Besylate salt of a btk inhibitor

(Cross-reference to related applications) The present invention claims priority to US Provisional Application No. 61/372349, which was filed on August 10, 2010, which is incorporated herein by reference in its entirety.

(Field of the present invention) The present invention provides salt forms and compositions useful as inhibitors of protein kinases.

(Background of invention) The search for new therapeutic agents has recently been significantly facilitated by a better understanding of the structure of disease-related enzymes and other biomolecules. One important class of enzymes that has been extensively studied is protein kinases.

Protein kinases constitute a large family of structurally related enzymes involved in the regulation of various signal transduction processes in the cell. Retaining its structure and catalytic function, protein kinases are thought to originate from a common ancestral gene. Almost all kinases contain 250-300 similar amino acid catalytic domains. Kinases can be classified into multiple families according to the substrates they phosphorylate (eg, protein-tyrosine, protein-serine / threonine, lipid, etc.).

In general, protein kinases mediate intracellular signal transduction by affecting the transfer of phosphate groups from nucleoside triphosphates to protein receptors, which are involved in signaling pathways. These phosphorylation events act as molecular on / off switches that can regulate or control the biological function of the target protein. These phosphorylation events are ultimately triggered in response to various extracellular and other stimuli. Examples of such stimuli are environmental and chemical stress signals (eg, osmotic shock, heat shock, UV light, bacterial endotoxin and H).₂O₂), Cytokines (eg, interleukin-1 (IL-1) and tumor necrosis factor α (TNF-α)), growth factors (eg, granulocyte macrophage colony stimulator (GM-CSF) and fibroblast growth factor (eg) FGF))) is included. Extracellular stimulation affects one or more cellular responses associated with cell growth, migration, differentiation, hormone secretion, transcription factor activation, muscle contraction, glucose metabolism, regulation of protein synthesis and regulation of the cell cycle. Can exert.

Many diseases are associated with abnormal cell responses caused by protein kinase-mediated events such as those described above. These disorders include, but are not limited to, autoimmune disorders, inflammatory disorders, bone disorders, metabolic disorders, neurological and neurodegenerative disorders, cancer, cardiovascular disease, allergies and asthma, Alzheimer's disease. Also includes hormone-related diseases. Therefore, it is still necessary to find a protein kinase inhibitor that is useful as a therapeutic agent.

<p num="0007"> It has been found here that the novel salt forms and compositions thereof of the present invention are useful as inhibitors of one or more protein kinases and exhibit desirable characteristics for that purpose. In general, this salt form and its pharmaceutically acceptable composition are useful in treating or reducing the severity of various diseases or disorders, as described in detail herein.<u style="single">In a preferred embodiment of the present invention, for example, the following items are provided.</u><u style="single">(Item 1)</u><u style="single"> Less than:</u><chemistry num="9"><img id="000002" he="39" wi="75" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry><u style="single">Compound 2.</u><u style="single">(Item 2)</u><u style="single"> The compound according to item 1, which is in solid form.</u><u style="single">(Item 3)</u><u style="single"> The compound according to item 2, which is crystalline.</u><u style="single">(Item 4)</u><u style="single"> The compound according to item 3, which is a crystalline solid substantially free of amorphous compound 2.</u><u style="single">(Item 5)</u><u style="single"> The compound according to item 1, which is substantially free of impurities.</u><u style="single">(Item 6)</u><u style="single"> The solid form according to item 2, which is form P1.</u><u style="single">(Item 7)</u><u style="single"> Item 6. The solid form of item 6, having one or more peaks selected from peaks at about 6.21, about 9.48 and about 13.29 ° 2θ in the PXRD.</u><u style="single">(Item 8)</u><u style="single"> 7. The solid form of item 7, having at least two peaks selected from peaks at about 6.21, about 9.48 and about 13.29 ° 2θ in the PXRD.</u><u style="single">(Item 9)</u><u style="single"> The solid form of item 6, having a PXRD substantially similar to that shown in FIG.</u><u style="single">(Item 10)</u><u style="single"> The solid form according to item 2, which is form P22.</u><u style="single">(Item 11)</u><u style="single"> 10. The solid form of item 10 having one or more peaks selected from peaks at about 7.29, about 8.38 and about 11.12 ° 2θ in its PXRD.</u><u style="single">(Item 12)</u><u style="single"> The solid form of item 11 having at least two peaks selected from peaks at about 7.29, about 8.38 and about 11.12 ° 2θ in its PXRD.</u><u style="single">(Item 13)</u><u style="single"> The solid form of item 10, having a PXRD substantially similar to that shown in FIG.</u><u style="single">(Item 14)</u><u style="single"> A composition comprising the compound according to item 1 and a pharmaceutically acceptable carrier or excipient.</u><u style="single">(Item 15)</u><u style="single"> A method of inhibiting BTK in a patient, which comprises administering to the patient the compound according to item 1 or a composition thereof.</u><u style="single">(Item 16)</u><u style="single"> A method of treating a BTK-mediated disorder in a patient, comprising administering to the patient the compound or composition thereof according to item 1.</u><u style="single">(Item 17)</u><u style="single"> 16. The method of item 16, wherein the disorder is selected from autoimmune disease, heteroimmune disease, inflammatory disease, cancer, bone and joint disease or thromboembolic disorder.</u><u style="single">(Item 18)</u><u style="single"> The disorders include rheumatoid arthritis, multiple sclerosis, diabetes, B-cell chronic lymphocytic leukemia, acute lymphocytic leukemia, hairy cell leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, multiple myeloma, colorectal cancer, pancreatic cancer, bone. 17. The method of item 17, selected from cancer, metastasis to bone, osteoporosis, irritable bowel syndrome, Crohn's disease, systemic erythematous leukemia or disorders associated with renal transplantation.</u></p>

<figref num="1">Figure 1 shows the FT-Raman spectrum of compound 2 form P1 (3400-100 cm).^-1).</figref><figref num="2">FIG. 2 represents the PXRD pattern of compound 2 form P1.</figref><figref num="3">FIG. 3 represents TG-FTIR of compound 2 form P1.</figref><figref num="4">FIG. 4 represents a DSC thermogram of Compound 2 form P1 showing cooling step 2.</figref><figref num="5">FIG. 5 represents a DSC thermogram of compound 2 form P1 showing a four-step heating and cooling process.</figref><figref num="6">FIG. 6 shows the PXRD pattern of compound 2 form P22 compared to the PXRD pattern of compound 2 form P1.</figref><figref num="7">Figure 7 shows the FT-Raman spectrum of compound 2 form P22 (3400-100 cm).^-1).</figref><figref num="8">FIG. 8 represents TG-FTIR of compound 2 form P22.</figref><figref num="9">FIG. 9 shows Compound 2 Form P22 prepared according to Example 3 below.¹Represents 1 H NMR.</figref><figref num="10">FIG. 10 represents the DSC thermogram of compound 2 form P22.</figref>

Schematic description of a particular aspect of the invention: US Publication No. 20100029610, published February 4, 2010 (Published '610, incorporated herein by reference in its entirety) is a member of TEC kinase, Bruton's tyrosine kinase (BTK). Specific 2,4-disubstituted pyrimidine compounds that covalently and irreversibly inhibit the activity of one or more protein kinases, including. Such compounds are Compound 1:

<chemistry num="1"><img id="000003" he="36" wi="55" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>including.

Compound 1 (N- (3- (5-fluoro-2- (4- (2-methoxyethoxy) phenylamino) pyrimidin-4-ylamino) phenyl) acrylamide) is designated compound number I-182 and is a compound. The synthesis of 1 is detailed in Example 20 published in '610.

Compound 1 is active in various assays and therapeutic models demonstrating covalent and irreversible inhibition of BTK (in enzyme and cellular assays). Notably, Compound 1 has been shown to inhibit B cell proliferation both in vitro and in vivo. Therefore, Compound 1 is useful in treating one or more disorders associated with the activity of BTK.

It is desirable to provide compound 1 in salt form that imparts features such as improved aqueous solubility, stability and ease of formulation compared to compound 1. Therefore, the present invention provides the besilate of compound 1.

According to one embodiment, the present invention describes Compound 2:

<chemistry num="2"><img id="000004" he="38" wi="75" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>The vesylate of Compound 1 represented by is provided.

Those skilled in the art will recognize that benzenesulfonic acid and compound 1 are ionic bonded to form compound 2. It is believed that compound 2 can exist in a variety of physical forms. For example, compound 2 may be in solution, suspension or solid form. In certain embodiments, Compound 2 is in solid form. When compound 2 is in solid form, the compound may be amorphous, crystalline or a mixture thereof. Typical solid forms will be described in more detail below.

In another embodiment, the invention provides compound 2 which is substantially free of impurities. As used herein, the term "substantially free of impurities" means a compound that is free of significant amounts of foreign matter. Such foreign matter may include excess benzenesulfonic acid, excess compound 1, residual solvent or any other impurities that may result from the preparation and / or isolation of compound 2. In certain embodiments, at least about 95% by weight of Compound 2 is present. In yet another embodiment of the invention, at least about 99% by weight of Compound 2 is present.

According to one embodiment, Compound 2 is present in an amount of at least about 97, 97.5, 98.0, 98.5, 99, 99.5, 99.8% by weight, where the percentage is based on the total weight of the composition. According to other embodiments, Compound 2 contains no more than about 3.0 HPLC area percent HPLC for the total area of the HPLC chromatogram, and in certain embodiments it is about 1.5 HPLC. Contains less than a percentage of the total organic impurities. In other embodiments, compound 2 contains only any single impurity of about 1.0% HPLC area percent or less; only any single impurity of about 0.6 HPLC area percent or less relative to the total area of the HPLC chromatogram and is specific. In the embodiment, it contains only any single impurity of about 0.5 HPLC area percent or less.

The structure shown for compound 2 is meant to include all tautomeric forms of compound 2. Furthermore, the structure shown herein also means containing compounds that differ only in the presence of atoms enriched with one or more isotopes. For example, replacing hydrogen with deuterium or tritium, or replacing carbon¹³C-or¹⁴Compounds having the structure of the present invention except that they are replaced with C-enriched carbon are within the scope of the present invention.

Solid form of compound 2: It is known that compound 2 can exist in various solid forms. Such forms include polymorphs, solvates, hydrates and amorphous. All such forms are considered in the present invention. In certain embodiments, the present invention provides compound 2 as a mixture of one or more solid forms selected from polymorphic phase, solvates, hydrates and amorphous compound 2.

As used herein, the term "polymorphic phase" refers to a different (unsolvated form) crystal structure in which a compound can crystallize. The term "solvate" as used herein refers to a crystal form in which a stoichiometric or non-stoichiometric solvent is incorporated into the crystal structure. Similarly, the term "hydrate" refers to a crystalline form in which a stoichiometric or non-stoichiometric amount of water is incorporated into the crystal structure.

In certain embodiments, compound 2 is a crystalline solid. In other embodiments, compound 2 is a crystalline solid that is substantially free of amorphous compound 2. As used herein, the term "substantially free of amorphous compound 2" means that the compound does not contain a significant amount of amorphous compound 2. In certain embodiments, at least about 95% by weight of crystalline compound 2 is present. In yet another embodiment of the invention, at least about 99% by weight of crystalline compound 2 is present.

In certain embodiments, Compound 2 is in neat crystalline form and therefore does not contain any water or solvent incorporated into the crystal structure. Compound 2 has been found to be present in at least one distinct neat (ie, anhydrous) crystalline form or polymorphic phase. In some embodiments, the invention provides a polymorphic form of compound 2, referred to herein as form P1. In certain embodiments, the present invention provides a polymorphic phase form of Compound 2, referred to herein as Form P22.

In certain embodiments, the present invention provides form P1 of Compound 2. According to one embodiment, the form P1 of Compound 2 has a powder X-ray diffraction pattern substantially similar to that shown in FIG. According to another embodiment, Form P1 of Compound 2 is characterized by having one or more peaks selected from peaks at about 6.21, about 9.48 and about 13.29 ° 2θ in its powder X-ray diffraction pattern. And. In some embodiments, Form P1 of Compound 2 is characterized by having two or more peaks selected from peaks at about 6.21, about 9.48 and about 13.29 ° 2θ in its powder X-ray diffraction pattern. .. Form P1 of Compound 2 is characterized by having all three peaks selected from peaks at about 6.21, about 9.48 and about 13.29 ° 2θ in its powder X-ray diffraction pattern.

When used in connection with the ° 2θ value, the term about as used herein refers to the described value ± 0.1 ° 2θ. The method for preparing form P1 of compound 2 is described below.

In certain embodiments, the present invention provides form P22 of Compound 2. According to one embodiment, the form P22 of Compound 2 has a powder X-ray diffraction pattern substantially similar to that shown in FIG. According to another embodiment, Form P22 of Compound 2 is characterized by having one or more peaks selected from peaks at about 7.29, about 8.38 and about 11.12 ° 2θ in its powder X-ray diffraction pattern. And. In some embodiments, Form P22 of Compound 2 is characterized by having two or more peaks selected from peaks at about 7.29, about 8.38 and about 11.12 ° 2θ in its powder X-ray diffraction pattern. .. Form P22 of Compound 2 is characterized by having all three peaks selected from peaks at about 7.29, about 8.38 and about 11.12 ° 2θ in its powder X-ray diffraction pattern.

In some embodiments, embodiment P22 is characterized by a melting point of 194 ° C. The method for preparing Form P22 of Compound 2 is described below.

According to other embodiments, the present invention provides compound 2 as an amorphous solid. Amorphous solids are well known to those of skill in the art and are generally prepared by methods such as lyophilization, melting and precipitation from supercritical fluids, among others.

General method of providing compound 2: Compound 1 is prepared according to the method detailed in '610 Publication , which is incorporated herein by reference in its entirety . Compound 2 is prepared from compound 1 according to the scheme below.

<chemistry num="3"><img id="000005" he="38" wi="127" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> As shown in the schematic scheme above, compound 2 is prepared from compound 1 by mixing compound 1 with benzenesulfonic acid to form its vesylate. Therefore, another aspect of the invention is compound 2:

<chemistry num="4"><img id="000006" he="41" wi="75" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>Is a method for preparing Compound 1:

<chemistry num="5"><img id="000007" he="37" wi="54" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>And the steps to provide With the step of combining compound 1 with benzenesulfonic acid in a suitable solvent; With the step of isolating compound 2 at the option Provide a method including.

A suitable solvent can solubilize one or more of the reaction components, or a suitable solvent can facilitate agitation of one or more suspensions of the reaction components. Examples of suitable solvents useful in the present invention are protic and aprotic solvents or mixtures thereof. In certain embodiments, suitable solvents include ethers, esters, alcohols, ketones or mixtures thereof. In certain embodiments, suitable solvents are methanol, ethanol, isopropanol or acetone, the solvent being anhydrous or in combination with water or heptane. In other embodiments, suitable solvents include tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, glyme, diglyme, methyl t-butyl ether, t-butanol, n-butanol and acetonitrile. In other embodiments, the suitable solvent is absolute ethanol. In some embodiments, the suitable solvent is MTBE.

According to other embodiments, the present invention relates to compound 2:

<chemistry num="6"><img id="000008" he="39" wi="75" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>Is a method for preparing Compound 1:

<chemistry num="7"><img id="000009" he="38" wi="54" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry>With the step of combining with a suitable solvent and optionally heating to form the solution; With the step of adding benzenesulfonic acid to the solution; With the step of isolating compound 2 at the option Provide a method including.

As outlined above, compound 1 is optionally heated and dissolved in a suitable solvent. In certain embodiments, compound 1 is dissolved at about 50 to about 60 ° C. In other embodiments, compound 1 is dissolved at about 50 to about 55 ° C. In yet another embodiment, compound 1 is dissolved at the boiling temperature of the solvent. In other embodiments, compound 1 is dissolved without heating.

In certain embodiments, about 1 equivalent of benzenesulfonic acid is added to compound 1 to give compound 2. In another embodiment, less than 1 equivalent of benzenesulfonic acid is added to compound 1 to give compound 2. In yet another embodiment, more than 1 equivalent of benzenesulfonic acid is added to compound 1 to give compound 2. In yet another embodiment, about 0.9 to about 1.1 equivalents of benzenesulfonic acid is added to compound 1 to give compound 2. In another embodiment, about 0.99 to about 1.01 equivalents of benzenesulfonic acid is added to compound 1 to give compound 2.

It will be appreciated that benzenesulfonic acid can be added in any suitable form to a mixture of compound 1 and a suitable solvent. For example, benzenesulfonic acid can be added in solid form or as a solution or suspension in a suitable solvent. The suitable solvent may be the same suitable solvent as that combined with Compound 1, or a different solvent. According to one embodiment, benzenesulfonic acid is added in solid form. In certain embodiments, benzenesulfonic acid is combined with a suitable solvent prior to addition to compound 1. According to another embodiment, benzenesulfonic acid is added as a solution in a suitable solvent. In other embodiments, suitable solvents for dissolving benzenesulfonic acid are polar protic or aprotic solvents. Such solvents include water, alcohols, ethers and ketones. Examples of such solvents include water, methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, glyme, diglyme, methyl t-butyl ether, t-butanol, n-butanol and acetonitrile. In certain embodiments, a suitable solvent is selected from those described above, which is anhydrous. According to one embodiment, benzenesulfonic acid is dissolved in MTBE.

In certain embodiments, the resulting compound 2 containing mixture is cooled. In another embodiment, the compound 2 containing mixture is cooled below 20 ° C.

In certain embodiments, compound 2 precipitates from the mixture. In another embodiment, compound 2 crystallizes from the mixture. In other embodiments, compound 2 crystallizes from solution following seed addition to solution (ie, adding crystals of compound 2 to solution).

Crystalline compound 2 is precipitated from the reaction mixture or some or all of the solvent is removed by methods such as evaporation, distillation, filtration (eg nanofiltration, ultrafiltration), reverse osmosis, absorption and reaction. It can be produced by doing so, by adding an antisolvent such as heptane, by cooling, or by various combinations of these methods.

As outlined above, compound 2 is optionally isolated. It will be appreciated that compound 2 can be isolated by any suitable physical means known to those of skill in the art. In certain embodiments, the precipitated solid compound 2 is separated from the supernatant by filtration. In another embodiment, the precipitated solid compound 2 is separated from the supernatant by decanting the supernatant.

In certain embodiments, the precipitated solid compound 2 is separated from the supernatant by filtration.

In certain embodiments, the isolated compound 2 is dried in air. In another embodiment, the isolated compound 2 is optionally dried under reduced pressure at high temperature.

Use, prescription and administration Pharmaceutically acceptable composition According to other embodiments, the present invention provides a composition comprising Compound 2 and a pharmaceutically acceptable carrier, adjunct or vehicle. The amount of Compound 2 in the compositions of the invention is such that it is effective in measurable inhibition of at least one of protein kinases, especially TEC kinases or variants thereof, in a biological sample or patient. is there. In certain embodiments, the amount of Compound 2 in the composition of the invention is effective in measurable inhibition of at least one of the TEC kinases or variants thereof in a biological sample or patient. The amount. In certain embodiments, the compositions of the invention are formulated for administration to a patient in need of such composition. In some embodiments, the compositions of the invention are formulated for oral administration to a patient.

As used herein, the term "patient" means an animal, preferably a mammal, most preferably a human.

The term "pharmaceutically acceptable carrier, adjunct or vehicle" refers to a non-toxic carrier, adjunct or vehicle that does not negate the pharmacological activity of the compound formulated with it. The pharmaceutically acceptable carriers, auxiliaries or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, aluminas, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffers. Substances such as phosphate, glycine, sorbic acid or potassium sorbate, partial glyceride mixture of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc Includes salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

The compositions of the invention can be administered by oral, parenteral, inhalation spray, topical, transrectal, nasal, oral, vaginal or implantable reservoirs. As used herein, the term "parenteral" refers to subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Is included. The composition is preferably administered orally, intraperitoneally or intravenously. The form of the sterile injection of the composition of the invention may be an aqueous or oily suspension. These suspensions can be formulated according to techniques known in the art with suitable dispersants or wetting and suspending agents. The sterile injectable formulation may be a sterile injection or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer solution and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as solvents or suspension vehicles.

To that end, any non-irritating fixed oil containing synthetic mono or diglycerides can be used. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially those of their polyoxyethylated type. It is useful for. These oils or suspensions also include carboxymethyl cellulose or long chain alcohol diluents or dispersants such as similar dispersants commonly used in the preparation of pharmaceutically acceptable dosage forms containing emulsions and suspensions. Can include. Other commonly used surfactants such as Tweens, Spans, and other emulsifiers or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solids, liquids or other dosage forms. Can be used for prescription.

The pharmaceutically acceptable compositions of the present invention can be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions or solutions. .. For tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also usually added. Diluents useful for oral administration in the form of capsules include lactose and dried corn starch. If an aqueous suspending agent is required for oral use, the active ingredient is mixed with an emulsifier and suspending agent. Certain sweeteners, flavors or colorants can also be added if desired.

Alternatively, the pharmaceutically acceptable compositions of the present invention can be administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycol.

The pharmaceutically acceptable compositions of the present invention may also be administered topically, especially if the treatment target comprises areas or organs that are easily reachable by topical use, such as diseases of the eye, skin or lower intestinal tract. it can. Suitable topical formulations are readily prepared for each of these areas or organs.

Topical use for the lower intestinal tract can be performed with a rectal suppository formulation (see above) or a suitable enema formulation. Topical transdermal patches can also be used.

For topical use, the provided pharmaceutically acceptable compositions can be formulated with a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsified wax and water. Alternatively, the provided pharmaceutically acceptable composition may be formulated with a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Can be done. Suitable carriers include, but are not limited to, mineral oils, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The pharmaceutically acceptable compositions provided for use with the eye are fine powders in isotonic pH-controlled sterile saline with or without preservatives such as benzylalconium chloride. It can be formulated as a suspension or preferably as a solution in isotonic pH-controlled sterile saline. Alternatively, a pharmaceutically acceptable composition for ocular use can be formulated with an ointment such as petrolatum.

The pharmaceutically acceptable compositions of the present invention can also be administered by nasal aerosol or inhalation. Such compositions are prepared by techniques well known in the field of formulation and formulation technology, such as benzyl alcohol or other suitable preservatives, absorption enhancers that promote bioavailability, fluorocarbons and / or other conventional solubilization agents. It can be prepared as a solution in physiological saline using an agent or a dispersant.

In some embodiments, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration.

The amount of the compound of the invention that can be mixed with the carrier material to give a single dosage form composition will vary depending on the host being treated and the particular mode of administration. In certain embodiments, the provided compositions are formulated such that a dosage of Compound 2 in the range 0.01-100 mg / kg body weight / day is administered to a patient accepting these compositions.

Specific medication and treatment regimens for any particular patient include the activity, age, weight, overall health, gender, diet, dosing time, excretion rate, drug combination, and physician of the specific compound used. It should also be understood that it depends on various factors, including the judgment and severity of the specific disease being treated. The amount of compound 2 in the composition also depends on the specific compound in the composition.

Use of compounds and pharmaceutically acceptable compositions Compound 2 and compositions described herein are generally useful for inhibiting the protein kinase activity of one or more enzymes. Examples of kinases that are inhibited by compound 2 and compositions described herein and for which the methods described herein are useful include BTK and other TECs including ITK, TEC, BMX and RLK or variants thereof. Includes kinases.

Bruton's tyrosine kinase (BTK), a member of TEC kinase, is a major signaling enzyme expressed in all hematopoietic cell types except T lymphocytes and natural killer cells. BTK plays an essential role in the B cell signaling pathway that links cell surface B cell receptor (BCR) stimulation with the intracellular response downstream.

BTK is a major regulator of B cell development, activation, signal transduction and survival (Kurosaki, Curr Op Imm, 2000, pp. 276-281; Schaeffer and Schwartzberg, Curr Op Imm 2000, pp. 282-288). .. In addition, BTK also produces several other hematopoietic cell signaling pathways, such as Toll-like receptors (TLRs) and cytokine receptor-mediated TNF-α production in macrophages, and IgE receptors (Fc_epsilon_RI) in mast cells. It plays a role in signaling, inhibition of Fas / APO-1 apoptosis signaling in line B lymphocyte-like cells, and collagen-stimulated platelet aggregation. For example, CA Jeffries et al. (2003), Journal of Biological Chemistry 278: 26258-26264; NJ Horwood et al. (2003), The Journal of Experimental Medicine 197: 1603-1611; Iwaki et al. (2005), Journal of Biological Chemistry Vol. 280 (No. 48): pp. 40261-40270; Vassilev et al. (1999), Journal of Biological Chemistry Vol. 274 (No. 3): pp. 16461656 and Quek et al. (1998), Current Biology Vol. 8 (1998) No. 20): 1137 ~ 1140.

Patients with BTK mutations have significant impairments in B cell development, resulting in a near-complete lack of mature B lymphocytes and plasma cells, significantly lower Ig levels and marked inhibition of the humoral response to recall antigens. (Vihinen et al., Frontiers in Bioscience Vol. 5, outlined on pages 917-928). In addition, BTK-deficient mice have lower peripheral B cell numbers and significantly lower serum levels of IgM and IgG3. BTK deletion in mice has a significant effect on anti-IgM-induced B cell proliferation and inhibits the immune response to thymus-independent type II antigens (Ellmeier et al., J Exp Med, Vol. 192, pp. 1611-1623 (Ellmeier et al., J Exp Med, Vol. 192: 1611-1623). the year of 2000)). BTK also plays a very important role in mast cell activation via the high affinity IgE receptor (Fc_epsilon_RI). Mast cells in BTK-deficient mice have reduced degranulation and reduced production of pro-inflammatory cytokines following Fc_epsilon_RI cross-linking (Kawakami et al., Journal of Leukocyte Biology) Volume 65: pp. 286-290).

Compound 2 is an inhibitor of BTK and is therefore useful in the treatment of one or more disorders associated with BTK activity. Thus, in some embodiments, the invention provides a method for treating a BTK-mediated disorder, comprising administering to a patient in need thereof compound 2 or a pharmaceutically acceptable composition thereof. To do.

As used herein, the term "BTK-mediated" disorder or condition means any disease or other adverse condition in which BTK or a variant thereof is known to play a role. Thus, another embodiment of the invention relates to treating or reducing the severity of one or more diseases in which BTK or variants thereof are known to play a role. Specifically, the invention is a method of treating or reducing the severity of a disease or condition selected from proliferative or autoimmune disorders, the present invention for patients in need thereof. The present invention relates to a method comprising administering Compound 2 or a composition according to.

In some embodiments, the present invention provides a method of treating or reducing the severity of one or more diseases and conditions associated with BTK. In some embodiments, the disease or condition is an autoimmune disease such as inflammatory bowel disease, arthritis, acne, rheumatoid arthritis, psoriasis arthritis, osteoarthritis, Still disease, juvenile arthritis, diabetes, severe Ankylosing spondylitis, Hashimoto thyroiditis, Ord's thyroiditis, Graves' disease, Schegren's syndrome, multiple sclerosis, Gillan-Valley syndrome, acute disseminated encephalomyelitis, Addison's disease, opsoclonus- myoclonus syndrome), ankylosing spondylitis (ankylosing) spondylosis), antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, celiac disease, good pasture syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, Reiter syndrome, Takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis, systemic alopecia, Behcet's disease, chronic fatigue, autonomic neuropathy, endometriosis, interstitial cystitis , Neuromuscular tonicity, scleroderma or chronic vulvodynia. In some embodiments, the disease or condition is an immune-mediated disease, including hyperproliferative disease or rejection of transplanted organs or tissues and acquired immunodeficiency syndrome (AIDS, also known as HIV).

In some embodiments, the present invention is a method for treating or reducing the severity of one or more BTK-related diseases and conditions, wherein the disease or condition includes them. To host-induced transplant disease, transplantation, blood transfusion, anaphylaxis, allergies (eg, plant pollen, latex, drugs, food, insect toxins, animal hair, animal indulgence, yellowtail or cockroach calyx), but not limited to A method selected from heterologous immune conditions or diseases including allergies), type I hypersensitivity, allergic conjunctivitis, allergic rhinitis and atopic dermatitis.

In some embodiments, the present invention is a method for treating or reducing the severity of one or more BTK-related diseases and conditions, wherein the disease or condition is inflammatory. Diseases such as asthma, epididymitis, eyelid inflammation, epididymitis, bronchitis, epididymitis, cervical inflammation, cholangitis, cholecystitis, colitis, conjunctivitis, cystitis, lacrimal adenitis, dermatomyositis, encephalitis , Endocarditis, Endocolitis, Enterocolitis, Small intestinal colitis (enterocolitis), Epididymitis, Epididymitis, Myositis, Cohesive histitis, Gastric inflammation, Gastroenteritis, Hepatitis, Sweat gland abscess, Laryngitis , Pneumonitis, meningitis, myelitis, myocolitis, myitis, nephritis, ovarian inflammation, epididymitis, osteoitis, ear inflammation, pancreatitis, parotid inflammation, peritonitis, peritonitis, pharyngitis, pneumonitis, veins Flame, pneumonitis, pneumonia, enterocolitis, prostatic inflammation, nephritis nephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, vaginitis, vaginitis, vasculature Provide a method to choose from flame or vaginitis.

In some embodiments, the present invention is a method for treating or reducing the severity of one or more diseases and conditions associated with BTK, wherein the disease or condition is selected from cancer. Provide a way to be done. In one embodiment, the cancer is a B cell proliferative disorder, such as diffuse large B cell lymphoma, follicular lymphoma, chronic lymphoma, chronic lymphoma, acute lymphoma, preB cell lymphoma. Leukemia, lymphoplasmacytic lymphoma / Waldenstram macroglobulinemia, splenic marginal zone lymphoma, multiple myeloma (also known as plasmocellular myeloma), non- Hodgkin lymphoma, Hodgkin lymphoma, plasmacytoma, extranodal marginal zone B cell lymphoma, nodal marginal zone B cell lymphoma), mantle cell lymphoma, mediastinal (thymic) large B cell lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma (Primary effusion lymphoma), Berkit lymphoma / leukemia or lymphoma-like granulomatosis. In some embodiments, the cancer is breast cancer, prostate cancer or mast cell cancer (eg, mastocytoma, mast cell leukemia, mast cell sarcoma, systemic mastocytoma). In one embodiment, the cancer is a bone cancer. In another embodiment, the cancer is of primary origin and has metastasized to bone.

In some embodiments, the invention is limited to, but not limited to, rheumatoid arthritis, seronegative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis and Reiter's disease), Bechet's disease, Sjogren's disease. Treat or treat one or more BTK-related diseases or conditions, including syndromes, systemic sclerosis, osteoporosis, bone cancer and bone and joint diseases including metastasis to bone. Provide a way to mitigate.

In some embodiments, the present invention is a method for treating or reducing the severity of one or more BTK-related diseases and conditions, wherein the disease or condition is thromboembolic. Disorders (thromboembolic disorder), such as myocardial infarction, angina, restenosis after angioplasty, restenosis after angioplasty, restenosis after aortic coronary bypass, restenosis after aortic coronary bypass, cerebral infarction , Transitory ischemia, peripheral arterial occlusive disorder, pulmonary embolism or deep venous thrombosis.

In some embodiments, the invention treats or treats one or more diseases and conditions associated with BTK, including infectious and non-infectious inflammatory events and autoimmune and other inflammatory diseases. Provide methods for reducing severity. These autoimmune and inflammatory diseases, disorders and syndromes include inflammatory pelvic disease, urethritis, skin tan, sinusitis, pneumonia, encephalitis, meningitis, myocarditis, Nephritis, myelitis, myitis, hepatitis, gastrointestinal inflammation, enteritis, dermatitis, gingitis, cystitis, pancreatitis, chorocystitus, agammaglobulinemia, psoriasis, allergies, Crohn's disease, irritable bowel syndrome, ulcerative Colonitis, Sjogren's disease, tissue transplant rejection, hyperacute rejection of transplant organs, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), autoimmune polyglandular disease (autoimmune) Autoimmune polyglandular Also known as syndrome)), autoimmune alopecia, malignant anemia, glomerulonephritis, dermatitis, multiple sclerosis, sclerosis, vasculitis, autoimmune hemolytic and thrombocytopenic status, good Pasture syndrome, atherosclerosis, Addison's disease, Parkinson's disease, Alzheimer's disease, type I diabetes, septic shock, systemic lupus erythematosus (SLE), rheumatoid arthritis, psoriatic arthritis, juvenile arthritis, osteoarthritis , Chronic idiopathic thrombocytopenic purpura, Waldenström macroglobulinemia, severe myasthenia, Hashimoto thyroiditis, atopic dermatitis, degenerative joint disease, leukoplakia, autoimmune pituitary dysfunction, Giran -Includes Valley Syndrome, Bechet's disease, scleraderma, mycobacterial sarcoma, acute inflammatory reactions (such as acute respiratory distress syndrome and ischemic / reperfusion injury) and Graves' disease.

In some embodiments, the invention presents rheumatoid arthritis, multiple sclerosis, diabetes, B-cell chronic lymphocytic leukemia, acute lymphocytic leukemia, hairy cell leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, multiple myeloma, bone. Treat one or more BTK-related diseases and conditions selected from cancer, colorectal cancer, pancreatic cancer, bone metastasis, osteoporosis, hypersensitivity bowel syndrome, Crohn's disease, leukemia and renal transplantation Or provide a method for reducing its severity.

BTK is a member of the TEC kinase, which shares a common cysteine at the equivalent position of Cys481 in Btk, which is also capable of irreversible inhibition, as described in the '610 publication. Therefore, examples of kinases that are inhibited by compound 2 and compositions described herein and for which the methods described herein are useful include ITK, TEC, BMX and RLK or variants thereof, other than BTK. Includes additional TEC kinases.

The activity of Compound 2 as an inhibitor of TEC kinase or variants thereof can be assayed in vitro, in vivo or in a cell line. Assays in vitro include assays that determine the phosphorylation activity and / or subsequent functional outcome of activated TEC kinase or variants thereof or inhibition of ATPase activity. An alternative assay in vitro quantifies the ability of Compound 2 to bind to TEC kinase. Inhibitor binding involves labeling the inhibitor with a radioisotope prior to binding, isolating the inhibitor / TEC kinase (ie, TEC, BTK, ITK, RLK and BMX complex) and the amount of radiolabel bound. Can be measured by determining. Detailed conditions for assaying Compound 2 as an inhibitor of TEC kinase or variants thereof are detailed in the '610 publication.

Protein tyrosine kinases are a class of enzymes that catalyze the transfer of phosphate groups from ATP or GTP to tyrosine residues located on protein substrates. Receptor tyrosine kinases act to transmit signals from the outside to the inside of cells by activating secondary transmission effectors through phosphorylation events. Various cellular processes are facilitated by these signals, including proliferation, carbohydrate utilization, protein synthesis, angiogenesis, cell growth, and cell survival.

The terms "treatment," "treat," and "treating" as used herein refer to a disease or disorder as described herein, or one or more of them. It refers to reversing, alleviating, delaying, or stopping the progression of symptoms. In some embodiments, the procedure can be given after the onset of one or more symptoms. In other embodiments, the procedure can be given before symptoms are present. For example, treatment can be applied to an individual susceptible to the disease prior to the onset of symptoms (eg, in the light of history and / or in the light of genetic or other susceptibility factors). Treatment can be continued even after the symptoms have resolved, for example to prevent or delay its recurrence.

The TEC family of non-receptor tyrosine kinases, referred to herein as "TEC kinases," plays a central role in signal transduction by antigen receptors such as the TCR, BCR and Fc receptors (Miller A et al., Review, Current Opinion). in Immunology Vol. 14; pp. 331-340 (2002). TEC kinases are essential for T cell activation. Three members of the family, Itk, Rlk and Is activated downstream of antigen receptor involvement in T cells and transmits signals to downstream effectors, including PLC-γ. Deletion of Itk and Rlk together in mice results in intracellular parasites (Toxoplasma) It results in significant inhibition of TCR responses, including proliferation, cytokine production and immune responses to gondii)) (Schaeffer et al., Science Vol. 284; pp. 638-641 (1999)). Intracellular signaling following TCR involvement occurs in ITK / RLK-deficient T cells; inositol trisphosphate production, calcium recruitment and MAP kinase activation are all reduced. Tec kinase is also essential for B cell growth and activation.

TEC kinases contain five family members, which are mainly expressed in hematopoietic cells. That is, their members are TEC, BTK, ITK (also known as TSK and EMT), RLK (also known as TXK) and BMX (also known as ETK). Other related TEC kinases have been found in Drosophila melanogaster, zebrafish (Danio rerio), skate (Raja eglanteria) and sea urchin (sea urchin) (Anthocidaris crassis pina).

Compound 2 is an inhibitor of one or more TEC kinases and is therefore useful in treating one or more disorders associated with the activity of one or more TEC kinases. Thus, in certain embodiments, the present invention provides a method for treating a TEC-mediated disorder comprising the step of administering Compound 2 or a pharmaceutically acceptable composition thereof to a patient in need thereof.

As used herein, the term "TEC-mediated state" means any disease or other adverse condition in which TEC kinase is known to play a role. Such conditions include this specification and Melcher, M et al., "The Role of TEC Family Kinases in Inflammatory Processes", Anti-Inflammatory &amp; Anti-Allergy Agents in Medicinal. Includes those described in Chemistry, Vol. 6, No. 1, pp. 61-69 (February 2007). Therefore, another embodiment of the invention relates to treating or reducing the severity of one or more diseases in which TEC kinase is known to play a role. Specifically, the present invention describes acquired immunodeficiency syndrome (AIDS) (also known as HIV), which includes autoimmune, inflammatory, proliferative and hyperproliferative disorders and rejection of transplanted organs or tissues. A method of treating or reducing the severity of a disease or condition selected from among immunologically mediated diseases, including administration of Compound 2 or a composition thereof to a patient in need thereof. Provide a method including.

In some embodiments, the invention is limited to, but is not limited to, bronchial, allergic, intrinsic, extrinsic and dusty asthma, especially chronic or refractory asthma (eg, delayed asthma airway hyperresponsiveness). Methods for treating or reducing the severity of one or more diseases and conditions associated with TEC kinase, including airway diseases, including reversible obstructive airway diseases, including asthma and bronchitis. provide. In some embodiments, the invention relates to chronic rhinitis, including acute rhinitis, allergic, atrophic rhinitis and dry dairy rhinitis, hypertrophic rhinitis, purulent rhinitis, dry rhinitis and drug-induced rhinitis; grouped, fibrotic. And membranous rhinitis including pseudomembranous rhinitis and scrofoulous rhinitis, seasonal rhinitis including neurogenic rhinitis (pollinosis) and vasomotor neuronasitis, sarcoidosis, farmer's lung and related diseases, pulmonary fibrosis and How to treat or reduce the severity of one or more TEC kinase-related diseases and conditions, including conditions characterized by inflammation of the nasal mucus membrane, including idiopathic stromal pneumitis. I will provide a.

In some embodiments, the invention is limited to, but not limited to, rheumatoid arthritis, serum reaction negative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis and Reiter's disease), Behcet's disease, Sjogren's syndrome, systemic. To treat or reduce the severity of one or more diseases and conditions associated with TEC kinase, including bone and joint disorders, including sclerosis, osteoporosis, bone cancer and bone metastasis. Provide a method.

In some embodiments, the invention is limited to, but not limited to, psoriasis, systemic sclerosis, atopic dermatitis, contact dermatitis and other eczema dermatitis, seborrheic dermatitis ( seborrhoetic dermatitis), squamous moss, scoliosis, bullous vesicle, epidermal vesicular disease, urticaria, dermatitis, vasculitis, erythema, cutaneous eosinophilias, vegetation, alopecia areata Provides methods for treating or reducing the severity of one or more TEC kinase-related diseases and conditions, including skin diseases and disorders, including alopecia, areata and spring catal. To do.

In some embodiments, the invention is limited to, but not limited to, celiac disease, proctitis, eosinophilic gastroenteritis, obesity cytosis, pancreatitis, Crohn's disease, ulcerative colitis, effects away from the intestinal tract. To treat or reduce the severity of one or more diseases and conditions associated with TEC kinase, including gastrointestinal disorders and disorders, including food-related allergies, such as Crohn's disease, proctitis and eczema. Provide a method.

In some embodiments, the invention is limited to, but not limited to, multiple sclerosis, atherosclerosis, lupus erythematosus, systemic lupus erythematosus, Hashimoto thyroiditis, severe myasthenia, Diabetes type I, nephrotic syndrome, lupus erythematosus (eosinophilia fascitis), high IgE syndrome, epilepsy, cesarly syndrome and idiopathic thrombocytopenic purpura, restenosis following angiogenesis, tumors (eg, eg Treatment or severity of one or more diseases and conditions associated with TEC kinase, including diseases and disorders of other tissues including leukemia, lymphoma and prostatic cancer) and atherosclerosis and systemic diseases. Provide a method for reducing.

In some embodiments, the invention is limited to, for example, rejection of acute and chronic allogeneic transplantation following transplantation of kidney, heart, liver, lung, bone marrow, skin and corneum; and chronic vs. host. Provided are methods for treating or reducing the severity of one or more diseases and conditions associated with TEC kinase, including rejection of allografts, including sex graft disease.

In some embodiments, the present invention is, and requires, a method of treating or reducing the severity of one or more of the diseases or conditions associated with TEC kinase as described above. It relates to a method comprising administering to a patient the compound 2 or composition according to the invention.

Compound 2 and its compositions according to the methods of the invention treat or severely treat cancer, autoimmune disorders, neurodegenerative or neurological disorders, schizophrenia, bone-related disorders, liver or heart disorders. It can be administered using any amount and any route of administration that is effective in reducing severity. The exact amount required will vary from subject to subject depending on the subject's species, age and general condition, severity of infection, specific drug, method of administration and the like. Compound 2 and compositions of the present invention are preferably formulated in dosage unit form to facilitate administration and dosing uniformity. As used herein, the term "medication unit form" refers to a physically discrete unit of drug suitable for the patient being treated. However, it will be appreciated that the total daily dose of the compounds and compositions of the present invention will be determined by the attending physician, within sound medical judgment. Specific effective dose levels for any particular patient or organism are the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient Age, weight, overall health, gender and diet; time of administration, route and rate of excretion of the specific compound used; duration of treatment; medications used in combination with or at the same time as the specific compound used It will depend on a variety of factors, including, as well as similar factors well known in the medical arts.

The pharmaceutically acceptable compositions of the present invention may be oral, transrectal, parenteral, intracapsular, transvaginal, intraperitoneal, topical (powder, ointment or) depending on the severity of the infection being treated. It can be administered to humans and other animals by drop), orally, or by oral or nasal spray. In certain embodiments, the compounds of the invention are about 0.01 mg / kg to about 50 mg / kg, preferably about 1 mg / kg to about 25 mg / kg per subject body weight per day to obtain the desired therapeutic effect. At the dosing level, it can be administered orally or parenterally once or multiple times daily.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to Compound 2, the liquid dosage form is an inert diluent commonly used in the art, such as water or other solvents, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol. , 1,3-butylene glycol, dimethylformamide, oil (specifically, cotton seed oil, lacquer oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acids Solubilizers and emulsifiers such as esters and mixtures thereof can be included. In addition to the Inactive Diluent, the oral composition can also include auxiliary agents such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and fragrances.

Injectable formulations, such as sterile aqueous or oily injectable suspensions, can be formulated by techniques known in the art with suitable dispersants or wetting agents and suspending agents. The sterile injectable formulation may be a non-toxic parenteral acceptable diluent or solution in a solvent for sterile injection, suspension or emulsion, eg, solution in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as solvents or suspension vehicles. For that purpose, any sterile fixed oil containing synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid are used in the formulation of injectable substances.

Injectable formulations are, for example, filtered through a bacterial retention filter or mixed with a sterile agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or sterile injectable vehicle prior to use. Can be sterilized by.

It is often desirable to delay the absorption of the compound by subcutaneous or intramuscular injection in order to sustain the action of compound 2 of the invention. This can be achieved by using a suspension of crystalline or amorphous material with low solubility in water. The absorption rate of a compound depends on its dissolution rate. Therefore, its rate depends on the crystal size and crystal morphology. Alternatively, delaying the absorption of the compound form administered parenterally is achieved by dissolving or suspending it in an oily vehicle. The injectable depot form is made by forming a microencapsulating matrix of compounds in a biodegradable polymer such as polylactide-polyglycolide. The rate of release of the compound can be controlled depending on the ratio of the compound to the polymer and the properties of the specific polymer used. Examples of other biodegradable polymers include poly (orthoester) and poly (acid anhydride). Depot injection formulations are also prepared by incorporating the compound into liposomes or microemulsions that are compatible with living tissue.

Compositions for transrectal or vaginal administration melt compound 2 of the present invention solid at ambient temperature but liquid at body temperature, thus melting in the rectum or vaginal cavity to release the active compound. It can be prepared by mixing with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol or suppository wax.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, compound 2 is subjected to at least one pharmaceutically acceptable inert excipient or carrier, such as sodium citrate or calcium dibasic phosphate, and / or a) filler or bulking agent, such as starch. , Lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose and acacia, c) moisturizers such as glycerol, d) disintegrants such as agar, carbonic acid. Calcium, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) dissolution retarders such as paraffin, f) absorption enhancers such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and Glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form can also include buffers.

Similar types of solid compositions can also be used as fillers in soft and hard gelatin capsules with excipients such as lactose or lactose and high molecular weight polyethylene glycol. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical art. They can optionally contain opalescent agents and, optionally, in a delayed form, are made of a composition that releases the active ingredient alone or preferentially in a particular portion of the intestinal tract. You may. Examples of implantable compositions that can be used include polymeric substances and waxes. Similar solid compositions can also be used as fillers in soft and hard gelatin capsules with excipients such as lactose or lactose and high molecular weight polyethylene glycol.

Compound 2 may be in the form of microencapsulation with one or more excipients as described above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells such as enteric coatings, release control coatings and other coatings well known in the pharmaceutical art. In such a solid dosage form, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also include other substances besides the inert diluent, such as tableting lubricants such as magnesium stearate and microcrystalline cellulose, and other tableting aids, as is commonly practiced. Can be done. In the case of capsules, tablets and pills, the dosage form can also include buffers. They can optionally contain opalescent agents, and in a optionally delayed form, in a composition that releases only the active ingredient or preferentially releases it in a particular portion of the intestinal tract. It may be made. Examples of implantable compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of the compounds of the invention include ointments, pastes, creams, lotions, gels, powders, liquids, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and optionally any required preservative or buffer. Ophthalmic formulations, ear drops and eye drops are also conceivable and are also within the scope of the present invention. In addition, the present invention considers the use of transdermal patches, which have the additional advantage of controlled release of compounds into the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable vehicle. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate can be controlled by providing a rate control membrane or by dispersing the compound in a polymer matrix or gel.

According to one embodiment, the invention is a method of inhibiting protein kinase activity in a biological sample, comprising contacting the biological sample with a compound of the invention or a composition comprising the compound. Regarding the method.

According to another embodiment, the invention is a method of inhibiting the activity of a TEC kinase or variant thereof in a biological sample, wherein the biological sample is contacted with Compound 2 or a composition comprising said Compound. It relates to a method including a step of causing. In certain embodiments, the present invention is a method of irreversibly inhibiting the activity of a TEC kinase or variant thereof in a biological sample, wherein the biological sample is combined with Compound 2 or a composition comprising said Compound. It relates to a method including a step of contacting.

The term "biological sample" as used herein is not limited to these; cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, Includes feces, semen, tears or other body fluids or extracts thereof.

Inhibition of the activity of protein kinases in biological samples, ie protein kinases selected from TEC kinases or variants thereof, is useful for a variety of purposes known to those of skill in the art. Examples of such objectives include, but are not limited to, blood transfusions, organ transplants, storage of biological samples and biological assays.

Another embodiment of the invention relates to a method of inhibiting protein kinase activity in a patient, comprising the step of administering compound 2 or a composition comprising said compound to said patient.

According to another embodiment, the invention is a method of inhibiting the activity of one or more of a patient's TEC kinase or variant thereof, wherein the patient is administered with Compound 2 or a composition comprising said Compound. Regarding methods including steps. According to certain embodiments, the present invention is a method of irreversibly inhibiting the activity of one or more of a patient's TEC kinase or variant thereof, the composition comprising the compound 2 or the compound in said patient. The present invention relates to a method including a step of administering. In another embodiment, the invention is a method for treating a disorder mediated by one or more of TEC kinases or variants thereof in a patient in need thereof, wherein the patient is given Compound 2 or Provided are methods that include the step of administering the pharmaceutically acceptable composition. Such obstacles are described in detail herein.

All features of each aspect of the invention apply to all other aspects with the necessary modifications.

The following examples are shown so that the inventions described herein can be better understood. It should be understood that these examples are for illustrative purposes only and are not considered to limit the invention in any manner.

As shown in the examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. While this general method demonstrates the synthesis of a particular compound of the invention, the following general methods and other methods known to those of skill in the art are described herein in all compounds and subordinate to each of these compounds. It will be understood that it can be applied to the categories and species of.

<p num="0110"> General procedure The powder X-ray diffraction pattern was obtained with a Bruker D8 Advance equipped with Cu-Kα radiation and a LynxEye detector. The powder sample was placed on a polished silicon sample holder with zero background and rotated during the measurement. The measurements were performed as follows: 40 kV / 40 mA tube output, 0.02 ° 2θ step size, 37 second step time and 2.5-50 ° 2θ scan range.</p><p num="0111"> Proton nuclear magnetic resonance (¹1 H NMR) spectra were obtained with a Bruker model DPX-300MHz NMR spectrometer.¹The 1 H NMR spectrum was recorded with a 30 ° excitation pulse at 300.13 MHz with a pulse delay of 1 second and 16 scans. Deuterated DMSO was used as the solvent.</p><p num="0112"> DSC data were obtained by Perkin Elmer DSC 7 using a closed gold crucible. The sample was filled and dried under nitrogen. The sample was heated at -50 ° C to 250 ° C and 10 K / min with this device.</p><p num="0113"> TG-FTIR data was obtained using a Netzsch Thermo-Microbalance TG209 equipped with a Bruker FT-IR spectrometer Vector22. Samples were measured by heating in an aluminum crucible (with micropores) in a nitrogen atmosphere at 25-250 ° C, 10 K / min.</p><p num="0114"> (Example 1) Preparation of compound 2 (form P1)</p><p num="0115"><chemistry num="8"><img id="000010" he="37" wi="124" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></chemistry> Compound 1 is prepared according to the method described in detail in Example 20 of Publication '610, which is incorporated herein by reference in its entirety.</p><p num="0116"> The besilate of compound 1, i.e. compound 2, was prepared as follows. Compound 1 was added to MTBE under nitrogen to form a slurry and the mixture was heated to 50-55 ° C. Benzenesulfonic acid in MTBE was added and the resulting mixture was stirred for 1 hour. The mixture was cooled to 0-5 ° C and stirred for 1 hour. The resulting solid was collected by filtration and then dried under vacuum at 65-70 ° C to give compound 2. The characterization of the material obtained demonstrated that Compound 2 was crystalline. This crystal form is referred to as form P1.</p><p num="0117"> The FT-Raman spectrum for compound 2 and form P1 is shown in FIG.</p><p num="0118"> Figure 2 shows the PXRD for compound 2 and form P1. Table 1 below shows the X-ray diffraction peaks observed for Form P22 of Compound 2, each value at ° 2θ.</p><p num="0119"><tables num="1"><img id="000011" he="72" wi="102" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Figure 3 shows the TG-FTIR for compound 2 and form P1. The resulting thermogram shows a loss of about 0.9% by weight dichloromethane (residual solvent) at 130-160 ° C.</p><p num="0120"> DSCs for compound 2, form P1 are shown in FIGS. 4 and 5.</p><p num="0121"> (Example 2) Solubility of compound 2 Solubility of Compound 2 at room temperature was measured by manual dilution in 17 solvents and a mixture of 2 solvents in combination with visual observation. The results are summarized in Table 2 below.</p><p num="0122"><tables num="2"><img id="000012" he="97" wi="138" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> (Example 3) Preparation of compound 2 (form P22) Compound 2 (82.2 mg) was suspended in methyl ethyl ketone (6 mL) and the suspension was heated to 68 ° C with the addition of 8 mL of methyl ethyl ketone. A clear solution was obtained, which was heated to 75 ° C. The solution was cooled to 5 ° C. and the solvent was partially evaporated to give a white precipitate. The obtained solid was recovered by centrifugal filtration to obtain Form P22. This substance was characterized. The results are as follows.</p><p num="0123"> The PXRD pattern of compound 2 form P22 is shown in FIG. 6 in comparison with form P1. The FT-Raman spectrum of morphology P22 is shown in FIG. The TG-FTIR spectrum of morphology P22 is shown in FIG. Form P22¹1 H NMR is shown in FIG. This is consistent with the structure of compound 2 which has a 1: 1 ratio to besilate. The DSC thermogram is shown in Figure 10. This indicates a single endothermic event at 193.7 ° C.</p><p num="0124"> Table 3 below shows the X-ray diffraction peaks observed for Form P22 of Compound 2, each value at ° 2θ.</p><p num="0125"><tables num="3"><img id="000013" he="87" wi="104" file="JP6068340B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p>