Salt and crystal forms of activin receptor-like kinase inhibitor

Abstract

To provide solid forms of activin receptor-like kinase-2 (ALK2) inhibitor having a property of allowing storage for extended periods of time with minimal absorption of water, decomposition or transformation into other solid forms, a property of being suitable for formulation, and a property of allowing easy absorption following administration to a subject.SOLUTION: The invention provides a succinate salt of Compound (I) represented by the structural formula in the figure, in which the molar ratio between Compound (I) and succinic acid is 1:1.5. Preferably, the succinate salt is in a single crystalline form. The single crystalline form is characterized by a variety of properties and physical measurements. Methods of preparing specific crystalline forms are also disclosed.SELECTED DRAWING: None

Description

Related Applications

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a priority application of U.S. Provisional Application No. 62 / 885,977, filed August 13, 2019. The entire contents of the aforementioned application are incorporated herein by reference. [Background technology]

[0002] Activin receptor-like kinase-2 (ALK2) is a kinase that encodes the activin A receptor type I gene (A ALK2 is a kinase inhibitor in the bone morphogenetic protein (BMP) pathway. It is a serine / threonine kinase that acts as a Inhibition of ALK2 and mutant ALK2 Agents include fibrodysplasia ossificans progressiva (FOP); e.g., major surgical intervention, trauma (head injury) trauma or blast injury), prolonged immobilization, or severe burns ossification (HO); diffuse intrinsic pontine glioma (DIPG), a rare form of brain cancer; and chronic Several diseases, including anemia associated with chronic inflammatory, infectious, or neoplastic diseases, It has the potential to treat.

[0003] No. 10,233,186, the entire teachings of which are incorporated herein by reference. (2013) disclose potent and highly selective inhibitors of ALK2 and mutant forms of ALK2. The inhibitors disclosed in U.S. Pat. No. 10,233,186 (referred to herein as "compounds ( The structure of one of them, designated "I"), is shown below:

[0004] [ka] The successful development of a pharmaceutically active agent such as Compound (I) typically requires facile post-synthetic modification. properties that allow for easy isolation and purification, properties that are amenable to large-scale manufacturing, water absorption, degradation, or It has the property of being able to be stored for a long period of time with minimal conversion to other solid forms, and is suitable for formulation. certain properties, as well as the ability to be easily absorbed after administration to a subject (e.g., soluble in water and gastric juices). The identification of a solid form having the properties (i.e., the solubility of the solid form is 0.01%) is required. Summary of the Invention

[0005] The free base of Compound (I) is physically unstable in moist environments and decomposes when exposed to water. It has now been found that compound (I) tends to become rubbery. It has proven difficult to isolate when prepared on a large scale.

[0006] Also, 1.5:1 succinate (i.e., sesquisuccinate), 1:1 hydrochloride (1:1 The 1:1 hydrochloride and 1:1 fumarate salts (1:1 fumarate salts) were synthesized under well-defined conditions. It has now also been found that it can be crystallized to provide a non-hygroscopic crystalline form ( (See Examples 2-7.) These three salts also have good solubility in water and simulated gastric fluid. It has a high melting point (see Table 2) and is suitable for large-scale synthesis. The 1.5:1 succinate salt exists as a single polymorph and undergoes a thermal transition below its melting point. Since the cellulose acylates do not have any morphological changes, they have the additional advantage of exhibiting a high degree of dimensional stability (see Example 2). (See .4.) The notation "1:1" refers to the ratio of an acid (hydrochloric acid or fumaric acid) to a compound ( The notation "1.5:1" refers to the molar ratio between the acid (succinic acid) and the compound (I The molar ratio between the two carboxylic acid groups on succinic acid and the three carboxylic acid groups on compound (I) is Because of the four basic nitrogen atoms, several possible stoichiometries are possible. For example, Compound (I) forms both a 1:1 hydrochloride and a 2:1 hydrochloride salt. The acid salt is referred to herein as "1:1 Compound (I) HCl" and the 1.5:1 succinate salt is It is referred to herein as "1.5:1 Compound (I) sesquisuccinate salt."

[0007] Compound (I) HCl, Compound (I) fumarate, and Compound (I) The sesquisuccinate salt was identified from a screen of salts with 13 different acids (investigation (See Example 1.) From this salt screen, only eight crystalline forms were identified. The crystalline salts were prepared from benzenesulfonic acid, benzoic acid, fumaric acid, HCl (1 molar equivalent and 2 molar equivalents), maleic acid, salicylic acid, and succinic acid. Therefore, the besylate, maleate, and 2:1HCl have low crystallinity and are stable in a humid environment. It has been found that benzoates are not suitable due to their low water instability (deliquescence). It has been found to be unsuitable due to its solubility and high mass loss upon melting. is preferred due to its low solubility in water, high mass loss upon melting, and potential for polymorphism. It was found that there was no

[0008] In one aspect, the present disclosure provides a succinate salt of Compound (I), wherein Compound (I) and The molar ratio between succinic acid and hydroxybenzoic acid is 1:1.5. As mentioned above, this salt is also referred to herein as "1.5:1 Compound (I) Sesqui-Succinate" It is also called.

[0009] In another aspect, the present disclosure provides an HCl salt of Compound (I), wherein Compound (I) and The molar ratio between the HCl acid is 1:1. As noted above, this salt is also referred to herein as " It is also referred to as "1:1 Compound (I) HCl salt."

[0010] In yet another aspect, the present disclosure provides a fumarate salt of Compound (I), wherein Compound The molar ratio between (I) and fumaric acid is 1:1. This salt is also referred to herein as "1:1 It is also called "Compound (I) fumarate."

[0011] In another aspect, the present disclosure provides a 1.5:1 Compound (I) sesquisuccinate salt (or 1:1 salt). Compound (I) HCl salt or 1:1 Compound (I) fumarate salt) and a pharmaceutically acceptable carrier or a diluent.

[0012] The present disclosure provides a method of treating or ameliorating fibrodysplasia ossificans progressiva in a subject, comprising: and administering to a subject in need thereof a pharmaceutically effective amount of a salt or corresponding pharmaceutical composition disclosed herein. The method comprises administering a composition comprising:

[0013] The present disclosure provides a method of treating or ameliorating diffuse intrinsic pontine glioma in a subject, and administering to a subject in need thereof a pharmaceutically effective amount of a salt or corresponding pharmaceutical composition disclosed herein. The method comprises administering a composition comprising:

[0014] The present disclosure also provides a method of inhibiting aberrant ALK2 activity in a subject in need thereof. administering to a subject of interest a pharmaceutically effective amount of a salt or corresponding pharmaceutical composition disclosed herein. The method also includes:

[0015] The present disclosure also relates to a method for producing a compound of the present disclosure, comprising administering to a subject a salt or a compound thereof in any of the methods of the present disclosure described above. Also provided is a use of the pharmaceutical composition comprising the compound of the present disclosure described herein. and a salt of the present disclosure or a pharmaceutical composition thereof comprising the same for use in any of the methods of In another embodiment, a composition is provided for any of the methods of the present disclosure described. The present invention also provides a use of a salt of the present disclosure or a pharmaceutical composition comprising the same for the manufacture of a medicament for can be. [Brief explanation of the drawings]

[0016] [Figure 1] 1 shows the X-ray powder diffraction (XRPD) pattern of 1.5:1 Compound (I) sesquisuccinate salt. [Figure 2] 1 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) thermograms of 1.5:1 Compound (I) sesquisuccinate salt. [Figure 3] 1.5:1 H-nuclear magnetic resonance spectroscopy (H-NMR) of Compound (I) sesquisuccinate. [Figure 4] 1 shows the DVS isotherm of 1.5:1 Compound (I) sesquisuccinate salt. [Figure 5] 1 shows the XRPD patterns of 1.5:1 Compound (I) sesquisuccinate salt (Form A) before (bottom) and after (top) DVS measurement. [Figure 6] 1 shows the XRPD patterns of 1.5:1 Compound (I) sesquisuccinate salt (Form A) at varying humidity. From bottom to top, XRPD diffractograms were acquired in situ for varying humidity steps of 40% RH, 60% RH, 90% RH, 40% RH, 0% RH, and again 40% RH. [Figure 7] 1 shows the variable temperature XRPD patterns of 1.5:1 Compound (I) sesquisuccinate salt (Form A). From bottom to top, XRPD diffractograms were acquired in situ for variable temperature steps of ambient conditions, 40° C., 60° C., 80° C., 100° C., 120° C., 140° C., 160° C., and again at 25° C. [Figure 8] FIG. 1 shows the XRPD pattern of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). [Figure 9] 1 shows TGA and DSC thermograms of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). [Figure 10] 1 shows the H-NMR of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). [Figure 11] 1 shows the DVS isotherm of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). [Figure 12] 1 shows the XRPD patterns of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A) before (bottom) and after (top) DVS measurement. The extra peak observed after DVS is indicated by the arrow. [Figure 13] 1 shows the XRPD patterns of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A) at varying humidity. From bottom to top, each XRPD diffractogram was acquired in situ for varying humidity steps of ambient conditions, 40% RH, 90% RH, 0% RH, and again 40% RH. [Figure 14] 1 shows the variable temperature XRPD patterns of 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). From bottom to top, XRPD diffractograms were acquired in situ for variable temperature steps of ambient conditions, 50° C., 100° C., 160° C., and again at 25° C. [Figure 15] 1 shows the XRPD patterns of anhydrous 1:1 Compound (I) crystalline HCl salt (Form D) observed during initial screening (bottom) and scale-up (top). [Figure 16] FIG. 1 shows TGA and DSC thermograms of anhydrous 1:1 Compound (I) crystalline HCl salt (Form D). [Figure 17] 1 shows the H-NMR of anhydrous 1:1 Compound (I) crystalline HCl salt (Form D). [Figure 18] 1 shows XRPD patterns of anhydrous 1:1 Compound (I) crystalline HCl salt (Form G) observed during screening (bottom), scale-up (wet) (middle), and drying (top). [Figure 19]1 shows TGA and DSC thermograms of anhydrous 1:1 Compound (I) crystalline HCl salt (Form G). [Figure 20] 1 shows the H-NMR of anhydrous 1:1 Compound (I) crystalline HCl salt (Form G). [Figure 21] 1 shows the XRPD patterns of the anhydrous 1:1 Compound (I) crystalline HCl salt (Form I) observed during initial screening (bottom) and scale-up (top). [Figure 22] 1 shows TGA and DSC thermograms of anhydrous 1:1 Compound (I) crystalline HCl salt (Form I). [Figure 23] FIG. 1 shows the H-NMR of anhydrous 1:1 Compound (I) crystalline HCl salt (Form I). [Figure 24] 1 shows the DVS isotherm of the free base of Compound (I). [Figure 25] FIG. 1 shows the XRPD pattern of 2:1 Compound (I) crystalline HCl salt (Form B). [Figure 26] 1 shows the XRPD patterns of the anhydrous 1:1 Compound (I) crystalline fumarate salt (Form A) observed during initial screening (bottom) and scale-up (top). [Figure 27] 1 shows TGA and DSC thermograms of anhydrous 1:1 Compound (I) crystalline fumarate salt (Form A). [Figure 28] 1 shows the H-NMR of anhydrous 1:1 Compound (I) crystalline fumarate salt (Form A). [Figure 29] FIG. 1 shows the XRPD pattern of 1:1 Compound (I) crystalline fumarate salt (Form C). [Figure 30] FIG. 1 shows the XRPD pattern of 1:1 Compound (I) crystalline fumarate salt (Form D). DETAILED DESCRIPTION OF THE INVENTION

[0017] The present disclosure provides a novel succinate salt of Compound (I) (i.e., 1:1.5 sesquisuccinate salt). ), a novel hydrochloride salt of compound (I) (i.e., 1:1 hydrochloride), and a novel fumarate salt ( i.e., 1:1 fumarate), as well as each of the polymorphic forms listed above.

[0018] "Hydrated form" refers to a solid or crystalline form of Compound (I) in the form of a free base or salt. In this case, water is an integral part of the solid or crystal of the free base compound (I) or the corresponding salt. They are combined in a stoichiometric ratio (e.g., a molar ratio of Compound (I):water of 1:1 or 1:2). "Unhydrated form" refers to a mixture of water and the free base of Compound (I) or Compound (I) and the water is substantially absent in the solid form (e.g. (less than 10% by weight by Karl Fischer analysis) form. Suitable solid forms include hydrated and unhydrated forms.

[0019] As used herein, "crystalline" means that the individual molecules are highly uniform and regular in three dimensions. It refers to a solid that has a crystalline structure and has an original configuration.

[0020] The disclosed crystalline Compound (I) salts may be crystals of one single crystalline form or different single crystalline forms. The single crystalline form means that Compound (I) is a single crystal or a mixture of crystals of different crystalline forms. Alternatively, it means that each crystal is a plurality of crystals having the same crystalline form.

[0021] With respect to the crystalline forms of Compound (I) disclosed herein, the ratio of 1.5:1 Compound (I) At least a certain weight percentage of the salt is in single crystalline form. The certain weight percentages include 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, Contains 96%, 97%, 98%, 99%, 99.5%, 99.9% or compounds (I) 70% to 75% by weight, 75% to 80% by weight, or 80% to 85% by weight of salt , 85% to 90% by weight, 90% to 95% by weight, 95% to 100% by weight, 70 Weight percentages of up to 80 wt%, 80-90 wt%, and 90-100 wt% are in single crystal form All values ​​and ranges between these values ​​and ranges are intended to be encompassed by the present disclosure. Please understand that this is possible.

[0022] A crystalline Compound (I) salt is a compound having a specified percentage of Compound (I) salt in one particular crystalline form. When defined as such, the remainder is amorphous and / or one or more of the specified characteristics. Examples of single crystalline forms include those discussed herein. The 1.5:1 compound (I) sesquioxane is characterized by one or more of the following properties: Succinate (Form A), 1:1 Compound (I) HCl salt (Forms A, D, G, and I), and Compound (I) 1:1 fumarate (Forms A, C, and D) .

[0023] Compound (I) has a chiral center. The salts and polymorphs of compound (I) disclosed herein are I) is relative to the other stereoisomers (i.e., the weight of that stereoisomer relative to the weight of all stereoisomers). Proportion by weight of stereoisomers) at least 80%, 90%, 99%, or 0.9% pure by weight.

[0024] The crystalline Compound (I) salts disclosed herein are those that exhibit highly crystalline materials, A strong characteristic XRPD with a sharp peak and flat baseline corresponding to the position of the angular peak at The XRPD patterns disclosed herein are shown in Figure 1. , obtained from a copper radiation source (Cu Kα1, λ=1.5406 Å).

[0025] Characterization of the 1.5:1 Compound(I) Sesquisuccinate Crystalline Form In one embodiment, the 1.5:1 Compound (I) sesquisuccinate has a solubility of 8.5 degrees 2θ, 15 degrees Characterized by an X-ray powder diffraction pattern including peaks at 0.4° and 21.3° ± 0.2° In another embodiment, Form A has a 2θ of 4. 3°, 8.5°, 14.0°, 15.4°, and 21.3°±0.2°. Characterized by an X-ray powder diffraction pattern containing at least three peaks (or four peaks). In another embodiment, Form A has an average molecular weight of 4.3°, 8.5°, 14.0°, 15°, 2θ. Characterized by an X-ray powder diffraction pattern including peaks at 0.4° and 21.3° ± 0.2° In yet another embodiment, Form A has angles of 4.3°, 6.7°, 8.5°, 1° 2θ. The peaks were at 2.8°, 14.0°, 15.4°, 17.0°, and 21.3° ± 0.2°. In yet another embodiment, Form A is characterized by an X-ray powder diffraction pattern comprising: 2θ: 4.3°, 6.7°, 8.5°, 12.8°, 14.0°, 15.4°, 15.7 °, 16.6°, 17.0°, 18.1°, 19.4°, 19.8°, 20.1°, 20 0.7°, 21.3°, 22.3°, 25.0°, 29.1°, and 34.4°±0.2 In yet another embodiment, the compound is characterized by an X-ray powder diffraction pattern comprising peaks: Form A is characterized by an X-ray powder diffraction pattern substantially similar to that shown in FIG.

[0026] For any given crystalline form, the position of the angular peaks varies with temperature fluctuations, sample misalignment, It is well known in the field of crystallography that there may be slight differences due to factors such as the presence or absence of an internal standard. In this disclosure, the variability of the angular peak position is ±0.2 in 2θ. Therefore, the relative peak intensities for a given crystalline form depend on the crystalline form in the sample preparation for XRPD analysis. This variability can vary due to differences in crystallite size and non-random crystallite orientation. It is well known in the art that the present invention may account for the above factors without precluding the clear identification of the state of affairs. It is knowledge.

[0027] In another embodiment, Form A as the 1.5:1 Compound (I) sesquisuccinate salt is 17 It is characterized by a peak phase transition temperature by differential scanning calorimetry (DSC) of 7±2°C.

[0028] Characterization of the crystalline form of 1:1 Compound (I) hydrochloride In one embodiment, Form A has an average refractive index of 12.9°, 17.0°, 19.0°, 21.1° 2θ. , and at least three peaks (or four peaks) selected from 22.8°±0.2 In another embodiment, the 1:1 compound is characterized by an X-ray powder diffraction pattern comprising: The hydrochloride salt of compound (I) exhibited the following 2θ angles: 12.9°, 17.0°, 19.0°, 21.1°, and 22°. Single crystalline material characterized by an X-ray powder diffraction pattern containing peaks at 0.8°±0.2°. In another embodiment, Form A has a molecular weight of 12.9° 2θ, 13.8° 2θ, and 14.9° 2θ. ±°, 15.1°, 17.0°, 19.0°, 19.6°, 21.1°, and 22.8° The compound is characterized by an X-ray powder diffraction pattern including a peak at 0.2. In the 2θ phase, Form A exhibits 5.7°, 10.1°, 12.6°, 12.9°, and 13.8°. , 15.1°, 17.0°, 19.0°, 19.6°, 20.3°, 21.1°, 22. 1°, 22.8°, 23.4°, 24.0°, 24.8°, 25.5°, 26.1°, and It is characterized by an X-ray powder diffraction pattern including peaks at 28.6°±0.2° and 28.6°±0.2°. In another embodiment, Form A has an X-ray powder diffraction pattern substantially similar to that of FIG. It can be characterized as follows.

[0029] In another embodiment, Form A as the 1:1 Compound (I) hydrochloride salt has a differential It is characterized by the peak phase transition temperature by differential scanning calorimetry (DSC).

[0030] In one embodiment, the 1:1 Compound (I) hydrochloride salt has an angle of 10.8°, 16.9°, 18° in 2θ. At least three peaks selected from 0.8°, 22.1°, and 24.7°±0.2° ( a single crystalline form characterized by an X-ray powder diffraction pattern containing one or four peaks In another embodiment, the 1:1 Compound (I) hydrochloride has a 2θ of 10 Includes peaks at 0.8°, 16.9°, 18.8°, 22.1°, and 24.7° ±0.2° Form D as a single crystalline form characterized by its X-ray powder diffraction pattern.

[0031] In another embodiment, Form D has angles of 10.8°, 13.3°, 16.9°, 18.8° 2θ. by an X-ray powder diffraction pattern containing peaks at 22.1°, 24.7°±0.2° In yet another embodiment, Form D is characterized by an angle of 10.8°, 13.1° 2θ, 13.3°, 16.6°, 16.9°, 17.4°, 18.8°, 20.8°, 22.1 Characterized by an X-ray powder diffraction pattern containing peaks at 24.7°±0.2° and 24.7°±0.2° In yet another embodiment, Form D has an X-ray powder diffraction pattern substantially similar to that of FIG. It is characterized by

[0032] In another embodiment, Form D as the 1:1 Compound (I) hydrochloride salt has a differential It is characterized by the peak phase transition temperature by differential scanning calorimetry (DSC).

[0033] In one embodiment, the 1:1 Compound (I) hydrochloride salt has an angle of 10.2°, 12.8°, 16° in 2θ. At least three angles selected from the following: 0.7°, 17.4°, 18.4°, and 22.5° ±0.2° Characterized by an X-ray powder diffraction pattern containing one or four peaks In another embodiment, the 1:1 Compound (I) hydrochloride salt is Form G as a single crystalline form. , 10.2°, 12.8°, 16.7°, 17.4°, 18.4°, and 22.5° in 2θ A single crystalline form characterized by an X-ray powder diffraction pattern containing peaks at ±0.2° In another embodiment, Form G has a molecular weight of 10.2°, 12.8°, 13.6°, 14.4°, 15.2°, 16.8°, 17.8°, 18.8°, 19.8°, 20.8°, 21.8°, 22.8°, 23.8°, 24.8°, 25.8°, 26.8°, 27. 16.7°, 17.4°, 18.4°, 21.3°, 22.0°, 22.5°, and 24 It is characterized by an X-ray powder diffraction pattern containing peaks at 0.3°±0.2°. In an embodiment, Form G has angles of 10.2°, 12.8°, 14.9°, 16.7° in 2θ, 17.4°, 18.4°, 20.5°, 21.3°, 22.0°, 22.5°, and 24 It is characterized by an X-ray powder diffraction pattern containing peaks at 0.3°±0.2°. In this embodiment, Form D is characterized by an X-ray powder diffraction pattern substantially similar to FIG. be assigned.

[0034] In another embodiment, Form G as the 1:1 Compound (I) hydrochloride salt is at 175±4° C. and 1 Characterized by a peak phase transition temperature by differential scanning calorimetry (DSC) of 97±4°C .

[0035] In one embodiment, the 1:1 Compound (I) hydrochloride salt has an angle of 5.4°, 8.2°, 16.3° in 2θ. At least three angles selected from the following: 16.5°, 18.4°, and 21.5°±0.2 A single peak (or four peaks) characterized by an X-ray powder diffraction pattern In another embodiment, the crystalline form is Form I. In another embodiment, the 1:1 Compound (I) hydrochloride is 2 θ 5.4°, 8.2°, 16.3°, 16.5°, 18.4°, and 21.5°±0. as a single crystalline form, characterized by an X-ray powder diffraction pattern containing the peaks of 2 In another embodiment, Form I has a refractive index of 5.4°, 8.2°, 13.1°, 2θ. X-ray powder containing peaks at 16.3°, 16.5°, 18.4°, and 21.5° ±0.2 In yet another embodiment, Form I is characterized by a diffraction pattern of 5. 4°, 8.2°, 10.2°, 13.1°, 16.3°, 16.5°, 17.1°, 18 The X-ray powder diffraction pattern included peaks at 0.4°, 21.5°, and 21.8° ± 0.2°. In yet another embodiment, Form I is characterized by X substantially similar to FIG. It is characterized by a X-ray powder diffraction pattern.

[0036] In another embodiment, Form I as the 1:1 Compound (I) hydrochloride salt is at 187±4° C. and 2 Characterized by a peak phase transition temperature by differential scanning calorimetry (DSC) of 00±4°C .

[0037] Characterization of the crystalline form of 2:1 Compound (I) hydrochloride salt In one embodiment, the 2:1 Compound (I) hydrochloride salt has an angle of 10.6°, 17.0°, 18° in 2θ. At least three peaks selected from 0.3°, 20.9°, and 21.1°±0.2° ( a single crystalline form characterized by an X-ray powder diffraction pattern containing one or four peaks In one embodiment, the 2:1 Compound (I) hydrochloride has a 2θ of 10. X including peaks at 6°, 17.0°, 18.3°, 20.9°, and 21.1° ± 0.2 Form B is a single crystalline form characterized by a X-ray powder diffraction pattern. In an embodiment, Form B as the 2:1 Compound (I) hydrochloride salt has an average molecular weight of 10.6° in 2θ, 12. 7°, 15.8°, 17.0°, 18.3°, 18.9°, 20.9°, 21.1°, and It is characterized by an X-ray powder diffraction pattern including peaks at 22.0°±0.2° and 22.0°±0.2°. In yet another embodiment, Form B as the 2:1 Compound (I) hydrochloride salt has an angle of 7.8 degrees 2θ, 8.6°, 10.6°, 11.9°, 12.7°, 13.3°, 15.4°, 15.8° , 16.5°, 17.0°, 18.3°, 18.9°, 19.7°, 20.9°, 21. 1°, 22.0°, 22.6°, 24.5°, 26.7°, 27.1°, 28.9°, and It is characterized by an X-ray powder diffraction pattern including peaks at 29.7°±0.2° and 29.7°±0.2°. In yet another embodiment, Form B as the 2:1 Compound (I) hydrochloride salt is substantially as shown in FIG. They are characterized by similar X-ray powder diffraction patterns.

[0038] Characterization of 1:1 Compound(I) fumarate crystalline forms In one embodiment, the 1:1 Compound (I) fumarate salt is Form A as a single crystalline form, 2 θ can be selected from 5.7°, 15.3°, 16.9°, 22.4°, and 23.0°±0.2 The X-ray powder diffraction pattern contains at least three peaks (or four peaks) that correspond to the In one embodiment, the 1:1 Compound (I) fumarate salt has a molecular weight of 5.7 in 2θ. X-rays containing peaks at 15.3°, 16.9°, 22.4°, and 23.0°±0.2° Form A as a single crystalline form characterized by a powder diffraction pattern. In this embodiment, Form A has an average azimuth angle of 5.7°, 7.5°, 9.8°, 10.3°, 12.3° 2θ. , 15.3°, 16.9°, 17.5°, 22.4°, and 23.0° ± 0.2 peaks. In yet another embodiment, Form A is characterized by an X-ray powder diffraction pattern comprising: are 5.7°, 7.5°, 9.8°, 10.3°, 11.2°, 12.3°, and 14° in 2θ. .8°, 15.3°, 16.2°, 16.9°, 17.2°, 17.5°, 18.3°, 18.8°, 19.9°, 20.7°, 21.5°, 22.4°, 23.0°, 23.5 It is characterized by an X-ray powder diffraction pattern including peaks at 25.8°±0.2° and 25.8°±0.2°. In yet another embodiment, Form A has an X-ray powder diffraction pattern substantially similar to that of FIG. It is characterized by

[0039] In another embodiment, Form A as a 1:1 Compound (I) fumarate salt is at 224±2° C. It is characterized by peak phase transition temperatures by differential scanning calorimetry (DSC).

[0040] In one embodiment, the 1:1 Compound (I) fumarate salt has an angle of 6.3°, 9.0°, 13° in 2θ. At least three peaks selected from 0.5°, 18.9°, and 22.5°±0.2° ( a single crystalline form characterized by an X-ray powder diffraction pattern containing one or four peaks In one embodiment, the 1:1 Compound (I) fumarate salt has a 6 .times.2θ angle. X including peaks at 0.3°, 9.0°, 13.5°, 18.9°, and 22.5° ±0.2 Form C is a single crystalline form characterized by a X-ray powder diffraction pattern. In embodiments, Form C has an average molecular weight of 4.5°, 6.3°, 9.0°, 13.5°, 14.7°, 2θ. °, 18.9°, 19.7°, 21.0°, 22.5°, and 23.6° ±0.2 In yet another embodiment, the form C is 4.5°, 6.3°, 7.4°, 9.0°, 13.5°, 14.7°, and 16° in 2θ. .2°, 16.8°, 17.4°, 17.8°, 18.4°, 18.9°, 19.7°, 21.0°, 22.5°, 23.6°, 25.5°, 26.2°, 27.5°, and 28 It is characterized by an X-ray powder diffraction pattern containing peaks at 0.3°±0.2°. In this embodiment, Form C is characterized by an X-ray powder diffraction pattern substantially similar to that of FIG. be assigned.

[0041] In one embodiment, the 1:1 Compound (I) fumarate salt has an angle of 4.6°, 11.0°, 1 At least three peaks selected from 8.5°, 20.5°, and 21.0°±0.2 (or four peaks) in an X-ray powder diffraction pattern. In one embodiment, the 1:1 Compound (I) fumarate salt has a molecular weight of 1:1 at 2θ of 1:1. Includes peaks at 4.6°, 11.0°, 18.5°, 20.5°, and 21.0° ± 0.2°. Form D is a single crystalline form characterized by an X-ray powder diffraction pattern containing In another embodiment, Form D has angles of 4.6°, 11.0°, 15.1°, 18.5° in 2θ, X-ray powder containing peaks at 19.4°, 20.5°, 21.0°, and 25.0° ±0.2 In yet another embodiment, Form D is characterized by a diffraction pattern at 2θ of 4. 6°, 11.0°, 12.0°, 14.3°, 15.1°, 18.5°, 19.4°, 2 Includes peaks at 0.5°, 21.0°, 22.8°, 23.6°, and 25.0° ±0.2. In yet another embodiment, Form D is characterized by an X-ray powder diffraction pattern comprising: Characterized by an X-ray powder diffraction pattern substantially similar to that shown in FIG.

[0042] In one embodiment, the 1:1 Compound (I) fumarate salt is a monocrystalline form admixed with Form D. where Form C has peaks at 6.3°, 9.0°, 13.5°, and 1° 2θ. At least three peaks (or four peaks) selected from 8.9° and 22.5°±0.2 Form D is characterized by an X-ray powder diffraction pattern containing a peak at 2θ of 4.6 11.0°, 18.5°, 20.5°, and 21.0°±0.2. Characterized by an X-ray powder diffraction pattern containing at least three peaks (or four peaks). It can be done.

[0043] In one embodiment, the 1:1 Compound (I) fumarate salt is a monocrystalline form admixed with Form D. where Form C has peaks at 6.3°, 9.0°, 13.5°, and 1° 2θ. Characterized by an X-ray powder diffraction pattern containing peaks at 8.9° and 22.5° ± 0.2° The angles of curvature of the 2θ plane are 4.6°, 11.0°, 18.5°, 20.5°, and 21. It is characterized by an X-ray powder diffraction pattern including a peak at 0°±0.2°.

[0044] In one embodiment, the 1:1 Compound (I) fumarate salt is a monocrystalline form admixed with Form D. where Form C has angles of 4.5°, 6.3°, 9.0°, 13° 2θ. 0.5°, 14.7°, 18.9°, 19.7°, 21.0°, 22.5°, and 23.6 Form D is characterized by an X-ray powder diffraction pattern containing peaks at 2θ ±0.2°. 4.6°, 11.0°, 15.1°, 18.5°, 19.4°, 20.5°, 21.0 It is characterized by an X-ray powder diffraction pattern containing peaks at 25.0°±0.2° and 25.0°±0.2°. do.

[0045] In one embodiment, the 1:1 Compound (I) fumarate salt is a monocrystalline form admixed with Form D. Form C as, where Form C exhibits 4.5°, 6.3°, 7.4°, and 9. 0°, 13.5°, 14.7°, 16.2°, 16.8°, 17.4°, 17.8°, 1 8.4°, 18.9°, 19.7°, 21.0°, 22.5°, 23.6°, 25.5° , 26.2°, 27.5°, and 28.3°±0.2°. Form D is characterized by the following 2θ angles: 4.6°, 11.0°, 12.0°, and 14. 3°, 15.1°, 18.5°, 19.4°, 20.5°, 21.0°, 22.8°, 2 Characterized by an X-ray powder diffraction pattern containing peaks at 3.6° and 25.0° ± 0.2° It can be done.

[0046] Pharmaceutical Compositions The pharmaceutical compositions of the present disclosure may comprise a salt of Compound (I) as described herein, or a crystalline form thereof. and one or more pharmaceutically acceptable carrier(s) or diluent(s). The term "pharmaceutically acceptable carrier" refers to any subject composition or component thereof. Liquid or solid fillers, diluents, excipients, solvents, etc., involved in carrying or transporting ingredients. a pharmaceutically acceptable material, composition, or vehicle, such as a solvent, or encapsulating material, Each carrier must be compatible with the subject composition and its components and must be It must be "acceptable" in the sense of not being harmful to the body. Some examples of materials that may serve are: (1) lactose, glucose, and sucrose; (2) sugars such as corn starch and potato starch; 3) Sodium carboxymethylcellulose, ethyl cellulose, cellulose acetate, etc. cellulose and its derivatives, (4) powdered tragacanth, (5) malt, (6) gelatin, ( 7) Talc, (8) Cocoa butter and suppository wax and other excipients, (9) Peanut oil, cottonseed (10) Oils such as safflower oil, sesame oil, olive oil, corn oil, and soybean oil; Glycols such as propylene glycol, (11) glycerin, sorbitol, mannitol and polyols such as polyethylene glycol, (12) ethyl oleate and lauric acid esters such as ethyl acetate, (13) agar, (14) magnesium hydroxide and aluminum hydroxide (15) alginic acid, (16) pyrogen-free water, (17) isotonic saline solution, (18) Ringer's solution, (19) ethyl alcohol, (20) phosphate buffer solution, and (2 1) Other non-toxic compatible substances used in pharmaceutical formulations.

[0047] The compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, or intravenously. The compositions may be administered intravenously, bucally, intravaginally, or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, and intrasternal In some embodiments, the injection or infusion techniques include intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. The compositions of the present disclosure may be administered orally, intraperitoneally, or intravenously. The spray form may be an aqueous or oily suspension. These suspensions may be prepared by dispersing the liquid in a suitable dispersing agent. or using wetting agents and suspending agents according to techniques known in the art. Sterile injectable preparations may also be prepared, for example, as solutions in 1,3-butanediol. It may be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride. In addition, sterile, fixed oils are conventionally used as a solvent or suspending medium. I can.

[0048] For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of injectables. Likewise, natural pharmaceutically acceptable oils such as olive oil or castor oil, among others, are also suitable. These oil solutions or suspensions are useful in polyoxyethylated versions of these compounds. It also contains carbamazepines commonly used in formulating pharmaceutically acceptable dosage forms, including emulsions and suspensions. a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersant; It may contain: Tweens, Spans, and other emulsifiers or bioavailability enhancers Other commonly used surfactants, such as may also be used for formulation purposes.

[0049] Pharmaceutically acceptable compositions of the present disclosure include capsules, tablets, aqueous suspensions, or aqueous solutions. It may be orally administered in any orally acceptable dosage form, including but not limited to: For tablet use, commonly used carriers include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient may be added to an aqueous suspension containing an emulsifying agent. and suspending agents. If desired, certain sweetening, flavoring, or coloring agents may be added. may also be added.

[0050] Alternatively, the pharmaceutically acceptable compositions of the present disclosure may be administered in the form of suppositories for rectal administration. These allow the drug to be solid at room temperature but liquid at rectal temperature, and Therefore, by mixing with a suitable non-irritating excipient that will melt in the rectum and release the drug Such materials include cocoa butter, beeswax, and polyethylene glycols. Included.

[0051] The pharmaceutically acceptable compositions of the present disclosure also are useful for treating, among other conditions, conditions in which the therapeutic target is the eye, skin, or underarm. involving areas or organs readily accessible by topical application, including diseases of the upper intestinal tract; Suitable topical formulations are readily available for each of these areas or organs. Topical application for the lower intestinal tract may be effected in a rectal suppository formulation (see above) or This can be accomplished in a suitable enema formulation. Topical transdermal patches may also be used.

[0052] For topical application, the pharmaceutically acceptable composition may be suspended or sprayed in one or more carriers. Alternatively, the composition may be formulated in a suitable ointment containing the active components dissolved therein. Carriers for topical administration of the compound include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, and the like. Contains coal, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water Alternatively, a pharmaceutically acceptable composition may comprise one or more A suitable lotion containing the active ingredients suspended or dissolved in a number of pharmaceutically acceptable carriers. The composition may be formulated in a ointment or cream. Suitable carriers include mineral oil, monostearate, Sorbitan, Polysorbate 60, Cetyl Esters Wax, Cetearyl Alcohol, 2 -including but not limited to octyldodecanol, benzyl alcohol, and water do not have.

[0053] The pharmaceutically acceptable compositions of the present disclosure may also be administered by nasal aerosol or inhalation. Such compositions may be prepared and used in accordance with techniques well known in the art of pharmaceutical formulation. benzyl alcohol or other suitable preservative, to enhance bioavailability Absorption enhancers, fluorocarbons, and / or other conventional solubilizing or dispersing agents may be used to It may be prepared as a solution in saline.

[0054] The amount of the compound of the present disclosure that may be combined with the carrier to produce a composition in a single dosage form is Determined by the host being treated, the particular mode of administration, and the person administering the single dosage form. It will vary depending on other factors.

[0055] dosage The toxicity and therapeutic efficacy of the salts of Compound (I) described herein, or crystalline forms thereof, are This can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. LD 50 is the dose that is lethal to 50% of the population. 50 is cured in 50% of the population The dose ratio between toxic and therapeutic effects (LD 50 / ED 50 ) is cured Salts of Compound (I) or crystalline forms thereof that exhibit large therapeutic indices are preferred. The salts of Compound (I) described herein, or crystalline forms thereof, may be used to treat toxic side effects. However, to minimize the possibility of damage to uninfected cells, such salts or bonds may be used. Designing a delivery system that targets the crystalloid form to the site of affected tissue, thereby reducing side effects Care should be taken to ensure that:

[0056] Data obtained from cell culture assays and animal studies are used to evaluate drug candidates for use in humans. The dosage ranges of such salts or crystalline forms may be used in formulating the dosage ranges. ED with little or no toxicity 50 The circulating concentration of the antibody may fall within a range including: The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. With respect to any salt of Compound (I) described herein, or a crystalline form thereof, The appropriate dose can be estimated initially from cell culture assays. When the IC 50 (i.e., the concentration of test compound that achieves half-maximal inhibition of symptoms). It may also be formulated in animal models to achieve a circulating plasma concentration range encompassing This information can be used to more accurately determine useful doses in humans. Levels may be measured, for example, by high performance liquid chromatography.

[0057] The specific dosage and treatment regimen for any particular subject will depend on the specific compound used. activity of the compound, age, weight, general health, sex, diet, time of administration, rate of excretion, drug composition The combination of these factors, as well as the judgment of the treating physician and the severity of the particular condition being treated, It should also be understood that this will depend on a variety of factors, including but not limited to: The amount of a salt or crystalline form of Compound (I) of the present disclosure in a composition may also be determined by the amount of a particular The results will also depend on the compound.

[0058] Treatment method "Subject" refers to a mammal, preferably a human, but also to any animal in need of veterinary treatment, For example, companion animals (e.g., dogs, cats, etc.), livestock animals (e.g., cattle, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). .

[0059] A regimen for "treating" a subject with an effective amount of a compound of the present disclosure may consist of a single administration or Alternatively, it may involve a series of applications, for example, a 1:1 Compound (I) fumarate salt. and 1:1 Compound (I) maleate may be administered at least once a week. However, in another embodiment, the compound is administered from about once per week to about once per day for a given treatment. The length of treatment may depend on the severity of the disease, the age of the subject, and the specificity of the method disclosed herein. The amount of treatment or effect will depend on various factors, such as the concentration and activity of the compound or compounds, or a combination thereof. The effective dose of a compound used for treatment or prophylaxis will vary over the course of a particular treatment or prophylaxis regimen. It will also be understood that dosages may be increased or decreased depending on the dosage. may result from or be revealed by known standard diagnostic assays. In some cases, chronic administration may be required.

[0060] Mutations in ALK2 cause the kinase to become inappropriately activated, leading to a variety of diseases. Compound (I), its salts and crystalline forms disclosed herein inhibit the activity of mutated ALK2 genes. For example, by inhibiting a mutant ALK2 gene that results in the expression of an ALK2 enzyme with an amino acid modification. In another aspect, Compound (I), its salts and crystalline forms disclosed herein are It inhibits both wild-type (WT) and mutant ALK2 proteins. For purposes of disclosure, the sequence information for ALK2 is available from the National Center for Biotechnology Information (NCBI). BI) webpage (https: / / www.ncbi.nlm.nih.gov / ) ACVR1 activin A receptor type 1 [Homo sapiens (human)] ;Found under Entrez Gene ID (NCBI):90. It is FOP, Also known as ALK2, SKR1, TSRI, ACTRI, ACVR1A, and ACVRLK2 and the sequence information is incorporated herein by reference.

[0061] In one embodiment, the present disclosure provides a method of inhibiting aberrant ALK2 activity in a subject. and administering to a subject in need thereof a pharmaceutically effective amount of Compound (I) as described herein, or The method comprises administering a salt, crystalline form, or pharmaceutical composition. In embodiments, the aberrant ALK2 activity is determined by the following mutations: L196P, PF197-8L, R202I, R 206H, Q207E, R258S, R258G, R325A, G328A, G328V , G328W, G328E, G328R, G356D, and R375P The ALK2 gene expression pathway results in the expression of the ALK2 enzyme with several amino acid modifications. In one embodiment, the ALK2 enzyme is a cytoplasmic gene encoding the amino acid modification Has R206H.

[0062] Compound (I), or a salt thereof, as described herein, due to its activity against ALK2, The crystalline forms or pharmaceutical compositions are used to treat conditions associated with aberrant ALK2 activity. The subject can be treated. In one embodiment, the condition associated with aberrant ALK2 activity is Fibrodysplasia ossificans progressiva. The diagnosis of FOP is made by the congenital anomaly of the big toe (hallux valgus) and It is based on the presence of fibrous nodules in the soft tissues, which transform into heterotopic bone. These soft tissue lesions may occur in the head, neck, or back. Approximately 97% of patients with FOP have ACVR1 (ALK2) They share the same c.617G>A;R206H mutation in the gene. Genetic testing is available through Pennsylvania (Kaplan et all, Pediatrics 2008,121(5):e1295-e1300) .

[0063] Other common congenital anomalies include thumb malformations, short and wide femoral necks, osteochondromas of the tibia, and The cervical fused facet joints are used for infant crawling. FOP is often misdiagnosed. Most likely (approximately 80% are cancer or fibromatosis), and subjects undergo inappropriate diagnostic procedures such as biopsies. This frequently exacerbates the disease and causes permanent disability.

[0064] In some embodiments, the present disclosure provides a method for treating or ameliorating fibrodysplasia ossificans progressiva in a subject. The method comprises administering to a subject in need thereof a pharmaceutically effective amount of a compound described herein. (I), or a salt, crystalline form, or pharmaceutical composition thereof. Provide.

[0065] In one embodiment, the condition associated with aberrant ALK2 activity is fibrodysplasia ossificans progressiva. (FOP), and the subjects were L196P, PF197-8L, R202I, R206H, Q207E, R258S, R258G, R325A, G328A, G328W, G328 E, G328R, G356D, and R375P They have a mutation in the ALK2 gene that results in the expression of the ALK2 enzyme with amino acid modifications. In one aspect of this embodiment, the ALK2 enzyme has the amino acid modification R206H.

[0066] The present disclosure relates to Compound (I), or a salt, crystalline form, or pharmaceutical composition as described herein. In one embodiment, the method includes a method for identifying and / or diagnosing a subject for treatment with an agent, the method comprising: The present disclosure provides a method for detecting a condition associated with aberrant ALK2 activity, e.g., FOB, in a subject. a. obtaining a sample, e.g., plasma, from a subject, e.g., a human subject; b. One or more mutations in the ALK2 gene as described herein are present in the sample In another embodiment, the present disclosure provides a method for detecting abnormalities in a subject. The present invention provides a method for diagnosing a condition associated with abnormal ALK2 activity, the method comprising: a. obtaining a sample from a subject; and b. detecting the presence of a nucleotide sequence as described herein using the detection methods described herein. detecting whether one or more mutations in the ALK2 gene are present in the sample; c. Diagnosing a subject with the condition if the presence of one or more mutations is detected. Methods for detecting mutations include hybridization-based methods, amplification-based methods, and Width-based methods, microarray analysis, flow cytometry analysis, DNA sequencing sequencing, next-generation sequencing (NGS), primer extension, PCR, in situ hybridization These include, but are not limited to, immunoblotting, dot blotting, and Southern blotting. In one embodiment, the present disclosure provides a method for detecting a disease associated with aberrant ALK2 activity in a subject. The present invention provides a method for diagnosing and treating a condition, the method comprising: a. obtaining a sample from a subject; and b. one or more mutations in the ALK2 gene as described herein are present in the sample. to detect the presence or absence of one or more mutations in a sample; diagnosing the subject with the condition and administering an effective amount of a compound (I) described herein, if or administering a salt, crystalline form, or pharmaceutical composition thereof to the diagnosed subject. In some embodiments, the disclosure provides a method for treating a condition associated with aberrant ALK2 activity in a subject. a. a subject has a gene encoding one of the ALK2 genes as described herein; or determining whether or not a patient has multiple mutations, and b. receiving information that the patient has one or more of the following: Identifying subjects who are responsive to a plurality of compounds or pharmaceutical compositions; and c. administering to a subject an amount of Compound (I), or a salt, crystalline form, or pharmaceutical composition Includes.

[0067] In one embodiment, the condition associated with aberrant ALK2 activity is a brain tumor, e.g., a glioma. In one embodiment, the glial tumor is a diffuse intrinsic pontine glioma (DIPG). In an embodiment, the present disclosure provides a method for treating or administering to a subject a diffuse intrinsic pontine glioma. The method for treating or ameliorating a disease comprising administering to a subject in need thereof a pharmaceutically effective amount of a compound described herein. The method comprises administering Compound (I), or a salt, crystalline form, or pharmaceutical composition. Provide the law.

[0068] In one embodiment, the condition associated with aberrant ALK2 activity is diffuse intrinsic pontine glioma. The target is R206H, G328V, G328W, G328E, and G356D. and (c) providing expression of the ALK2 enzyme having an amino acid modification selected from one or more of the following: In one aspect of this embodiment, the ALK2 enzyme is It has an acid modification R206H.

[0069] In one embodiment, the condition associated with aberrant ALK2 activity is inflammation, cancer, or a chronic disease. It is anemia associated with the disease.

[0070] In one embodiment, the condition associated with aberrant ALK2 activity is a condition caused by trauma or surgery. Induced heterotopic ossification.

[0071] In certain embodiments, the compounds of the present disclosure are useful in treating the disease to be treated, e.g., FOP. and a second therapeutic agent useful in treating the The drugs are co-administered (either as separate dosage forms administered together or after each other). In one embodiment, the compounds of the present disclosure are administered in combination with steroids (e.g., prednisone) or orally. It is co-administered with other anti-allergy drugs such as marizumab.

[0072] In certain embodiments, the compounds of the present disclosure are useful in treating the disease to be treated, e.g., FOP. The compound is co-administered with an RAR-γ agonist or an antibody against activin to treat In embodiments, the RAR-γ agonist to be co-administered is palovarotene (pal In one embodiment, the activin to be co-administered is The corresponding antibody is REGN2477.

[0073] In certain embodiments, compounds of the present disclosure target mast cells, making them useful in treating FOP. In certain embodiments, the compounds of the present disclosure are co-administered with a therapeutic agent that includes a KIT inhibitor. In some embodiments, the co-administration may be with a mast cell inhibitor, including but not limited to, a steroid or anti-inflammatory drug. The mast cell inhibitor to be used is cromolyn sodium (or sodium cromoglycate). um), brentuximab (ADCETRIS®), ibrutinib (IMBR UVICA®), omalizumab (XOLAIR®), anti-leukotriene antihistamines (e.g., montelukast (SINGULAIR®) or zileuton) (ZYFLO® or ZYFLO CR®), and a KIT inhibitor (e.g., imatinib (GLEEVEC®), midostaurin (PKC412 A), masitinib (MASIVET® or KINAVET®), avapritinib, DCC-2618, and PLX9486).

[0074] The following examples are intended to illustrate and not to limit the scope of the present disclosure in any way. It is not intended to be limiting. [Example]

[0075] [Table 1] [Table 2] [Table 3]

[0076] Analysis conditions X-ray powder diffraction (XRPD) Powder X-ray diffraction was performed using a Rigaku Mini equipped with a Lynxeye detector in reflectance mode. The procedure was performed using a Flex 600 or a Bruker D8 Advance (i.e. The sample was a silicon zero return wafer (zero Typical scans were performed at 40 kV and 15 mA. High resolution: 4 to 30 degrees 2θ using a step size of 0.05 degrees over 5 minutes. The energy scan was performed at 40 kV and 15 mA over 30 minutes using a step size of 0.05 degrees. Typical parameters for XRPD are listed below: [Table 4]

[0077] Thermogravimetric analysis and differential scanning calorimetry (TGA and DSC) Thermogravimetric analysis and differential scanning calorimetry were performed using a Mettler Toledo TGA / DSC 3+The desired amount of sample was placed in an airtight container with a small hole. Weigh directly into the aluminum pan. Typical sample mass for the measurement is 5-10 mg. The typical temperature range is 30°C to 300°C at a heating rate of 10°C per minute (27 The protection and purge gas was nitrogen (20-30 mL / min and 50-100 m Typical parameters for DSC / TGA are listed below: [Table 5]

[0078] Differential scanning calorimetry (DSC) 1–5 mg of material was weighed into an aluminum DSC pan and non-hermetically sealed with an aluminum lid. The sample dish was then placed in a TA Instruments Q2000 (equipped with a cooler). Once a stable heat flow response was obtained at 30°C, the sample and reference material were loaded at 10°C. The sample was heated to 300°C at a rate of 1 / min and the resulting heat flow response was monitored. The instrument was calibrated for temperature and heat flow using a sodium reference standard. With the help of the ERSAL Analysis 2000 software, the temperature of the thermal event The values ​​are referred to as the onset and peak temperatures measured according to the manufacturer's specifications. The analysis was carried out with: Method gas: N2 at 60.00 mL / min.

[0079] 1 H-nuclear magnetic resonance spectroscopy ( 1 H-NMR) Proton NMR was performed on a Bruker Avance 300 MHz spectrometer. was dissolved in 0.75 mL of deuterated solvent in a 4 mL vial and analyzed in an NMR tube (Wilma d 5mm thin-walled 8", 200MHz, 506-PP-8). Typical measurements are usually , 16 scans. Typical parameters for NMR are listed below. [Table 6]

[0080] Dynamic Vapor Sorption (DVS) Dynamic vapor sorption (DVS) was performed using a DVS Intrinsic 1. Sample was loaded into the sample pan and suspended from the microbalance. A typical sample mass for DVS measurements is The desired relative humidity is provided by nitrogen gas bubbled through distilled water. The material was held at each level for a minimum of 5 minutes, with a variation of <0.002% of the weight between measurements (60-second interval). Only when there was a change or 240 minutes had elapsed was the next humidity level advanced. Such a measurement involves the following steps: 1- Equilibrate at 50% RH 2-50%~2%. (50%, 40%, 30%, 20%, 10%, and 2%) a. Each humidity level is maintained for a minimum of 5 minutes and a maximum of 60 minutes. The pass criterion is less than 0.002%. It's a change 3-2%~95% (2%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 95%) a. Each humidity level is maintained for a minimum of 5 minutes and a maximum of 60 minutes. The pass criterion is less than 0.002%. It's a change 4-95%~2% (95%, 80%, 70%, 60%, 50%, 40%, 30%, 20 %, 10%, 2%) a. Each humidity level is maintained for a minimum of 5 minutes and a maximum of 60 minutes. The pass criterion is less than 0.002%. It's a change 5-2%~50% (2%, 10%, 20%, 30%, 40%, 50%) a. Each humidity level is maintained for a minimum of 5 minutes and a maximum of 60 minutes. The pass criterion is less than 0.002%. It's a change

[0081] High-performance liquid chromatography (HPLC) Agilent 1220 Infinity LC: High performance liquid chromatography ( HPLC) was performed using an Agilent 1220 Infinity LC. The flow rate range is 0.2-5.0 mL / min, and the operating pressure range is 0-600 bar. The temperature range is from 5°C above ambient temperature to 60°C, and the wavelength range is from 190 to 600 nm. [Table 7]

[0082] Karl Fischer titration Karl Fischer titration for determining water content is performed using a current generating cell with a diaphragm and a double Mettler Toledo C20S coulometric KF titrator equipped with a platinum pin electrode The test was carried out using Aquastar™ C in both the anode and cathode compartments. The ombiCoulomat fritless reagent was used. Approximately 0.03-0.10 g The sample was dissolved in the anode compartment and heated until the potential of the solution dropped below 100 mV. A 1 wt% Hydranal water standard is used for verification before sample analysis.

[0083] Microscopic observation Optical microscopy was performed using 2.5x, 10x, 20x, and 40x objectives and polarizers. The images were taken using a Zeiss AxioScope A1 equipped with the built-in A Captured with a xiocam 105 digital camera and provided by Zeiss Process using N2 (blue edition) software.

[0084] Example 1: Combinatorial salt screening 1.1.Salt screening According to the Marvin Sketch software prediction, the free base of compound (I) is This compound has multiple pKa values ​​of 8.95, 3.57, and 2.86. It has three basic nitrogens with pKa values. The theoretical logP is 2.98.

[0085] Salt screening was performed using 13 different counterions. All counterions were HCl was also tested using 2.2 equivalents of counterion, Acids were tested using 0.5 equivalents of counter ions. A list of counter ions is provided in Table 1.

[0086] A stock solution of compound (I) was prepared in absolute EtOH (20 wt %, density 0.8547 g Stock solutions of all counterions were also prepared in EtOH. The stock solution of ion is prepared to be 0.02 g / mL, and the counter ion of the liquid is 10% by volume. It was prepared as follows.

[0087] Salt formation was carried out in a 2 mL vial at room temperature. 25 mg of Compound (I) (145.6 1 μL of stock solution) and 1.1 equivalents of counterion were added to each vial. In the case of HCl, 1.1 and 2.2 equivalents of counter ion were added. The solvent was evaporated overnight with stirring at 30°C, then placed under vacuum at 50°C. Allowed to dry completely for 4 hours.

[0088] For screening, approximately 25 volumes of solvent (0.625 mL) were added to each vial. The three solvents chosen were EtOH, EtOAc, and IPA:water (9:1 by volume). Once the solvent was added, the mixture (or solution) was heated to 45°C and 1. The mixture was held for 5 hours, cooled to room temperature, and stirred overnight. Once a slurry was formed, it was analyzed for XRPD analysis. The solid was filtered off.

[0089] XRPD analysis was performed in three stages: wet cake XRPD was performed on all samples. (If a solid is observed). The specific solid is then left on the XRPD plate and subjected to vacuum The solid was dried at 50° C. for at least 3 hours. XRPD of the unique dried solid was then performed. The solid was then exposed to >90% relative humidity for 1 day, and the resulting XRP The humid environment was created by placing a beaker of saturated potassium sulfate solution in a sealed container. All XRPD patterns were generated by comparing the XRPD patterns of the counterions and known ions. The patterns were compared with those of the isolated molecules.

[0090] Solids formed in the first three screening solvents (EtOH, EtOAc, IPA:water) If not, the cap was opened and the solvent was evaporated with stirring at 30°C. Evaporate to dryness by placing under 50 °C for 3-4 hours, and add a second round of solvent. (IPOAc, MBK, MtBE). If no solid was formed in the second round of solvents. The solvent was evaporated to dryness again and DEE was added. [Table 8]

[0091] Benzene sulfonic acid (BSA), benzoic acid, fumaric acid, HCl (1 and 2 equivalents), maleic acid Crystalline solids were observed when screening with carboxylic acid, salicylic acid, and succinic acid. One unique XRPD pattern was obtained from BSA, benzoic acid, HCl (2 equivalents), salicylic acid, and succinic acid. Multiple patterns were observed with HCl (1 equivalent) and fumaric acid. Two patterns were observed for maleic acid, both of which deliquesced upon exposure to humidity. Of these, benzoic acid, fumaric acid, HCl (1 equivalent), salicylic acid, and succinic acid were used. The solid resulting from the leaning did not deliquesce upon exposure to humidity.

[0092] Characterize crystalline salts to determine melting point, crystallinity, stability upon dry and wet exposure, and solubility in water The compounds were evaluated for feasibility based on their polymorphism, polymorphism, and counterion acceptability.

[0093] In view of acceptable physicochemical properties, the mono-HCl salt, succinate salt, and fumarate salt are also preferred. For comparison, the free base was also included for further characterization. .

[0094] Benzoate was not chosen due to its low solubility in water and high mass loss upon melting. Salicylates have low solubility in water, high mass loss upon melting, and the potential for polymorphism. The besylate, maleate, and bis-HCl salts were not selected due to their low yield. It was not selected due to its crystallinity and instability in humid environments (deliquescence).

[0095] The free base sample showed an onset of melting at 116.19°C in DSC. The rum had a gradual mass loss of 0.16 wt.% before melting and a gradual mass loss of 0.05 wt.% upon melting. The solid was finely divided by microscopic observation. Karl Fisher's free base -Titration indicated a water content of 0.37% by weight.

[0096] The free base has high solubility in many organic solvent systems (in most organic solvents tested, High solubility in simulated fluids (>200 mg / mL at room temperature, 0.08 mg / mL in water, fasting Approximately 17 mg / mL in simulated gastric fluid and approximately 7 mg / mL in simulated intestinal fluid under fasting conditions), and acceptable dissolution (onset 116°C) and exhibited low residual solvent (<0.20 wt% by thermogravimetric analysis). The disadvantage of the free base is that it is polymorphic (four patterns were observed during limited screening). It is physically unstable in humid environments (>90% relative humidity) and becomes sticky within 4 days. The results are that it becomes a rubber and becomes rubbery in water. It was also shown that the free base would be difficult to isolate as a crystalline solid on a preparative scale. .

[0097] The mono-HCl salt exhibits high melting (onset temperature of 203°C) and is a hydrate (channel hydrate). It has high crystallinity by X-ray powder diffraction. It has high solubility in water and simulated fluids ( >30 mg / mL in water and fasted simulated gastric fluid, and approximately 7 mg / mL in fasted simulated intestinal fluid Disadvantages of the mono-HCl salt include its sensitivity to added equivalents (as little as 1.3 molar equivalents). These include the formation of a bis-HCl salt at even low HCl levels, and sensitivity to drying.

[0098] The succinate salt showed only one pattern during screening and was stable upon dry and wet exposure. It is less hygroscopic than the mono-HCl salt and the free base, and has high solubility in water and simulated fluids. (>22 m / mL in all fluids), high melting (onset 173°C), and thermal gravity during melting The quantitative analysis indicated an acceptable mass loss (0.27 wt%).

[0099] Fumarate exhibits high solubility in water and simulated fluids (>15 m / mL in all fluids). However, a hypothetical hydrate, designated Form B, was stable upon dry and wet exposure. (anhydrous) exhibited a high melting point (onset 221°C).

[0100] A summary of the physicochemical properties of the free base and selected salts is provided in Table 2 below. [Table 9]

[0101] 1.2. Wet exposure of the free base The crystalline form of the free base was exposed to high humidity (RH > 90%) overnight. The humid environment was maintained in a sealed container. This was created by placing a beaker of saturated aqueous potassium sulfate in the vessel.

[0102] The solid remained in the same crystalline form after overnight exposure to humidity, but retained some crystallinity. After XRPD analysis, the same sample was placed in a humid environment again. After one week, the sample was It was observed that the solid had deliquesced on the plate. A second experiment was started under the same conditions. It became darker and tackier. An XRPD was taken of the sample on day 6. The peak intensities were A low baseline shift was observed, indicating increased amorphous content.

[0103] Example 2: 1.5:1 Preparation of a Crystalline Form of Compound (I) Sesquisuccinate Salt (Form A) and characterization 2.1. Preparation Method A: Compound (I) in its free base form was weighed into a 4 mL vial and 1.1 equivalents of succinic acid was added. EtOH (15 vol) was then added at room temperature. The solid dissolved in the solution. The slurry was heated to 45°C and held under stirring for 2 hours, then allowed to cool to room temperature. The solids still remained in solution, so the samples from the screening The solution was seeded with the starting succinate. The seeds were retained and a white slurry quickly formed. The slurry was stirred overnight at room temperature. Before filtration, the slurry was a medium-thick beige / off-white color. The slurry was filtered, washed twice with 2 volumes of EtOH, and then evaporated under vacuum. It was dried overnight at 50°C. The purity by HPLC was 99.79 area %. The solid was analyzed by XRPD (see Figure 1 and Table 3), TGA-DSC (Figure 2), 1 HNM R (Figure 3), and was further characterized by single-crystal X-ray crystallography.

[0104] The initial mass loss by TGA was 0.19 wt%, followed by 0.30 wt% upon melting. %. See Figure 2. The DSC thermogram shows a melting point with an onset of 172.9°C. , followed by decomposition of the sample above 200°C. [Table 10]

[0105] Method B: Compound (I) free base and 1.6 equivalents of succinic acid under several different solvent conditions Salt formation was carried out using approximately 30 mg of the free base weighed into a 2 mL vial and 0 volume of solvent was added. In all solvents except MtBE, the free base was soluble at room temperature. Succinic acid was then added as a stock solution in EtOH, with each solvent composition containing approximately 40% by volume of EtOH. The solution / slurry was stirred at room temperature until precipitation was observed, and then The solids were collected for XRPD analysis. A summary of the solids obtained from the salt formation experiments is provided in Table 4. Provide. [Table 11]

[0106] Method C: Amorphous Slurry Approximately 30 mg of Compound (I) sesquisuccinate was dissolved in a 2 mL vial to obtain an amorphous A glass-like solid was produced. Solvent (450 μL) was added to each vial along with a stir bar at room temperature. In all cases, a glass-like solid adhered to the bottom of the vial, so a spatula was used to remove it. In many cases, the solids were loosened immediately after being mixed. A brown slurry was formed. When precipitation was observed, the slurry was sent for XRPD analysis. The earliest time point for sampling was approximately 30 minutes after the addition of the solvent. The results and observations from the rally experiment are summarized in Table 5. [Table 12]

[0107] Method D: Amorphous Vapor Diffusion Approximately 10 mg of amorphous Compound (I) sesquisuccinate was placed in a 4 mL vial. Each 4 mL vial was then placed into a 20 mL vial containing 3 mL of solvent and sealed. The vials were kept at room temperature over the weekend before the solids were sampled for XRPD. Most solids range in appearance from light beige glass (crushed from amorphous foam) to white / The amorphous solid, exposed to a moist atmosphere (water as the solvent), turned into a yellowish-white powder. A summary of the solids obtained from the amorphous vapor diffusion experiments is outlined in Table 6. [Table 13]

[0108] In polymorph screening for Compound (I) sesquisuccinate, an amorphous solid was prepared. Including the experiments utilized, over 10 crystallization or salt formation methods have been used to produce solids. Throughout the polymorph screening of Compound (I) sesquisuccinate, crystalline Form A and non-crystalline Form B were obtained. Only crystalline solids were observed.

[0109] A sample of the amorphous solid (Compound (I) sesquisuccinate) was heated to 140°C, followed by room temperature. The resulting solid was crystalline Form A by XRPD.

[0110] Amorphous solid (compound (I) sesquisuccinate) was exposed to RH 75% / 40°C for 1 week. The solid changed appearance from a light beige-yellow solid to a hard yellow glass. XRPD of the compound showed crystalline Form A.

[0111] Form A was found to be crystalline with a melting onset of 173°C and exhibited excellent crystalline properties upon dry and wet exposure. It was stable upon dew and exhibited high solubility in water and simulated fluids (>22 mg in all fluids). / mL.

[0112] Method E: Intermediate 6-(5-(4-ethoxy-1-isopropylpiperidin-4-yl)pyridine -2-yl)-4-(piperazin-1-yl)pyrrolo[1,2-b]pyridazine (3.5 kg, 7.8 mol) (which is disclosed in U.S. Pat. No. 10,233,186) R)-Tetrahydrofuran-3-yl 1H-imidazole-1-carboxylate (1. The mixture was dissolved in isopropyl acetate (IPAc, 2.75 volumes) containing 2 equivalents of methyl methylpropional. The mixture was heated and stirred until complete conversion. The reaction mixture was treated with aqueous ammonia (2 volumes). Upon quenching, additional IPAc (4 volumes) was added. Phase separation, water washing, and distillation yielded This gave a solution of Compound (I) in anhydrous IPAc (approximately 3.5 kg in 3 volumes). While heating, succinic acid in ethanol (1.45 equivalents in 10 volumes) was added. The mixture is heated to 75-85°C for 30 minutes. After cooling to 70-75°C, the solution is added with Compound (I). The suspension was seeded with sesquisuccinate and cooled to 10° C. over 8 hours. The suspension was isolated by filtration. and washed with ethanol (2×3 volumes) to obtain Compound (I) in the form of sesquisuccinate salt. I got an A.

[0113] 2.2. Wet exposure The sesquisuccinate salt obtained in Example 2.1 was exposed to 75% relative humidity at 40°C for 1 week. The samples were placed in 4 mL vials coated with Kimwipe® and then washed for 3-4 min. The 20 mL vial was placed in a 20 mL vial containing 4 mL of saturated aqueous NaCl. It was sealed and kept at 40° C. After one week, the solid was collected for XRPD analysis. Form A is It was physically stable by XRPD after 1 week under humid conditions.

[0114] DVS The DVS showed a mass change of 0.59 to 0.60 wt.% at 25°C and 2 to 95% relative humidity. Of this mass change, 0.34 to 0.35 wt% accounts for over 80%. The XRPD after DVS measurement showed Form A remained (Figure 5).

[0115] DVS was also performed on the free base of Compound (I) (Figure 24), which showed that the free base reacted with 2- At 95% relative humidity, the reversible mass change was 0.88-0.92 wt%. A mass change of 0.46 to 0.53 wt% occurred at relative humidity above 70%.

[0116] 2.4.VT-XRPD and VH-XRPD Possibility of Compound (I) Form A as the sesquisuccinate salt as determined using XRPD Experiments under variable humidity show that no changes in the crystalline structure are observed with humidity ( See Figure 6).

[0117] Variable Temperature Analysis of Compound (I) Form A as Sesquisuccinate Salt Performed Using XRPD In experiments at 160°C, no change in the crystalline structure was observed below 160°C (i.e., the melting point). It can be seen that the

[0118] Example 3: Preparation and Characterization of 1:1 Compound (I) Crystalline HCl Salt Monohydrate 3.1. Preparation Method A: First, 25 to 35 mg of Compound (I) free base was weighed into a 2 mL vial. , solvent was added to the vial (25 vol or 5 vol), followed by 0.9, 1.1, 1.5, 2.2, and 3.5 molar equivalents of HCl stock solution were added.

[0119] First, add IPA:water (9:1 by volume) to make a total of 25 volumes (including the volume of the HCl stock solution). Initially, everything formed a solution. The 1.1 equiv. experiment showed precipitation overnight. All others remained in solution, which may be due to differences in solvent composition. Therefore, the remaining solution (0.9, 1.5, 2.2, and 3.5 equivalents) was heated at 50°C in air. and then evaporated to dryness under active vacuum at 50 °C for approximately 3 h. Additional experiments with 1.1 equiv. was prepared in a similar manner by adding 5 volumes of IPA and the appropriate amount of HCl stock solution. , followed by evaporation to dryness under low vacuum at 50°C and then under active vacuum at 50°C for approximately 3 hours.

[0120] To the evaporated solid was added 25 volumes of EtOAc and allowed to stir at room temperature overnight. A slurry was formed at 100°C. The color of the slurry changed from a white slurry (0.9 equiv.) to a bright white slurry (0.9 equiv.). The reaction mixture varied from a light / dark yellow slurry (over 1.5 equiv).

[0121] The slurry was then filtered and the solid collected for XRPD analysis. The salt formed in equivalent amounts showed Form A (mono HCl) by XRPD (Figure 8). and the use of 1.5 equivalents produces a mixture of Form A (mono-HCl) and Form B (bis-HCl). The use of 2.2 and 3.5 equivalents resulted in Form B (bis-HCl).

[0122] Compound (I) Form A as a crystalline HCl salt was confirmed by TGA-DSC (FIG. 9). 1 H NMR (Figure 10) and further characterized by single crystal X-ray crystallography (Table 7). The DSC showed an onset of melting at 202.86°C (Figure 9). The TGA thermogram A mass loss of 2.81 wt. % occurred before melting (associated with a very broad endotherm in the DSC). The Karl Fischer spectroscopy of the HCl salt showed a step mass loss of 0.44 wt.% upon melting. A water titration indicated a water content of 3.17 wt.%, indicating that the resulting crystalline HCl salt was The theoretical amount of water in the monohydrate of the HCl salt is 3.0% by weight. is. [Table 14] Method B: The free base of compound (I) (4.3 kg) was dissolved in isopropyl acetate (5.5 volumes) and isopropyl acetate (1.5 volumes). The mixture was heated to reflux and HCl (in water) was dissolved in propyl alcohol (2.5 volumes). 16.5% (w / w), 0.95 equivalents) was added over 0.75 hours. Reflux for 1 hour After allowing to stand, the solution was cooled to 20-25°C over 2 hours and held for 0.5 hours. was isolated by filtration and washed with a mixture of IPAc, IPA, and water to give a 1:1 compound ( I) Form A was obtained as a crystalline HCl salt monohydrate.

[0123] 3.2. Wet exposure Form A as the HCl salt monohydrate was placed at 40°C / 75% relative humidity for 1 week. A 1g sample was placed in a 4mL vial coated with Kimwipe®. The vial was placed in a 20 mL sealed vial containing saturated aqueous sodium chloride solution. After one week of wet exposure, some small peak shifts were observed in the XRPD. Peak shifts were also observed in the XRPD patterns of some long-term slurries. This means that Form A as the HCl salt monohydrate is a channel hydrate, and the peak shift is due to the presence of water. This indicates that this may be due to fluctuations in the amount of

[0124] On the day of sampling after one week of humid exposure (RH 75% and 40°C), the samples were collected in a laboratory with low humidity (RH 75%). H<25%).

[0125] After standing in the sealed vial (ambient conditions) for 14 days, the solid was sampled again.

[0126] The solid was determined by XRPD to be 1:1 Compound (I) crystalline HCl salt monohydrate (Form A). Ta.

[0127] 3.3. DVS Form A as HCl Salt Monohydrate DVS was performed on Form A as the HCl salt monohydrate (Figure 11). The mass change was 1.30 to 1.43 wt % at a relative humidity of 2 to 95%. Mass changes of 0.99 to 1.11 wt% occurred at relative humidities below 20%.

[0128] After the DVS measurement, the sample was analyzed by XRPD (Figure 12). All peaks of Form A as a solvate were present, but extra peaks were observed.

[0129] 3.4.VT-XRPD and VH-XRPD Variable humidity analysis performed on Form A as the HCl salt monohydrate using XRPD. The experiment at 0% RH is shown in Figure 13. The peaks at about 10 and 13° (2θ) The small shift in the higher angle direction observed (i.e., smaller d-spacing) is due to the water damage. This is consistent with a contraction of the crystal structure after precipitation and is therefore consistent with a channel hydrate.

[0130] A variable temperature experiment carried out using XRPD is shown in Figure 14. The observed change in g above 100°C is essentially due to the thermal expansion of the unit cell and the simultaneous This confirms that the shrinkage is due to the removal of molecules. The crystalline structure is due to the freezing of bound water / crystallization water. It does not appear to collapse and / or reconstruct as it does in the presence of This is consistent with the formation of a hydrate.

[0131] 3.5. Exposure to dry conditions and rehumidification The 1:1 Compound (I) crystalline HCl salt monohydrate obtained from Example 3.1 was dried under various conditions. The samples were exposed to the conditions and then analyzed by XRPD.

[0132] The conditions were as follows: (1) room temperature in a vial containing P2O5 at 50 °C; 2) 60°C under vacuum, and (3) heated to 140°C in the DSC.

[0133] In all three cases, new XRPD patterns were observed, which were consistent with the 1:1 compound ( I) The anhydrous form of the HCl salt was identified. The relative humidity in the laboratory was adjusted by re-using the sample on the bench. DVS shows no significant mass loss until the relative humidity drops below 20%. I didn't.

[0134] At the 7 day mark, the HCl salt exposed to P2O5 at 50 °C was sampled for XRPD. XRPD immediately after sampling indicated Form D. The sample was stored on the bench (22-23°C, The solid was allowed to stand (RH 28%) for 2.25 hours and analyzed by XRPD. The solid had converted to Form A. After leaving it on the bench overnight, the same sample was analyzed by XRPD and the XRPD showed It remained as Form A. The XRPD pattern is shown in FIG.

[0135] Example 4: Preparation and characterization of anhydrous 1:1 Compound (I) crystalline HCl salt (Form D) 4.1. Preparation The anhydrous 1:1 crystalline HCl salt of Compound (I) (Form D) was dissolved in water containing phosphorus pentoxide at 50°C. It was prepared by prolonged drying of Form A (monohydrate) in a sealed vial. 100 mg of Form A (monohydrate) obtained from Example 4.1 was placed in a dry environment for 4 days. The 4 mL open vial containing the sample was transferred to a 20 mL sealed vial containing P2O5 at 50 °C. The vial was left for 2 days before being sampled, which was identified by XRPD as a new crystalline form. (Form D) (Figure 15).

[0136] Form D converted to Form A upon exposure to ambient conditions (22°C, RH 35%) for 2.25 hours. Therefore, characterization of Form D with minimal exposure to ambient conditions was possible. was carried out.

[0137] The DSC thermogram of the Form D sample showed an endotherm onset at 202.4°C (Figure 16). TGA of the Form D sample showed a total mass loss of 1.10 wt% (Figure 16). The product also exhibits a DSC endotherm with an onset at 202-203°C after dehydration.

[0138] Karl Fischer titration indicated a water content of 0.72 wt% for the Form D sample. The Form D sample exhibited a cubic morphology under microscopic observation. This morphology was consistent with that of the starting material (Form A). The purity of the Form D sample was 98.82 area percent by HPLC. It was percent.

[0139] Form D is a morphology that changes upon humid exposure (overnight at RH>90% and 1 week at RH74% / 40°C). A (monohydrate).

[0140] Compound (I) Form D as a crystalline HCl salt 1 Further characterization was performed by H NMR. (Figure 17). [Table 15]

[0141] Example 5: Preparation and characterization of anhydrous 1:1 Compound (I) crystalline HCl salt (Form G) 5.1. Preparation From IPA solution and from amorphous slurry in EtOAc and MtBE (low crystallinity) Form G was observed during rapid cooling in IPA. Approximately 200 mg of raw HCl salt was weighed into a 20 mL vial and diluted with 60 mL of IPA was added with stirring at 50°C. The solid dissolved and the solution was poured into a beaker of ice water (0°C). The solution was seeded with a sample of Form G at 0°C. The seed crystals were retained, but the thick slurry No solids were formed. The sample was transferred to a -20°C freezer where the solids were allowed to settle over the weekend. He made him the lord.

[0142] The resulting slurry appeared to be spongy. Filtration was extremely slow. The solid was somewhat sticky. The collected solid was quite wet due to insufficient filtration. The collected solid was dried under vacuum overnight at 50°C. The solids obtained were less crystalline than those observed during screening (Figure 18). .

[0143] The DSC thermogram of Form G shows two endotherms with onsets at 163.1°C and 189.6°C. , followed by decomposition (Figure 19). The TGA thermogram shows a 2.6 A gradual initial mass loss of 2 wt. % was followed by a smaller mass loss (0.35 wt. %) during the endothermic event. The stand-alone DSC showed 0.07 wt.% and 0.07 wt.%. The combined DSC-TG The data agree well with the A data and show a broad endotherm between 80 and 130°C. The sample was heated to above a broad endotherm in SC and then cooled to room temperature. No changes were observed in the

[0144] Karl Fischer titration indicated a water content of 2.79 wt% for the sample.

[0145] Microscopic examination of the Form G sample showed solid clumps and some irregular / fine particles. Form G The purity of the sample was 98.89 area percent by HPLC.

[0146] Form G samples partially converted to Form A overnight in a high humidity environment (RH>90%). Form G was stable after 1 week of humidity exposure (RH 75% / 40°C) (XRPD). twist). [Table 16]

[0147] Example 6: Preparation and Characterization of Anhydrous 1:1 Compound (I) Crystalline HCl Salt (Form I) 6.1. Preparation Salt formation experiments were carried out in anhydrous solvent systems (MtBE:IPA and cyclohexane:IPA). Form I was observed when salt formation was carried out in cyclohexane:IPA. Form I was scaled up. Approximately 200 mg of Compound (I) free base was added to a 4 mL flask. The mixture was weighed into a vial and 15 volumes of cyclohexane was added to form a slurry. One molar equivalent of HCl was added as a 0.55 M solution in IPA over 30 minutes. The Cl solution was dispensed dropwise into three aliquots. After the first aliquot, a yellow slurry The rubber formed and subsequently rubberized. The rubber remained upon addition of the final two aliquots. The vials were then heated to 45°C for 1 hour and seeded with the Form I sample. After seeding, the sample was allowed to cool to room temperature. After seeding, a white solid was observed. The sample was primarily a white slurry with some yellow gum stuck to the vial wall. The slurry was filtered and washed twice with 2 volumes of cyclohexane.

[0148] The DSC thermogram of the Form I sample shows an endotherm with an onset at 180.5°C, followed by an endotherm at 198°C. The TGA thermogram shows a small endotherm with an onset of 2.34 wt.% before melting (Figure 22). % and a mass loss of 0.26 wt. % upon melting.

[0149] The stand-alone DSC data agrees well with the combined DSC-TGA data and also shows an endotherm between 90 and 120 °C. Form I samples were heated to 150°C in the DSC and subsequently run at room temperature for XRPD analysis. The mixture was cooled to room temperature. No changes were observed in the XRPD pattern. All peaks is shifted to slightly higher 2theta, which must be due to sample misalignment. do.

[0150] Karl Fischer titration indicated a water content of 2.64 wt % for sample Form I.

[0151] Microscopic observation showed fines (needles) and aggregates. The purity of Form I was 99% by HPLC. The area percent was .51.

[0152] X-ray powder diffraction (XRPD) patterns of the anhydrous 1:1 Compound (I) crystalline HCl salt (Form I) The pattern is shown in Figure 21.

[0153] Form I samples partially converted to Form A overnight in a high humidity environment (RH>90%). . [Table 17]

[0154] Example 7: Preparation and Characterization of Anhydrous 1:1 Compound (I) Crystalline Fumarate Salt (Form A) 7.1. Preparation The free base of Compound (I) was weighed into a 4 mL vial and 1.1 equivalents of fumaric acid was added. Form A as the fumarate salt was scaled up by adding 100 ml of ethyl acetate to the toluene solution. (15 vol) was added at room temperature. The solids mostly dissolved (the slurry became very thin). ), then the solids settled to form a thick white slurry. An additional 5 volumes of EtOAc was added. The slurry was heated to 45°C and kept under stirring for 2 hours. The slurry was stirred overnight at room temperature. The resulting slurry was a thick white slurry. The slurry was filtered and washed twice with 2 volumes of EtOAc. The solid was then dried under vacuum overnight at 50° C. The resulting solid was analyzed by XRPD (FIG. 26 and Table 2). 11), TGA-DSC (Figure 27), 1 1 H NMR (Figure 28) Further characterization was performed. [Table 18]

[0155] Example 8: Preparation and characterization of anhydrous 1:1 Compound (I) crystalline fumarate salt (Form C) 8.1. Preparation To the free base of Compound (I) in a 4 mL vial was added 1.1 equivalents of fumaric acid. IPAc (15 vol) was added at room temperature. The solids were mostly dissolved (the slurry was very The solid then precipitated as an off-white slurry. The slurry was heated to 45°C. The slurry was heated and stirred for 2 hours, then allowed to cool to room temperature. Before filtration, the slurry was a thick white slurry. The slurry was filtered and The solid was washed twice with 100 ml of IPAc and then dried under vacuum overnight at 50° C. was further slurried in EtOH and EtOAc and characterized by XRPD ( See Figure 29 and Table 12). [Table 19]

[0156] Example 9: Preparation and Characterization of Anhydrous 1:1 Compound (I) Crystalline Fumarate Salt (Form D) 9.1. Preparation To the free base of Compound (I) in a 4 mL vial was added 1.1 equivalents of fumaric acid. IPAc (15 vol) was added at room temperature. The solids were mostly dissolved (the slurry was very The solid then precipitated as an off-white slurry. The slurry was heated to 45°C. The slurry was heated and stirred for 2 hours, then allowed to cool to room temperature. Before filtration, the slurry was a thick white slurry. The slurry was filtered and The solid was washed twice with 100 ml of IPAc and then dried under vacuum overnight at 50° C. was further slurried in a mixture of IPA:water (95:5 by volume) and characterized by XRPD. The efficacy was assessed (see Figure 30 and Table 13). [Table 20]

Claims

1. The following structural formula: 【Chemical 1】 A succinate salt of compound (I) represented by the formula: and the molar ratio between the succinate and the hydroxybenzoate is 1:1.

5.

2. 2. The succinate salt of claim 1, wherein the succinate salt is crystalline.

3. 2. The succinate salt of claim 1, wherein the succinate salt is in a single crystalline form.

4. 4. The succinic acid salt of claim 1, wherein the succinate salt is unsolvated. salt.

5. The succinate salt exhibits peaks at 8.5°, 15.4°, and 21.3°±0.2° 2θ. Form A as a single crystalline form characterized by an X-ray powder diffraction pattern comprising The succinate salt according to any one of claims 1 to 4.

6. The succinate salt exhibits the following symmetries in 2θ: 4.3°, 8.5°, 14.0°, 15.4°, and 21. 3°±0.2° by an X-ray powder diffraction pattern containing at least three peaks selected from 5. The method according to claim 1, wherein the monocrystalline form is Form A, characterized by the following: The succinate salt described.

7. The succinate salt exhibits the following symmetries in 2θ: 4.3°, 8.5°, 14.0°, 15.4°, and 21. A single crystalline form characterized by an X-ray powder diffraction pattern containing peaks at 3°±0.2°. The succinate salt according to any one of claims 1 to 4, which is in Form A.

8. The succinate salt exhibits the following chromaticities in 2θ: 4.3°, 6.7°, 8.5°, 12.8°, 14.0°, X-ray powder diffraction pattern including peaks at 15.4°, 17.0°, and 21.3°±0.2 Any one of claims 1 to 4, which is Form A as a single crystalline form, characterized by:

2. The succinate salt according to claim 1.

9. The succinate salt exhibits the following chromaticities in 2θ: 4.3°, 6.7°, 8.5°, 12.8°, 14.0°, １５．４°、１５．７°、１６．６°、１７．０°、１８．１°、１９．４°、１９．８ °, 20.1°, 20.7°, 21.3°, 22.3°, 25.0°, 29.1°, and A single crystal characterized by an X-ray powder diffraction pattern including a peak at 34.4°±0.2°.

5. The succinate salt according to any one of claims 1 to 4, which is in form A as an crystalline form.

10. The succinate salt has a peak phase transition temperature by differential scanning calorimetry (DSC) of 177±2°C. Any one of claims 2 to 9, which is Form A as a single crystalline form, characterized by:

2. The succinate salt according to claim 1.

11. At least 90% by weight of the succinate is in the single-crystalline form A, Claims 5 to 10 The succinate according to any one of the above.

12. The hydrochloride of the compound (I) represented by the following structural formula, 【Chemistry 2】 where the molar ratio between the compound (I) and hydrochloric acid is 1:

1. The above hydrochloride.

13. The hydrochloride according to Claim 12, wherein the salt is crystalline.

14. The hydrochloride according to Claim 12, wherein the salt is in a single-crystalline form.

15. The hydrochloride according to Claim 13 or 14, wherein the salt is a monohydrate.

16. The hydrochloride according to Claim 13 or 14, wherein the salt is not solvated.

17. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including at least three peaks selected from 2θ at 12.9°, 17.0°, 19.0°, 21.1°, and 22. 8° ± 0.

2. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 12.9°, 17.0°, 19.0°, 21.1°, and 22.

18. 8° ± 0.

2. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 12.9°, 13.8°, 15.1°, 17.**. 0°, 19.0°, 19.6°, 21.1°, and 22.8° ± 0.

2.

19. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 12.9°, 13.8°, 15.1°, 17.0°, 19.**. 0°, 19.6°, 21.1°, and 22.8° ± 0.

2. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 12.9°, 13.8°, 15.1°, 17.0°, 19.**. 0°, 19.6°, 21.1°, and 22.8° ± 0.

2.

20. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 5.7°, 10.1°, 12.6°, 12.9°, 13.8°, １５．１°、１７．０°、１９．０°、１９．６°、２０．３°、２１．１°、２２．１ °, 22.8°, 23.4°, 24.0°, 24.8°, 25.5°, 26.1°, and 28.6° ± 0.

2. The hydrochloride according to Claim 15, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including peaks at 2θ of 5.7°, 10.1°, 12.6°, 12.9°, 13.8°,

21. The hydrochloride according to any one of Claims 17 to 20, which is in the single-crystalline form A characterized by a peak phase transition temperature by a differential scanning calorimeter (DSC) at 207 ± 2°C. The hydrochloride according to any one of Claims 17 to 20, which is in the single-crystalline form A characterized by a peak phase transition temperature by a differential scanning calorimeter (DSC) at 207 ± 2°C. The hydrochloride according to any one of Claims 17 to 20, which is in the single-crystalline form A characterized by a peak phase transition temperature by a differential scanning calorimeter (DSC) at 207 ± 2°C.

22. The hydrochloride according to any one of Claims 17 to 20, which is in the single-crystalline form A characterized by an X-ray powder diffraction pattern including at least three peaks selected from 2θ at 5.4°, 8.2°, 16.3°, 16.5°, 18.4°, and 21.5° ± 0.

2. 12, 13, and 14, which are Form I as a single crystalline form, characterized by 16. The hydrochloride salt according to any one of claims 15 and 16.

23. The hydrochloride salt exhibits an average angle of 5.4°, 8.2°, 16.3°, 16.5°, 18.4°, and and 21.5°±0.2, characterized by an X-ray powder diffraction pattern including peaks at 21.5°±0.

2.

16. The compound according to any one of claims 12, 13 and 15, which is in form I as a crystalline form. Hydrochloride salt.

24. The hydrochloride salt exhibits an average angle of 5.4°, 8.2°, 13.1°, 16.3°, 16.5°, and 1° 2θ. Characterized by an X-ray powder diffraction pattern including peaks at 8.4° and 21.5°±0.2°.

16. Any of claims 12, 13, and 15, wherein the monocrystalline form is Form I.

2. The hydrochloride salt according to claim 1.

25. The hydrochloride salt exhibits an average angle of 5.4°, 8.2°, 10.2°, 13.1°, 16.3°, and 1° 2θ. The peaks at 6.5°, 17.1°, 18.4°, 21.5°, and 21.8°±0.2° were included. Form I as a single crystalline form, characterized by an X-ray powder diffraction pattern including:

16. The hydrochloride salt of any one of claims 12, 13 and 15.

26. The hydrochloride salt exhibited differential scanning calorimetry (DSC) peaks of 187±4°C and 200±4°C.

22. Form I as a single crystalline form characterized by a phase transition temperature 26. The hydrochloride salt according to any one of claims 1 to 25.

27. at least 90% by weight of said hydrochloride salt is a single crystalline form selected from Form A and Form I; The hydrochloride salt according to any one of claims 17 to 26,

28. The following structural formula: 【Chemistry 3】 wherein the compound (I) and the fumaric acid salt are and the molar ratio between is 1:

1.

29. 29. The fumarate salt of claim 28, wherein the salt is crystalline.

30. 29. The fumarate salt of claim 28, wherein the salt is in a single crystalline form.

31. The fumarate salt exhibits the following angular positions in 2θ: 5.7°, 15.3°, 16.9°, 22.4°, and 23° . An X-ray powder diffraction pattern containing at least three peaks selected from the range of 0°±0.2° 30. The fumaric acid of claim 29, in form A as a single crystalline form, characterized by: salt.

32. The fumarate salt exhibits the following angular positions in 2θ: 5.7°, 15.3°, 16.9°, 22.4°, and 23° . Single crystalline, characterized by an X-ray powder diffraction pattern including peaks at 0°±0.2° 30. The fumarate salt of claim 29, in the form of Form A.

33. The fumarate salt exhibits an average angle of 5.7°, 7.5°, 9.8°, 10.3°, 12.3° in 2θ. Peaks at 15.3°, 16.9°, 17.5°, 22.4°, and 23.0°±0.2 Form A as a single crystalline form characterized by an X-ray powder diffraction pattern comprising 30. The fumarate salt of claim 29.

34. The fumarate salt exhibits an average angle of 5.7°, 7.5°, 9.8°, 10.3°, 11.2° 2θ, １２．３°、１４．８°、１５．３°、１６．２°、１６．９°、１７．２°、１７．５ °、１８．３°、１８．８°、１９．９°、２０．７°、２１．５°、２２．４°、２３ The X-ray powder diffraction pattern includes peaks at 0°, 23.5°, and 25.8°±0.2°.

30. The fumaric acid of claim 29, in form A as a single crystalline form, characterized by: salt.

35. The fumarate salt has a peak phase transition temperature by differential scanning calorimetry (DSC) of 224±2° C. Any of claims 31 to 34, which is form A as a single crystalline form, characterized by The fumarate salt according to any one of claims 1 to 10.

36. The fumarate salt exhibits the following symmetries in 2θ: 6.3°, 9.0°, 13.5°, 18.9°, and 22. 5°±0.2° with an X-ray powder diffraction pattern containing at least three peaks selected from 30. The fumarate salt of claim 29, which is Form C as a single crystalline form, characterized by: 。

37. The fumarate salt exhibits the following symmetries in 2θ: 6.3°, 9.0°, 13.5°, 18.9°, and 22. A single crystalline form characterized by an X-ray powder diffraction pattern containing peaks at 5°±0.2°.

30. The fumarate salt of claim 29, in the form of Form C.

38. The fumarate salt exhibits an average refractive index of 4.5°, 6.3°, 9.0°, 13.5°, 14.7° in 2θ. Peaks at 18.9°, 19.7°, 21.0°, 22.5°, and 23.6°±0.2 Form C as a single crystalline form characterized by an X-ray powder diffraction pattern comprising 30. The fumarate salt of claim 29.

39. The fumarate salt exhibits an average angle of 4.5°, 6.3°, 7.4°, 9.0°, 13.5°, 1 ４．７°、１６．２°、１６．８°、１７．４°、１７．８°、１８．４°、１８．９° 、１９．７°、２１．０°、２２．５°、２３．６°、２５．５°、２６．２°、２７． It is characterized by an X-ray powder diffraction pattern containing peaks at 28.5° and 28.3°±0.2°.

30. The fumarate salt of claim 29, wherein the fumarate salt is Form C as a single crystalline form.

40. The fumarate salt exhibits an average angle of 4.6°, 11.0°, 18.5°, 20.5°, and 21° 2θ. . An X-ray powder diffraction pattern containing at least three peaks selected from the range of 0°±0.2° 35. The fumaric acid of claim 34, which is form D as a single crystalline form, characterized by: salt.

41. The fumarate salt exhibits an average angle of 4.6°, 11.0°, 18.5°, 20.5°, and 21° 2θ. . Single crystalline, characterized by an X-ray powder diffraction pattern including peaks at 0°±0.2° 35. The fumarate salt of claim 34, in the form of Form D.

42. The fumarate salt exhibits a 2θ angle of 4.6°, 11.0°, 15.1°, 18.5°, 19.4° An X-ray powder diffraction pattern including peaks at 20.5°, 21.0°, and 25.0°±0.2° 35. The compound of claim 34, which is form D as a single crystalline form characterized by a Fumarate.

43. The fumarate salt exhibits a refractive index of 4.6°, 11.0°, 12.0°, 14.3°, 15.1° in 2θ. °, 18.5°, 19.4°, 20.5°, 21.0°, 22.8°, 23.6°, and A single crystal characterized by an X-ray powder diffraction pattern including a peak at 25.0°±0.2°.

35. The fumarate salt of claim 34, which is Form D as an crystalline form.

44. Form C as the fumarate salt exhibits the following symmetries in 2θ: 4.6°, 11.0°, 18.5°, 20. X-ray powder diffraction containing at least three peaks selected from the angles 21.0°±0.2° and 21.5°.

37. The fumarole of claim 36, which is mixed with Form D characterized by a folding pattern. Acid salts.

45. Form C as the fumarate salt exhibits the following symmetries in 2θ: 4.6°, 11.0°, 18.5°, 20. It is characterized by an X-ray powder diffraction pattern containing peaks at 21.0°±0.2° and 21.5°.

38. The fumarate salt of claim 37, wherein the fumarate salt is admixed with Form D, which is

46. Form C as the fumarate salt exhibits the following symmetries in 2θ: 4.6°, 11.0°, 15.1°, 18. X including peaks at 5°, 19.4°, 20.5°, 21.0°, and 25.0°±0.2 39. The compound of claim 38, wherein the compound is blended with Form D characterized by a 1-ray powder diffraction pattern Fumarate salt of.

47. Form C as the fumarate salt exhibits the following symmetries in 2θ: 4.6°, 11.0°, 12.0°, 14. ３°、１５．１°、１８．５°、１９．４°、２０．５°、２１．０°、２２．８°、２ characterized by an X-ray powder diffraction pattern including peaks at 3.6° and 25.0°±0.2°.

40. The fumarate salt of claim 39, wherein said salt is admixed with Form D.

48. At least 90% by weight of the fumarate salt is selected from Form A, Form C, and Form D.

48. The fumarate salt of any one of claims 31 to 47, in a single crystalline form.

49. A salt according to any one of claims 1 to 48 and a pharmaceutically acceptable carrier or diluent.

10. A pharmaceutical composition comprising:

50. A method of treating or ameliorating fibrodysplasia ossificans progressiva in a subject, comprising: A pharmaceutically effective amount of a salt or compound according to any one of claims 1 to 48 is administered to the subject in need thereof.

50. The method comprising administering the pharmaceutical composition of claim 49.

51. The subject is L196P, PF197-8L, R202I, R206H, Q207E, R258S, R258G, R325A, G328A, G328W, G328E, G328 and an amino acid modification selected from one or more of R, G356D, and R375P.

51. The method of claim 50, wherein the ALK2 gene has a mutation that results in expression of the ALK2 enzyme. How to post.

52. 52. The method of claim 51, wherein the ALK2 enzyme has the amino acid modification R206H. 。

53. A method of treating or ameliorating diffuse intrinsic pontine glioma in a subject, comprising administering to said subject a therapeutically effective amount of acetaminophen. administering to the subject in need thereof a pharmaceutically effective amount of at least one of the compounds according to any one of claims 1 to 48 50. The method comprising administering a compound or pharmaceutical composition of claim 49.

54. The subject has R206H, G328V, G328W, G328E, and G356D cysts. and (iii) producing an ALK2 enzyme having an amino acid modification selected from one or more of the following: , the method of claim 53, wherein the patient has a mutation in the ALK2 gene.

55. 55. The method of claim 54, wherein the ALK2 enzyme has the amino acid modification R206H. 。

56. A method of inhibiting abnormal ALK2 activity in a subject, said subject in need thereof. a pharmaceutically effective amount of at least one compound according to any one of claims 1 to 48 50. The method of claim 49, comprising administering the pharmaceutical composition of claim 49.

57. The abnormal ALK2 activity is , Q207E, R258S, R258G, R325A, G328A, G328V, G32 One or more of 8W, G328E, G328R, G356D, and R375P A mutation in the ALK2 gene that results in expression of the ALK2 enzyme having an amino acid modification selected from the group consisting of:

57. The method of claim 56, wherein the mutation is caused by a mutation.

58. 58. The method of claim 57, wherein the ALK2 enzyme has the amino acid modification R206H. 。

59. The subject has fibrodysplasia ossificans progressiva or diffuse intrinsic pontine glioma.

59. The method according to any one of claims 56 to 58.

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