EA040127B1 - CRYSTALLINE FORMS OF COMPOUNDS AND THEIR APPLICATION FOR THE TREATMENT OF LEUKEMIA - Google Patents

Abstract

The present description relates to crystalline forms of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2a]pyrazin-8-amine monomesylate and succinate, which is a Syk inhibitor, and to their use in the treatment of leukemia. The present description also provides pharmaceutical compositions that contain said crystalline forms.

Description

Technical field

The present description relates to crystalline forms of the compounds and their use in the treatment of leukemia. The present description also relates to pharmaceutical compositions containing such crystalline forms.

State of the art

Protein kinases are the largest family of human enzymes and include over 500 proteins. Spleen tyrosine kinase (Syk) is a member of the Syk tyrosine kinase family and regulates early B cell development as well as mature B cell activation, signaling and survival.

Inhibition of Syk activity may be useful in the treatment of allergic disorders, autoimmune diseases, and inflammatory diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis, multiple vasculitis, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease ( COPD), adult respiratory distress syndrome (ARDS), and asthma. In addition, Syk has been reported to play an important role in B cell receptor mediated ligand-independent tonicity signaling, which is known to be an important survival signal in B cells. Thus, inhibition of Syk activity may also be useful in the treatment of certain types of cancer, including B-cell lymphoma and leukemia. US Pat. Nos. 8,455,493 and 8,440,667 describe Syk inhibitors and the contents of those patents are hereby incorporated by reference in their entirety.

There is an ongoing need to provide compounds that are effective inhibitors of Syk, including compounds having desirable pharmacokinetic properties for use as therapeutic agents for the treatment of cancer and other diseases.

Brief description of the invention

In a first aspect of the present invention, 6-(6aminopyrazin-2-yl)-K-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] crystalline Form I is provided. pyrazine-8-amine monomesylate, characterized by X-ray powder diffraction pattern peaks at about 19.7, about 17.3, about 17.9, about 21.6, and about 25.8°2θ±0.2°2θ.

In some embodiments, said diffraction pattern contains peaks at 6.0, 6.2, 8.6, and 9.6°2θ±0.2°2θ.

In a second aspect of the present invention, 6-(6aminopyrazin-2-yl)-K-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] crystalline Form II is provided. pyrazine-8-amine monomesylate, characterized by X-ray powder diffraction pattern peaks at about 17.3, about 25.1, about 20.4, about 19.6, and about 18.5°2θ±0.2°2θ.

In some embodiments, said diffraction pattern contains peaks at 6.1,6.9, 11.0, and 13.6°2θ±0.2°2θ.

In a third aspect of the present invention, 6-(6aminopyrazin-2-yl)-K-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine succinate, characterized by X-ray powder diffraction pattern peaks at about 16.5, about 24.5, about 17.7, about 28.4, and about 21.8°2θ±0.2°2θ.

In some embodiments, said diffraction pattern contains peaks at 8.0 and 8.3°2θ±0.2°2θ.

In a fourth aspect of the present invention there is provided crystalline form II of 6-(6aminopyrazin-2-yl)-K-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine succinate, characterized by X-ray powder diffraction pattern peaks at about 25.0, about 16.3, about 22.0, about 7.9, and about 7.6°2θ±0.2°2θ.

In a fifth aspect of the present invention, there is provided a pharmaceutical composition comprising a crystalline form according to the previous aspects of the invention and at least one pharmaceutically acceptable carrier.

In a sixth aspect of the present invention, there is provided the use of a crystalline form according to the previous aspects of the invention or a pharmaceutical composition according to the fifth aspect of the invention for the treatment of leukemia in a subject.

In some embodiments, said disease or condition is acute myeloid leukemia.

In some embodiments, said subject is a human.

In some embodiments, said compound, or a pharmaceutically acceptable salt or co-crystal thereof, is administered intravenously, intramuscularly, parenterally, nasally, or orally.

In some embodiments, said compound, or a pharmaceutically acceptable salt or co-crystal thereof, is administered orally once a day.

In some embodiments, said compound, or a pharmaceutically acceptable salt or co-crystal thereof, is administered orally twice a day.

In a seventh aspect of the present invention, there is provided the use of a crystalline form according to the previous aspects of the invention for the preparation of a medicament for the treatment of a disease or condition according to the previous embodiments.

Additional aspects and embodiments of the present description are given in this document.

Brief description of the drawings

In FIG. 1 shows the results of XRPD analysis of Form I of the monomesylate salt of the compound of Example 2.

In FIG. 2 shows the results of NMR analysis of Form I of the monomesylate salt of the compound of Example 2.

In FIG. 3 shows the results of the DSC analysis for Form I of the monomesylate salt of the compound according to Example 2.

In FIG. 4 shows the results of TGA analysis for Form I of the monomesylate salt of the compound of Example 2.

In FIG. 5 shows the results of XRPD analysis of Form II of the monomesylate salt of the compound of Example 2.

In FIG. 6 shows the results of NMR analysis of Form II of the monomesylate salt of the compound of Example 2.

In FIG. 7 shows the results of DSC analysis for Form II of the monomesylate salt of the compound of Example 2.

In FIG. 8 shows the results of TGA analysis for Form II of the monomesylate salt of the compound of Example 2.

In FIG. 9 shows the results of XRPD analysis of Form I succinate of the compound of Example 2.

In FIG. 10 shows the results of NMR analysis for Form I succinate of the compound according to Example 2.

In FIG. 11 shows the results of DSC analysis for Form I succinate of the compound according to Example 2.

In FIG. 12 shows the results of TGA analysis for Form I succinate of the compound according to Example 2.

In FIG. 13 shows the results of XRPD analysis for Form II succinate of the compound of Example 2.

In FIG. 14 shows the results of NMR analysis for form II of the compound according to example 2.

In FIG. 15 shows the results of DSC analysis for Form II succinate of the compound according to Example 2.

In FIG. 16 shows the results of TGA analysis for Form II of the compound according to Example 2.

Detailed description of the invention

Surprisingly, it was found that the claimed crystalline forms have the preferred properties that make them attractive for use according to the present description. The compounds, in addition to being Syk inhibitors, have desirable solubility and pharmacokinetic properties. These results are particularly striking given the properties of compounds with comparable parameters of a similar basic structure.

The following description sets forth illustrative methods, parameters, and the like. However, it should be noted that such a description is not intended to limit the scope of the present description, but rather is presented as a description of illustrative implementations.

Definitions.

For the purposes of this specification, the following words and phrases are generally intended to have the meanings given below, unless the context in which they are used indicates otherwise.

The term therapeutically effective amount, or pharmaceutically effective amount, refers to an amount that is sufficient for effective treatment, as defined below, when administered to a subject (eg, a mammal, such as a human) in need of such treatment. The therapeutically or pharmaceutically effective amount will vary depending on the subject and the disease state being treated, the weight and age of the subject, the severity of the disease state, the route of administration, and the like, which can be readily determined by one of ordinary skill in the art. For example, a therapeutically effective amount, or a pharmaceutically effective amount, of a crystalline form is an amount sufficient to modulate Syk expression or activity and thereby treat a subject (e.g., human) suffering from a condition or to alleviate or alleviate existing symptoms of the condition. For example, a therapeutically or pharmaceutically effective amount may be an amount sufficient to reduce a symptom of a disease or condition responsive to inhibition of Syk activity.

- 2 040127

The term polymorph refers to the different crystal structures of a crystalline compound. Different polymorphs may result from differences in crystal packing (packing polymorphism) or differences in packing between different conformers of the same molecule (conformational polymorphism).

The term inhibition means a reduction, such as a significant reduction, of a biological activity or process relative to the original activity. Inhibition of Syk activity refers to a decrease in Syk activity as a direct or indirect response to the presence of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 relative to Syk activity in the absence of such compound or a pharmaceutically acceptable salt or co-crystal thereof. . Said decrease in activity may be due to the compound's direct interaction with Syk, or due to the interaction of the compound(s) described herein with one or more other factors which in turn affect Syk activity. For example, the presence of the compound(s) can decrease Syk activity by binding directly to Syk, by causing another factor to decrease Syk activity (directly or indirectly), or by (directly or indirectly) reducing the amount of Syk present in a cell or organism. In some embodiments, inhibition of Syk activity may be compared for the same subject prior to treatment, or may be compared with other untreated subjects.

Inhibition of Syk activity also refers to the observed inhibition of Syk activity in a standard biochemical assay for Syk activity, such as the ATP hydrolysis assay described in Example 12 below.

In some embodiments, a compound described herein, e.g., Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, inhibits Syk kinase activity with an IC ₅₀ value of less than or equal to 1 μM, such as 0.1 nM to 1 µM or 1 nM to 1 µM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC ₅₀ value less than or equal to less than 500 nM, such as 0.1 to 500 nM, or 1 to 500 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an IC ₅₀ value less than or equal to less than 200 nM, such as 0.1 to 200 nM, or 1 to 200 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an IC ₅₀ value less than or equal to less than 100 nM, such as 0.1 to 100 nM, or 1 to 100 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an IC ₅₀ value less than or equal to 50 nM, such as 0.1 to 50 nM, or 1 to 50 nM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC50 value less than or equal to 20 nM, such as 0.1 to 20 nM or 1 to 20 nM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC50 value less than or equal to 10 nM, such as 0.1 to 10 nM or 1 to 10 nM. In some embodiments, the IC ₅₀ value is determined as described in the study according to Example 12.

Inhibition of B cell activity refers to a decrease in B cell activity as a direct or indirect response to the presence of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 relative to B cell activity in the absence of such compound, or a pharmaceutically acceptable salt or co-crystal thereof. The decrease in activity may be due to direct interaction of the compound with Syk, or one or more factors in turn affect B cell activity.

Inhibition of B cell activity also refers to the observed inhibition of CD86 expression in the standard assay. In some embodiments, a compound described herein has an IC ₅₀ value less than or equal to 10 μM, such as 1 nM to 10 μM, or 10 nM to 10 μM. In some embodiments, the compound has an IC ₅₀ value less than or equal to less than 1 μM, such as 1 nM to 1 μM, or 10 nM to 1 μM. In some embodiments, the compound has an IC ₅₀ value less than or equal to 500 nM, such as 1 to 500 nM or 10 to 500 nM.

The term B cell activity also includes activation, redistribution, rearrangement, or capping of one or more different B cell membrane receptors or membrane-bound immunoglobulins, such as IgM, IgG, and IgD. Most B cells also have membrane receptors for the Fc portion of IgG, either in the form of antigen-antibody complexes or aggregated IgG. B cells also contain membrane receptors for activated complement components such as C3b, C3d, C4 and C1q. These various membrane receptors and membrane-bound immunoglobulins are characterized by membrane mobility and can undergo redistribution and capping, which can initiate signal transduction.

B cell activity also includes the synthesis and production of antibodies or immunoglobulins. Immunoglobulins are synthesized by a series of B cells and have common structural features and structural units. There are five classes of immunoglobulins, i.e. IgG, IgA, IgM, IgD and IgE, based on structural differences in their heavy chains, including amino acid sequence and polypeptide chain length. Antibodies to a particular antigen may be defined for all or several classes of immunoglobulins and may be limited to one class or subclass of immunoglobulins. Autoantibodies, or autoimmune antibodies, may likewise belong to one or more classes of immunoglobulins. For example, rheumatoid factors (anti-IgG antibodies) are most commonly identified as IgM immunoglobulin, but they can also be composed of IgG or IgA.

In addition, B cell activity is also thought to involve a series of events leading to clonal expansion (proliferation) of B cells from progenitor B lymphocytes and differentiation into antibody-producing plasma cells, which occurs in conjunction with antigen binding and signaling. cytokines from other cells.

The term inhibition of B cell proliferation refers to inhibition of the proliferation of abnormal B cells, such as cancerous B cells, such as lymphoma B cells, or inhibition of normal, non-diseased B cells. The term inhibition of B cell proliferation means any significant reduction in the number of B cells, either in vitro or in vivo. Thus, inhibition of B cell proliferation in vitro is any significant reduction in the number of B cells in an in vitro sample when it is brought into contact with Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 of compared with an equivalent sample that was not brought into contact with the compound(s).

Inhibition of B cell proliferation also refers to the observed inhibition of B cell proliferation in a standard thymidine study for B cell proliferation, such as a study known in the art. In some embodiments, the compounds described herein, for example, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, have an IC ₅₀ value less than or equal to 10 μM, such as from 1 nM to 10 µM or 10 nM to 10 µM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC ₅₀ value less than or equal to less than 1 μM, such as 1 nM to 1 μM, or 10 nM to 1 μM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC ₅₀ value less than or equal to 500 nM, such as 1 to 500 nM or 10 to 500 nM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC50 value less than or equal to 200 nM, such as 1 to 200 nM or 10 to 200 nM. In some embodiments, the compound or a pharmaceutically acceptable salt or co-crystal thereof has an IC50 value less than or equal to 100 nM, such as 1 to 100 nM or 10 to 100 nM.

The term reduction in basophil activation refers to the ability of the compounds described herein to reduce basophil activation. Basophil activation has been implicated, for example, in the inflammatory and autoimmune diseases described herein, and such reduction in basophil activation is required for the compounds described herein, for example Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 example 2. Basophil activation can be assessed by determining the expression of CD63 by basophils, for example, by testing CD63 in human whole blood cells (25% blood), such as the test described in example 9 below.

In some embodiments, a compound described herein, e.g., Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, has an EC ₅₀ value in an appropriate CD63 assay less than or equal to 10 μM, such as 1 nM to 10 µM or 10 nM to 10 µM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an EC50 value less than or equal to less than 1 μM, such as 1 nM to 1 μM, or 10 nM to 1 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an EC ₅₀ value less than or equal to 500 nM, such as 1 to 500 nM, or 10 to 500 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an EC ₅₀ value less than or equal to 200 nM, such as 1 to 200 nM, or 10 to 200 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an EC ₅₀ value less than or equal to 150 nM, such as 1 to 150 nM, or 10 to 150 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an IC50 value less than or equal to 100 nM, such as 1 to 100 nM, or 10 to 100 nM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has an EC50 value less than or equal to 75 nM, such as 1 to 75 nM, or 10 to 75 nM. In some embodiments, the EC50 value is determined as described in the study of Example 9.

The term kinetic solubility refers to the study of the solubility of a compound in a suitable buffer, such as phosphate buffer, pH 7.4 at a given temperature, eg 37°C. In one case, the kinetic solubility is measured at 37°C in phosphate buffer at pH 7.4,

- 4 040127 for example, through the study described in example 10.

In some embodiments, the compounds described herein, for example, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, have a kinetic solubility at 37° C. in phosphate buffer pH 7.4 greater than or equal to 10 μM, such as from 10 to 500 μM or from 10 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 20 μM, such as 20 to 500 μM or 20 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 30 μM, such as 30 to 500 μM or 30 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 40 μM, such as 40 to 500 μM or 40 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 50 μM, such as 50 to 500 μM or 50 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37° C. in pH 7.4 phosphate buffer greater than or equal to 60 μM, such as 60 to 500 μM or 60 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 70 μM, such as 70 to 500 μM or 70 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37° C. in pH 7.4 phosphate buffer greater than or equal to 80 μM, such as 80 to 500 μM or 80 to 250 μM. In some embodiments, the compound, or a pharmaceutically acceptable salt or co-crystal thereof, has a kinetic solubility at 37°C in pH 7.4 phosphate buffer greater than or equal to 90 μM, such as 90 to 500 μM or 90 to 250 μM. In some embodiments, kinetic solubility is determined by the assay described in Example 10.

Stability in human hepatocytes is a measure of the stability of compounds with respect to metabolism by human hepatocytes and is measured as the estimated hepatic plasma clearance of compounds in l/h/kg. Estimated clearance in hepatocytes can be determined, for example, by the assay described in Example 11.

In some embodiments, the compounds described herein, for example, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2, have an estimated liver plasma clearance of less than or equal to 0.50 l/h/ kg, such as 0.005 to 0.50 l/h/kg or 0.01 to 0.50 l/h/kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.40 L/h/kg, such as 0.005 to 0.40 L/h/kg or 0.01 to 0.40 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.30 L/h/kg, such as 0.005 to 0.30 L/h/kg or 0.01 to 0.30 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.20 L/h/kg, such as 0.005 to 0.20 L/h/kg or 0.01 to 0.20 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.10 L/h/kg, such as 0.005 to 0.10 L/h/kg or 0.01 to 0.10 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.09 L/h/kg, such as 0.005 to 0.09 L/h/kg or 0.01 to 0.09 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.08 L/h/kg, such as 0.005 to 0.08 L/h/kg or 0.01 to 0.08 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.07 L/h/kg, such as 0.005 to 0.07 L/h/kg or 0.01 to 0.07 L/h/kg. kg. In some embodiments, the compound has an estimated liver plasma clearance less than or equal to 0.06 L/h/kg, such as 0.005 to 0.06 L/h/kg or 0.01 to 0.06 L/h/kg. kg. In some embodiments, estimated clearance in hepatocytes is determined by the assay described in Example 11.

An allergy, or allergic disorder, refers to an acquired hypersensitivity to a substance (allergen). Allergic conditions include eczema, allergic rhinitis or runny nose (coryza), hay fever, bronchial asthma, hives (rash) and food allergies, as well as other atopic conditions.

Asthma refers to a disorder of the respiratory system characterized by inflammation, constriction of the airways, and increased airway response to inhaled agents. Asthma is often, but not exclusively, associated with atopic or allergic symptoms.

By significant is meant any detectable change that is statistically significant in a standard parametric test of statistical significance, such as the Student's t-test, in which p<0.05.

A disease susceptible to inhibition of Syk activity is a disease

- 5 040127 in which the inhibition of Syk kinase provides a therapeutic effect, such as alleviating symptoms, reducing the progression of the disease, delaying the onset of the disease, or inhibiting the aberrant activity of certain cell types (monocytes, B cells and mast cells).

The term subject refers to an animal, such as a mammal, that is or will be the subject of treatment, observation, or experiment. The methods described herein can be applied to both human therapy and veterinary applications. In some embodiments, the subject is a mammal, in some embodiments, the subject is a human, and in some embodiments, the subject is selected from cats and dogs. The term subject in need, or human in need thereof, refers to a subject, such as a human, who may have or is suspected of having diseases or conditions that are beneficially affected by some treatment, such as treatment with Form I or II of the monomesylate salt of the compound according to Example 2 or Form I or II of the succinate compound of Example 2 described herein. These terms include a subject who may be determined to be at risk or suspected of having a disease or condition such that treatment would prevent the development of the disease or condition.

Treatment, or administration of treatment, is an approach to obtain preferred or desired outcomes, including clinical outcomes. Preferred or desired clinical outcomes may include one or more of the following:

(i) suppressing the disease or condition (eg, reducing the severity of one or more symptoms that occur due to the disease or condition, and/or reducing the degree of the disease or condition);

(ii) slowing or inhibiting the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and/or preventing or delaying the spread (e.g., metastasis) of the disease or condition ); and/or (iii) alleviating the disease, i. e. providing an action that causes regression of clinical symptoms (for example, alleviating a disease state, providing partial or complete remission of a disease or condition, enhancing the effect of another drug, delaying the progression of a disease, increasing quality of life and / or prolonging survival time).

Delaying the development of a disease or condition means delaying, preventing, slowing down, inhibiting, stabilizing and/or postponing the development of a disease or condition. This delay may be of varying duration depending on the history of the disease or condition and/or the subject being treated. A method that delays the development of a disease or condition is a method that reduces the likelihood of developing a disease or condition in a certain period of time and/or reduces the degree of a disease or condition in a certain period of time compared to when the method is not used. Such comparisons are usually based on clinical studies using a statistically significant number of subjects. The development of a disease or condition can be detected using standard methods such as routine physical examination, mammography, imaging, or biopsy. Development may also refer to the progression of a disease or condition, which may not be initially detectable, and includes occurrence, recurrence (relapse), and onset.

In many cases, the compounds according to the present description are able to form acidic and/or basic salts due to the presence of amino and/or carboxyl groups or groups similar to them.

Pharmaceutically acceptable salts include, for example, inorganic acid salts and organic acid salts. Examples of salts may include hydrochloride, phosphate, diphosphate, hydrobromide, sulfate, sulfinate, nitrate, malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate (mesylate), benzenesulfonate (besylate), p-toluenesulfonate ( tosylate), 2-hydroxyethylsulfonate, benzoate, salicylate, stearate and alkanoate (such as acetate, HOOC-(CH ₂ ) _p -COOH, where n is 0-4). In addition, when the compounds described herein are obtained as an acid addition salt, the free base can be obtained by alkalizing the acid salt solution. Conversely, if the product is a free base, an addition salt, in particular a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid in accordance with conventional techniques for preparing acid addition salts from basic compounds. Those skilled in the art will recognize various synthetic procedures that can be used to prepare non-toxic, pharmaceutically acceptable addition salts.

Form I and II of the monomesylate salt of the compound of Example 2 and Form I and II of the succinate of the compound of Example 2 may also be a pharmaceutically acceptable co-crystal or co-crystal salt. The terms co-crystal or co-crystal salt as used herein means a crystalline material composed of two or more unique solids at room temperature, each with its own distinctive physical characteristics,

- 6 040127 such as structure, melting point, as well as heats of fusion, hygroscopicity, solubility and stability. A co-crystal or co-crystal salt can be obtained according to a known co-crystalization method. The terms co-crystal (or co-crystal) or co-crystal salt also refer to a multi-component system in which there is a host API molecule or molecules, such as form I or II of the monomesylate salt of the compound according to example 2 or form I or II of the succinate of the compound according to example 2, and the guest molecule or molecules (or co-formers). In particular embodiments, said pharmaceutically acceptable co-crystal of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 with a co-forming molecule is in a crystalline form selected from malonic acid co-crystal, succinic acid co-crystal, co-crystal with decanoic acid, salicylic acid co-crystal, vanillic acid co-crystal, maltol co-crystal or glycolic acid co-crystal. The co-crystals may have improved properties compared to the original form (ie, free molecule, zwitterion, etc.) or salt of the parent compound. Improved properties may include increased solubility, increased dissolution, increased bioavailability, increased dose response, reduced hygroscopicity, crystalline form of a compound that is generally amorphous, crystalline form of a compound that is difficult to form a salt or a non-salt forming compound, reduced diversity of form diversity, more desirable morphology, and etc.

The term crystalline forms and the corresponding terms used herein refer to various crystalline modifications of a given substance, including, but not limited to, polymorphs, solvates, hydrates, co-crystals and other molecular complexes, as well as their salts, solvates of salts, hydrates of salts, other molecular salt complexes and their polymorphs. Crystalline forms of the substance can be obtained using a number of methods known in the art. Such methods include, but are not limited to, melt recrystallization, melt cooling, solvent recrystallization, recrystallization in confined spaces such as, for example, nanopores or capillaries, recrystallization on surfaces or matrices, such as, for example, polymers, recrystallization in the presence of additives such as co-crystal auxiliary molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, pulverization, and solvent droplet pulverization, for example.

As used herein, a pharmaceutically acceptable excipient is a pharmaceutically acceptable excipient which includes, without limitation, any and all carriers, diluents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional vehicle or agent is incompatible with the active ingredient, it is intended to be used in therapeutic compositions. Additional active ingredients may also be included in the compositions.

The term carrier refers to an adjuvant or carrier, which includes, but is not limited to, diluents, disintegrants, settling inhibitors, surfactants, glidants, binders, lubricants, and the like, with which the compound is administered. Carriers are generally described herein as well as in Remington's Pharmaceutical Sciences, E.W. Martin. Examples of carriers include, but are not limited to, aluminum monostearate, aluminum stearate, carboxymethylcellulose, sodium carboxymethylcellulose, crospovidone, glyceryl isostearate, glyceryl monostearate, hydroxyethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxyoctacosanylhydroxystearate, hydroxypropylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, lactose monohydrate, magnesium nitrate, microcrystalline cellulose, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 188, poloxamer 237, poloxamer 407, povidone, silicon dioxide, colloidal silicon dioxide, silicone, silicone adhesive 4102 and silicone emulsion. However, it should be understood that the carriers chosen for pharmaceutical compositions and the amounts of such carriers in the composition may vary depending on the method of preparation of the formulation (examples are dry granular formulation, solid dispersion formulation).

The term diluent generally refers to a substance that is used to dilute the compound of interest prior to delivery. Diluents may also serve to stabilize compounds. Examples of diluents may include starch, saccharides, disaccharides, sucrose, lactose, polysaccharides, cellulose, cellulose ethers, hydroxypropyl cellulose, sugar alcohols, xylitol, sorbitol, maltitol, microcrystalline cellulose, calcium or sodium carbonate, lactose, lactose monohydrate, dibasic calcium phosphate, cellulose, compressible sugars, dibasic calcium phosphate dehydrate, mannitol, microcrystalline cellulose and tricalcium phosphate.

The term disintegrant generally refers to a substance which, when added to a solid preparation, facilitates its disintegration or decomposition after administration and causes the active ingredient to be released as efficiently as possible to ensure its rapid dissolution. Examples of disintegrants may include corn starch, sodium starch glycolate, sodium croscarmellose, crospovidone, microcrystalline cellulose, modified corn starch, sodium carboxymethyl starch, povidone, pregelatinized starch, and alginic acid.

The term precipitation inhibitors generally refers to a substance that prevents or inhibits the precipitation of an active agent from a supersaturated solution. One example of a precipitation inhibitor includes hydroxypropyl methylcellulose (HPMC).

The term surfactants generally refers to a substance that reduces the surface tension between liquid and solid, which can improve the wetting of the active agent or improve the solubility of the active agent. Examples of surfactants include poloxamer and sodium lauryl sulfate.

The term glidant generally refers to substances used in tablet and capsule formulations to improve flow properties during tablet compression and to provide an anti-blocking effect. Examples of glidants may include colloidal silica, talc, fumed silica, starch, starch derivatives, and bentonite.

The term "binder" generally refers to any pharmaceutically acceptable film that can be used to bind the active and inert components of the carrier to maintain the associated and separate parts. Examples of binders may include hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone, copovidone and ethylcellulose.

The term lubricant generally refers to a substance that is added to a powder mixture to prevent the compacted powder mass from sticking to the equipment during the tableting or encapsulation process. The lubricant can help push the tablet out of the mold and can improve the flow of the powder. Examples of lubricants may include magnesium stearate, stearic acid, silica, fats, calcium stearate, polyethylene glycol, sodium stearyl fumarate, or talc, as well as solubilizers such as fatty acids, including lauric acid, oleic acid, and C ₈ /C fatty acid. _1o .

Connections.

In this document and elsewhere in the present description, compounds are provided, such as those given in the Brief Description and Examples sections.

The names of the compounds provided herein are given using ChemBioDraw Ultra 12.0, and one skilled in the art will appreciate that the structure of a compound may be named and identified using other conventional nomenclature and symbol systems, including CAS and IUPAC.

Exemplary compounds of the present invention are listed in Table A below. The names of the compounds shown in Table A are given using ChemBioDraw Ultra 12.0 and it should be understood that other names may be used to identify compounds with the same structure. Other compounds or radicals may be named using conventional names, or systematic or non-systematic names. Compound names may also be given using other systems of nomenclature and symbols that are commonly recognized in the field of chemistry, including, for example, the nomenclature and symbol systems of the Chemical Abstract Service (CAS) and the International Union of Pure and Applied Chemistry ( IUPAC). Any ambiguity in the designation of compounds can be overcome by referring to the structure when it is presented.

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Table A

Illustrative Connections

Structure Name °L τ 1 Η ₂ Ν XJ 6-(6-amino-5-methylpyrazin-2-yl)-M-(4-(4(oxetan-3-yl)piperazin-1yl)phenyl)imi dazo[1,2-a]pyrazin-8-amine Λ Λ Λ Λ .Λ Λ 0 0 0 0 0 0 0 { 0 Ρ ¹ 0 $ 0 ε* i / ύ- ^s 0! U® 6-(6-Aminopyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazine8-amine (K)-(4-(4-((6-(6-Aminopyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)phenyl)morpholin-2yl)methanol 6-(6-aminopyrazin-2-yl)-5-methyl-M-(4-(4(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)-2-(4-(oxetan-3yl)piperazin-1-yl)phenoxy )ethanol 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)phenyl)piperazin-1yl)methyl)propan-1,3 - diol 2-(5-((6-(6-amino-5-methylpyrazin-2yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4(oxetan-3-yl)piperazin- 1-yl)phenoxy)ethanol

In one embodiment, the present invention provides 6-(6-aminopyrazin-2-yl)-N(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazin-8-amine mesylate, e.g. -8-amine monomesylate. For example, the present invention provides Form I 6-(6-aminopyrazin-2-yl)-N-(4-(4(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine monomesylate, which may have XRPD peaks at about 19.7, about 17.3, about 17.9, about 21.6, and about 25.8 (degrees 2-theta). Also proposed is Form II of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine monomesylate, which may have XRPD peaks at about 17.3, about 25.1, about 20.4, about 19.6, and about 18.5 (degrees 2-theta).

In one embodiment, the present invention provides 6-(6-aminopyrazin-2-yl)-K(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine succinate. For example, the present invention provides Form I 6-(6-aminopyrazin-2-yl)-K-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazine -8-amine succinate, which may have XRPD peaks at about 16.5, about 24.5, about 17.7, about 28.4, and about 21.8 (degrees 2-theta). The present invention also provides Form II 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine succinate, which may have XRPD peaks at about 25.0, about 16.3, about 22.0, about 7.9, and about 7.6 (degrees 2-theta).

The term about as used in relation to XRPD peaks means, for example, ±0.2, ±0.1, ±0.05 (degrees 2-theta), and the like.

The compounds described herein, for example Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2, provide distinct advantages as Syk inhibitors. The compounds described herein are inhibitors of Syk kinase activity, as defined, for example, as inhibition of Syk kinase activity in a biochemical assay, or as a reduction in basophil activation, as measured by CD63 expression, as described in the Examples section. The compounds described herein also have desirable properties for drug use, including kinetic solubility at 37° C. in phosphate buffer pH 7.4 and low clearance levels in hepatocytes. These features provide Syk inhibitors for the treatment of disease having pharmacokinetic characteristics that provide a therapeutic window such that the compounds can be effective at lower doses than currently known compounds. In this regard, the proposed compounds provide effective doses with minimal off-target activity, which can reduce undesirable side objects, reduce the chance of drug interaction and increase the chances of subject compliance with a given therapeutic regimen.

In some embodiments, the compounds described herein, for example Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2, are effective for one or more of inhibiting Syk kinase activity or reducing basophil activation as measured by by CD63 expression. For example, the compound inhibits Syk kinase activity with an IC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 100 nM, less than or equal to 50 nM, less than or equal to 20 nM or less, or equal to 10 nm according to a suitable assay for Syk kinase activity, such as the assay described in Example 12; and/or reduces CD63 expression activity with an EC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 150 nM, less than or equal to 100 nM, or less than or equal to 75 nM as determined by a suitable assays to determine CD63 expression in basophils, such as the assay described in Example 9.

In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 are effective in both inhibiting Syk kinase and reducing CD63 expression. For example, the compound inhibits Syk kinase activity with an IC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 100 nM, less than or equal to 50 nM, less than or equal to 20 nM or less, or equal to 10 nm according to a suitable assay for Syk kinase activity, such as the assay described in Example 12; and reduces CD63 expression with an EC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 150 nM, less than or equal to 100 nM, or less than or equal to 75 nM in a suitable study to determine expression of CD63 in basophils, such as the study described in example 9.

In some embodiments, in addition to one or more of the properties of Syk kinase inhibition and reduction of basophil activation as determined by CD63 expression, including both Syk kinase inhibition and reduction of basophil activation as measured by CD63 expression, Form I or II of the monomesylate salt of the compound according to Example 2 or Form I or II of the succinate compound of Example 2 have desirable properties for drug use, including one or more of kinetic solubility and low hepatocyte clearance levels. In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 have one or more of the desirable properties of kinetic solubility and low levels of clearance in hepatocytes, including kinetic solubility at 37° C. in phosphate buffer. pH 7.4, greater than or equal to 10 μM, greater

- 10 040127 or equal to 20 µM, greater than or equal to 30 µM, greater than or equal to 40 µM, greater than or equal to 50 µM, greater than or equal to 60 µM, greater than or equal to 70 µM, greater than or equal to 80 µM, or greater or equal to 90 µM by data from a suitable determination of kinetic solubility, such as the study described in example 10; and/or estimated clearance in hepatocytes less than or equal to 0.50 l/h/kg, less than or equal to 0.40 l/h/kg, less than or equal to 0.30 l/h/kg, less than or equal to 0.20 l/h/kg, less than or equal to 0.10 l/h/kg, less than or equal to 0.09 l/h/kg, less than or equal to 0.08 l/h/kg, less than or equal to 0.07 l/ h/kg or less than or equal to 0.06 l/h/kg as determined by a suitable determination of the estimated clearance in hepatocytes, such as the study described in example 11.

In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 have the desired kinetic solubility property and low levels of clearance in hepatocytes, including kinetic solubility at 37° C. in phosphate buffer pH 7, 4 greater than or equal to 10 μM, greater than or equal to 20 μM, greater than or equal to 30 μM, greater than or equal to 40 μM, greater than or equal to 50 μM, greater than or equal to 60 μM, greater than or equal to 70 μM, greater than or equal to 80 μM, or greater than or equal to 90 μm according to a suitable definition of kinetic solubility, such as the study described in example 10; and estimated clearance in hepatocytes less than or equal to 0.50 l/h/kg, less than or equal to 0.40 l/h/kg, less than or equal to 0.30 l/h/kg, less than or equal to 0.20 l/ h/kg, less than or equal to 0.10 l/h/kg, less than or equal to 0.09 l/h/kg, less than or equal to 0.08 l/h/kg, less than or equal to 0.07 l/h/kg kg or less than or equal to 0.06 l/h/kg as determined by a suitable determination of the estimated clearance in hepatocytes, such as the study described in example 11.

In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 are effective in both inhibiting Syk kinase and reducing CD63 expression, and also have the desired kinetic solubility property and low clearance levels in hepatocytes. For example, the compound inhibits Syk kinase activity with an IC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 100 nM, less than or equal to 50 nM, less than or equal to 20 nM or less, or equal to 10 nm according to a suitable assay for Syk kinase activity, such as the assay described in Example 12; and causes a decrease in CD63 expression with an EC ₅₀ value less than or equal to 1 μM, less than or equal to 500 nM, less than or equal to 200 nM, less than or equal to 150 nM, less than or equal to 100 nM, or less than or equal to 75 nM according to an appropriate study for determining the expression of CD63 in basophils, such as the study described in example 10; and has a kinetic solubility at 37°C in phosphate buffer pH 7.4 greater than or equal to 10 μM, greater than or equal to 20 μM, greater or equal to 30 μM, greater or equal to 40 μM, greater than or equal to 50 μM, greater than or equal to 60 μM, greater than or equal to 70 μM, greater than or equal to 80 μM, or greater than or equal to 90 μM according to a suitable kinetic solubility determination, such as the assay described in Example 10; and has an estimated clearance in hepatocytes less than or equal to 0.50 l/h/kg, less than or equal to 0.40 l/h/kg, less than or equal to 0.30 l/h/kg, less than or equal to 0.20 l /h/kg, less than or equal to 0.10 l/h/kg, less than or equal to 0.09 l/h/kg, less than or equal to 0.08 l/h/kg, less than or equal to 0.07 l/h /kg or less than or equal to 0.06 l/h/kg according to a suitable determination of the estimated clearance in hepatocytes, such as the study described in example 11.

Application methods.

The present invention provides Form I and II of the monomesylate salt of the compound of Example 2 and Form I and II of the succinate of the compound of Example 2 for use in therapy. What is provided is a method of treating a subject, for example a mammal, such as a human, having a disease responsive to inhibition of Syk activity, comprising administering to a subject having such a disease or suspected of having such a disease, an effective amount of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II succinate of the compound of Example 2. In one aspect, a subject, such as a human, is administered a pharmaceutical composition comprising Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 and a pharmaceutically acceptable carrier. The present invention also provides Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 for use in such methods.

In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, may be administered to a subject (e.g., human) at risk for, or having a family history of, the disease or condition.

In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may also inhibit other kinases, thereby treating the disease, disease symptoms, and conditions associated with those kinases.

The treatments also include inhibition of Syk activity and/or inhibition of Syk activity

- 11 040127

B cells by inhibiting ATP binding or hydrolysis by Syk, or by some other mechanism in vivo in a subject suffering from a disease responsive to inhibition of Syk activity, by administering an effective concentration of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of succinate compounds according to example 2. An example of an effective concentration is a concentration effective to inhibit activity in vitro. The effective concentration can be determined experimentally, for example, by analyzing the concentration of the compound in the blood after administration to a human, or theoretically by calculating the bioavailability.

In some embodiments, the condition responsive to inhibition of Syk activity and/or B cell activity is cancer, an allergic disorder, and/or an autoimmune and/or inflammatory disease, and/or an acute inflammatory response.

Also provided is a method for inhibiting B cell activity in a subject in need thereof, comprising administering an effective amount of the Form I or II monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2.

Also provided is a method for inhibiting B cell proliferation in a subject in need thereof, comprising administering an effective amount of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2.

Also provided is a method of treating a subject having leukemia by administering an effective amount of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2. In some embodiments, a condition and disease that can be treated using Form I or The monomesylate salt II of the compound according to example 2 or the succinate form I or II of the compound according to example 2 are acute myeloid leukemia (AML).

Syk is a known inhibitor of apoptosis in lymphoma B cells. Mistaken apoptosis contributes to pathogenesis and drug resistance in leukemias and lymphomas. Thus, a method is also provided for causing or inducing apoptosis in cells expressing Syk, comprising contacting said cell with Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2. Also provided herein is a compound described herein, such as Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, for use in the preparation of a medicament for the treatment of the disease or condition described herein, such as leukemia.

Subjects.

Any of the proposed treatments may be used to treat a subject diagnosed or suspected of having an allergic disorder and/or an autoimmune and/or inflammatory disease and/or an acute inflammatory response or cancer.

In some embodiments of any of the methods provided herein, the subject is a human at risk of developing cancer (eg, a human who is genetically or otherwise predisposed to cancer) and who has or has not been diagnosed with cancer. In the context of the present description, a subject at risk is a subject at risk of developing cancer (eg, hematological malignancy). A subject may or may not have a detectable disease and may or may not have a detectable disease prior to treatment with the methods described herein. A subject at risk may have one or more so-called risk factors, which are measurable parameters correlated with the development of cancer, such as those described herein. A subject who has one or more of these risk factors is more likely to develop cancer than an individual who does not have these risk factors. These risk factors may include, for example, age, gender, race, diet, previous medical history, presence of a previous disease, genetic (eg, hereditary) factors, and environmental exposure. In some embodiments, a subject at risk of cancer includes, for example, a subject whose relatives have had the disease and those whose risk is determined by analysis of genetic or biochemical markers. A previous history of cancer may be a risk factor for cancer recurrence.

Also provided herein are methods of treating a subject (eg, human) having one or more symptoms associated with a cancer (eg, a hematologic malignancy). In some embodiments, the subject is in an early stage of cancer. In other embodiments, the subject is in an advanced stage of cancer.

Also provided herein are methods of treating a subject (eg, human) who is undergoing one or more conventional therapies for the treatment of cancer (eg, hematologic malignancy), such as chemotherapy, radiation therapy, immunotherapy, and/or surgery. Thus, in some of the above embodiments, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, is administered before, during, or after the administration of chemotherapy, radiation therapy,

- 12 040127 immunotherapy and/or surgery. In another aspect, the present disclosure provides methods for treating a subject (eg, human) that is resistant to cancer treatment or that has relapsed after cancer treatment (eg, hematologic malignancy). A subject resistant to cancer therapy is one that does not respond to a particular treatment and is also referred to as resistant. The cancer may be resistant to treatment from the start of treatment, or may become resistant during the course of treatment, for example, there may be some effect on the cancer after treatment, but not enough to be considered a remission or partial remission. A subject having a relapse means that the cancer has returned or its signs and symptoms have returned after a period of improvement, for example, after treatment has been shown to effectively reduce the cancer, such as after the subject has been in remission or partial remission. In some embodiments, the subject may be a human who (i) is refractory to at least one anti-cancer therapy, or (ii) relapses after treatment with at least one anti-cancer therapy, or is characterized by both (i) and ( ii). In some embodiments, the subject is resistant to at least two, at least three, or at least four cancer therapies (including, for example, standard or experimental chemotherapy).

In some embodiments, the subject is refractory to at least one, at least two, at least three, or at least four cancer therapies (including, for example, standard or experimental chemotherapy) selected from fludarabine, rituximab, obinutuzumab, alkylating agents, alemtuzumab and other chemotherapy treatments such as CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone); R-CHOP (rituximab-CHOP); hyperCVAD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine); R-hyperCVAD (rituximab-hyperCVAD); FCM (fludarabine, cyclophosphamide, mitoxantrone); R-FCM (rituximab, fludarabine, cyclophosphamide, mitoxantrone); bortezomib and rituximab; temsirolimus and rituximab; temsirolimus and Velcade (Velcade®); Iodine-131 tositumomab (Vexhag®) and CHOP; CVP (cyclophosphamide, vincristine, prednisone); R-CVP (rituximabCVP); ICE (ifosfamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR (fludarabine, cyclophosphamide, rituximab); FR (fludarabine, rituximab); D.T. PACE (dexamethasone, thalidomide, cisplatin, Adriamycin®, cyclophosphamide, etoposide); and idealisib.

Other examples of chemotherapy treatments (including standard and experimental chemotherapy) are described below. In addition, the treatment of some lymphomas is reviewed in Cheson, B.D., Leonard, J.P.: Monoclonal Antibody Therapy for B-Cell Non-Hodgkin's Lymphoma, The New England Journal of Medicine, 2008, 359(6), p. 613-626; and Wierda, W.G.: Current and Investigational Therapies for Patients with CLL, Hematology, 2006, p. 285-294. For charts of the occurrence of lymphoma in the United States, see Morton, L.M. et al.: Lymphoma Incidence Patterns by WHO Subtype in the United States, 1992-2001, Blood, 2006, 107(1), p. 265-276.

For example, the treatment of non-Hodgkin's lymphomas (NHL), especially those of B-cell origin, includes the use of monoclonal antibodies, standard chemotherapeutic approaches (eg, CHOP, CVP, FCM, MCP, etc.), radioimmunotherapy and combinations thereof, in particular the integration antibody therapy with chemotherapy. Examples of unconjugated monoclonal antibodies for the treatment of non-Hodgkin's lymphoma/B cell cancers include rituximab, alemtuzumab, human or humanized anti-CD20, limuiliximab, anti-TRAIL, bevacizumab, galiximab, epratuzumab, SGN-40, and anti-CD74. Examples of experimental antibody agents used in the treatment of non-Hodgkin's lymphoma/B cell cancers include ofatumumab, ha20, PRO131921, alemtuzumab, galiximab, SGN-40, CHIR-12.12, epratuzumab, limuiliximab, apolizumab, milatuzumab, and bevacizumab. Examples of standard chemotherapy regimens for non-Hodgkin's lymphoma/B-cell cancers include CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone), FCM (fludarabine, cyclophosphamide, mitoxantrone), CVP (cyclophosphamide, vincristine, and prednisone), MCP (mitoxantrone, chlorambucil, and prednisolone) , R-CHOP (rituximab plus CHOP), R-FCM (rituximab plus FCM), R-CVP (rituximab plus CVP), and R-MCP (R-MCP). Examples of radioimmunotherapy for non-Hodgkin's lymphoma/B cell cancers include yttrium-90 labeled ibritumomab tiuxetan and iodine 131 labeled tositumomab.

In another example, therapeutic therapies for mantle cell lymphoma (MCL) include a combination of chemotherapy such as CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone), hyperCVAD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine) and FCM (fludarabine). , cyclophosphamide, mitoxantrone). In addition, these regimens can be supplemented with the monoclonal antibody rituximab (Rituxan) to provide combination therapies using R-CHOP, hyperCVAD-R, and R-FCM. Other approaches include combining any of the above therapies with stem cell transplantation or treatment with ICE (ifosfamide, carboplatin and etoposide). Other approaches to the treatment of mantle cell lymphoma include immunotherapy, such as using monoclonal

- 13 040127 antibodies such as rituximab (Rituxan). Rituximab can be used to treat indolent B-cell cancer, including marginal zone cell lymphoma, MB, CLL, and small lymphocytic lymphoma. The combination of rituximab and chemotherapy agents is particularly effective. A modified approach is radioimmunotherapy, in which a monoclonal antibody is combined with a radioisotope particle. Examples are iodine-131-tositumomab (Bexxar®) and yttrium-90-ibritumomab thioxetane (Zevalin®). In another example, Bexxar (Bexxar®) is used in the treatment of sequential CHOP. cancers that are based on the genetic processing of an individual subject's tumor.An example of a lymphoma vaccine is the GTOP-99 vaccine (MyVax®).Other approaches to treat mantle cell lymphoma include autologous stem cell transplantation combined with high-dose chemotherapy or treatment of mantle cell lymphoma includes the administration of proteasome inhibitors such as Velcade® (bortezomib or PS-341) or anti-angiogenesis agents such as thalidomide, especially in combination with Rituxan.Another approach to treatment is the administration of drugs, which lead to the degradation of the Bcl-2 protein and increase the sensitivity of cancer cells to chemotherapy, such as the administration of oblimersen (Genasense) in combination with other chemotherapeutic agents. Another treatment approach involves the administration of mTOR inhibitors, which can lead to inhibition of cell growth and even cell death; a non-limiting example is temsirolimus (CCI-779) and temsirolimus in combination with Rituxan®, Velcade® or other chemotherapeutic agents.

Other recent therapies for MCL have been described (Nature Reviews, Jares, P., 2007). Such examples include Flavopiridol, PD0332991, R-roscovitine (Selicilib, CYC202), styrylsulfones, Obatoclax (GX15-070), TRAIL, anti-TRAIL antibodies DR4 and DR5, temsirolimus (CC1-779), everolimus (RAD001), BMS-345541 , Curcumin, Vorinostat (SAHA), Thalidomide, lenalidomide (Revlimid®, CC-5013), and Geldanamycin (17-AAG).

Examples of other therapies used to treat Waldenstrom's macroglobulinemia (WM) include perifosine, bortezomib (Velcade®), rituximab, sildenafil citrate (Viagra®), CC-5103, thalidomide, epratuzumab (humanized antibody hLL2- antiCD22), simvastatin, enzastaurin, campas-1H (campath-III), dexamethasone, DT PACE, oblimersen, antineoplaston A10, antineoplaston AS2-1, alemtuzumab, beta-alethine, cyclophosphamide, doxorubicin hydrochloride, prednisone, vincristine sulfate, fludarabine, filgrastim , melphalan, recombinant interferon-alpha, carmustine, cisplatin, cyclophosphamide, cytarabine, etoposide, melphalan, dolastatin 10, monoclonal antibody MN-14 with indium In-111, yttrium Y-90 humanized epratuzumab, antithymocyte globulin, busulfan, cyclosporine, methotrexate, mycophenolate mofetil, therapeutic allogeneic lymphocytes, yttrium Y-90 ibritumomab thioxetane, sirolimus, tacrolimus, carboplatin, thiotepa, paclitaxel, aldesleukin, recombinant inter feron-alpha, docetaxel, ifosfamide, mesnu, recombinant interleukin-12, recombinant interleukin-11, ABT-263 inhibitor of the Bcl-2 protein family, denileukin diphytotox, tanespymycin, everolimus, pegfilgrastim, vorinostat, alvocidib, ligand to recombinant flt3, recombinant human thrombopoietin, lymphokine-activated killer cells, amifostine trihydrate, aminocamptothecin, irinotecan hydrochloride, caspofungin acetate, clofarabine, epoetin alfa, nelarabine, pentostatin, sargramostim, vinorelbine ditartrate, WT-1 analog peptide vaccine, WT1 126-134 peptide vaccine, fenretinide, ixabepilone, oxaliplatin, CD19 monoclonal antibody, CD20 monoclonal antibody, omega-3 fatty acids, mitoxantrone hydrochloride, octreotide acetate, tositumomab and iodine 1-131 tositumomab, motoxafine gadolinium, arsenic trioxide, tipifarnib derived from autologous human tumor HSPPC-96, veltuzumab, bryostatin 1, pegylated liposomal doxorubicin hydrochloride and any of their bination.

Examples of therapeutic procedures used to treat WM include peripheral blood stem cell transplantation, autologous hematopoietic stem cell transplantation, autologous bone marrow transplantation, antibody therapy, biological therapy, enzyme inhibitor therapy, total body irradiation, stem cell infusion, assisted bone marrow ablation stem cell transplantation, in vitro treated peripheral blood stem cell transplantation, cord blood transplantation, enzyme immunoassay, pharmacological testing, low-level therapy, LET-cobalt-60-gamma-ray therapy, bleomycin, conventional surgery, radiotherapy, and non-myeloablative allogeneic hematopoietic stem transplantation cells.

Examples of other therapeutic agents used in the drug treatment of diffuse large B-cell lymphoma (DLBCL) (Blood, 2005, Abramson, J.) include cyclophosphamide, doxorubicin, vincristine, prednisone, anti-CD20 monoclonal antibodies, etoposide, bleomycin, many of the agents listed for macroglobulinemia, Waldenström, and any combination thereof such as ICE and R ICE.

- 14 040127

Examples of other therapies used to treat chronic lymphocytic leukemia (CLL) (Spectrum, 2006, Fernandes, D.) include idelicib (Zydelig®), chlorambucil (Leukeran), cyclophosphamide (Cyloxan, Endoxan, Endoxana, Cislostin) , Fludarabine (Fludara), Penstatin (Nipent), Cladribine (Leistarine), Doxorubicin (Adriamycin (Adriamycin®), Adriblastine), Vincristine (Oncovin), Prednisone, Prednisolone, Alemtuzumab (Campas (Campath), MabCampath), many of the agents listed for Waldenström's macroglobulinemia, combined chemotherapy and chemoimmunotherapy, including the general combination regimen: CVP (cyclophosphamide, vincristine, prednisolone); R-CVP (rituximab-CVP); ICE (ifosfamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR (fludarabine, cyclophosphamide, rituximab); and FR (fludarabine, rituximab).

In another aspect, a method is provided for sensitizing a subject (e.g., a human) that (i) is refractory to at least one chemotherapy treatment, or (ii) has a relapse following chemotherapy treatment, or is characterized by both (i) and (ii), said method comprising administering to said subject a Form I or II monomesylate salt of a compound of Example 2 or a Form I or II succinate of a compound of Example 2, or a pharmaceutical composition containing the same. A subject being sensitized is one responding to treatment involving administration of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2, or who has not developed resistance to such treatment.

In another aspect, the present disclosure provides methods for treating a subject (eg, human) for cancer with an underlying disease, said treatment also being effective in treating the associated disease. A cancer-related disease is a disease that occurs at the same time as cancer.

In some embodiments, the present disclosure provides methods for treating a subject (eg, a human) for chronic lymphocytic leukemia (CLL) with a comorbid disease, said disease also being effective in treating the comorbid disease. Many subjects with CLL will have one or more diseases, such as diseases affecting the blood pressure system, the vascular and cardiac systems, the endocrine and metabolic systems, the genitourinary system, the musculoskeletal system, the respiratory system, the neurological system, the upper and lower gastrointestinal systems. intestinal tract, mental system, ear, nose and throat systems, kidney system or liver system. Specific disease states of CLL include, but are not limited to, one or more other cancers (eg, breast, head and neck, lung, melanoma, non-Hodgkin's T-cell lymphoma, prostate, colon, small intestine, gynecological, and urinary pathways), hypertension, hyperlipidemia, coronary heart disease, peripheral vascular disease, cardiomyopathy, acquired heart disease, atrial fibrillation, cerebrovascular disease (eg, transient ischemic attack, stroke), chronic obstructive pulmonary disease, joint disease, peptic ulcer, inflammatory bowel disease , mental illness, thyroid disease, benign prostatic hyperplasia, diabetes mellitus, and osteoarthritis (Satram-Hoang et al., Journal of Cancer Therapy, 2013, 4:1321-1329; Thurmes et al., Leukemia&Lymphoma, 2008, 49(l ):49-56).

In some embodiments, a method of treating a comorbid CLL disease in a subject (e.g., a human) is provided, comprising administering to said subject a Form I or II monomesylate salt of a compound of Example 2, or a Form I or II succinate of a compound of Example 2, or a pharmaceutical composition containing them. In some embodiments, the comorbid disease is selected from the group consisting of one or more other cancers (e.g., breast, head and neck, lung, melanoma, non-Hodgkin's T-cell lymphoma, prostate, colon, small intestine, gynecological, and urinary tract), hypertension, hyperlipidemia, coronary artery disease, peripheral vascular disease, cardiomyopathy, acquired heart disease, atrial fibrillation, cerebrovascular disease (eg, transient ischemic attack, stroke), chronic obstructive pulmonary disease, joint disease, peptic ulcer, inflammatory disease bowel disease, mental illness, thyroid disease, benign prostatic hyperplasia, diabetes mellitus, and osteoarthritis.

Monotherapy.

Also provided are methods of treatment in which Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 is the only active agent given to the subject, and also included are methods of treatment in which Form I or II of the monomesylate salt the compounds according to example 2 or the succinate form I or II of the compound according to example 2 is given to the subject in combination with one or more additional active agents. Both monotherapy and combination therapy are contemplated and described for use in the methods detailed herein, such as for the treatment of any of the diseases or conditions detailed herein, and for use in any subject

- 15 040127 described in detail in this document.

Monotherapy.

In some embodiments, a method of treating cancer, an allergic disorder, and/or an autoimmune and/or inflammatory disease, and/or an acute inflammatory response comprises administering to a subject in need thereof an effective amount of a Form I or II monomesylate salt of a compound of Example 2 or Form I or II succinate compound according to example 2, and the specified subject is not being treated for the same disease or condition through another agent or another procedure.

In some embodiments in which Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 is administered as monotherapy to a subject diagnosed or suspected of having cancer, said subject may be a human, which (i) is resistant to at least one anti-cancer therapy, or (ii) relapses after treatment with at least one anti-cancer therapy, or is characterized by both (i) and (ii). In some embodiments, the subject is resistant to at least two, at least three, or at least four cancer therapies (including, for example, conventional or experimental chemotherapy). For example, in some embodiments, the subject may be a human who is (i) refractory to therapy with an anti-CD20 antibody, an alkylating agent (eg, bendamustine), a purine analog (eg, fludarabine), an anthracycline, or any combination thereof; (ii) has relapsed following treatment with an anti-CD20 antibody, an alkylating agent (eg, bendamustine), a purine analog (eg, fludarabine), an anthracycline, or any combination thereof, or is characterized by both (i) and (ii).

A human subject who is resistant to at least one anti-cancer therapy as described above may have received one or more therapies in the past. In some embodiments, such subjects received one, two, three, or four, or at least one, at least two, at least three, at least four, or at least five, or one to ten, one to nine, one to eight, one to seven, one to six, one to five, or one to four cancer therapies prior to initiation of treatment using the methods described herein (e.g., prior to administration of Form I or II monomesylate salt of the compound according to example 2 or form I or II of the succinate of the compound according to example 2 as monotherapy).

It should be understood that when a subject (e.g., a human) is receiving treatment with Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 as monotherapy, the subject may be receiving one or more other therapies that are not are cancer therapies.

In some embodiments, a method is provided for treating a cancer-related disease, including but not limited to CLL, in a subject (e.g., human) who has been diagnosed with cancer, such as CLL, the method comprising administering therapy to the subject for the treatment of the underlying disease in combination with I or II monomesylate salt of the compound according to example 2 or form I or II of the succinate of the compound according to example 2 or a pharmaceutical composition containing them. In some embodiments, said comorbid disease is selected from the group consisting of one or more other cancers (e.g., breast, head and neck, lung, melanoma, non-Hodgkin's T-cell lymphoma, prostate, colon, small intestine, gynecological, and urinary tract), hypertension, hyperlipidemia, coronary heart disease, peripheral vascular disease, cardiomyopathy, acquired heart disease, atrial fibrillation, cerebrovascular disease (eg, transient ischemic attack, stroke), chronic obstructive pulmonary disease, joint disease, peptic ulcer, inflammatory bowel disease, mental illness, thyroid disease, benign prostatic hyperplasia, diabetes mellitus, and osteoarthritis.

Pharmaceutical compositions and administration.

Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 is usually administered as a pharmaceutical composition. Thus, the present description provides pharmaceutical compositions that contain as an active ingredient one or more of the described compounds or their pharmaceutically acceptable salts, pharmaceutically acceptable co-crystals or pharmaceutically acceptable esters and one or more pharmaceutically acceptable carriers (vehicles), such as excipients , carriers, including inert solid diluents and excipients, diluents, including sterile aqueous solution and various organic solvents, penetration enhancers, solubilizers and adjuvants. Pharmaceutical compositions may be administered alone or in combination with other therapeutic agents. Such compositions are prepared in a manner well known in the pharmaceutical art (see, for example, Remington's Pharmaceutical Sciences, Mace Publishing

- 16 040127

Co., Philadelphia, PA, 17th ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd ed. (G.S.

Banker&C.T. Rhodes, eds.)).

Pharmaceutical compositions may be administered in single or multiple doses by any of the acceptable routes of administration for agents having similar uses, such as those described in patents and patent applications incorporated by reference, including rectal, buccal, intranasal, and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, by inhalation, or through an impregnated or coated device such as a stent, for example, or a cylindrical polymer inserted into an artery.

One type of administration is parenteral, in particular by injection. Forms in which Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may be included for administration by injection include aqueous or oily suspensions or emulsions with sesame oil, corn oil, cottonseed oil, or peanut oil. oil, as well as elixirs, mannitol, dextrose or sterile aqueous solution and similar pharmaceutical carriers. Also traditionally, aqueous solutions in saline can be used for injection. Ethanol, glycerin, propylene glycol, liquid polyethylene glycol, and the like can also be used. (and suitable mixtures thereof), cyclodextrin derivatives and vegetable oils. Proper fluidity can be maintained, for example, by applying a coating such as lecithin, by maintaining the desired particle size in the case of a dispersion, and by using surfactants. Prevention of the action of microorganisms can be carried out by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating a compound of the present disclosure in the required amount in an appropriate solvent with various other ingredients listed above, and if necessary followed by sterilization by filtration. Typically, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile carrier which contains the basic dispersion medium and the other necessary ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying methods, which allow the preparation of a powder of the active ingredient plus any additional desired ingredient from a pre-sterile filtered solution. In some embodiments, for parenteral administration, sterile injectable solutions are prepared containing a therapeutically effective amount, for example, from 0.1 to 1000 mg, of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2. however, it should be understood that the amount of compound actually administered will generally be determined by the attending physician in light of the relevant circumstances, including the condition being treated, the route of administration chosen, the compound actually administered and its relative potency, the age, weight and response of the particular subject, the severity of the subject's symptoms, and etc.

Oral administration is another route of administration of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2. Administration may be through an enteric-coated capsule or tablet or the like. In the preparation of pharmaceutical compositions which contain Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2, the active ingredient is usually diluted with an excipient and/or enclosed in such a carrier, which may be in the form of a capsule, sachet , paper or other container. When the excipient serves as a diluent, it may be in the form of a solid, semi-solid, or liquid material (as defined above) that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be presented in the form of tablets, pills, powders, lozenges, sachets (sachets), cachets (cachets), elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions and sterile packaged powders.

Some examples of suitable excipients in oral formulations include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic (gum acacia), calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup and methylcellulose. These compositions may additionally contain lubricating agents such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners and flavoring agents.

The pharmaceutical compositions described herein may be formulated to provide rapid, delayed or delayed release of the active ingredient after administration to a subject using procedures known in the art. Controlled release oral drug delivery systems include osmotic pump systems and dissolution systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are shown in US Pat. Such transdermal patches can be used to provide continuous or intermittent infusion of compounds according to the present description in controlled amounts. The design and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, for example, US Pat.

In some embodiments, the compositions described herein are formulated as unit dosage forms. The term unit dosage form refers to physically discrete units suitable as unit doses for humans and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect, in combination with a suitable pharmaceutical excipient (for example, in a tablet, capsule, ampoule). Compounds according to the present description is administered in a pharmaceutically effective amount. In some embodiments for oral administration, each dosage unit contains from about 1 mg to about 5000 mg, from about 1 mg to about 4000 mg, from about 1 mg to about 3000 mg, from about 1 mg to about 2000 mg, from about 2 mg to about 2000 mg, about 5 mg to about 2000 mg, about 10 mg to about 2000 mg, about 1 mg to about 1000 mg, about 2 mg to about 1000 mg, about 5 mg to about 1000 mg , from about 10 mg to about 1000 mg, from about 25 mg to about 1000 mg, from about 50 mg to about 1000 mg, from about 75 mg to about 1000 mg, from about 100 mg to about 1000 mg, from about 125 mg up to about 1000 mg, from about 150 mg to about 1000 mg, from about 175 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 225 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from approx. 400 mg to about 1000 mg, about 450 mg to about 1000 mg, about 500 mg to about 1000 mg, about 550 mg to about 1000 mg, about 600 mg to about 1000 mg, about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 750 mg to about 1000 mg, from about 800 mg to about 1000 mg, from about 850 mg to about 1000 mg, from about 900 mg to about 1000 mg, from about 950 mg to about 1000 mg, about 1 mg to about 750 mg, about 2 mg to about 750 mg, about 5 mg to about 750 mg, about 10 mg to about 750 mg, about 25 mg to about 750 mg, from about 50 mg to about 750 mg, from about 75 mg to about 750 mg, from about 100 mg to about 750 mg, from about 125 mg to about 750 mg, from about 150 mg to about 750 mg, from about 175 mg to about 750 mg, from about 200 mg to about 750 mg, from about 225 mg to about 750 mg, from about 250 mg to about 750 mg, from about 300 mg to about 750 mg, from about 350 mg to about 750 mg, from about 400 mg to about 750 mg, from about 450 mg to about 750 mg, from about 500 mg to about 750 mg, from about 550 mg to about 750 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, about 700 mg to about 750 mg, about 1 mg to about 500 mg, about 2 mg to about 500 mg, from about 5 mg to about 500 mg, from about 10 mg to about 500 mg, from about 25 mg to about 500 mg, from about 50 mg to about 500 mg, from about 75 mg to about 500 mg, from about 100 mg to about 500 mg, about 125 mg to about 500 mg, about 150 mg to about 500 mg, about 175 mg to about 500 mg, about 200 mg to about 500 mg, about 225 mg to about 500 mg , from about 250 mg to about 500 mg, from about 300 mg to about 500 mg, from about 350 mg to about 500 mg, from about 400 mg to approx. 500 mg, from about 450 mg to about 500 mg, from about 1 mg to about 400 mg, from about 2 mg to about 400 mg, from about 5 mg to about 400 mg, from about 10 mg to about 400 mg, from about 25 mg to about 400 mg, about 50 mg to about 400 mg, about 75 mg to about 400 mg, about 100 mg to about 400 mg, about 125 mg to about 400 mg, about 150 mg to about 400 mg, from about 175 mg to about 400 mg, from about 200 mg to about 400 mg, from about 225 mg to about 400 mg, from about 250 mg to about 400 mg, from about 300 mg to about 400 mg, from about 350 mg to about 400 mg, about 1 mg to about 300 mg, about 2 mg to about 300 mg, about 5 mg to about 300 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 75 mg to about 300 mg, about 100 mg to about 300 mg, about 125 mg to about 3 00 mg, about 150 mg to about 300 mg, about 175 mg to about 300 mg, about 200 mg to about 300 mg, about 225 mg to about 300 mg, about 250 mg to about 300 mg, from about 1 mg to about 250 mg, about 2 mg to about 250 mg, about 5 mg to about 250 mg, about 10 mg to about 250 mg, about

- 18 040127 mg to about 250 mg, from about 50 mg to about 250 mg, from about 75 mg to about 250 mg, from about 100 mg to about 250 mg, from about 125 mg to about 250 mg, from about 150 mg to about 250 mg, from about 175 mg to about 250 mg, from about 200 mg to about 250 mg, from about 225 mg to about 250 mg, from about 1 mg to about 225 mg, from about 2 mg to about 225 mg, from about 5 mg to about 225 mg, about 10 mg to about 225 mg, about 25 mg to about 225 mg, about 50 mg to about 225 mg, about 75 mg to about 225 mg, about 100 mg to about 225 mg, from about 125 mg to about 225 mg, from about 150 mg to about 225 mg, from about 175 mg to about 225 mg, from about 200 mg to about 225 mg, from about 1 mg to about 200 mg, from about 2 mg to about 200 mg, about 5 mg to about 200 mg, about 10 mg to about 200 mg, about 25 mg to about 200 mg, from about about 50 mg to about 200 mg, about 75 mg to about 200 mg, about 100 mg to about 200 mg, about 125 mg to about 200 mg, about 150 mg to about 200 mg, about 175 mg to about 200 mg, about 1 mg to about 175 mg, about 2 mg to about 175 mg, about 5 mg to about 175 mg, about 10 mg to about 175 mg, about 25 mg to about 175 mg, about 50 mg to about 175 mg, from about 75 mg to about 175 mg, from about 100 mg to about 175 mg, from about 125 mg to about 175 mg, from about 150 mg to about 175 mg, from about 1 mg to about 150 mg, from about 2 mg to about 150 mg, from about 5 mg to about 150 mg, from about 10 mg to about 150 mg, from about 25 mg to about 150 mg, from about 50 mg to about 150 mg, from about 75 mg to about 150 mg, from about 100 mg to about 150 mg, from about 125 mg to about 150 mg, from about 1 mg to about 125 mg, from about 2 mg to about 125 mg, from about 5 mg to about 125 mg, from about 10 mg to about 125 mg, from about 25 mg to about 125 mg, from about 50 mg to about 125 mg, from about 75 mg to about 125 mg, from about 100 mg to about 125 mg, about 1 mg to about 100 mg, about 2 mg to about 100 mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 25 mg to about 100 mg, about 50 mg to about 100 mg, or about 75 mg to about 100 mg Form I or II monomesylate salt of the compound of Example 2 or Form I or II succinate of the compound of Example 2.

In some oral embodiments, each dosage unit contains about 1 mg, about 2 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg , about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg or about 1000 mg Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2.

Doses for oral administration described above may be administered once daily (QD) or twice daily (BID). In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, or a pharmaceutical composition containing them, is administered orally in a unit dose of about 1 mg once a day, about 2 mg once a day. , about 5 mg once a day, about 10 mg once a day, about 15 mg once a day, about 20 mg once a day, about 25 mg once a day, about 30 mg once a day, about 35 mg once daily, approximately 40 mg once daily, approximately 45 mg once daily, approximately 50 mg once daily, approximately 75 mg once daily, approximately 100 mg once daily, approximately 125 mg once a day, about 150 mg once a day, about 175 mg once a day, about 200 mg once a day, about 225 mg once a day, about 250 mg once a day, about 300 mg once a day per day, about 350 mg once a day, about 400 mg once a day about 450 mg once a day, about 500 mg once a day, about 550 mg once a day, about 600 mg once a day, about 650 mg once a day, about 700 mg once a day, about 750 mg once a day, about 800 mg once a day, about 850 mg once a day, about 900 mg once a day, about 950 mg once a day, or about 1000 mg once a day. In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, or a pharmaceutical composition containing them, is administered orally at a unit dose of about 1 mg twice daily, about 2 mg twice daily, about 5 mg twice a day, about 10 mg twice a day, about 15 mg twice a day, about 20 mg twice a day, about 25 mg twice a day, about 30 mg twice a day, about 35 mg twice a day, about 40 mg twice a day, about 45 mg twice a day, about 50 mg twice a day, about 75 mg twice a day, about 100 mg twice a day, about 125 mg twice a day about 150 mg twice a day, about 175 mg twice a day, about 200 mg twice a day, about 225 mg twice a day, about 250 mg twice a day, about 300 mg twice a day 24 hours, approximately 350 mg twice daily, approximately 400 mg twice daily 19 040127 approximately 550 mg twice daily, approximately 600 mg twice daily, approximately 650 mg twice daily, approximately 700 mg twice daily about 750 mg twice a day, about 800 mg twice a day, about 850 mg twice a day, about 900 mg twice a day, about 950 mg twice a day, or about 1000 mg twice a day day.

In some parenteral embodiments, each dosage unit contains 0.1 mg to 1 g, 0.1 to 700 mg, or 0.1 to 100 mg of the Form I or II monomesylate salt of the compound of Example 2 or Form I or II succinate compound according to example 2.

However, for any of the dosage units described herein, it is understood that the amount of compound actually administered will generally be determined by the physician in light of the relevant circumstances, including the condition being treated, the chosen route of administration, the particular compound being administered and its relative potency, age, the weight and response of an individual subject, the severity of the subject's symptoms, and the like.

To obtain solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient to obtain a solid pre-formulated composition containing a homogeneous mixture of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2. When said That these pre-formulated compositions are homogeneous implies that the active ingredient is evenly distributed in the composition such that said composition can be readily divided into equal effective unit dosage forms such as tablets, pills and capsules.

The tablets or pills described herein may be coated or otherwise formulated into a dosage form that provides the advantage of prolonged action or protection from the acidic environment of the stomach. For example, a tablet or pill may contain an internal dosage component and an external dosage component, the latter in said form being applied to the former. The two components can be separated by an enteric layer which serves to resist degradation in the stomach and allows the internal component to pass intact into the duodenum or delays the release of the internal component. Various materials can be used for such enteric layers or coatings, including a range of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures and powders thereof. Liquid or solid compositions containing Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may contain suitable pharmaceutically acceptable excipients as previously described. Preferably, the compositions are administered orally or nasally via inhalation to produce a local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents can be sprayed using inert gases. Nebulized solutions can be inhaled directly from a nebulization device, or said nebulization device can be connected to an inhalation mask or intermittent positive pressure breathing device. Solution, suspension or powder compositions may be administered preferably orally or nasally from devices that deliver the formulation in an appropriate manner.

Dosing regimen.

In the methods provided herein, Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, or a pharmaceutical composition containing them, is administered in a therapeutically effective amount to achieve the desired purpose. The determination of a therapeutically effective amount is within the skill of the art, especially in light of the detailed description provided herein. In some embodiments (methods of treating cancer), a therapeutically effective amount of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) suppress, delay, slow down to some extent and preferably stop the infiltration of cancer cells into peripheral organs; (iv) suppress (eg, slow to some extent and preferably stop) tumor metastases; (v) inhibit tumor growth; (vi) delay the onset and/or re-emergence of a tumor; and/or (vii) alleviate to some extent one or more symptoms associated with cancer. In various embodiments, said amount is sufficient to alleviate, alleviate, reduce and/or delay one or more symptoms of cancer.

A therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the route of administration, which can be readily determined by some or those of ordinary skill in the art.

- 20 040127

The dosage regimen of the Form I or II monomesylate salt of the compound of Example 2 or the Form I or II succinate of the compound of Example 2 in the methods provided herein may vary, for example, depending on the indication, route of administration, and severity of the condition. Depending on the route of administration, a suitable dose may be calculated depending on body weight, body surface area and organ size. The final mode of administration is determined by the attending physician in accordance with good medical practice, taking into account various factors that modify the effect of drugs, for example, the specific activity of the compound, the type and severity of the disease state, the subject's ability to respond to treatment, age, condition, body weight, sex and the subject's diet; and the severity of any infection. Additional factors that may be taken into account include timing and frequency of administration, drug combinations, response sensitivity, and tolerability/response to therapy. Further refinement of dosages suitable for treatment, including any formulations mentioned herein, is done routinely by the practitioner without undue experimentation, especially in light of the dosage and study disclosures and pharmacokinetic data observed in humans in clinical trials. Appropriate doses can be established by using specific assays to determine the concentration of the agent in a body fluid or other sample in conjunction with dose-response data.

The selected formulation and route of administration may be tailored to the individual subject, the nature of the condition being treated in said subject, and the overall judgment of the attending physician. For example, the therapeutic index of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can be increased by modifying or derivatizing the compound to target cancer cells expressing a marker identifying said cells as such. For example, the compounds may be linked to an antibody that recognizes a marker that is selective or specific for cancer cells, such that the compounds are placed close to said cells to exert their effects locally, as previously described. See, for example, Pietersz et al., Immunol. Rev. 129:57 (1992); Trail et al., Science, 261:212 (1993); and Rowlinson-Busza et al., Curr. Opin. Oncol. 4:1142 (1992).

A therapeutically effective amount of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may be presented in single dose or multiple doses to achieve the desired end result of treatment. As used herein, the term dose refers to the total amount of active ingredient (e.g., Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2) that a subject (e.g., human) takes each time. The dose administered, for example, for oral administration as described above, may be administered once daily (QD), twice daily (BID), three times daily, four times daily, or more than four times daily. In some embodiments, a dose of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 is administered once daily. In some embodiments, a dose of the Form I or II monomesylate salt of the compound of Example 2 or the Form I or II succinate of the compound of Example 2 is administered twice daily.

In some embodiments, exemplary dosages of the Form I or II monomesylate salt of the compound of Example 2 or the Form I or II succinate salt of the compound of Example 2 for a human subject may be from about 1 mg to about 5000 mg, from about 1 mg to about 4000 mg, from about 1 mg to about 3000 mg, from about 1 mg to about 2000 mg, from about 2 mg to about 2000 mg, from about 5 mg to about 2000 mg, from about 10 mg to about 2000 mg, from about 1 mg to about 1000 mg, about 2 mg to about 1000 mg, about 5 mg to about 1000 mg, about 10 mg to about 1000 mg, about 25 mg to about 1000 mg, about 50 mg to about 1000 mg, from about 75 mg to about 1000 mg, from about 100 mg to about 1000 mg, from about 125 mg to about 1000 mg, from about 150 mg to about 1000 mg, from about 175 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 225 mg to n about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 750 mg to about 1000 mg, from about 800 mg to about 1000 mg, from about 850 mg to about 1000 mg, from about 900 mg to about 1000 mg, from about 950 mg to about 1000 mg, from about 1 mg to about 750 mg, from about 2 mg to about 750 mg, about 5 mg to about 750 mg, about 10 mg to about 750 mg, about 25 mg to about 750 mg, about 50 mg to about 750 mg, about 75 mg to about 750 mg, from about 100 mg to about 750 mg, from about 125 mg to about 750 mg, from about

150 mg to about 750 mg, about 175 mg to about 750 mg, about 200 mg to about

750 mg, from about 225 mg to about 750 mg, from about 250 mg to about 750 mg, from about

300 mg to about 750 mg, about 350 mg to about 750 mg, about 400 mg to about

- 21 040127

750 mg, from about 450 mg to about 750 mg, from about 500 mg to about 750 mg, from about 550 mg to about 750 mg, from about 600 mg to about 750 mg, from about 650 mg to about 750 mg, from about 700 mg to about 750 mg, about 1 mg to about 500 mg, about 2 mg to about 500 mg, about 5 mg to about 500 mg, about 10 mg to about 500 mg, about 25 mg to about 500 mg, from about 50 mg to about 500 mg, from about 75 mg to about 500 mg, from about 100 mg to about 500 mg, from about 125 mg to about 500 mg, from about 150 mg to about 500 mg, from about 175 mg to about 500 mg, about 200 mg to about 500 mg, about 225 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg to about 500 mg, about 350 mg to about 500 mg , from about 400 mg to about 500 mg, from about 450 mg to about 500 mg, from about 1 mg to about 400 mg, from about 2 mg to about 400 mg, from about 5 mg to about 400 mg, from about 10 mg to about 400 mg, from about 25 mg to about 400 mg, from about 50 mg to about 400 mg, from about 75 mg to about 400 mg, from about 100 mg to about 400 mg, about 125 mg to about 400 mg, about 150 mg to about 400 mg, about 175 mg to about 400 mg, about 200 mg to about 400 mg, about 225 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg to about 400 mg, about 350 mg to about 400 mg, about 1 mg to about 300 mg, about 2 mg to about 300 mg, about 5 mg to about 300 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 300 mg, about 75 mg to about 300 mg, about 100 mg to about 300 mg , from about 125 mg to about 300 mg, from about 150 mg to about 300 mg, from about 175 mg to about 300 mg, from about 200 mg to about 30 0 mg, about 225 mg to about 300 mg, about 250 mg to about 300 mg, about 1 mg to about 250 mg, about 2 mg to about 250 mg, about 5 mg to about 250 mg, about 10 mg to about 250 mg, about 25 mg to about 250 mg, about 50 mg to about 250 mg, about 75 mg to about 250 mg, about 100 mg to about 250 mg, about 125 mg to about 250 mg, about 150 mg to about 250 mg, about 175 mg to about 250 mg, about 200 mg to about 250 mg, about 225 mg to about 250 mg, about 1 mg to about 225 mg, about 2 mg to about 225 mg, about 5 mg to about 225 mg, about 10 mg to about 225 mg, about 25 mg to about 225 mg, about 50 mg to about 225 mg, about 75 mg to about 225 mg , from about 100 mg to about 225 mg, from about 125 mg to about 225 mg, from about 150 mg to about 225 mg, from about 175 mg to about 225 mg, from about 200 mg to about 225 mg, from about 1 mg to about 200 mg, from about 2 mg to about 200 mg, from about 5 mg to about 200 mg, from about 10 mg to about 200 mg, from about 25 mg to about 200 mg, from about 50 mg to about 200 mg, from about 75 mg to about 200 mg, from about 100 mg to about 200 mg, from about 125 mg to about 200 mg, from about 150 mg to about 200 mg, from about 175 mg to about 200 mg, about 1 mg to about 175 mg, about 2 mg to about 175 mg, about 5 mg to about 175 mg, about 10 mg to about 175 mg, about 25 mg to about 175 mg, about 50 mg to about 175 mg, about 75 mg to about 175 mg, about 100 mg to about 175 mg, about 125 mg to about 175 mg, about 150 mg to about 175 mg, about 1 mg to about 150 mg, about 2 mg to about 150 mg, about 5 mg to about 150 mg, about 10 mg to about 150 mg, about 2 5 mg to about 150 mg, about 50 mg to about 150 mg, about 75 mg to about 150 mg, about 100 mg to about 150 mg, about 125 mg to about 150 mg, about 1 mg to about 125 mg, from about 2 mg to about 125 mg, from about 5 mg to about 125 mg, from about 10 mg to about 125 mg, from about 25 mg to about 125 mg, from about 50 mg to about 125 mg, from about 75 mg to about 125 mg, about 100 mg to about 125 mg, about 1 mg to about 100 mg, about 2 mg to about 100 mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg , from about 25 mg to about 100 mg, from about 50 mg to about 100 mg, or from about 75 mg to about 100 mg.

In some embodiments, exemplary doses of the Form I or II monomesylate salt of the compound of Example 2 or the Form I or II succinate salt of the compound of Example 2 for a human subject may be about 1 mg, about 2 mg, about 5 mg, about 10 mg , about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg , about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1200 mg, about 1400 mg, about 1600 mg, about 1800 mg, about 2000 mg, about 2200 mg, about 2400 mg, approximately 2600 mg, approximately 2800 mg, approximately 3000 mg, about 3200 mg, about 3400 mg, about 3600 mg, about 3800 mg, about 4000 mg, about 4200 mg, about 4400 mg, about 4600 mg, about 4800 mg or about 5000 mg.

In other embodiments, provided methods comprise continuing treatment of a subject (e.g., human) by administering doses of the Form I or II monomesylate salt of the compound according to

- 22 040127 Example 2 or Form I or II of the succinate of the compound according to Example 2, at which clinical efficacy is achieved, or stepwise reduction of doses to a level at which efficacy can be maintained. In some embodiments, the methods include administering to a subject (e.g., a human) an initial daily dose of Form I or II of the monomesylate salt of a compound of Example 2, or Form I or II of a succinate of a compound of Example 2, from 100 to 1000 mg, and administering subsequent daily doses of Form I or II monomesylate salt of the compound according to example 2 or form I or II of the succinate of the compound according to example 2 for at least six days, with each subsequent dose increased by 50-400 mg. Thus, it should be understood that the dose of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may be increased by stepwise escalation until clinical efficacy is achieved. Increases of about 25 mg, about 50 mg, about 100 mg, or about 125 mg, or about 150 mg, or about 200 mg, or about 250 mg, or about 300 mg may be used to increase the dose. The dose can be increased daily, once every two days, two, three, four, five or six times a week or once a week.

The frequency of dosing will depend on the pharmacokinetic parameters of the compound being administered, the route of administration, and the particular disease being treated. The dose and frequency of dosing may also depend on pharmacokinetic and pharmacodynamic data, as well as data on toxicity and therapeutic efficacy. For example, information on the pharmacokinetics and pharmacodynamics of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can be collected from preclinical in vitro and in vivo studies and then validated in humans during a course of clinical trials. research. Thus, for Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 used in the methods provided herein, the therapeutically effective dose can be initially estimated from the results of biochemical studies and/or studies on cells. Further, the dose can be determined in animal models to achieve the desired range of circulating concentrations at which Syk expression or activity is modulated. When conducting research in humans, the following information will be obtained regarding appropriate dosage levels and duration of treatment for various diseases and conditions.

The toxicity and therapeutic efficacy of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD ₅₀ (lethal dose for 50 % of the population) and ED ₅₀ (therapeutically effective dose in 50% of the population). The ratio of doses that provide toxic and therapeutic effects is called the therapeutic index, which is usually expressed as the ratio LD ₅₀ /ED ₅₀ . Compounds with high therapeutic indices, ie. where the toxic dose is substantially higher than the effective dose are preferred. Data from such cell culture studies and additional studies in animal models can be used to determine the dosage range for use in humans. Doses of such compounds are preferably within a range of circulating concentrations including the ED ₅₀ with little or no toxicity.

Administration of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can be done after a meal. The term post-meal, or variants thereof, refers to the consumption or ingestion of food in solid or liquid form or calories in a suitable form prior to or during administration of compounds or pharmaceutical compositions containing them. For example, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may be administered to a subject (eg, human) within minutes or hours of ingestion of calories (eg, food). In some embodiments, Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 may be administered to a subject (e.g., human) within 5-10 minutes, about 30 minutes, or about 60 minutes after caloric intake. .

Products and kits.

Compositions (including, for example, formulations and unit doses) containing Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 can be prepared, placed in an appropriate container, and labeled for the treatment of the indicated condition. Accordingly, an article of manufacture is also provided, such as a container containing a unit dosage form of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 and a label containing instructions for use of the compounds. In some embodiments, the product is a container containing a unit dosage form of Form I or II of the monomesylate salt of the compound of Example 2 or Form I or II of the succinate of the compound of Example 2 and at least one pharmaceutically acceptable carrier. Said product may be a bottle, vial, ampoule, single use applicator or the like containing a pharmaceutical component.

- 23 040127 the position proposed in the present description. Said container may be made from a variety of materials, such as glass or plastic, and in one aspect also includes a label on or associated with the container indicating directions for use in the treatment of cancer or inflammatory conditions. It should be understood that the active ingredient may be packaged in any material capable of improving chemical and physical stability, such as an aluminum foil bag. In some embodiments, the diseases or conditions indicated on the label may include, for example, the treatment of cancer.

Any pharmaceutical composition provided herein may be used in articles as if each and every composition were specifically and individually indicated for use in the article.

Also provided are kits containing a pharmaceutical composition containing Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2. For example, the kit may contain unit dosage forms of Form I or II of the monomesylate salt of the compound of Example 2, or Forms I or II succinate of the compound according to example 2 and a leaflet containing instructions for using the composition for the treatment of a medical condition. In some embodiments, the kit contains a single dosage form of Form I or II of the monomesylate salt of the compound of Example 2, or Form I or II of the succinate of the compound of Example 2, and at least one pharmaceutically acceptable carrier. Instructions for use of the kit may be for the treatment of cancer, including, for example, hematological malignancy. In some embodiments, the instructions are for use of the pharmaceutical composition for the treatment of cancer, such as leukemia or lymphoma, including relapsed and refractory leukemia or lymphoma. In some embodiments, instructions for use of the kit may be for the treatment of a hematologic cancer selected from the group consisting of small cell lymphocytic lymphoma, non-Hodgkin's lymphoma, indolent non-Hodgkin's lymphoma, refractory indolent non-Hodgkin's lymphoma, mantle cell lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, marginal zone cell lymphoma, immunoblastic large cell lymphoma, lymphoblastic lymphoma, splenic marginal zone B-cell lymphoma (+/-villous lymphocytes), nodular marginal zone cell lymphoma (+/-monocytoid B cells), extranodal B-cell lymphoma - cells of the marginal zone of the type of lymphoid tissue associated with the mucosa, cutaneous T-cell lymphoma, extranodal T-cell lymphoma, anaplastic large cell lymphoma, angioimmunoblastic T-cell lymphoma, mycosis fungoides, B-cell lymphoma, diffuse large B-cell lymphoma, To mediastinal large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, small uncleaved cell lymphoma, Burkitt's lymphoma, multiple myeloma, plasmacytoma, acute lymphocytic leukemia, T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, juvenile myelomonocytic leukemia, minimal residual disease, hairy cell leukemia, primary myelofibrosis, secondary myelofibrosis, chronic myeloid leukemia, myelodysplastic syndrome, myeloproliferative disease, and Waldenström's macroglobulinemia. In one embodiment, the instructions for use of the kit may be for the treatment of chronic lymphocytic leukemia or non-Hodgkin's lymphoma. In one embodiment, NHL is diffuse large B cell lymphoma, mantle zone cell lymphoma, follicular lymphoma, small cell lymphocytic lymphoma, lymphoplasmacytic lymphoma, and marginal zone cell lymphoma. In one embodiment, the hematologic malignancy is indolent non-Hodgkin's lymphoma. In some embodiments, the diseases or conditions listed on the label may include, for example, the treatment of cancer.

In some instances, the instructions are for use of a pharmaceutical composition for the treatment of a solid tumor, wherein said solid tumor is derived from a cancer selected from the group consisting of pancreatic cancer, genitourinary cancer, bladder cancer, colorectal cancer, colon cancer, breast cancer. , prostate cancer, kidney cancer, hepatocellular cancer, thyroid cancer, gallbladder cancer, lung cancer (eg, non-small cell lung cancer, small cell lung cancer), ovarian cancer, cervical cancer, gastrointestinal cancer, endometrial cancer, cancer esophagus, head and neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain tumor (eg, glioma, anaplastic oligodendroglioma, adult glioblastoma multiforme, and adult anaplastic astrocytoma), bone cancer, soft tissue sarcoma, retinoblastoma, neuroblastoma, peritoneal effusions, malignant pleural effusions, mesothelioma, tumors Wilms, trophoblastic neoplasms, hemangiopericytoma, Kaposi's sarcoma, myxoid carcinoma, round cell carcinoma, squamous cell carcinoma, squamous cell carcinoma of the esophagus, oral cavity carcinoma, adrenal cortex cancer, tumors producing adrenocorticotropic hormone.

- 24 040127

In some instances, the instructions are directed to the use of the pharmaceutical composition for the treatment of an allergic disorder and/or an autoimmune and/or inflammatory disease and/or an acute inflammatory reaction. In some embodiments, the instructions are directed to the use of a pharmaceutical composition for the treatment of an autoimmune disease. In some embodiments, the instructions are directed to the use of a pharmaceutical composition for the treatment of an autoimmune disease selected from the group consisting of systemic lupus erythematosus, myasthenia gravis, rheumatoid arthritis, acute disseminated encephalomyelitis, idiopathic thrombocytopenic purpura, multiple sclerosis, Sjögren's syndrome, psoriasis, autoimmune hemolytic anemia , asthma, ulcerative colitis, Crohn's disease, irritable bowel syndrome and chronic obstructive pulmonary disease. In some embodiments, the autoimmune disease is selected from the group consisting of asthma, rheumatoid arthritis, multiple sclerosis, chronic obstructive pulmonary disease, and systemic lupus erythematosus.

Any pharmaceutical composition provided herein may be used in kits as if each and every composition were specifically and individually indicated for use in the kit.

Synthesis.

Compounds according to the present description can be obtained using the methods disclosed in the present description and their routine modifications, which will be apparent in view of this description, and methods well known in the art. Conventional and well known synthetic methods may be used in addition to those provided herein. The synthesis of typical compounds described above can be carried out as described in the following examples. If available, reagents can be purchased from, for example, Sigma Aldrich or other chemical suppliers.

General methods of synthesis.

Exemplary embodiments of the compounds of the present disclosure may be synthesized using the general reaction schemes described below. It will be apparent, in view of the description given herein, that these general schemes can be modified by replacing the starting materials with other materials having similar structures to obtain products that, accordingly, will differ. Descriptions of the synthesis are provided to provide numerous examples of how starting materials can be modified to provide appropriate products. For a desired product for which substituent groups have been identified, the necessary starting materials can generally be determined by selection. Starting materials are generally obtained from commercial sources or synthesized using published methods. For the synthesis of compounds that are embodiments of the present disclosure, the selection of the structure of the compound to be synthesized will provide the form of each substituent group. Based on the type of final product, as a rule, as a result of a simple selection, taking into account the examples given in the present description, the type of necessary starting materials will be obvious.

Synthesis reaction parameters.

Compounds of the present invention can be prepared from readily available starting materials using, for example, the following general methods and techniques. It should be understood that when typical or preferred process conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given, other process conditions may also be used unless otherwise noted. . Optimum reaction conditions may vary depending on the specific reagents or solvents used, but such conditions can be determined by one of ordinary skill in the art using standard optimization procedures.

In addition, as will be apparent to those skilled in the art, commonly used protecting groups may be necessary for certain functional groups to prevent unwanted reactions. Suitable protecting groups for various functional groups, as well as suitable conditions for protecting and deprotecting specific functional groups, are well known in the art. For example, numerous protecting groups are described in Greene, T/W., Wuts, G.M. (1999), Protecting Groups in Organic Synthesis, 3rd edition, Wiley, New York; and references given in the specified source.

In addition, the compounds according to the present description may contain a chiral center. Accordingly, if necessary, such compounds can be obtained or isolated in the form of pure stereoisomers, i.e. individual enantiomers, or as stereoisomeric enriched mixtures. All such stereoisomers (and enriched mixtures) are included within the scope of this specification unless otherwise noted. Pure stereoisomers (or enriched mixtures) can be obtained using, for example, optically active starting materials or stereoselective reagents well known in the art. Alternatively, racemic mixtures of such compounds can be separated using, for example, chiral column chromatography, chiral resolving agents

- 25 040127 item, etc.

The starting materials for the following reactions are generally known compounds, or they may be prepared by known techniques or obvious modifications thereof. For example, many starting materials are available from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA). Other materials may be prepared by the methods, or obvious modifications thereof, described in standard references such as Fieser an Fieser's Reagents for Organic Synthesis, vol. 1-15 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, vol. 1-5; and supplements thereto (Elsevier Science Publishers, 1989) Organic Reactions, vol. 1-40 (John Wiley and Sons, 1991); March's Advanced Organic Chemistry (John Wiley and Sons, 5th edition, 2001); and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

The terms solvent, inert organic solvent, or inert solvent refer to a solvent that is inert under the reaction conditions described in connection with it (including, for example, benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, methylene chloride (or dichloromethane), diethyl ether, methanol, pyridine, etc.). Unless otherwise indicated, the solvents used in the reactions according to the present description are inert organic solvents, and these reactions are carried out in an atmosphere of inert gas, preferably nitrogen.

The term "sufficient amount" means the addition of an amount sufficient to achieve the stated function, for example to bring the volume of the solution to the desired value (ie 100%).

The following examples are provided to demonstrate preferred embodiments of the present disclosure. It will be appreciated by those skilled in the art that the techniques described in the following examples reflect the techniques disclosed by the inventor of the present invention for the proper practice of the present invention and thus may be considered as constituting the preferred embodiments of the invention. However, those skilled in the art, in light of the present disclosure, will appreciate that, without departing from the scope and spirit of the present disclosure, a large number of changes to the specific disclosed embodiments could be made and similar or analogous results would still be obtained.

List of abbreviations and abbreviations.

Reduction Meaning °C Degrees Celsius anal. Analytical ATP Adenosine 5'-triphosphate ATX II Anemonia sulcata toxin ASON Acetic acid ACN Acetonitrile CAN Cerium ammonium nitrate CDI 1, G-carbonyldiimidazole CHO Chinese hamster ovary CONC. Concentrated d doublet DABCO 1,4-Diazabicyclo[2,2.2]octane DAST (diethylamino)sulfur trifluoride dd Doublet of doublets DCE 1,2-dichloroethane DXM dichloromethane DEAD Diethylazodicarboxylate DIPEA Ν,Ν-diisopropylethylamine DMAP 4-dimethylaminopyridine

- 26 040127

DME 1,2-dimethoxyethane DMF Dimethylformamide DMSO Dimethyl sulfoxide dppf 1,1'-Bis(diphenylphosphino)ferrocene EA Ethanol ECF extracellular fluid EDTA Ethylenediaminetetraacetic acid EGTA Ethylene glycoltetraacetic acid equiv./equiv. Equivalents IES Electrospray ionization ace Acetate Et Ethyl EtOAc ethyl acetate G grams HEPES (4-(2-Hydroxyethyl)-1-piperazineethane sulfonic acid) HATU 2-(7-Aza-1 H-benzotriazol-1-yl)-1,1,3,3tetramethyluronium hexafluorophosphate hERG human ether-a-go-go Related Gene HMDS Hexamethyldisilazane (azide) HPLC High Performance Liquid Chromatography H Watch Hz Hertz IPA Isopropyl alcohol IC50 Half maximum inhibitory concentration IMR-32 Human neuroblastoma cell line J Interaction constant Kg Kilogram kHz KHz LAH lithium ammonium hydride LCMS/LCMS Liquid chromatography-mass spectrometry

- 27 040127

M Molar M Multiplet m/z mass-to-charge ratio M+ Peak by mass M+N Peak by mass plus hydrogen mCPBA 3-chloroperoxybenzoic acid Me Methyl MeOH methanol Mg Milligram MHz Megahertz min/m Minute ml Milliliter mm millimolar mmol Millimole

Nmol nanomol mOsmol Milliosmol MRM Magnetic resonance microscopy MS Mass spectroscopy MS Millisecond mV Millivolt microwaves Microwave And. Normal mole mole NMP A-methylpyrrolidinone NMR Nuclear magnetic resonance pA picoamps Ph Phenyl ppm parts per million prey. Preparative q.s. Quantity sufficient to achieve the stated function RF RP Retention factor Reversed phase rt/rt Room temperature C Second S Singlet SEM 2-(Trimethylsilyl)ethoxymethyl T Triplet TV Tonic block TEA Triethylamine TFU Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layer chromatography TMS Trimethylsilyl TTX Tetrodotoxin UDB Block according to application wt wild type Δ chemical shift mcg microgram µl microliter µM micromolar Mkm Micrometer mole Micromol

- 28 040127

Examples

Obtaining common intermediates.

Intermediate connection 1.01. Preparation of tert-butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate IV and tert-butyl-4 -(4-(oxetan-3-yl)piperazin-1yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]pyrazin-8-yl)carbamate V.

1-(4-Nitrophenyl)-4-(oxetan-3-yl)piperazine I.

In a 500 mL round bottom flask, 1-(oxetan-3-yl)piperazine (3.02 g, 21.26 mmol) potassium carbonate (5.87 g, 42.52 mmol) and 1-fluoro-4-nitrobenzene (3 00 g, 21.26 mmol) was combined with acetonitrile (33 ml) and stirred under nitrogen overnight at 100°C. The resulting mixture was diluted with water (100 ml) and subjected to extraction with DCM (100 mlx3), dried over anhydrous sodium carbonate, filtered and the filtrate was concentrated. The resulting residue was dissolved in a minimum amount of DCM using an ultrasonicator and extracted with hexane. The resulting precipitate was filtered, washed with hexane and dried to give the title compound I.

4-(4-(Oxetan-3-yl)piperazin-1-yl)aniline II.

In a hydrogenation vessel, 1-(4-nitrophenyl)-4-(oxetan-3-yl)piperazine I (4.70 g, 17.85 mmol) was dissolved in MeOH (26 ml) and DCM (5 ml) to the maximum quantity. Pd/C (10%) (2.85 g, 2.68 mmol) was added and the resulting reaction mixture was kept under nitrogen. The reaction mixture was stirred in a Parr hydrogenator at 45 psi (about 310 kPa). After 15 minutes, the reaction mixture was completely re-pressurized to 45 psi (about 310 kPa) and stirred for another 1 hour. The resulting material was filtered through celite, washed with 25% MeOH/DCM, and concentrated to give the title compound II.

6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine III.

To 4-(4-(oxetan-3-yl)piperazin-1-yl)aniline II (2.00 g, 8.57 mmol) was added Hunig's base (3.29 ml) and 6,8-dibromimidazo[1, 2-a]pyrazine (2.37 g, 8.57 mmol) in DMF (43 ml). The reaction mixture was stirred at 85° C. in a pressure tube overnight. The resulting material was quenched with saturated sodium bicarbonate solution, subjected to extraction with DCM (120 mlx3), the organic layers were combined and washed with water (120 mlx3), dried over anhydrous sodium carbonate and concentrated. The crude material was purified on a 120 g Isco column and eluted using a 0-60% step gradient (10% MeOH/DCM). The desired fractions were combined and concentrated to give the title compound III.

tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate IV.

6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine III (1000 mg, 2.33 mmol) , di-tert-butyl dicarbonate (1016.72 mg, 4.66 mmol) and N,N-dimethylpyridin-4-amine (21.34 mg, 0.17 mmol) were stirred in DCM (1.01 ml) and heated with reflux at 65°C for 3 h. The resulting reaction mixture was diluted with 100 ml DCM, washed with H ₂ O (x3), dried, filtered and concentrated. The resulting crude material was dissolved in a minimum amount

- 29 040127

DCM was loaded onto a prefilled silica bed and eluted on a 40 g column using 0-30% MeOH/DCM, 20 column volumes. The desired fractions were combined and concentrated to give the title compound. This compound was used in example 2.

tert-Butyl-4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]pyrazin-8yl)carbamate V.

In a 350 ml pressure tube tert-butyl-6-bromimidazo[1,2a]pyrazin-8-yl(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate IV (8150 mg, 15.39 mmol), 1,1,1,2,2,2 hexabutyl distannan (11.67 ml, 23.09 mmol), tetrakis(triphenylphosphine)palladium (889.43 mg, 0.77 mmol) and tetrabutylammonium iodide (5686.03 mg, 15.39 mmol) was combined in dioxane (62 ml) and heated to 110° C. overnight. According to LCMS, no starting material remained. The reaction mixture was absorbed on celite and eluted on a 160 g alumina column using a 0-10-20-30-100% gradient (50% EtOAc/Hex-Hex), stopping at 50% for 10-15 column volumes, total 50-60 column volumes to give the title compound V. This compound was used in Examples 1 and 2.

Intermediate 1.02. Preparation of tert-butyl-(6-bromo-5-methylimidazo[1,2-a]pyrazin-8yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate X.

6-Methylpyrazine-2-amine VI.

To a solution of anhydrous zinc(II) chloride (26.3 g, 193 mmol) in THF (150 ml) at 0°C was added 3 M methylmagnesium bromide in diethyl ether (129 ml) dropwise over a period of 1 h Next, [1,3-bis(diphenylphosphino)propane]nickel(II) chloride (2.08 g, 3.85 mmol) was added and the resulting mixture was allowed to warm to room temperature. A solution of 6-chloro-2-aminopyrazine (5.00 g, 38.6 mmol) in anhydrous THF (25 mL) was added to the above mixture, and the reaction mixture was stirred under nitrogen at reflux for 6 hours. the mixture was cooled to room temperature, then to 0°C and carefully quenched with saturated aqueous ammonium chloride (50 ml). The organic layer was separated and dried over sodium sulfate. The drying agent was filtered off and the filtrate was concentrated under reduced pressure to give crude 6-methylpyrazine-2-amine VI, which was used in the next step without purification.

1H NMR (400 MHz, CDC13) δ 7.63 (s, 1H), 7.53 (s, 1H), 4.96 (bs, 2H), 2.16 (s, 3H).

3,5-Dibromo-6-methylpyrazin-2-amine VII.

To a solution of 6-methylpyrazin-2-amine VI (2.00 g, 18.3 mmol) in THF (40 ml) at 10°C was added N-bromosuccinimide (6.70 g, 37.6 mmol) in portions over 15 min and the mixture was allowed to warm to room temperature while stirring. After 2 hours the reaction mixture was concentrated under reduced pressure and the resulting residue was purified by column chromatography (silica, gradient, hexanes-EtOAc) to give 3,5-dibromo-6-methylpyrazine-2-amine VII.

1H NMR (400 MHz, CDC13) δ 4.93 (bs, 2H), 2.38 (s, 3H).

- 30 040127

6,8-Dibromo-5-methylimidazo[1,2-a]pyrazine VIII.

A mixture of 2-bromo-1,1-diethoxyethane (3.21 mL, 20.7 mmol) and 48% aqueous hydrobromic acid (1.0 mL) was stirred at reflux for 2 hours. The resulting reaction mixture was then cooled to room temperature and treated with sodium bicarbonate until gas evolution ceased. The resulting mixture was filtered and the filtrate was diluted with ethanol (15 ml). To this mixture was added 3,5-dibromo-6-methylpyrazin-2-amine VII (3.00 g, 11.2 mmol) was added and the resulting reaction mixture was stirred at reflux for 16 hours. After that, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to a volume of approximately 10 ml. The resulting suspension was filtered and the filter residue was washed with cold ethanol (5 ml). Next, the filter residue was taken up in water (50 ml) and the pH was adjusted to ~8 with potassium carbonate. The resulting suspension was filtered and the filter cake was dried under vacuum to constant weight to give 6,8-dibromo-5-methylimidazo[1,2a]pyrazine VIII.

¹ H NMR (400 MHz, CDCL) δ 7.90 (s, 1H), 7.72 (s, 1H), 2.74 (s, 3H).

6-Bromo-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine IX.

Compound BC was prepared from 6,8-dibromo-5-methylimidazo[1,2-a]pyrazine VIII using the method described for the preparation of 6-bromo-N-(4-(4-(oxetan-3-yl)piperazine- 1-yl)phenyl)imidazo[1,2a]pyrazin-8-amine III in the example of Intermediate 1.01.

tert-Butyl-(6-bromo-5-methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)carbamate X.

Compound X was prepared from 6-bromo-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazine-8-amine IX using the method described for the preparation of tert-butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate IV in the example with intermediate connection 1.01. This compound was used in example 4.

Synthesis according to examples 1-7.

Example 1 Preparation of 6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazine -8-amine (1).

2-Bis(tert-butoxycarbonyl)amino-6-bromo-3-chloropyrazine XI.

6-Bromo-3-chloropyrazin-2-amine (2000 mg, 9.59 mmol) was dissolved in DCM (48 ml) and then triethylamine (3.99 ml, 28.78 mmol), di-tert-butyl dicarbonate ( 4188.12 mg, 19.19 mmol) and N,N-dimethylpyridin-4-amine (87.91 mg, 0.72 mmol). The resulting reaction mixture was allowed to stir at room temperature overnight. The resulting crude material was washed with water, dried, filtered and concentrated. The resulting crude material was dissolved in a minimum amount of DCM and placed on a prefilled silica filler (25 g) and eluted on a 40 g column using 0-30% MeOH/DCM. The title compound XI was isolated and identified by LCMS and NMR. The product was a mixture of mono - and bis-boc-protected substances, mainly by NMR observed bis-boc protected substance.

- 31 040127 tert-Butyl-tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperazin-1 yl)phenyl)amino)imidazo[1,2- a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate XII.

tert-Butyl 4-(4-(Oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]pyrazin8-yl)carbamate V (1000 mg, 1.4 mmol), 2-bis (tert-butoxycarbonyl) amino-6-bromo-3-chloropyrazine XI (552 mg, 1.35 mmol) and PdCl ₂ (PPh ₃ ) ₂ (142.77 mg, 0.20 mmol) in 1,4-dioxane (11.27 ml) was subjected to microwave irradiation for 20 min at 140°C. The reaction mixture was absorbed on Celite and eluted on a 40 g Gold Isco column using 0-10-100% (30% MeOH/DCM), 20 column volumes. Fractions 34-39 were collected and concentrated. Based on NMR results, the title compound XII was identified and isolated.

tert-Butyl-(6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3yl)piperazin-1-yl )phenyl)carbamate XIII.

Microwave Vial ,2-a]pyrazin-6-yl)3-chloropyrazin-2-yl)carbamate XII (300 mg, 0.44 mmol), methylboronic acid (794.39 mg, 13.27 mmol), tetrakis(triphenylphosphine)palladium (51.12 mg, 0.04 mmol) and 2 M Na ₂ CO ₃ (0.44 ml) were combined in DME (1.77 ml) and subjected to microwave radiation for 20 min at 150°C. The resulting reaction mixture was treated with 25% MeOH/DCM and water. The organic layers were combined, dried, filtered and concentrated. The resulting crude material was placed on silica and eluted on a 40 g Gold column using 0-5-15-25-50% (30% MeOH/DCM), 45 column volumes. The required fractions were concentrated to give tert-butyl-(6-(6-amino-5methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl) piperazin-1-yl)phenyl)carbamate XIII as a minor product and the desired final compound 1 as an inseparable mixture (208 mg total) which was reacted with TFA.

6-(6-Amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2а]pyrazin-8-amine (1).

To a solution of tert-butyl-6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl(4-(4-(oxetan-3yl))piperazin-1-yl )phenyl)carbamate XIII (48 mg, 0.09 mmol) and 6-(6-amino-5-methylpurazin-2-yl)-N-(4-(4(oxetan-3-yl)piperazin-1-yl )phenyl)imidazo[1,2-a]pyrazine-8-amine (1.160 mg, 0.35 mmol) in DCM (2.5 ml) was added with TFA (0.16 ml, 2.15 mmol). Additional TFA (0.48 ml, 6.5 mmol) was added to the resulting reaction mixture to ensure that the reaction was complete. Next, the reaction mixture was cooled to 0°C and quenched with saturated NaHCO ₃ solution, then extraction was carried out with DCM (5 mlx3) and the combined organic layers were washed with water (5 mlx2), brine (5 mlx1), dried (Na ₂ SO ₄ ) with obtaining a crude product. This crude product was absorbed onto silica and eluted on a 24 g Gold Isco column using 0-15-25-40-100% (30% MeOH/DCM). The desired fractions were combined and concentrated to give the desired compound.

LCMS-ESI+ (m/z): [M+H]+: 458.22.

1H NMR (300 MHz, d ₆ -DMSO) δ 9.48 (s, 1H), 8.54 (s, 1H), 8.41 (s, 1H), 8.11 (s, 1H), 7, 95 (d, 2H), 7.6 (s, 1H), 6.98 (d, 2H), 6.2 (s, 2H), 4.58-4.45 (dt, 4H), 3.3 (m, 1H), 3.14 (t, 4H), 2.50-2.4 (dt, 4H), 2.33 (s, 1H).

Alternatively, Compound XII may be taken directly to this step and similarly deprotected to give the 5-chloropyrazine analog.

- 32 040127

Example 2 Preparation of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine (2).

2-Bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV.

To a mixture of 6-bromopyrazine-2-amine (5 g, 28.7 mmol) and di-tert-butyl dicarbonate (25.09 g, 114.94 mmol) was added DCM (10 mL) followed by DMAP (0.351 g, 29 mmol). The resulting reaction mixture was heated to 55°C for 1 h, cooled to RT, the resulting reaction mixture was partitioned between water and DCM, purified on silica and concentrated to give 2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV.

LCMS-ESI+ (m/z): [M+H]+: 374.14.

1H NMR (DMSO) δ 8.84 (d, 2H), 1.39 (s, 18H).

tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan-3-yl )piperazin-1-yl)phenyl)carbamate XVI - route with chemical transformation A.

tert-Butyl 4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]pyrazin-8yl)carbamate V (215 mg, 0.291 mmol) combined with 2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV (217.58 mg, 0.581 mmol), bis(triphenylphosphine)palladium(II) dichloride (30.61 mg, 0.044 mmol) and 1,4-dioxane (5 ml). The resulting reaction mixture was stirred in a microwave reactor at 120° C. for 30 minutes. The reaction mixture was quenched with saturated KF solution, subjected to extraction with EtOAc, purified on silica gel and eluted with EtOAc. The desired fractions were combined and concentrated to give 100 mg (46% yield) tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl )(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)carbamate XVI.

LCMS-ESI+ (m/z): [M+H]+: 744.4.

1H NMR (300 MHz d6-DMSO) δ 9.37 (s, 1H), 9.18 (s, 1H), 8.77 (s, 1H), 8.33 (d, 1H), 7.87 ( d, 1H), 7.28-7.25 (d, 2H), 6.92-6.89 (d, 2H), 4.55-4.41 (m, 4H), 3.4 (m, 1H), 3.14-3.11 (m, 4H), 2.37-2.34 (m, 4H), 1.37 (s, 18H), 1.3 (s, 9H).

- 33 040127 tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan- 3-yl)piperazin-1-yl)phenyl)carbamate XVI - route with chemical transformation B.

Stage 1. In a dry round-bottom flask with a capacity of 250 ml, 2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV (1.0 g, 1.0 eq., 2.67 mmol), KOAc (790 mg, 8.02 mmol , 3.0 eq.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (750 mg, 2 .94 mmol, 1.1 eq.), Pd(dba) (171 mg, 0.187 mmol, 0.07 eq.) and X-phos (128 mg, 0.267 mmol, 0.1 eq.), followed by 1 ,4-dioxane (25 ml), the resulting solution was sonicated for 5 minutes and then purged with N ₂ gas for 5 minutes. Next, the flask with its contents was placed in an N2 atmosphere and heated at 110°C for 90 min. After complete conversion to pinacolboronate, according to LCMS, the reaction mixture was removed from the heat source and allowed to cool to RT. After cooling, the reaction mixture was filtered through Celite and the filter cake was washed with 3x20 ml EtOAc. Further, the resulting solution was concentrated to obtain a dark red-orange syrup containing N,N-6ucBoc-6(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine XV , which was used directly in the next step.

Step 2: Freshly prepared N,N-bisBoc-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine2-amine XV (2.67 mmol based on 100% conversion , 2.0 eq. in terms of bromide) was dissolved in 20 ml of 1,2-dimethoxyethane, to this solution was added tert-butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-( 4(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate IV (707 mg, 1.34 mmol, 1.0 eq.), Na ₂ CO ₃ (283 mg, 2.67 mmol, 2.0 equiv.), Pd(PPh ₃ ) ₄ (155 mg, 0.134 mmol, 0.1 equiv.) and water (10 ml) and the resulting solution was degassed for 5 min using gaseous N ₂ . Next, the resulting reaction mixture was placed under N2 atmosphere and heated at 110°С for 90 min. LCMS showed complete disappearance of the bromide starting material, the reaction mixture was removed from the heat source and allowed to cool to RT. The resulting reaction mixture was diluted with 100 ml of water and 100 ml of 20% MeOH/DCM and the organic layer was isolated, subjected to extraction with 1x sat. NaHCO ₃ , 1x brine and then dried over Na ₂ SO ₄ . The resulting solution was then filtered and concentrated to an orange-red solid. Next, the resulting sample was suspended in warm MeOH, sonicated, and then filtered, washing with 2x20 ml of cold MeOH, after which the resulting cream-colored solid was dried under high vacuum overnight to obtain 905 mg of tert-butyl-(6-(6- (bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XVI .

6-(6-Aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8amine (2).

To a solution of tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2а]pyrazin-8-yl)(4-(4-(oxetan-3- yl)piperazin-1-yl)phenyl)carbamate XVI (200 mg, 0.269 mmol) in DCM (2 ml) was added with TFA (0.5 ml, 6.578 mmol). The resulting reaction mixture was stirred at RT for 16 h, saturated sodium bicarbonate solution was added, extraction was carried out with EtOAc and purification on silica gel eluting with 5% MeOH/EtOAc, 20% MeOH/EtOAc. The desired fractions were combined and concentrated to give the title compound 2.

LCMS-ESI+ (m/z): [M+H] ⁺ : 444.2.

¹ H NMR (300 MHz d ₆ -DMSO) δ 9.5 (s,lH), 8.588 (s, 1H), 8.47 (s, 1H), 8.12 (d, 1H), 7.95- 7.92 (d, 2H), 7.88 (s, 1H), 7.62 (s, 1H), 6.99-6.96 (d, 2H), 6.46 (s, 2H), 4 .57-4.53 (m, 2H), 4.48-4.44 (m, 2H), 3.43 (m, 1H), 3.15-3.12 (m, 4H), 2.41 -2.38 (m, 4H).

- 34 040127

Example 2.1. Alternative synthesis method.

Di-tert-butyl-{6-[8-({4-[4-(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[1,2-a]pyrazin-6yl]pyrazin- 2-yl}imidodicarbonate.

Di-tert-butyl-(6-bromopyrazin-2-yl)imidodicarbonate (18.5 kg, 1.41 eq., 49 mol), bis(pinacolato)diboron (13.8 kg, 1 .56 eq., 54 mol), potassium propionate (11.9 kg, 3.02 eq., 106 mol) and bis(di-tert-butyl (4-dimethylaminophenyl)phosphine)dichloropalladium (1.07 kg, 0. 0043 eq., 1.5 mol) followed by degassed toluene (173 L). The resulting mixture was degassed, then heated at 65°C until the reaction was considered complete (0% tert-butyl-2((6-bromopyrazin-2-yl)(tert-butoxycarbonyl)amino)-2-oxoacetate) over ultra performance liquid chromatography (UPLC) data. After completion of the reaction, the resulting mixture was cooled to 23°C. After cooling, 6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2a]pyrazin-8-amine (15.0 kg, 1.00 eq., 35 mol) and the mixture was degassed. Next, a degassed aqueous potassium carbonate solution prepared using water (54 L) and potassium carbonate (20.6 g, 4.26 eq., 149 mol) was added to the reaction mixture, and the contents of the reactor were degassed. The contents of the reactor were heated at 65°C until the reaction was considered complete (1% 6-bromo-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazine -8-amine) according to HPLC. After completion of the reaction, the resulting mixture was cooled to 24°C.

The cooled mixture was concentrated and then diluted with dichloromethane (300 L), transferred to a 1900 L reactor and rinsed with dichloromethane (57 L). N-acetyl-L-cysteine (3.8 kg) was added and the resulting mixture was shaken for 15 hours. Water (135 L) was then added and the resulting mixture was filtered and rinsed with dichloromethane (68 L). The resulting organic layer was isolated and washed with brine prepared using water (68 L) and sodium chloride (7.5 kg).

The resulting organic layer was filtered on a polishing filter, then concentrated and tert-butyl methyl ether (89.9 kg) was slowly added while maintaining the temperature at 31°C. The resulting material was cooled to 0°C and aged, then filtered, rinsed with tert-butyl methyl ether (32.7 kg) and dried at 40°C to give 17.2 kg of di-tert-butyl-{6-[8-({ 4-[4(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[1,2-a]pyrazin-6-yl]pyrazin-2yl}imidodicarbonate.

LCMS-ESI+ (m/z): [M+H]+: 644.3.

Ή NMR (400 MHz, CDCl ₃ ) δ 9.43 (s, 1H), 8.58 (s, 1H), 8.53 (s, 1H), 8.02 (s, 1H), 7.84 ( m, 2H), 7.63 (d, 1H), 7.61 (d, 1H), 7.04 (m, 2H), 4.71 (m, 4H), 3.59 (m, 1H), 3.27 (m, 4H), 2.55 (m, 4H), 1.46 (s, 18H).

6-(6-Aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8amine succinate (Example 2 ).

To a suspension of di-tert-butyl-{6-[8-({4-[4-(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[1,2a]pyrazin-6-yl] pyrazin-2-yl}imidodicarbonate (225 g, 0.35 mol, 1 mol equiv.) in water (12 parts) was added a solution of sulfuric acid (3.1 parts, 6.99 mol, 20 mol equiv.) in water (5 parts). The resulting reaction mixture was heated to approximately 40° C. and stirred at this temperature for approximately 4 hours, during which the reaction was considered to be complete. The resulting reaction

- 35 040127 the mixture was cooled to approximately 22°C, acetone (3 parts) was added and a solution of sodium carbonate (4.1 parts, 8.75 mol, 25.0 mol-eq.) in water (15 parts) was added. The resulting suspension was filtered and the resulting wet filter cake was washed in parts with water (4x1 part) and then with tert-butyl methyl ether (4 parts). The wet filter cake (free base according to example 2) was dried at approximately 60°C. To a suspension of dry free base according to example 2 in 2-propanol (2.3 parts) was added a solution of succinic acid (based on the isolated free base according to example 2: 0.43 parts, 1.6 mol-equiv.) in 2-propanol (15 parts). The resulting suspension was heated to approximately 40°C, stirred at this temperature for approximately 2 hours and then cooled to approximately 22°C, after which it was stirred for approximately 16 hours. The resulting suspension was filtered at approximately 22°C, the wet filter cake was washed 2-propanol (5 parts) and dried at approximately 60°C to obtain the product.

LCMS-ESI+ (m/z): [M+H]+: 620.65.

¹ H NMR (400 MHz d ₆ -DMSO) δ 12.2 (broad s, 1.5H), 9.58 (s, 1H), 8.63 (s, 1H), 8.50 (s, 1H ), 8.15 (s, 1H), 7.95 (d, 2H), 7.90 (s, 1H), 7.64 (s, 1H), 7.00 (d, 2H), 6.50 (s, 2H), 4.52 (dd, 4H), 3.45 (m, 1H), 3.19 (m, 4H), 2.40 (m, 10H).

Example 3 Preparation of (R)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)phenyl)morpholin-2-yl)methanol (3).

(R)-(4-(4-((6-Bromimidazo[1,2-a]pyrazin-8-yl)amino)phenyl)morpholin-2-yl)methanol XVII.

Into a 250 ml round-bottom flask equipped with a condenser,

6,8-dibromimidazo[1,2-a]pyrazine (2000 mg, 7.22 mmol) and 30 ml of isopropanol was added, followed by N,N-diisopropylethylamine (2.52 ml, 14.44 mmol) and (R )-(4-(4-aminophenyl)morpholin-2-yl)methanol (1504.12 mg, 7.22 mmol). The resulting reaction mixture was heated to reflux (oil bath 95° C.) overnight. The reaction mixture was cooled, the precipitate was collected by filtration and washed in isopropanol and then in hexane to give the desired compound XVII.

(R)-tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate XVIII.

(R)-(4-(4-((6-bromimidazo[1,2-a]pyrazin-8yl)amino)phenyl)morpholin-2-yl)methanol XVII (2.80 g , 6.9 mmol) and DCM was added, followed by triethylamine (2.9 ml, 2.1 g, 20.8 mmol), DMAP (63 g, 0.52 mmol) and di-tert-butyl dicarbonate (3. 8 g, 17.3 mmol). The resulting reaction mixture was stirred overnight, then diluted with DCM and water, separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by chromatography: ISCO 40 g silica with 25 g silica loading, eluting with 0-100% EtOAc/hexane to give the compound

- 36 040127 XVIII.

(R)-tert-Butyl-(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6(tributylstannyl)imidazo[1,2-a]pyrazin-8-yl)carbamate XIX .

(R)-tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate XVIII was reacted according to the method , analogous to the example of Intermediate 1.01 to give (R)-tert-butyl(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6-(tributylstannyl)imidazo[ 1,2a]pyrazin-8-yl)carbamate XIX.

(R)-tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)(4-(2-((( tert-Butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate XX.

(R)-tert-butyl-(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6(tributylstannyl)imidazo[1,2-a]pyrazin-8-yl)carbamate XIX was reacted with 2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV according to a method similar to the method designated as chemical transformation reaction A described in example 2, to obtain the desired compound (R)-tert-butyl-(6-(6 -(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate XX.

(R)-(4-(4-((6-(6-Amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)phenyl)morpholin-2yl)methanol (3).

(R)-tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2yl)imidazo[1,2-a]pyrazin-8-yl)(4-(2 -(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate XX (460 mg, 0.56 mmol) in DCM and TFA (1.29 ml, 16.85 mmol) was added. After stirring for ~5 h, the reaction was partially complete. Added an additional 10 eq. TFA and stirred overnight followed by concentration under reduced pressure. 10% MeOH/DCM (~100 ml) and saturated aqueous sodium bicarbonate were added and stirred for 15 min, separation, extraction with ~100 ml 10% MeOH/DCM. The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered, concentrated under reduced pressure and dried under vacuum. The resulting solid was triturated with DCM, the solids were collected by filtration and dried under vacuum to give compound 3.

LCMS-ESI+ (m/z): [M+H] ⁺ : 419.2.

1H NMR (300 MHz d ₆ -DMSO) δ 9.57 (s, 1H), 8.59 (s, 1H), 8.47 (s, 1H), 8.13 (d, J=1.2 Hz , 1H), 8.06-7.90 (m, 2H), 7.87 (s, 1H), 7.62 (d, J= 1.1 Hz, 1H), 7.05-6.93 ( m, 2H), 6.49 (s, 2H), 4.78 (t, J=5.8 Hz, 1H), 3.98-3.87 (m, 1H), 3.71-3.36 (m, 7H), 2.63 (td, J=11.7, 3.4Hz, 1H), 2.37 (dd, J=12.1, 10.5Hz, 1H).

The corresponding (S)-isomer or racemic mixture of compounds was prepared in a similar manner using (S)-(4-(4-aminophenyl)morpholin-2-yl)methanol or the racemic mixture of (4-(4aminophenyl)morpholin-2-yl)methanol , respectively, at the first stage.

Example 4 Preparation of 6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazine -8-amine (4).

tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)-5-methylimidazo[1,2-a]pyrazin8-yl)(4-(4-(oxetan-3 -yl)piperazin-1 -yl)phenyl)carbamate XXI.

tert-Butyl-(6-bromo-5-methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)carbamate X was reacted with XV according to the methods of chemical transformation B, described in example 2, to obtain the desired compound XXI.

6-(6-Aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2а]pyrazin-8-amine (4).

From the compound tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)-5methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetane -3-yl)piperazin-1-yl)phenyl)carbamate XXI was deprotected in a manner analogous to that described in Example 2 to give the desired compound 4.

LCMS-ESI ⁺ (m/z): [M+H] ⁺ : 458.32.

¹ H NMR (300 MHz, d ₆ -DMSO) δ 9.28 (s, 1H), 8.28 (s, 1H), 8.04 (s, 1H), 7.89 (d, 2H), 7 .83 (s, 1H), 7.7 (s, 1H), 6.91 (d, 2H), 6.46 (s, 2H), 4.6-4.4 (dt, 4H), 3, 43 (m, 1H), 3.1 (t, 4H), 2.49 (s, 3H), 2.4 (t, 4H).

Example 5 Preparation of 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4(oxetan-3-yl) piperazin-1-yl)phenoxy)ethanol (5).

2-(2-(2-Fluoro-5-nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII.

A mixture of 2-fluoro-5-nitrophenol (4 g, 25 mmol), 2-(2-bromoethoxy)tetrahydro-2H-pyran (4.4 ml, 28 mmol) and potassium carbonate (4.2 g, 30 mmol) in DMF (50 ml) was stirred at 50°C for 16 hours. The resulting reaction mixture was cooled to room temperature, diluted with EtOAc and H2O. The aqueous layer was separated and subjected to extraction with EtOAc. The combined organic extracts were washed with H2O (5x to remove DMF) and brine and dried over sodium sulfate. The resulting residue was purified by column chromatography on an ISCO Rf column (40 g column) eluting with a gradient of 100% hexane - 1:1 hexane:EtOAc to give 2-(2-(2-fluoro-5nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII .

1-(4-Nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4-(oxetan-3-yl)piperazine XXIII.

A mixture of 2-(2-(2-fluoro-5-nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII (1550mg, 5.43mmol), 1-(oxetan-3-yl)piperazine (772mg, 5.43 mmol) and potassium carbonate (1126.41 mg, 8.15 mmol) in NMP (6 ml) were stirred at 100° C. for 8 hours. The aqueous layer was separated and subjected to extraction with

- 38 040127

EtOAc. The combined organic extracts were washed with H2O (5x to remove NMP) and brine and dried over sodium sulfate. The resulting residue was purified by column chromatography on an ISCO Rf column (24 g column) eluting with a gradient of 100% DCM - 60:35:5

DCM:Et ₂ O:MeOH to give 1-(4-nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4(oxetan-3-yl)piperazine XXIII .

4-(4-(Oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline XXIV.

To a suspension of 1-(4-nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4-(oxetan-3yl)piperazine XXIII (2100 mg, 5.1 mmol) in ethanol (50 ml) was added 10% Pd/C (moisture 50%, dry weight 390 mg) in a 500 ml Parr hydrogenation bottle. The bottle was evacuated, pressurized with hydrogen gas to 50 psi (about 345 kPa) and shaken at RT for 2 hours in a Parr hydrogenator. The resulting reaction mixture was filtered and washed with ethanol. The filtrate was concentrated in vacuo to give 4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline XXIV.

6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2yl)oxy)ethoxy)phenyl)imidazo[1 ,2-a]pyrazine-8-amine XXV.

To a solution of 4-(4-(oxetan-3-yl)piperazin-1 -yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline XXIV (619 mg, 2, 17 mmol) and 6,8-dibromimidazo[1,2-a]pyrazine (601 mg, 2.2 mmol) in IPA (15 ml) was added N,N-diisopropylethylamine (0.95 ml, 5.43 mmol). The resulting mixture was stirred at 110° C. for 16 hours. DCM (10 ml) and saturated aqueous NaHCO ₃ (15 ml) were then added. The resulting aqueous layer was separated and subjected to extraction with DCM (2x10 ml). The combined organic extracts were washed with brine (10 ml) and dried over sodium sulfate. The resulting residue was purified by column chromatography on an ISCO Rf column (24 g column) eluting with a gradient of 100% DCM - 60:35:5 DCM:Et ₂ O:MeOH to give 6-bromo-N-(4-(4(oxetane- 3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)imidazo[1,2a]pyrazin-8-amine XXV.

tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2((tetrahydro-2Н -pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI.

6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2yl)oxy)ethoxy)phenyl)imidazo[1 ,2-a]pyrazin-8-amine XXV (1.2 g, 2.4 mmol) was reacted according to a method similar to the method described under the heading described in the example of intermediate compound 1.01 (conversion of III to IV) to give tert -butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H- pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI.

tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan-3-yl )piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVII.

tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2((tetrahydro-2Н -pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI was reacted with XV according to the chemical transformation B methods described in Example 2 to give the desired compound tert-butyl-(6-(6-(bis(tert- butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan-3-yl)piperazin-1-yl)-3-(2-(( tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVII.

2-(5-((6-(6-Aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxetan-3-yl)piperazin-1yl )phenoxy)ethanol (5).

From the compound tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)(4-(4-(oxetan-3 - yl)piperazin-1 -yl)-3-(2-((tetrahydro-2H-pyran-2yl)oxy)ethoxy)phenyl)carbamate XXVII (313 mg, 0.35 mmol) was deprotected by a method similar to that described in Example 2 to give 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)-2-(4-(oxetan-3-yl )piperazin-1-yl)phenoxy)ethanol (5).

LCMS-ESI+ (m/z): [M+H] ⁺ : 504.3.

1H NMR (300 MHz, d ₆ -DMSO) δ 9.52 (s, 1H), 8.61 (s, 1H), 8.51 (s, 1H), 8.14 (d, J=1.1 Hz, 1H), 7.89 (s, 1H), 7.81 (d, J=2.3 Hz, 1H), 7.74-7.60 (m, 2H), 6.90 (d, J = 8.6 Hz, 1H), 6.47 (s, 2H), 5.74 (s, 1H), 4.86-4.76 (m, 1H), 4.50 (dt, J=25, 6, 6.3 Hz, 4H), 4.04 (t, J=5.1 Hz, 2H), 3.73 (q, J=5.1 Hz, 2H), 3.51-3.42 ( m, 1H), 3.02 (s, 4H), 2.40 (s, 4H).

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Example 6 Preparation of 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)phenyl)piperazin-1-yl)methyl) propane-1,3-diol (6).

dhm

Oxetane-3-carbaldehyde XXVIII.

In a round bottom flask equipped with a stir bar, oxetan-3-ylmethanol (2.00 g, 22.7 mmol) was dissolved in DCM (50 mL) and Dess-Martin periodinan (10.67 g, 28.38 mmol) was added in one portion. . The reaction mixture was stirred at RT overnight. The resulting solids were filtered through Celite and washed with DCM (3 mlx5).

The filtrate was removed and concentrated in vacuo and the resulting crude oxetane-3carbaldehyde XXVIII was used directly in the next step.

1-(4-Nitrophenyl)-4-(oxetan-3-ylmethyl)piperazine XXIX.

In a round-bottom flask lowered with a stirrer arm, oxetane-3-carbaldehyde XXVIII (0.977 g, 11.35 mmol), 1-(4-nitrophenyl)piperazine (1.18 g, 5.68 mmol) in DCM (100 ml) were added and HOAc (1.70 g, 28.38 mmol) in DCM (2 ml). After 5 min NaBH(OAc) ₃ (24.06 g, 113.05 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours. Most of the volatiles were removed in vacuo. DCM (200 ml) was added followed by saturated aqueous NaHCO ₃ (20 ml) and the resulting mixture was stirred for 20 minutes. The organic phase was separated and washed with saturated aqueous NaHCO ₃ (20 mlx3), brine (20 mlx1), dried over Na ₂ SO ₄ , filtered and the solvents were removed under vacuum. The resulting residue was passed through a silica gel column (MeOH:DCM=0:100 to 5:95 to 25:75) to give the desired compound XXIX.

4-(4-(Oxetan-3-ylmethyl)piperazin-1-yl)aniline XXX.

To a round-bottom flask equipped with a stir bar was added 1-(4-nitrophenyl)-4-(oxetan-3ylmethyl)piperazine XXIX (3.20 g, 11.54 mmol), ethanol (60 ml) and water (60 ml). Iron (4.51 g, 80.77 mmol) and ammonium chloride (4.32 g, 80.77 mmol) were added, after which the reaction mixture was heated at 80°C for 1 h, and then filtered through celite and washed with DCM (5 mlx5). The resulting filtrate was subjected to extraction with DCM (20 mlx3). The combined organic extracts were washed with water (20 mlx2), brine (20 mlx1), dried over Na ₂ SO ₄ and concentrated in vacuo.

- 40 040127

The desired 4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)aniline XXX was obtained.

6-Bromo-N-(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine XXXI.

4-(4-(oxetan-3-ylmethyl)piperazin1-yl)aniline XXX (1.19 g, 4.81 mmol), 6,8-dibromimidazo[1,2-a ]pyrazine (1.33 g, 4.81 mmol), isopropanol (24.1 ml) and diisopropylethylamine (1.37 g, 10.58 mmol) and the resulting reaction mixture was heated at 100° C. overnight. Most of the solvents were removed under vacuum and DCM (200 ml) was added to the resulting mixture. The resulting solution was washed with H ₂ O (20 mlx2), brine (20 mlx1), dried over Na ₂ SO ₄ , filtered and the solvents were removed under vacuum. The resulting residue was passed through a silica gel column (MeOH:DCM=5:95) to give the desired compound XXXI, 0.692 g as a light red solid.

tert-Butyl-(6-bromimidazo[1,2-a]pyrazin-8-yl)(4-(1-(oxetan-3-ylmethyl)piperidin-4yl)phenyl)carbamate XXXII.

6-Bromo-N-(4-(4-(oxetan-3ylmethyl)piperazin-1-yl)phenyl)imidazo[1,2-а]pyrazin-8-amine XXXI (560 mg, 1.27 mmol), DCM (11 ml), di-tert-butyl dicarbonate (414.4 mg, 1.90 mmol) and triethylamine (640.5 mg, 6.33 mmol). The reaction mixture was heated at 50° C. overnight. DCM (200 ml) was added and the resulting solution was washed with water (20 mlx2), brine (20 mlx1), dried over Na ₂ SO ₄ , filtered and the solvents removed under vacuum. Column chromatography afforded the desired compound XXXII as a yellow solid.

tert-Butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4(oxetan-3-ylmethyl )piperazin-1-yl)phenyl)carbamate XXXIII.

tert-Butyl-(6-bromimidazo[1,2а]pyrazin-8-yl)(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)carbamate XXXII (150 mg, 0.276 mmol), N,N-bisBoc-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine XV (255.8 mg, 0.607 mmol) in DME (2.3 ml), Pd(PPh ₃ ) ₄ (16.0 mg, 0.14 mmol), Na ₂ CO ₃ aqueous solution (1.0 N, 0.91 ml, 0.91 mmol) and DME (2 ml). The resulting mixture was heated at 75°C for 2 h, then DCM (200 ml) was added, the resulting mixture was washed with water (30 mlx3), brine (30 mlx1), dried over MgSO ₄ , filtered and the solvents were removed under vacuum. Purification on a silica gel column (MeOH:DCM=5:95) provided the desired compound XXXIII.

2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)phenyl)piperazin-1yl)methyl)propan-1, 3-diol (6).

To a solution of tert-butyl-(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2а]pyrazin-8-yl)(4-(4-(oxetan-3- ylmethyl)piperazin-1-yl)phenyl)carbamate XXXIII (250 mg, 0.33 mmol) in DCM (30 mL) was added TFA (940.3 mg, 8.25 mmol). The resulting mixture was stirred at room temperature overnight. Additional TFA (752.2 mg, 6.60 mmol) was added and stirring was carried out at room temperature overnight. Most of the solvents were removed under vacuum, DCM (200 ml) and saturated aqueous NaHCO ₃ (30 ml) were added and the resulting mixture was stirred for 30 min. The organic phase was separated, washed with saturated aqueous NaHCO ₃ (20 mlx4), brine (20 mlx1). The aqueous phase was subjected to extraction with DCM (30 mlx2). The combined organic phases were washed with brine (20 mlx1), dried over Na ₂ SO ₄ , filtered and the solvents were removed under vacuum. The resulting crude material was purified on an ISCO column using MeOH:DCM=0:100 to 5:95 to 7.5:92.5 to 25:75 to elute the desired compounds. Two compounds were obtained: the first is the oxetane compound, and the second is the desired compound 6.

LCMS-ESI ⁺ (m/z): [M+H] ⁺ : 476.

¹ H NMR (300 MHz, d ₆ -DMSO) δ 9.51 (s, 1H), 8.60 (s, 1H), 8.49 (s, 1H), 8.14 (d, J=1, 5 Hz, 1H), 7.95 (d, J=9 Hz, 2H), 7.90 (s, 1H), 7.64 (s, 1H), 6.99 (d, J=9 Hz, 2H ), 6.48 (s, 2H), 4.51 (br S, 2H), 3.43 (d, J=6 Hz, 4H), 3.12 (br m, 4H), 2.54 (broad m, 4H), 2.34 (d, J=7.2 Hz, 2H), 1.83 (m, 1H).

- 41 040127

Example 7 Preparation of 2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2(4-(oxetan- 3-yl)piperazin-1-yl)phenoxy)ethanol (7).

tert-Butyl-tert-butoxycarbonyl (6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperazin-1yl)-3-(2-((tetrahydro-2H-pyran- 2-yl)oxy)ethoxy)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3chloropyrazin-2-yl)carbamate XXXIV.

Tert-butyl-(6bromimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl) was added to a flask equipped with a reflux condenser. -3-(2-((tetrahydro-2H-pyran-2yl)oxy)ethoxy)phenyl)carbamate XXVI (prepared as described in Example 5) (352 mg, 0.52 mmol), 2-(bis-boc- amino)-3-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine (obtained in a manner analogous to that used in Example 2 to obtain Compound XV) (500 mg, 1.1 mmol), Pd(PPh ₃ ) ₄ (30 mg, 0.03 mmol) in sodium carbonate (1.6 ml, 1 M in H2O) and DME (4.8 ml). The resulting mixture was heated to reflux for 1 h. The reaction mixture was cooled to room temperature, diluted with DCM and H2O. The resulting aqueous layer was separated and subjected to extraction with DCM. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography on an ISCO Rf column (4 g column) eluting with a gradient of 100% DCM-100% 60/35/5 DCM/Et ₂ O/MeOH, the appropriate fractions were combined and concentrated to give the desired compound, tert-butyl -tert-butoxycarbonyl(6-(8((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2yl) hydroxy)ethoxy)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate XXXIV.

tert-Butyl-tert-butoxycarbonyl (6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperazin-1yl)-3-(2-((tetrahydro-2H-pyran- 2-yl)oxy)ethoxy)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3methylpyrazin-2-yl)carbamate XXXV.

tert-butyl-tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl))piperazin-1-yl)-3-(2-( (tetrahydro2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate XXXIV (258mg, 0.28mmol) ), methylboronic acid (503 mg, 8.4 mmol), Pd(PPh ₃ ) ₄ (32 mg, 0.03 mmol) in sodium carbonate (0.8 ml, 1 MB H2O) and DME (2.5 ml) . The resulting mixture was heated at 150°C for 20 min. The reaction mixture was cooled to room temperature, diluted with DCM and H2O. The resulting aqueous layer was separated and subjected to extraction with DCM. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography on an ISCO Rf column (4 g column) eluting with a gradient of 100% DCM-100% 75/18/7 DCM/Et ₂ O/MeOH to give the desired compound, t-butyl-t-butoxycarbonyl(6 -(8((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2yl)oxy)ethoxy)phenyl) amino)imidazo[1,2-a]pyrazin-6-yl)-3-methylpyrazin-2-yl)carbamate XXXV.

2-(5-((6-(6-Amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxetan-3yl)piperazine -1-yl)phenoxy)ethanol (7).

To a solution of tert-butyl-tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-42 040127 yl)piperazin-1-yl)-3-(2-(( tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1,2-a]pyrazin6-yl)-3-methylpyrazin-2-yl)carbamate XXXV (165 mg, 0.18 mmol) in DCM (2.2 ml) TFA (1.1 ml, 0.11 mmol) was added. The resulting mixture was stirred at RT for 16 h. The reaction mixture was diluted 9:1 with DCM:MeOH and H2O. The resulting aqueous layer was separated and subjected to extraction with 9:1 DCM:McOH. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography eluting with a gradient of 100% 75/18/7 DCM/Et ₂ O/MeOH-100% 70/20/10 DCM/E1 ₂ O/McOH to give the desired compound 2-(5-(( 6-(6amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxetan-3-yl)piperazin-1yl)phenoxy)ethanol ( 7.56 mg, 59%).

LCMS-ESI+ (m/z): [M+H] ⁺ : 518.2.

1H NMR (300 MHz, d ₆ -DMSO) δ 9.49 (s, 1H), 8.56 (s, 1H), 8.44 (s, 1H), 8.13 (d, J=1.1 Hz, 1H), 7.85-7.66 (m, 2H), 7.62 (d, J=1.1 Hz, 1H), 6.90 (d, J=8.6 Hz, 1H), 6.25 (s, 2H), 4.87-4.77 (m, 1H), 4.50 (dt, J=25.2, 6.3 Hz, 4H), 4.04 (t, J= 5.1 Hz, 2H), 3.74 (q, J=5.2 Hz, 2H), 3.51-3.39 (m, 1H), 3.10-2.95 (m, 4H), 2.45-2.35 (m, 4H), 2.34 (s, 3H).

Alternatively, compound XXXIV may be taken directly at this stage and similarly deprotected to give the 5-chloropyrazine analog.

Monomesylate and succinate forms.

Analysis of monomesylate (MSC) and succinate forms of the compound according to example 2, given in the present description, by X-ray powder diffraction (XRPD) was carried out on a diffractometer (PANanalytical XPERT-PRO, PANalytical B.V., Almelo, Netherlands) using copper radiation (Cu Κα, λ \u003d 1.5418 A). Samples for analysis were prepared by placing a powdered sample in the center of an aluminum holder equipped with a zero background plate. The generator operated at a voltage of 45 kV and a current of 40 mA.

Soller slits of 0.02 rad, an antiscattering slit of 1.0°, and a divergence slit were used. The sample rotation speed was 2 s. Scanning was performed from 2 to 40° 2-theta. Data analysis was performed with X'Pert Highscore, version 2.2c (PANalytical B.V., Almelo, Netherlands) and X'Pert data viewer software, version 1.2d (PANalytical B.V., Almelo, Netherlands). XRPD curves for Monomesylate Forms I and II were generated using the following instrument settings: 45 kV, 40 mA, Cu Κα, λ=1.5418 A, scan range 2, -40°, step size 0.0167°, count time 15.875 s . XRPD curves for succinate forms I and II were generated using the following instrument settings: 45 kV, 40 mA, Cu Κα, λ=1.5418 A, scan range 2, -40°, step size 0.0084°, count time 95.250 s .

1H NMR spectra of the monomesylate (MSC) and succinate forms of the compound according to Example 2 were obtained on a Varian 400-MR 400 MHz instrument with a 7620AS sample changer. The initial parameters for protons were as follows: spectral width from 14 to -2 ppm (6397.4 Hz); relaxation delay 1 s; exposure time 2.5559 s; number of scans or repetitions 8; temperature 25°C. Samples were prepared in dimethyl sulfoxide-d ₆ unless otherwise noted. Offline analysis was performed using the MNova software.

Example 8 16-(6-Aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Form monomesylate.

Form I salt of methanesulfonic acid (MSA) was obtained by dissolving 6-(6-aminopyrazin2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2- a]pyrazine-8-amine (Example 2) in 11 volumes of a mixture of acetone/H ₂ O (36:64 vol.%) with 1 mol equivalent of methanesulfonic acid (MSA) at room temperature. 19 volumes of acetone were added to the resulting solution over 1 h, and the contents of the reactor were stirred at room temperature overnight.

XRPD Analysis of Form I α-amine monomesylate was carried out as described above, and the diffraction curve shown in FIG. 1 with the peaks shown in the table below.

No. Pos. [°2-theta] Rel. INT. [%1 1 19.6606 100 2 17.2746 93.07 3 17.8971 69.96 4 21.6306 65.74 5 25.7805 59.16 6 18.7593 51.5 7 13.7252 48.77 8 15.7206 41.91 9 24.7364 38.09 10 18.4345 36.84

In one embodiment, Form I is 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetane-3

- 43 040127 yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine monomesylate can be characterized by XRPD peaks 19.7 (19.6606), 17.3 (17.2746), 17 .9 (17.8971), 21.6 (21.6306) and 25.8 (25.7805). In another embodiment, Form I 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2a]pyrazin-8 -amine monomesylate can be characterized by XRPD peaks of 19.7 (19.6606), 17.3 (17.2746), 17.9 (17.8971) and 21.6 (21.6306). In another embodiment, Form I 6-(6-aminopyrazin-2-yl)-N-(4-(4(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine monomesylate can have XRPD peaks of 6.0, 6.2, 8.6 and 9.6.

NMR Analysis of 6-(6-Aminopyrazin-2-yl)-N-(4-(4(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] Monomesylate Salt Form I Pyrazine-8-amine I was carried out as described above, resulting in the NMR spectrum shown in FIG. 2.

1H NMR (400 MHz, DMSO-d6) δ 10.57 (s, 1H), 9.60 (s, 1H), 8.62 (s, 1H), 8.47 (s, 1H), 8.17 (d, J=1.2 Hz, 1H), 8.03-7.96 (m, 2H), 7.90 (s, 1H), 7.69 (d, J=1.2 Hz, 1H) , 7.09 (d, J=9.0 Hz, 2H), 4.78 (p, J=8.0 Hz, 4H), 4.49 (m, 1H), 4.00-2.8 ( m, 10H), 2.32 (s, 3H).

Differential Scanning Calorimetry (DSC).

DSC analysis was performed for each of the compounds according to the examples provided herein using a TA Instruments Q2000 DSC instrument. The sample was placed in an aluminum DSC crucible and the exact weight was recorded. The crucible was covered with a lid and then either crimped or hermetically sealed. The same cell was heated while purging with nitrogen at a rate of 10°C/min to a final temperature of 300°C. Indium was used as a calibration standard.

DSC analysis results for form I of 6-(6-aminopyrazin-2-yl)-N(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] monomesylate salt ]pyrazine-8-amine carried out as described above are shown in FIG. 3.

Thermogravimetric analysis (TGA).

TGA was performed for each of the compounds as described herein using a TA Instruments Q5000 TGA. Each sample was placed in an aluminum sample crucible and placed in a thermogravimetric analysis oven. The oven was heated under nitrogen at a rate of 10°C/min to a final temperature of 300°C. The TGA oven was calibrated using the Curie point method using magnets.

TGA analysis results for 6-(6-aminopyrazin-2-yl)-N-(4(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a monomesylate salt Form I ]pyrazine-8-amine carried out as described above are shown in FIG. 4.

Example 9 Form II amine monomesylate.

Form II of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine monomesylate salt obtained by drying Form I of the monomesylate salt of 6-(6aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine (Example 8) in a vacuum oven at ~40°C with N2 purge.

XRPD Analysis of Form II 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8 α-amine monomesylate was carried out as described above, resulting in the diffraction curve shown in FIG. 5 with the peaks shown in the table below.

No. Pos. [°2 theta] Rel. INT. [%1 1 17.2698 100 2 25.1384 67.84 3 20.4423 63.66 4 19.5732 62.11 5 18.5264 50.36 6 17.7884 50.07 7 21.6273 45.52 8 15.2397 44 9 6.855 35.01 10 13.65 26

In one embodiment, Form II 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine monomesylate can be characterized by XRPD peaks of 17.3 (17.2698), 25.1 (25.1384), 20.4 (20.4423), 19.6 (19.5732) and 18.5 (18. 5264). In a further embodiment Form II 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8 -amine monomesylate can be characterized by XRPD peaks of 17.3 (17.2698), 25.1 (25.1384), 20.4 (20.4423) and 19.6 (19.5732). In another embodiment, Form II 6-(6aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8 α-amine monomesylate can be characterized by XRPD peaks of 6.1, 6.9, 11.0 and 13.6.

As a result of NMR analysis for form II of the monomesylate salt of 6-(6-aminopyrazin-2-yl)-N(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2- a]pyrazine-8-amine carried out as described above, the NMR spectrum shown in FIG. 6.

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1H NMR (400 MHz, DMSOL) δ 9.61 (s, 1H), 8.63 (s, 1H), 8.47 (s, 1H), 8.19 (d, J=1.2 Hz, 1H ), 8.02-7.95 (m, 2H), 7.91 (s, 1H), 7.72 (d, J=1.2 Hz, 1H), 7.13-7.06 (m, 2H), 4.85-4.72 (m, 4H), 4.53-4.45 (m, 1H),

4.30-2.75 (m, 10H), 2.34 (s, 3H).

DSC analysis results for form II of 6-(6-aminopyrazin-2-yl)-N(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] monomesylate salt ]pyrazine-8-amine carried out as described above are shown in FIG. 7.

TGA Analysis Results for Form II of 6-(6-Aminopyrazin-2-yl)-N(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a] Monomesylate Salt ]pyrazine-8-amine carried out as described above are shown in FIG. 8.

Example 10 Form I amine succinate.

Form I 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2a]pyrazin-8-amine succinate was prepared by dissolving 1.6 mol-eq. succinic acid in THF and subsequent addition of the resulting acid solution of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a ]pyrazine-8-amine. Next, the resulting substance was stirred at room temperature with a magnetic stirrer overnight.

XRPD Analysis of Form I α-amine succinate was carried out as described above, resulting in the diffraction curve shown in FIG. 9 with the peaks shown in the table below.

No. Pos. [°2-theta] Rel. INT. [%1 1 16.5 100 2 24.5 38.64 3 17.7 9.27 4 28.4 8.68 5 21.8 7.57 6 8.0 6.53 7 23.1 4.59 8 12.1 4.38 9 8.3 3.78 10 27.1 3.65

In one embodiment, Form I 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine succinate can be characterized by XRPD peaks of 16.5, 24.5, 17.7, 28.4 and 21.8. In another embodiment, Form I 6-(6-aminopyrazin-2-yl)-N-(4(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine succinate can be characterized by XRPD peaks of 16.5, 24.5, 8.0 and 8.3.

As a result of NMR analysis of form I 8-amine succinate carried out as described above, the NMR spectrum shown in FIG. 10.

1H NMR (400 MHz, DMSO-d6) δ 12.12 (s, 2H), 9.48 (s, 1H), 8.59 (s, 1H), 8.48 (s, 1H), 8.12 (d, J=1.1 Hz, 1H), 7.97-7.86 (m, 3H), 7.62 (d, J=1.1 Hz, 1H), 7.01-6.94 ( m, 2H), 6.45 (s, 2H), 4.55 (t, J=6.5 Hz, 2H), 4.46 (t, J=6.1 Hz, 2H), 3.49- 3.38 (m, 1H), 3.13 (t, J=4.9 Hz, 4H), 2.40 (s, 10H).

Results of DSC analysis for Form I 8-amine succinate carried out as described above are shown in FIG. eleven.

Results of TGA analysis for Form I 8-amine succinate carried out as described above are shown in FIG. 12.

Preparation of Form I of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetαn-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine The succinate was also repeated using IPA, acetone and 2-MeTHF as solvents.

Example 11 Form II amine succinate.

To the free base of 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine was added 10.0 parts of 2-propanol, followed by vigorous shaking to form a suspension. A separate solution of succinic acid (0.43 parts, 1.6 mol equiv.) in 2-propanol (15 parts) was prepared at ambient temperature and added to said suspension. Next, the resulting suspension was shaken at room temperature for about 1 day. To this suspension was added another solution of succinic acid (0.09 parts, 0.3 mol-eq.) in 2-propanol (3 parts) and the resulting suspension was shaken at room temperature for about two days. An additional solution of succinic acid (0.27 parts, 1.0 mol equiv.) in 2-propanol (8 parts) was prepared at room temperature, added to this suspension, and the resulting suspension was shaken for about 2 days. Next, the temperature of the contents was brought to 40°C and the resulting suspension was shaken for about two hours. The contents were then brought back to room temperature and shaken for about 16 hours.

Next, the resulting suspension was filtered, sprayed with 2-propanol (7.0 parts) and dried at 60°C.

XRPD Analysis of Form II 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8 α-amine succinate was carried out as described above, resulting in the diffraction curve shown in FIG. 13 with the peaks shown in the table below.

No. Pos. [°2 theta] Rel. INT. [%] 1 24.9821 100 2 16.3186 38.39 3 21.952 18.44 4 7.8958 17.62 5 7.5828 6.9 6 28.5998 6.52 7 11.3329 5.73 8 30.8568 5.48 9 28.0273 5.21 10 21.5026 4.73

In one embodiment, Form II 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine succinate can be characterized by XRPD peaks of 25.0 (24.9821), 16.3 (16.3186), 22.0 (21.952), 7.9 (7.8958) and 7.6 (7.5828) . In another embodiment, Form II 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2a]pyrazin-8 -amine succinate can be characterized by XRPD peaks of 25.0 (24.9821), 16.3 (16.3186), 7.9 (7.8958) and 7.6 (7.5828).

As a result of NMR analysis for Form II -8-amine succinate carried out as described above, the NMR spectrum shown in FIG. 14.

^! H NMR (400 MHz, DMSO-( ₁₆ ) δ 12.13 (s, 2H), 9.48 (s, 1H), 8.58 (s, 1H), 8.47z (s, 1H), 8 .12 (d, J=1.1 Hz, 1H), 7.97-7.86 (m, 3H), 7.62 (d, J=l.l Hz, 1H), 7.02-6, 94 (m, 2H), 6.45 (s, 2H), 4.55 (t, J=6.5 Hz, 2H), 4.46 (t, J=6.0 Hz, 2H), 3, 44 (p, J=6.3 Hz, 1H), 3.17-3.10 (m, 4H), 2.40 (s, 10H), 1.02 (d, J=6.1 Hz, 2H ).

Results of DSC analysis for Form II The 8-amine succinate carried out as described above are shown in FIG. 15.

TGA Analysis Results for Form II 8-amine succinate carried out as described above are shown in FIG. 16.

biological examples.

Example 12 Biochemical Assay of Syk at High Processing Rate.

Syk activity was determined using KinEASE (Cisbio), a time-resolved fluorescence resonance energy transfer (TR-FRET) immunoassay. In this study, Syk catalyzes the phosphorylation of an XE665-labeled peptide substrate. A europium-conjugated phosphotyrosine-specific antibody binds to the resulting phosphorylated peptide. Phosphorylated peptide production was quantified by TR-FRET using europium as donor and XL665 as acceptor in a 2-way endpoint study. Briefly, test compounds were serially diluted in DMSO and plated into small volume white non-binding 384-well Corning plates using an Echo 550 acoustic liquid dispenser (Labcyte®). Syk enzyme and substrates were dispensed into assay plates using Multi-Flo (Bio-Tek Instruments). The standard 5 µl reaction mixture contained 20 µM ATP, 1 µM biotinylated peptide, 0.015 HMf Syk in reaction buffer (50 mM Hepes, pH 7.0, 0.02% NaN ₃ , 0.1% bovine serum albumin, 0.1 mM orthovanadate, 5 mM MgCl ₂ , 1 mM DTT, 0.025% NP-40). After incubation for 30 minutes at room temperature, 5 µl of a termination and detection solution (1:200 solution of europium cryptate labeled peptibody to stop phosphorylation and 125 nM streptavidin-XE665 indicator in 50 mM Hepes pH 7.0 buffer for detection) was added. containing a sufficient amount of EDTA). The plate was then further incubated for 120 min at room temperature and read using an Envision 2103 multifunctional reader (PerkinElmer) with FRET excitation/emission/emission at 340/615/665 nm, respectively. Fluorescence intensity at emission wavelengths of 615 and 665 nm was expressed as a ratio (665/615 nm). The percentage of inhibition was calculated as follows:

% Inhibition=100 x (Sample Ratio-0% Inhibition Ratio)/(100% Inhibition Ratio-0 0% Inhibition Ratio),

-46040127 where 0.1% DMSO (0% inhibition) served as a negative control and μM K252α (100% inhibition) was used as a positive control.

The activity of the compounds according to examples 1-7 is presented in the following table, from which it can be seen that these compounds are Syk inhibitors with an IC ₅₀ of less than 50 nM.

example no.: connection name 1Seo in rel. Syk (nM) Ex-1: 6-(6-amino-5-methylpyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 6.2 Ex-2: 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3-yl)piperazin1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine 13.5 Etc. -3: (K)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)morpholin-2-yl)methanol 13.3 Ex-4: 6-(6-aminopyrazin-2-yl)-5-methyl-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 44 Ex-5: 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3-yl) piperazin-1-yl)phenoxy)ethanol 12.2 Ex-6: 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)piperazin-1-yl)methyl )propane-1,3-diol 14.5 Etc. -7: 2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)-2-(4-(oxetan-3- yl)piperazin-1yl)phenoxy)ethanol 8.7

Example 13 Whole blood CD63 basophil testing in a 384-well high-throughput biological screen (HTBS).

Syk activity was assessed in relation to reduced basophil activation as measured by CD63 expression in a human whole blood basophil assay (25% blood). Basophil activation was determined in human whole blood using the Flow CASTt kit (Buhlmann Laboratories AG, Baselstrasse, Switzerland) according to the manufacturer's protocol with minor modifications. Fresh whole blood in heparin was collected and delivered on the same day (AllCells, Emeryville, CA). Whole blood samples were incubated with either DMSO (final concentration 1%) or serially diluted compounds in DMSO for 60 minutes at 37°C. Basophils were activated with anti-FceRI monoclonal antibodies and stained with anti-CD63-FITC and αanti-CCR3-PE for 20 min at 37°C (per well: 50 µl of whole blood mixed with 113 µl of stimulation buffer, 8.5 µl of monoclonal anti-FceRI antibodies, 8.5 µl of CCR3-PE/CD63-FITC antibody dye). Cells were centrifuged at 1000xg for 18 min and 150 µl/well of supernatant was removed. The erythrocytes were lysed and the cells were fixed in 2 lysis steps: the cell pellets were resuspended in 150 μl/well of 1X lysis buffer, incubated at room temperature for 10 min, and the cell pellets were harvested by centrifugation at 1200 rpm for 5 min. Cells were washed twice with 150 µl/well wash buffer and resuspended in a final volume of 75 µl/well wash buffer for either immediate flow cytometric analysis or overnight incubation at 4° C. followed by flow cytometry analysis. Degranulation (activation of basophils) was determined by the expression of CD63 on the surface of CCR3-positive cells. The percentage of CD63-positive cells in the gated basophil population was determined and normalized to DMSO (negative control) and control compound (positive control). The activity of the compounds according to examples 1-7 is presented in the following table, from which it can be seen that these compounds are effective in reducing the activation of basophils with an EC ₅₀ of less than 200 nM.________________

example no.: connection name ECso in rel. CD63 (nM) Ex-1: 6-(6-amino-5-methylpyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 51 Ex-2: 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3-yl)piperazin1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine 80 Etc. -3: (K)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)morpholin-2-yl)methanol 63 Ex-4: 6-(6-aminopyrazin-2-yl)-5-methyl-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 157 Ex-5: 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3-yl) piperazin-1-yl)phenoxy)ethanol 120 Ex-6: 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)piperazin-1-yl)methyl )propane-1,3-diol 128 Etc. -7: 2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)-2-(4-(oxetan-3- yl)piperazin-1yl)phenoxy)ethanol 167

Example 14 Kinetic Solubility

The kinetic solubility of the compounds in phosphate buffer pH 7.4 was assessed. Subject

- 47 040127 testing compounds were dissolved in dimethyl sulfoxide at a concentration of 10 mm. Stock samples were diluted: 3 µl in 297 µl of phosphate buffer pH 7.4 (Dulbecco's phosphate-buffered saline (SigmaAldrich D8662), total molarity was 0.149 M and pH 7.43). Next, the samples were incubated for 24 h at 37°C with stirring, centrifuged, an aliquot was taken, and the amount of the compound was determined in it relative to the known standard concentration of 0.1 mM. The kinetic solubility of the compound according to examples 1-7 is presented in the following table, from which it can be seen that the compounds have a kinetic solubility at pH 7.4 greater than 90 μM._____________

example no.: connection name Solubility at pH 7.4 (µM) Ex-1: 6-(6-amino-5-methylpyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 95 Ex-2: 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine 95 Etc. -3: (K)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)phenyl)morpholin-2-yl)methanol 91 Ex-4: 6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 100 Ex-5: 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3-yl) piperazin-1-yl)phenoxy)ethanol 97 Ex-6: 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)piperazin-1-yl)methyl )propane-1,3-diol 99

Example 15 Stability study in human hepatocytes.

The stability of the compounds in human hepatocytes was evaluated as estimated hepatocyte clearance in l/h/kg. Compounds to be tested were diluted to a concentration of 200 μM (4 μl of stock 10 mM DMSO in 196 μl of ACN:H ₂ O (50:50). Propranolol was used as a positive control and buffer without hepatocytes alone as a 0% control. They were further diluted : 4 µl with 891 µl of KHV buffer (InVitroGRO catalog # Z99074) to make a 2X dosing solution 250 µl of 2X dosing solution with 250 µl of hepatocyte cells (1 x 10 ⁶ viable cells/ml per well) was added to each well of a 24-well plate ) or KHV for controls to give a final compound concentration of 1 μM during incubation.The final solvent concentration was 0.01% DMSO and 0.25% ACN.The culture plate was placed on a shaker and incubated at 37°C, 5% CO2. Samples were sampled at time points 0, 1, 3 and 6 hours The loss of the main compound was determined using LC-MS methods from a standard curve The activity of the compounds according to examples 1-7 is presented in the following th table, from which it can be seen that the clearance in hepatocytes was approximately 0.12 l / h / kg or less. _________________________

example no.: connection name Clearance in hepatocytes (l/h/kg) Etc. -1: 6-(6-amino-5-methylpyrazin-2-yl)- M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a!pyrazin- 8-amine 0.12 Ex-2: 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine 0.055 Etc. -3: (K)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2a1pyrazin-8-yl)amino)phenyl)morpholin-2-yl)methanol 0.09 Ex-4: 6-(6-aminopyrazin-2-yl)-5-methyl-M-(4-(4-(oxetan-3yl)piperazin-1-yl)phenyl)imidazo[1,2-a] pyrazine-8-amine 0.08 Ex-5: 2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3-yl) piperazin-1-yl)phenoxy)ethanol 0.07 Ex-6: 2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin8-yl)amino)phenyl)piperazin-1-yl)methyl )propane-1,3-diol 0.08 Ex-7: 2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2a]pyrazin-8-yl)amino)-2-(4-(oxetan- 3-yl)piperazin-1yl)phenoxy)ethanol 0.05

Example 16 Comparison with Known Syk Inhibitors.

The studies according to examples 8-11 were used to compare the compounds described herein with compounds known in the art. Data comparing compounds according to examples 1-7 with previously described compounds are shown in the following table. From these results, it is clear that the compounds described herein are desirable Syk inhibitors with enhanced Syk and CD63 activity compared to known compounds, improved kinetic solubility (at least about 9 times more soluble) and clearance in hepatocytes ( at least about 2 times lower ground clearance). Thus, the combination of enhanced Syk and CD63 inhibitory activity with improved kinetic solubility and clearance provides compounds that are expected to be effective in the treatment of the diseases described herein with improved pharmacokinetic properties.

- 48 040127

Connection name IC50 in rel. Syk (nM) IC50 in rel. CD63 (nM) Solubility at pH 7.4 (µM) Clearance in hepatocytes (units) Ex-1: 6-(6-amino-5methylpyrazin-2-yl)-M-(4-(4(oxetan-3-yl)piperazin-1 yl)phenyl)imidazo[1,2a]pyrazin-8- amine 6.2 51 95 0.12 Ex-2: 6-(6-aminopyrazin-2yl)-M-(4-(4-(oxetan-3yl)piperazin-1 yl)phenyl)imidazo[1,2a]pyrazin-8-amine 13.5 80 95 0.055 Ex-3: (R)-(4-(4-((6-(6aminopyrazin-2yl)imidazo[1,2-a]pyrazin-8yl)amino)phenyl)morpholin-2yl)methanol 13.3 63 91 0.09 Ex-4: 6-(6-aminopyrazin-2yl)-5-methyl-M-(4-(4-(oxetan-3yl)piperazin-1 yl)phenyl)imidazo[1,2a]pyrazin-8-amine 44 157 100 0.08 Ex-5: 2-(5-((6-(6aminopyrazin-2yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3yl)piperazin-1yl)phenoxy )ethanol 12.2 120 97 0.07 Ex-6: 2-((4-(4-((6-(6aminopyrazin-2yl)imidazo[1,2-a]pyrazin-8yl)amino)phenyl)piperazin-1 yl)methyl)propan-1, 3-diol 14.5 128 99 0.08 Ex-7: 2-(5-((6-(6-Amino-5methylpyrazin-2yl)imidazo[1,2-a]pyrazin-8yl)amino)-2-(4-(oxetan-3yl)piperazin- 1 yl)phenoxy)ethanol 8.7 167 But 0.05 Known compounds: 6-(5-Aminopyridin-3-yl)-M-(4morpholinophenyl)imidazo[1,2a]pyrazine-8-amine 31 101 5 0.68 6-(3-aminophenyl)-M-(3,4dimethoxyphenyl)imidazo[1,2а]pyrazine-8-amine 188 809 3 0.24 6-(5-amino-6-methylpyridin-3yl)-N-(4- morpholinophenyl)imidazo[1,2a]pyrazine-8-amine 16 250 5 0.80 6-(6-aminopyridin-3 -yl)-M(3,4- dimethoxyphenyl)imidazo[1,2а]pyrazine-8-amine 53 734 10 0.90

In the present description, references are made to various patents, patent publications and other types of publication (eg, journal articles). The contents of all patents, patent applications and publications mentioned in this specification are hereby incorporated by reference in their entirety for all purposes.

Claims (15)

CLAIM 1. Crystal form I amine monomesylate, characterized by X-ray powder diffraction peaks at angles 2θ equal to 19.7±0.2°, 17.3±0.2°, 17.9±0.2°, 21.6±0.2° and 25 .8±0.2°. 2. The crystalline form according to claim 1, characterized in that said diffraction pattern contains peaks at angles 2θ equal to 6.0±0.2°, 6.2±0.2°, 8.6±0.2° and 9 .6±0.2°. 3. Crystal form II amine monomesylate, characterized by X-ray powder diffraction peaks at angles 2θ equal to 17.3±0.2°, 25.1±0.2°, 20.4±0.2°, 19.6±0.2° and 18 .5±0.2°. 4. The crystalline form according to claim 3, characterized in that said diffraction pattern contains peaks at angles 2θ equal to 6.1±0.2°, 6.9±0.2°, 11.0±0.2° and 13 .6±0.2°. 5. Crystal form I 6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1- - 49 040127 yl)phenyl)imidazo[1,2-a]pyrazine-8-amine succinate, characterized by X-ray powder diffraction pattern peaks at 2θ angles equal to 16.5±0.2°, 24.5±0.2°, 17.7±0.2°, 28.4±0.2° and 21.8±0.2°. 6. A crystalline form according to claim 5, characterized in that said diffraction pattern contains peaks at 2θ angles of 8.0±0.2° and 8.3±0.2°. 7. Crystal form II amine succinate, characterized by X-ray powder diffraction peaks at angles 2θ equal to 25.0±0.2°, 16.3±0.2°, 22.0±0.2°, 7.9±0.2° and 7 .6±0.2°. 8. A pharmaceutical composition comprising a crystalline form according to any one of claims 1, 2, 3, 4, 5, 6 or 7 and at least one pharmaceutically acceptable carrier. 9. The use of a crystalline form according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 8 for the treatment of leukemia in a subject. 10. Use of the crystalline form or pharmaceutical composition of claim 9, wherein said disease or condition is acute myeloid leukemia. 11. Use of a crystalline form or pharmaceutical composition according to any one of claims 9, 10, wherein said subject is a human. 12. Use of a crystalline form or pharmaceutical composition according to any one of claims 9, 10, wherein said crystalline form or pharmaceutical composition is administered intravenously, intramuscularly, parenterally, nasally or orally. 13. The use of a crystalline form or pharmaceutical composition according to claim 12, wherein said crystalline form or pharmaceutical composition is administered orally once a day. 14. The use of a crystalline form or pharmaceutical composition according to claim 12, wherein said crystalline form or pharmaceutical composition is administered orally twice a day. 15. The use of a crystalline form according to any one of claims 1 to 7 for the preparation of a medicament for the treatment of a disease or condition according to any one of claims 9, 10. Analysis of Form I monomesylate salt of 6-(6-aminopyrazin-2-yl)-X-(4-(4-(oxetan3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine by XRPD method Fig. 1 - 50 040127 Analysis of Form II monomesylate salt of 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine by NMR fl (ppm) Fig. 2 - 51 040127 Analysis of Form II monomesylate salt of 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine by XRPD method Fig. 5 - 52 040127 Analysis of Form II monomesylate salt of 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan 3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin- 8-amine by NMR Fig. 6 - 53 040127 Analysis of 6-(6-aminopyrazin-2-yl)-X-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine succinate by XRPD Fig. 9 - 54 040127 Analysis of Form I 6-(6-aminopyrazin-2-yl)-M-(4-(4-(oxetan-3-yl)piperazin-1 yl)phenyl)imidazo[1,2-a]pyrazin-8-amine succinate by NMR 4 13 12 AND 10 9 V 7 6 5 4 3 2 1 0 -1 -2 fl (ppm) Fig. 10 - 55 040127 Analysis of Form II 6-(6-aminopyrazin-2-yl)-X-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine succinate XRPD method Fig. 13 - 56 040127 Form II Analysis NMR method H (ppm) Fig. 14 - 57 040127 Form II assay for 6-(6-aminopyrazin-2-yl)-X-(4-(4-(oxetan-3-yl)piperazin-1yl)phenyl)imidazo[1,2-a]pyrazin-8-amine succinate by TGA