.5±0.3 degrees, 11.2±0.3 degrees, 25.0±0.3 degrees, 27.7±0.3 degrees; (10) 9.5±0.3 degrees, 19.1±0.3 degrees, 24.5±0.3 degrees, 17.2±0.3 degrees, 18.4±0.3 degrees, 24.9±0.3 degrees, 21.4±0.3 degrees, 23.7±0.3 degrees, 25.6±0.3 degrees; (11) 10.4±0.3 degrees, 11.9±0.3 degrees, 16.9±0.3 degrees, 15.2±0.3 degrees, 19.5±0.3 degrees, 20.7±0.3 degrees, 7.6±0.3 degrees, 24.7±0.3 degrees, 26.5±0.3 degrees; (12) 14.5±0.3 degrees, 21.2±0.3 degrees, 23.0±0.3 degrees, 9.8±0.3 degrees, 17.5±0.3 degrees, 20.4±0.3 degrees, 10.1±0.3 degrees, 13.7±0.3 degrees, 22.6±0.3 degrees; (13) 8.0 ± 0.3 degrees, 10.3 ± 0.3 degrees, 18.9 ± 0.3 degrees, 14.0 ± 0.3 degrees, 16.1 ± 0.3 degrees, 24.3 ± 0.3 degrees, 15.4 ± 0.3 degrees, 18.5 ± 0.3 degrees, 22.9 ± 0.3 degrees; and (14) 11.9±0.3 degrees, 18.3±0.3 degrees, 20.3±0.3 degrees, 24.7±0.3 degrees, 27.3±0.3 degrees, 28.6±0.3 degrees, 14.6±0.3 degrees, 20.8±0.3 degrees, 23.3±0.3 degrees, ±0.3 degrees.

5. The crystalline form according to any one of claims 1 to 4, wherein The crystalline form exhibits a mass loss in (thermogravimetric) analysis selected from the group consisting of: (1) Up to 1.3% mass loss at 120°C; (2) up to 0.4% mass loss at 120°C; (3) up to 0.7% mass loss at 120°C; (4) up to 5.7% mass loss at 120°C; (5) Up to 9.7% mass loss at 120°C; (6) Up to 16.7% mass loss at 120°C; (7) Up to 14.2% mass loss at 120°C; (8) Up to 8.7% mass loss at 120°C; (9) Up to 1.8% mass loss at 120°C; (10) up to 6.8% mass loss at 120 °C; (11) Up to 4.1% mass loss at 120°C; (12) Up to 6.8% mass loss at 120 °C; (13) 0.6% mass loss up to 120°C; and (14) Up to 8.3% 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) 135.3°C (endothermic); (2) 134.6°C (endothermic); (3) 136.5°C (endothermic); (4) 127.7°C (endothermic); (5) 115.3°C (endothermic); (6) 99.4°C (endothermic); (7) 112.3°C (endothermic); (8) 121.9°C (endothermic); (9) 151.8°C (endothermic); (10) 91.8°C (endothermic), 122.4°C (endothermic); (11) 113.8°C (endothermic); .1℃(endothermic), 118.4℃(endothermic); (13) 155.7°C (endothermic); and (14) 125.5℃ (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) 0.5 (molar ratio of maleic acid to free base); (2) 1 (molar ratio of maleic acid to free base); (3) 1 (molar ratio of maleic acid to free base); (4) 1.5 (molar ratio of maleic acid to free base); (5) 1 (molar ratio of hydrochloric acid to free base); (6) 1 (molar ratio of hydrochloric acid to free base); (7) 1.2 (molar ratio of hydrochloric acid to free base); (8) 1.1 (molar ratio of sulfuric acid to free alkali); (9) 1 (moles of p-toluenesulfonic acid and free base); (10) 1 (molar ratio of methanesulfonic acid to free base); (11) 1.1 (molar ratio of oxalic acid to free base); (12) 1.1 (molar ratio of oxalic acid to free base); (13) 1 (molar ratio of hydrobromic acid to free base); and (14) 1 (molar ratio of hydrobromic acid to free base).

8. The crystalline form according to any one of claims 1 to 7, wherein The crystalline form comprises no residual solvent or comprises a residual solvent (wt %) selected from the group consisting of: 2.9 wt % ACN; and 0.5 wt% IPA.

9. The crystalline form according to any one of claims 1 to 8, wherein The crystalline form exhibits a purity rate selected from the group consisting of: 99.50 area % in LC-MS (liquid chromatography-mass spectrometry) analysis; and 98.37 area%.

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: 2.8 mg / mL for SGF, 0.17 mg / mL for FaSSIF, 0.11 mg / mL for FeSSIF, and 0.77 mg / mL for H2O; 5.0 mg / mL for SGF, 0.68 mg / mL for FaSSIF, 0.12 mg / mL for, and 3.1 mg / mL for H2O; 2.4 mg / mL for SGF, 0.21 mg / mL for FaSSIF, 0.12 mg / mL for FeSSIF, and 1.3 mg / mL for H2O.

11. The crystalline form according to any one of claims 1 to 10, wherein The crystalline form exhibits a hygroscopicity selected from the group consisting of 0.03%, 9.1% and 1.2% in a DVS (dynamic vapor sorption) analysis at 25°C and 80% RH (room humidity).

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 any solvate or hydrate thereof.

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 mixed with an equal molar amount of maleic acid in IPA and then slurried at room temperature for 3 days; (2) Compound HY-A was mixed with maleic acid in 5 mL IPAc and then slurried at room temperature for 3 days; (3) Grind compound HY-A for 3-5 min; (4) Using compound HY-A, solution vapor diffusion was performed in CH2Br2 / n-hexane at room temperature for 4 days; (5) Compound HY-A was mixed with an equimolar amount of HCl in IPAc and then slurried at room temperature for 3 days; (6) Compound HY-A was mixed with equimolar HCl in ACN / H2O (19:1, v / v) and then slurried at room temperature for 3 days; (7) Compound HY-A was mixed with HCl (molar ratio 1:2, FB / acid) in IPA and slurried at room temperature for 3 days, and then evaporated at room temperature; (8) Compound HY-A was mixed with an equimolar amount of H2SO4 in IPAc and then slurried at room temperature for 3 days; (9) Compound HY-A was mixed with an equimolar amount of p-toluenesulfonic acid in IPAc and then slurried at room temperature for 3 days; (10) Compound HY-A was mixed with methanesulfonic acid (molar ratio 1:2, FB / acid) in IPA and then slurried at room temperature for 3 days; (11) Compound HY-A was mixed with an equimolar amount of oxalic acid in IPAc and then slurried at room temperature for 3 days; (12) Compound HY-A was mixed with an equimolar amount of oxalic acid in MIBK and then slurried at room temperature for 3 days; (13) Compound HY-A was mixed with an equimolar amount of HBr in IPA and then slurried at room temperature for 3 days; and Compound HY-A was slurried with equimolar HBr in IPAc at room temperature for 3 days.

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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