EA042667B1 - IMPROVED FORMS OF THE SELECTIVE PI3K-DELTA INHIBITOR FOR PHARMACEUTICAL APPLICATIONS - Google Patents

Description

This application claims priority under Indian Patent Application Nos. 2596/CHE/2014 filed May 27, 2014 and 2597/CHE/2014 filed May 27, 2014, each of which is incorporated herein by reference in its entirety.

Field of invention

The present invention relates to solid phase forms of the p-toluenesulfonic acid salt (PTSA) of the selective inhibitor PI3K delta(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1Hpyrazolo[3,4 -d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one (TGR-1202). The present invention also relates to processes for the preparation of said compounds, pharmaceutical compositions containing them, and their use in the treatment of PI3K (phosphatidylinositol-3-kinase) mediated diseases or disorders.

State of the art

TGR-1202, whose chemical name is (S)-2-(1-(4-amino-3-(3-fluoro-4isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl) -6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one has the following chemical structure:

The preparation of TGR-1202 and its salts is described in International Publication No. WO 2014/006572 and US Patent Publication No. 2014/0011819, each of which is incorporated herein by reference in its entirety. TGR-1202 is a drug currently undergoing numerous clinical trials in the field of hematological malignancies.

WO 2014/006572 and US 2014/0011819 describe the synthesis of TGR-1202 (Example B1) and also disclose the therapeutic activity of this molecule in inhibiting, regulating and/or modulating PI3K signal transduction.

The essence of the invention

The present invention relates to novel solid phase p-toluenesulfonic acid (PTSA) salt forms of the selective inhibitor PI3K-delta(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)1 H-pyrazolo[ 3,4-d]pyrimidin-1 -yl)ethyl)-6-fluoro-3 -(3-fluorophenyl)-4H-chromen-4-one (TGR-1202) ((S)-2(1-(4 -amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3fluorophenyl)-4H-chromen-4- it is 4-methylbenzenesulfonate.

The present inventors have found that TGR-1202 PTSA salt particles having certain particle sizes exhibit improved solubility and pharmacokinetic characteristics when administered orally.

In one embodiment, the PTSA salt has a d(0.9) value of from about 5 to about 50 microns, such as from about 5 to about 25 microns, or from about 5 to about 15 microns. The PTSA salt may also have a d(0.5) value of from about 1 to about 10 µm, such as from about 2.0 to about 10 µm, from about 1 to about 5 µm, or from about 2.0 to about 5 µm. The PTSA salt may have a d(0.1) value of from about 0.5 to about 1.5 microns, such as from about 0.5 to about 1.0 microns.

In another embodiment, the PTSA salt of TGR-1202 has a d(0.5) value of from about 1 to about 10 microns, such as from about 2.0 to about 10 microns, from about 1 to about 5 microns, or from about 2.0 to about 5 µm. The PTSA salt may also have a d(0.1) value of from about 0.5 to about 1.5 microns, such as from about 0.5 to about 1.0 microns. The PTSA salt may have a d(0.9) value of from about 5 to about 50 microns, such as from about 5 to about 25 microns, or from about 5 to about 15 microns.

In yet another embodiment, the PTSA salt has a d(0.1) value of from about 0.5 to about 1.5 microns, such as from about 0.5 to about 1.0 microns. The PTSA salt may also have a d(0.9) value of from about 5 to about 50 microns, such as from about 5 to about 25 microns, or from about 5 to about 15 microns. The PTSA salt may have a d(0.9) value of from about 5 to about 50 microns, such as from about 5 to about 25 microns, or from about 5 to about 15 microns.

In another embodiment, the TGR-1202 PTSA salt has a particle size distribution (PSD) in which

- 1 042667 (i) the value of d(0.1) is from about 0.5 to about 1.5 microns, the value of d(0.5) is from about 2.0 to about 10 microns and the value of d(0.9 ) is from about 5.0 to about 50 microns;

(ii) the d(0.1) value is from about 0.5 to about 1.5 microns, the d(0.5) value is from about 2.0 to about 5.0 microns, and the d(0.9) value is from about 5.0 to about 50 microns;

(iii) the d(0.1) value is from about 0.5 to about 1.5 microns, the d(0.5) value is from about 2.0 to about 10 microns, and the d(0.9) value is from about 5.0 to about 25 microns; or (iv) the d(0.1) value is from about 0.5 to about 1.0 µm, the d(0.5) value is from about 2.0 to about 5 µm, and the d(0.9) value is from about 5.0 to about 15 microns.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle size distribution (PSD) in which the d(0.1) value is from about 0.5 to about 1.5 µm, the d(0.5) is from about 2.0 to about 10 microns; and the value of d(0.9) is from about 5.0 to about 50 microns.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle size distribution (PSD) in which the d(0.1) value is from about 0.5 to about 1.5 µm, the d(0.5) is from about 2.0 to about 5.0 microns; and the value of d(0.9) is from about 5.0 to about 50 microns.

In another embodiment, the PTSA salt of TGR-1202 has a particle size distribution (PSD) in which the d(0.1) value is from about 0.5 to about 1.0 µm, the d(0.5) is from about 2.0 to about 10.0 microns; and the value of d(0.9) is from about 5.0 to about 15 microns.

In another embodiment, the PTSA salt of TGR-1202 has a particle size distribution (PSD) in which the d(0.1) value is from about 0.5 to about 1.0 µm, the d(0.5) is from about 2.0 to about 5.0 microns; and the value of d(0.9) is from about 5.0 to about 15 microns.

In another embodiment, not more than 5%, not more than 3%, not more than 2%, or not more than 1% of the TGR-1202 PTSA salt particles have a particle size of less than 1.0 microns. At the same time, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of TGR-1202 PTSA salt particles have a particle size of less than 100 microns. In one particular embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value of about 5.0 to about 50 microns. In another embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 50 microns. In yet another embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value from about 5.0 to about 25 microns. In yet another embodiment, the PTSA salt of TGR-1202 has a d(0.1) value of about 0.5 to about 1.0 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 15 microns.

In another embodiment, not more than 5%, not more than 3%, not more than 2%, or not more than 1% of the TGR-1202 PTSA salt particles have a particle size of less than 1.0 microns. At the same time, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of TGR-1202 PTSA salt particles have a particle size of less than 50 microns. In one particular embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value of about 5.0 to about 50 microns. In another embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 50 microns. In yet another embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value from about 5.0 to about 25 microns. In yet another embodiment, the PTSA salt of TGR-1202 has a d(0.1) value of about 0.5 to about 1.0 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 15 microns.

In another embodiment, not more than 5%, not more than 3%, not more than 2%, or not more than 1% of the TGR-1202 PTSA salt particles have a particle size of less than 1.0 microns. At the same time, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of TGR-1202 PTSA salt particles have a particle size of less than 25 microns. In one particular embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value from about 5.0 to about 25 microns. In yet another embodiment, the PTSA salt of TGR-1202 has a d(0.1) value of about 0.5 to about 1.0 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 15 microns.

In another embodiment, not more than 5%, not more than 3%, not more than 2%, or not more than 1% of the TGR-1202 PTSA salt particles have a particle size of less than 1.0 microns. At the same time, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of the PTSA particles salts of TGR-1202 have a particle size of less than 15 microns. In one particular embodiment, the PTSA salt of TGR-1202 also has a d(0.1) value of about 0.5 to about 1.5 µm, a d(0.5) value of about 2.0 to about 10 µm, and a d(0.9) value from about 5.0 to about 25 microns. In yet another embodiment, the PTSA salt of TGR-1202 has a d(0.1) value of about 0.5 to about 1.0 µm, a d(0.5) value of about 2.0 to about 5.0 µm and a d(0.9) value of about 5.0 to about 15 microns.

In yet another embodiment, TGR-1202 PTSA salt exhibits an X-ray powder diffraction (XRPD) substantially as shown in FIG. 1 (hereinafter referred to as form A in this document).

In another embodiment, the TGR-1202 PTSA salt exhibits a differential scanning calorimetry (DSC) curve having a characteristic endothermic peak at about 154° C. (Form A).

In yet another embodiment, TGR-1202 PTSA salt exhibits an x-ray powder diffraction pattern with one or more peaks selected from 5.0, 10.1, 15.9, 16.1, 16.3, 20.0, 22.1 , and 24.4±0.05, 0.1, or 0.2° 2θ. For example, the salt may show an x-ray powder pattern having one or more peaks (e.g., 2, 3, 4, 5, 6, 7, or 8 peaks) selected from 5.01, 10.09, 15.91, 16.13, 16.34, 20.00, 22.06 and 24.42±0.05, 0.1, or 0.2° 2θ.

In yet another embodiment, TGR-1202 PTSA salt exhibits an x-ray powder diffraction pattern substantially as shown in FIG. 2 (hereinafter referred to as form B in this document).

In another embodiment, the TGR-1202 PTSA salt exhibits a differential scanning calorimetry (DSC) curve having a characteristic endothermic peak at about 146° C. (Form B).

In another embodiment, TGR-1202 PTSA salt exhibits an x-ray powder diffraction pattern having one or more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.05, 0.1, or 0.2° 2θ. For example, the salt may show an x-ray powder pattern having one or more peaks selected from 5.04, 10.13, 22.11, and 24.45±0.05, 0.1, or 0.2° 2θ.

Another embodiment is TGR-1202 PTSA salt form B having a particle size distribution (PSD) defined by a d(0.1) value of about 0.5 to about 1.5 µm, a d(0. 5) from about 2.0 to 10 µm and a d(0.9) value from about 5 to about 50 µm.

Another embodiment is TGR-1202 PTSA salt form B having a particle size distribution (PSD) defined by a d(0.1) value of about 0.5 to about 1.5 µm, a d(0. 5) from about 2.0 to about 5 µm and a d(0.9) value from about 5 to about 25 µm.

Another embodiment is TGR-1202 PTSA salt form B having a particle size distribution (PSD) defined by a d(0.1) value of about 0.5 to about 1.0 µm, a d(0. 5) from about 2.0 to about 5 µm and a d(0.9) value from about 5 to about 15 µm.

The present invention further provides a pharmaceutical composition comprising a TGR-1202 PTSA salt according to any of the embodiments described herein (e.g., having a particle size distribution as defined in any of the embodiments herein) and a pharmaceutically acceptable excipient. substance.

The present invention further provides a pharmaceutical composition comprising TGR-1202 PTSA salt form B crystal as described in any embodiment herein and a pharmaceutically acceptable excipient.

The present invention further provides a pharmaceutical composition comprising TGR-1202 PTSA salt Form B crystal having a particle size distribution as defined in any embodiment herein and a pharmaceutically acceptable excipient.

Another embodiment provides a TGR-1202 PTSA salt having an x-ray powder pattern substantially as shown in FIG. 2 suitable for use in a pharmaceutical composition for the treatment of a PI3K associated disease, disorder or condition, for example a proliferative disease such as cancer.

Another embodiment provides a TGR-1202 PTSA salt having a particle size distribution as defined in any embodiment herein, suitable for use in a pharmaceutical composition for the treatment of a PI3K-associated disease, disorder, or condition, e.g., a proliferative disease such as cancer.

The present invention further provides a pharmaceutical composition comprising TGR-1202 PTSA salt crystalline Form B having a particle size distribution as defined in any embodiment herein and a pharmaceutically acceptable excipient. The pharmaceutical composition may further contain one or more additional active ingredients such as other active agents (such as anti-cancer agents and the active agents described below).

The invention further provides a pharmaceutical composition comprising a TGR-1202 PTSA salt having a particle size distribution as defined in any embodiment herein and a pharmaceutically acceptable excipient. The pharmaceutical composition may further contain one or more additional active ingredients such as other active agents (such as anti-cancer agents and the active agents described below).

One embodiment is a solid pharmaceutical composition for oral administration (such as a tablet) containing the PTSA salt of TGR-1202 as described herein and one or more solubilizers (such as hydroxypropyl betadex). The weight ratio of PTSA salt to solubilizer (such as hydroxypropyl betadex) can be from about 1.5:1 to about 1:1.5. The composition may further contain one or more excipients selected from suspending agents (such as hydroxypropyl cellulose), dispersing agents (such as microcrystalline cellulose), disintegrants (such as croscarmellose sodium), lubricants (such as magnesium stearate), and any combination of the foregoing. materials.

In additional embodiments, the PTSA salt, as defined in any embodiment herein, has an enantiomeric excess (EI) of at least about 60%, at least about 75%, at least about 80%, at least at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% .

Another embodiment is a TGR-1202 crystalline PTSA salt, wherein (i) the crystalline salt exhibits an x-ray powder pattern having one or more peaks (e.g., 2, 3, or 4 peaks) selected from 5.0, 10.1, 22 ,1, and 24.5±0.2° 2θ, and (ii) the crystalline salt is substantially free of other solid state forms of the PTSA salt.

Another embodiment is a TGR-1202 crystalline PTSA salt, wherein (i) the crystalline salt exhibits an x-ray powder pattern having one or more peaks (e.g., 2, 3, or 4 peaks) selected from 5.0, 10.1, 22 ,1, and 24.5±0.2° 2θ, and (ii) the crystalline salt contains less than 5% (eg, less than 4%, less than 3%, less than 2%, or less than 1%) of other solid state forms of the PTSA salt.

In another aspect, the present invention relates to a process for preparing the crystalline PTSA salt of TGR-1202 1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one). In one embodiment, the method comprises removing a solvent from a mixture of TGR-1202 PTSA salt and an ethereal solvent such as di-tert-butyl ether, diethyl ether, diethylene glycol diethyl ether, diisopropyl ether, dimethoxyethane, dimethoxymethane, ethyl-tert-butyl ether, methoxyethane, 2-(2methoxyethoxy) ethanol, methyl tert-butyl ether, 2-methyltetrahydrofuran, tetrahydrofuran and tetrahydropyran. The mixture is preferably stirred (eg for 3, 6, 12 or more hours) until the solvent is removed. The solvent can be removed by drying, in one preferred embodiment, the method includes mixing, filtering and drying a mixture of TGR-1202 PTSA salt and an ethereal solvent. The mixture is preferably a suspension, which, for example, can be obtained by suspending the PTSA salt of TGR-1202 in an ethereal solvent.

In another aspect, the present invention relates to a method for inhibiting PI3K delta in a patient in need thereof by administering to the patient an effective amount of TGR-1202 PTSA salt as described herein (e.g., TGR-1202 PTSA salt form B crystalline) having a distribution particle size, as defined in any embodiment herein).

Yet another embodiment is a method of treating, preventing, and/or inhibiting a PI3K protein kinase mediated disease, disorder or condition (such as cancer or other proliferative disease or disorder) in a patient by administering to the patient an effective amount of TGR-1202 PTSA salt as described in herein (for example, PTSA-salt TGR-1202 crystal Form B).

Another embodiment is a method of treating a PI3K-associated disease, disorder, or condition in a patient by administering to the patient an effective amount of a TGR-1202 PTSA salt as described herein (e.g., TGR1202 PTSA crystalline Form B). In one embodiment, the amount of TGR1202 PTSA crystalline Form B administered is sufficient to treat a PI3K-associated disease, disorder, or condition by inhibiting PI3K delta.

Another embodiment of the present invention provides a method for treating a proliferative disease by administering to a patient in need of such treatment an effective amount of

PTSA salt of TGR-1202 as described herein (eg, crystalline form B of PTSA salt of TGR-1202). In one embodiment, the amount of crystalline form B of TGR-1202 PTSC salt administered is sufficient to treat a proliferative disease by inhibiting

PI3K-delta.

Another embodiment of the present invention provides a method of treating a proliferative disease by administering to a patient in need of such treatment an effective amount of a TGR-1202 PTSA salt as described herein (e.g., TGR-1202 PTSA crystalline Form B), in combination (simultaneously or sequentially) with at least one other anticancer agent. In one embodiment, the amount of TGR-1202 PTSA salt (eg, Form B of TGR-1202 PTSA salt) administered is sufficient to treat (or improve treatment of) a proliferative disease by inhibiting PI3K delta.

Another embodiment is a method of treating a PI3K-associated disease, disorder, or condition in a patient, comprising administering to a patient in need of such treatment an effective amount of a TGR-1202 PTSA salt as described herein (e.g., PTSA salt form B crystalline TGR-1202) optionally mixed with at least one pharmaceutically acceptable excipient. In specific embodiments, the composition comprises a therapeutically effective amount of a TGR-1202 PTSA salt form B crystalline according to any embodiment herein (e.g., a TGR-1202 PTSA salt form B crystalline form having a particle size distribution defined in any embodiment herein) for the treatment of a PI3K-associated disease, disorder, or condition.

Specific embodiments provide a method of treating cancer in a patient, comprising administering to the patient a pharmaceutical composition comprising a TGR-1202 PTSA salt as described herein (e.g., TGR-1202 PTSA salt form B), optionally mixed with at least one pharmaceutically acceptable excipient. In specific embodiments, the composition comprises a therapeutically effective amount of a TGR-1202 PTSA salt form B crystalline according to any embodiment herein (e.g., a TGR-1202 PTSA salt form B crystalline form having a particle size distribution defined in any implementation in this document) for the treatment of cancer in a patient.

Crystalline Form B of TGR-1202 PTSA salt according to any embodiment herein (e.g., Crystalline Form B of TGR-1202 PTSA salt having a particle size distribution defined in any embodiment herein) is suitable for treatment various cancers, including but not limited to the following:

carcinoma, including carcinoma of the bladder, breast, colon, kidney, liver, lung (including small cell lung cancer), esophagus, gallbladder, uterus, ovary, testis, larynx, mouth, gastrointestinal tract (e.g., esophagus, stomach, pancreas), brain, cervix, thyroid, prostate, blood, and skin (including squamous cell carcinoma);

hematopoietic tumors of lymphoid origin, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma;

hematopoietic tumors of myeloid origin, including acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia;

tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma;

tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma and schwannomas; and other tumors including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosa, keratoacanthoma, follicular thyroid cancer, and Kaposi's sarcoma.

The PTSA salt of TGR-1202 described herein (including crystalline Form B) as an apoptosis modulator is useful in the treatment, prevention, and inhibition of cancer (including, but not limited to, the types mentioned herein).

The PTSA salt of TGR-1202 described herein (including crystalline Form B) is useful in cancer chemoprevention. Chemoprophylaxis includes inhibiting the development of invasive cancer by blocking the initiating mutagenic event, blocking the progression of precancerous cells that have already been damaged, or by inhibiting tumor recurrence. The PTSA salt, including Form B, is also useful in inhibiting angiogenesis and tumor metastasis. One embodiment of the invention is a method of inhibiting angiogenesis or tumor metastasis in a patient by administering an effective amount of TGR-1202 PTSA salt (such as TGR-1202 PTSA salt form B).

Another embodiment of the present invention is a method of treating an immune system related disease (e.g., an autoimmune disease), an inflammation related disease or disorder (e.g., asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel disease, glomerulonephritis, neuroinflammatory diseases, multiple sclerosis, uveitis and immune system disorders), cancer or other proliferative disease, liver disease or dysfunction, or kidney disease or dysfunction. The method includes administering an effective amount of one or more compounds of the present invention.

Examples of immune disorders that can be treated with the compounds of the present invention include, without limitation, psoriasis, rheumatoid arthritis, vasculitis, inflammatory bowel disease, dermatitis, osteoarthritis, asthma, inflammatory muscle disease, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ, bone marrow, stem cells and other cells and tissues), graft rejection, graft-versus-host disease, lupus erythematosus, inflammatory disease, type I diabetes, idiopathic pulmonary fibrosis (IPF) (or cryptogenic fibrosing alveolitis (CFA or idiopathic fibrosing interstitial pneumonia), pulmonary fibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (eg, Hashimoto's thyroiditis and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, multiple sclerosis, cystic fibrosis, primary chronic biliary hepatitis cirrhosis, allergic conjunctivitis and atopic dermatitis.

Another embodiment is a method of treating leukemia in a patient by administering a therapeutically effective amount of a compound of the present invention. For example, the compounds of the present invention are effective in the treatment of chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), multiple myeloma (MM), small lymphocyte lymphoma (LML), and indolent non-Hodgkin's lymphoma (indolent NHL). ).

In the above treatments, one or more additional active agents may be administered with a TGR-1202 PTSA salt, such as TGR-1202 PTSA salt form B, as described in any embodiment herein. For example, the PTSA salt of TGR-1202, as described in any embodiment herein, is suitable for use in combination (either co-administered or sequentially) with known anti-cancer therapies such as chemotherapy, radiation therapy, biological therapy, bone marrow transplantation. , stem cell transplantation, or any other anticancer therapy, or with one or more cytostatic, cytotoxic, or anticancer agents, or targeted therapy, alone or in combination, such as, but not limited to, for example, DNA interacting agents, such as fludarabine, cisplatin, chlorambucil, bendamustine, or doxorubicin; alkylating agents such as cyclophosphamide; topoisomerase II inhibitors such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents such as paclitaxel, docetaxel or epothilones (eg ixabepilone), natural or synthetic; hormonal agents such as tamoxifen; thymidylate synthase inhibitors such as 5-fluorouracil; antimetabolites such as methotrexate; other tyrosine kinase inhibitors such as Iressa and OSI-774; angiogenesis inhibitors; epidermal growth factor (EGF) inhibitors; vascular endothelial growth factor (VEGF) inhibitors; cyclin-dependent protein kinase (CDK) inhibitors; SR inhibitors; c-Kit inhibitors; Her1/2 inhibitors and monoclonal antibodies directed against growth factor receptors such as Erbitux (EGF) and Herceptin (Her2); CD20 monoclonal antibodies such as rituximab, publicstumab (TGR-1101), ofatumumab (HuMax; Intracel), ocrelizumab, veltuzumab, GA101 (obinutuzumab), AME-133v (LY2469298, Applied Molecular Evolution), ocaratuzumab (Mentrik Biotech), PRO131921, tositumomab, hA20 (Immunomedics, Inc.), ibritumomab-tiuxetan, BLX-301 (Biolex Therapeutics), reditux (Dr. Reddy's Laboratories), and PRO70769 (described in WO 2004/056312); other B cell-targeting monoclonal antibodies such as belimumab, atacicept, or chimeric proteins such as blisibimob and BR3-Fc; other monoclonal antibodies such as alemtuzumab; CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone); R-CHOP (rituximab-CHOP); hyperCV_AD (hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine); R-hyperCV-AD (rituximab-hyperCV_AD); FCM (fludarabine, cyclophosphamide, mitoxantrone); R-FCM (rituximab, fludarabine, cyclophosphamide, mitoxantrone); bortezomib and rituximab; temsirolimus and rituximab; temsirolimus and velcade (Velcade®); iodine-131 tositumomab (Bexxar®) and CHOP-CVP (cyclophosphamide, vincristine, prednisone); R-CVP (rituximab-CVP); ICE (ifosfamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR (fludarabine, cyclophosphamide, rituximab); FR (fludarabine, rituximab); and D.T. PACE (dexamethasone, thalidomide, cisplatin, adriamycin, cyclophosphamide, etoposide); and other protein kinase modulators.

Crystalline Form B of the PTSA salt of TGR-1202, as described in any embodiment herein, is also suitable for use in combination (either co-administered or sequentially) with one or more steroidal anti-inflammatory drugs, non-steroidal anti-inflammatory drugs (NSAIDs), or immunoselective anti-inflammatory derivatives (ImSAIDs).

Brief description of graphic materials

Fig. 1 is the X-ray powder diffraction spectrum of TGR-1202 PTSA salt,

- 6 042667 obtained by the method described in example 1 (form A).

Fig. 2 is an X-ray powder diffraction spectrum of TGR-1202 PTSA salt obtained by the method described in Example 2 (Form B).

Fig. 3 is a DSC curve of TGR-1202 PTSA salt prepared according to the method described in Example 1 (Form A).

Fig. 4 is a DSC curve of TGR-1202 PTSA salt prepared by the method described in Example 2 (Form B).

Detailed Description of the Invention

In this document, the following definitions of terms shall apply unless otherwise noted.

Useful ranges of physical quantities, such as molecular weight, or chemical properties, such as chemical formulas, are intended to include all combinations and subcombinations of the ranges of values, and particular embodiments. The term roughly, when referring to a number or range of numerical values, means that said number or range of numerical values are approximations within experimental variability (or within the statistical error of experiment) and thus the number or range of numerical values may differ, for example , by a value ranging from 1 to 15% of the specified number or range of numerical values. The term comprising (and related terms such as contain or contains or having or including) includes such embodiments, for example, such an embodiment of any combination of substances, compositions, methods or processes, etc., which consists of or consists essentially of described symptoms.

The following abbreviations and terms have the meanings given throughout this document: PI3-K = phosphoinositide-3-kinase; and AIDS = acquired immunodeficiency syndrome.

Abbreviations used in this document have the meanings of chemistry and biology unless otherwise noted.

The term cell proliferation refers to the phenomenon whereby the number of cells changes due to division. The term also encompasses cell growth that changes cell morphology (eg, increases in size) in response to a proliferative signal.

The terms co-administration, administration in combination with, and their grammatical equivalents, as used herein, encompass the administration of two or more agents to an animal such that both agents and/or their metabolites are present in the animal simultaneously. Co-administration includes simultaneous administration of separate compositions, administration of separate compositions at different times, or administration of a composition in which both agents are present.

The term effective amount or therapeutically effective amount refers to that amount of a compound described herein that is sufficient to achieve the intended purpose of use, including, but not limited to, the treatment of a disease. A therapeutically effective amount may vary depending on the intended use (in vitro or in vivo), or the subject and disease state being treated, such as the weight and age of the subject, the severity of the disease state, route of administration, and the like, and may be easily determined by one of ordinary skill in the art. The term also refers to a dose that will induce a specific response in target cells, such as reduced platelet adhesion and/or cell migration. The specific dosage will vary depending on the specific compounds selected, the administration regimen to be followed, whether administration is in combination with other compounds, the time of administration, the tissue being administered, and the physical delivery system in which it is carried out.

As used herein, the terms treatment and treat refer to an approach aimed at obtaining beneficial or desirable results, including, but not limited to, therapeutic benefit and/or prophylactic benefit. By therapeutic benefit is meant the eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved by eradicating or alleviating one or more of the physiological symptoms associated with the underlying disorder so that an improvement in the patient's condition is observed, despite the fact that the patient may remain affected by the underlying disorder. For prophylactic benefit, the compositions may be administered to a patient at risk for developing a particular disease, or to a patient reporting one or more physiological symptoms of a disease, even though the disease may not be diagnosed.

Therapeutic effect, as used herein, encompasses a therapeutic benefit and/or a prophylactic benefit, as described above. A prophylactic effect includes slowing down or eliminating the manifestations of the disease or condition, slowing down or eliminating the onset of symptoms of the disease or condition, slowing down, stopping or reversing the disease or condition, or any combination thereof.

The term subject or patient refers to an animal such as a mammal, for example,

- 7 042667 people. The methods described herein may be suitable for use in both human therapy and veterinary medicine. In some embodiments, the patient is a mammal, and in some embodiments, the patient is a human. For veterinary purposes, the terms subject and patient include, but are not limited to, farm animals including cattle, sheep, pigs, horses, and goats; domestic animals such as dogs and cats; exotic animals and/or animals in zoos; laboratory animals, including mice, rats, rabbits, guinea pigs and hamsters; and birds such as chickens, turkeys, ducks and geese.

Radiation therapy refers to exposing a patient, using methods and compositions known to the practitioner, with radiation sources such as alpha particle emitting radionuclides (e.g. actinium and thorium radionuclides), low linear energy transfer (LET) radiation sources (i.e. e. beta emitters), conversion electron sources (eg, strontium-89 and samarium-153-EDTMP), or high energy radiation, including, but not limited to, x-rays, gamma rays, and neutrons.

Signal transduction is the process by which stimulatory or inhibitory signals are transmitted into and within a cell in order to elicit an intracellular response. A signal transduction pathway modulator refers to a compound that modulates the activity of one or more cellular proteins identified as belonging to one particular signal transduction pathway. The modulator can enhance (agonist) or suppress (antagonist) the activity of the signaling molecule.

The term selective inhibition or selectively inhibit, when used in relation to a biologically active agent, refers to the ability of the agent to selectively reduce the target signaling activity compared to extraneous signaling activity, through direct or indirect interaction with the target.

As used herein, the terms selective PI3 kinase inhibitor delta and selective PI3 kinase inhibitor δ generally refer to a compound that inhibits the activity of a PI3 kinase δ isoenzyme or iso(en)zyme δ more effectively than other PI3K families (alpha , beta and gamma). For example, a selective inhibitor of PI3 kinase δ may refer to a compound that exhibits a 50% inhibitory concentration (IC50) with respect to delta PI3 kinase type I, which is at least 10 times, at least 20 times, at least 50-fold, at least 100-fold, or even less than the IC50 value of the inhibitor relative to the rest of the type I PI3 kinases (i.e., alpha, beta, and gamma).

Inhibition of PI3 kinase δ can provide a therapeutically beneficial effect in the treatment of various conditions, for example, conditions characterized by an inflammatory response, including, but not limited to, autoimmune diseases, allergic diseases, and arthritic diseases. Importantly, inhibition of PI3-kinase δ function does not appear to affect biological functions such as viability and fertility.

An inflammatory response, as used herein, is characterized by redness, fever, swelling, and pain (ie, inflammation) and is typically associated with tissue damage or destruction. The inflammatory response is usually a localized defensive response caused by tissue damage or destruction aimed at destroying, weakening, or sequestering (sequestering) both the noxious agent and the damaged tissue. Inflammatory responses are particularly associated with leukocyte influx and/or leukocyte (eg, neutrophilic leukocyte) chemotaxis. Inflammatory responses can be caused by infection with pathogenic organisms and viruses, non-infectious causes such as trauma or reperfusion after myocardial infarction or stroke, immune responses to foreign antigens, and autoimmune diseases. Inflammatory responses amenable to treatment with the methods and compounds of the invention encompass conditions associated with specific defense responses as well as conditions associated with non-specific defense responses.

Therapeutic methods of the invention include methods for treating conditions associated with inflammatory cell activation. Inflammatory cell activation refers to the induction by a stimulus (including, but not limited to, cytokines, antigens, or autoantibodies) of a proliferative cellular response, the production of soluble mediators (including, cytokines, oxygen radicals, enzymes, prostanoids, or vasoactive amines), or cell surface expression of novel or an increased amount of mediators (including, but not limited to, major histocompatibility complex antigens or cell adhesion molecules) in inflammatory cells including, but not limited to, monocytes, macrophages, T-lymphocytes, B-lymphocytes, granulocytes (polymorphonuclear leukocytes, including neutrophils , basophils and eosinophils) mast cells, dendritic cells, Langerhans cells, and endothelial cells). Those skilled in the art will appreciate that activation of one or a combination of these phenotypes in such cells may contribute to the initiation, maintenance, or exacerbation of an inflammatory state.

Autoimmune disease, as used herein, refers to any group

- 8 042667 disorders in which tissue damage is associated with humoral or cell-mediated responses to the components of the organism itself.

Graft rejection, as used herein, refers to an immune response directed against a transplanted tissue (including organs or cells (e.g., bone marrow) characterized by loss of function of the transplanted and surrounding tissues, pain, swelling, leukocytosis, and thrombocytopenia).

Allergic disease, as used herein, refers to any symptoms, tissue damage, or loss of tissue function as a result of an allergy.

Arthritic disease, as used herein, refers to any disease characterized by inflammatory lesions of the joints, which can be attributed to various etiologies.

Dermatitis, as used herein, refers to any of a large family of skin diseases characterized by inflammation of the skin, which can be attributed to various etiologies.

The term particle size distribution for a powder or particulate material, or particles dispersed in a liquid, as used herein, means a list of values or a mathematical function that defines the relative amounts of particles present, sorted by size. The values d(0.1), d(0.5) and d(0.9) indicate that 10%, 50% and 90% of the measured particles were less than or equal to the specified value. For example, the values d(0.1)=3, d(0.5)=10 and d(0.9)=100 mean that 10% of the particles were smaller than or equal to 3 µm, 50% were smaller than or equal to 10 µm and 90% had dimensions less than or equal to 100 µm.

The terms d(0.1), d(0.5), and d(0.9) refer to the mesh sizes of a single abstract sieve through which 10, 50, or 90% of the total sample particles pass. Thus, d(0.1)=2-100 µm indicates that the upper limit of the particle size range representing the smallest 10% of the sample particles is 2 to 100 µm. Thus, 10% of the total number of particles have a particle size not exceeding the value of d(0.1), that is, in this case, they have a maximum size in the range from 2 to 100 μm.

Pharmaceutical compositions

The present invention provides a pharmaceutical composition comprising the PTSA salt of TGR-1202 as described herein and one or more pharmaceutically acceptable excipients. For example, the PTSA salt of TGR-1202 may have the particle size distribution described herein and/or the crystal lattice characteristics described herein.

In one aspect, the present invention provides a pharmaceutical composition comprising a TGR-1202 PTSA salt form B crystal according to any embodiment herein (e.g., a TGR-1202 PTSA salt form B crystal having a particle size distribution defined in any embodiment herein), and one or more pharmaceutically acceptable excipients. In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of TGR-1202 PTSA crystalline Form B according to any embodiment herein (e.g., TGR-1202 PTSA crystalline Form B having a particle size distribution defined in any implementation in this document). The pharmaceutical composition may contain one or more additional active ingredients, as described in this document.

Excipients may be selected from diluents, fillers, salts, disintegrants, binders, lubricants, glidants, dispersing agents, suspending agents, wetting agents, sustained release matrices, coloring agents, flavoring agents, buffers, stabilizers, solubilizers, and their combinations.

The pharmaceutical compositions of the present invention may be administered alone or in combination with one or more other active agents. If necessary, the compounds and other agent(s) of the present invention may be mixed to form a formulation, or both components may be formulated as separate formulations for their combined use, either alone or simultaneously.

The PTSC salts of TGR-1202 and the pharmaceutical compositions described herein may be administered by any route that allows delivery of TGR-1202 to the site of exposure, such as orally, buccally, intranasally, topically (e.g., transdermally), intraduodenally, parenterally (including intravenously, intraarterially, intramuscularly, intravascularly, intraperitoneally, or by injection or infusion), intradermal, intramammary, intrathecal, intraocular, retrobulbar, intrapulmonary (eg, aerosolized drugs), or subcutaneously (including the administration of a depot for long-term release, for example, under capsule of the spleen, into the brain, or into the cornea), sublingually, anally, rectally, vaginally, or by surgical implantation (eg, under the capsule of the spleen, into the brain, or into the cornea).

The compositions may be administered in solid, semi-solid, liquid or gaseous form, or may be in the form of a dry powder such as a lyophilized form. Pharmaceutical compositions may be packaged in forms suitable for delivery, including, for example, solid dosage forms such as capsules, sachets, cachets, gelatin capsules, paper bags, tablets, suppositories, granules, pills, lozenges, and lozenges. The type of packaging will generally depend on the desired route of administration. Implantable sustained release compositions are also contemplated, such as those for transdermal administration.

The amount of TGR-1202 PTSA salt administered will depend on the mammal being treated, the severity of the disorder or condition, the rate of administration, the characteristics of the compound, and the judgment of the prescribing physician. However, an effective dose is in the range of from about 0.001 to about 100 mg per kg of body weight per day, preferably from about 1 to about 35 mg/kg/day, in single or multiple doses. For a 70 kg human, this would be from about 0.05 to 7 g/day, preferably from about 0.05 to about 2.5 g/day. An effective amount of the TGR-1202 PTCA salt described herein may be administered in single or multiple doses (eg, two or three times a day).

The PTSA salt of TGR-1202 described herein can be used in combination with one or more anti-cancer agents (eg, chemotherapeutic agents), therapeutic antibodies, and radiation therapy.

The TGR-1202 PTSA salt composition described herein may be formulated or administered in combination with other agents that ameliorate the symptoms of inflammatory conditions such as encephalomyelitis, asthma, and other diseases described herein. Such agents include non-steroidal anti-inflammatory drugs (NSAIDs).

Preparation of various pharmaceutical compositions known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition, McGraw Hill, 2003; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (The Pharmaceutical Press, London, 1999), which are all incorporated by reference herein in their entirety.

In one embodiment, the PTSA salt of TGR-1202 described herein is administered at a dose selected to create a blood concentration of the compound in the range of about 20 to 5000 ng/mL and maintain that concentration over a period of about 6 to 24 hours after administration. In another specific embodiment, the dose size and frequency is chosen to achieve a blood concentration of the compound in the range of about 50 to 2500 ng/mL and maintain that concentration for a period of about 6 to 24 hours from administration. In some embodiments, the dose size and frequency is chosen to achieve a blood concentration of TGR-1202 having a value in the range of about 100 to 1500 ng/ml after administration. In some embodiments, the dose size and frequency is chosen to achieve a blood concentration of TGR-1202 having a value in the range of about 100 to 750 ng/mL over a period of about 6 to 24 hours from administration. In further embodiments, the dose size and frequency is selected to achieve a plasma _Cmax of TGR-1202 PTSA salt of at least about 300 ng/mL and not greater than about 10,000 ng/mL.

Methods of treatment

The invention also provides methods of using the compounds and pharmaceutical compositions of the present invention for the treatment of disease states, including, but not limited to, diseases associated with dysfunction of one or more types of PI3 kinases.

Detailed descriptions of conditions and disorders mediated by PI3 δ kinase activity are provided, for example, in International Publication Nos. WO 2001/81346, WO 2010/123931, WO 2010/0111432 and WO 2010/057048, and US Publication No. US 2005/043239 , each of which is incorporated herein by reference in its entirety.

Methods of treatment provided herein include administering to a subject a therapeutically effective amount of a compound of the invention. In one embodiment, the present invention provides a method for treating an inflammatory disorder, including autoimmune diseases, in a mammal. The method includes administering to said mammal a therapeutically effective amount of a compound of the present invention.

It should be understood that the methods of treatment according to the invention are suitable for use in the field of human medicine and in veterinary medicine. Thus, the individual being treated may be a mammal, preferably a human, or other animals. For veterinary purposes, individuals include, but are not limited to, farm animals, including cattle, sheep, pigs, horses, and goats; domestic animals such as dogs and cats; exotic animals and/or animals in zoos; laboratory animals, including mice, rats, rabbits, guinea pigs and hamsters; and birds such as chickens, turkeys, ducks and geese.

- 10 042667

The invention also relates to a method for treating a hyperproliferative disorder in a subject, which comprises administering to said mammal a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof. In some embodiments, said method relates to the treatment of cancer, such as acute myeloid leukemia, thymus, brain, lung, squamous cell, skin, eye, retinoblastoma, intraocular melanoma, oral and oropharyngeal, urinary bladder, stomach, pancreas, bladder, breast, cervix, head, neck, kidney, liver, ovary, prostate, colorectal cancer, cancer of the esophagus, testis, gynecological cancer, cancer of the thyroid gland, CNS, paraneoplastic syndromes ( PNS), AIDS-associated cancers (eg, lymphoma and Kaposi's sarcoma), or virus-induced cancer. In some embodiments, the method relates to the treatment of a noncancerous hyperproliferative disorder such as benign hyperplasia of the skin (eg, psoriasis), restenosis, or prostate (eg, benign prostatic hypertrophy (BPH)).

Examples

The examples and preparations below further illustrate and exemplify preparations for the compounds and compositions of the invention. It should be understood that the scope of the present invention is not limited in any way by the scope of the following examples and preparations.

X-ray diffraction patterns were obtained using an X'PertPRO MPD diffractometer equipped with a Cu LFF X-ray tube (45 kV, 40 mA) using the following measurement parameters:

scan mode: continuous;

PSD mode (coordinate detector): scanning;

PSD length: 2.12° 2θ;

scan range: 2.5-40° 2θ;

step size: 0.017/step;

step scan time: 12.065 s;

deflecting gap type: automatic;

monochromatic incident beam: no;

rotation: yes

DSC spectra were obtained using a DSCQ2000 V24.11 Build 124 instrument.

Example 1 Preparation of the PTSA salt of TGR-1202 (Form A).

7100 g of TGR-1202 was loaded into a reactor containing 56.8 L of acetone and stirred at ambient temperature. Added 4680 g of p-toluosulfonic acid and the reaction mixture was heated at a temperature of 60-65°C for about 6 hours the Solvent was evaporated under reduced pressure to obtain a wet residue. The wet residue was degassed and left to cool to <20°C. Approximately 142 L of diethyl ether was then added and the resulting mixture was stirred overnight, then filtered to give a solid, which was washed with diethyl ether and dried in vacuo to give a solid. The solid was taken up in diethyl ether, stirred for 6 hours, and then filtered to give a solid, which was then dissolved in 56.8 L of acetone, filtered through a HiFlow pad, and concentrated under reduced pressure. The resulting mass of residue was stirred with water overnight, then filtered and dried under vacuum to give 6600 g of TGR-1202 PTSA salt. HPLC: 99.21% and chiral purity 99.64:0.36 (S:R).

- 11 042667

Example 2 Preparation of the PTSA salt of TGR-1202 (Form B).

1000 g of TGR-1202 was loaded into a reactor containing 8 L of acetone and stirred at ambient temperature. Then 666 g of p-toluosulfonic acid was added and the reaction mixture was heated at a temperature of 60-65°C for about 6 hours. The solvent was evaporated under reduced pressure to obtain a wet residue. The wet residue was degassed and left to cool to <20°C. Approximately 20 L of diethyl ether was added and the resulting mixture was stirred overnight then filtered to give a solid which was washed with diethyl ether and dried in vacuo to give a solid which was then dried under vacuum to give 1150 g of TGR-1202 PTSA salt. HPLC: 99.33% and chiral purity: 99.61:0.39 (S:R).

In table. 1 shows the X-ray diffraction peaks and relative peak intensities for the products of Examples 1 and 2.

__________________________________________________Table 1

Example 1 Example 2 2-theta Rel. INT. (%) 2-theta Rel. INT. (%) 5.0111 90.45 5.0368 99.91 6.1947 17.54 6.2245 23.13 7.3344 28.14 7.3722 27.98 7.8955 23.93 7.9120 15.31 8.4677 5.77 8.4862 3.10 10.0914 95.66 10.1351 100.00 12.2407 38.58 12.2720 25.41 13.0311 18.40 12.4494 28.74 13.3639 18.53 13.0607 18.06 15.9118 43.52 13.3947 6.18 16.1260 62.05 15.9313 18.53 16.3442 43.31 16.1429 30.80 17.3580 33.21 16.3476 17.15 17.7652 36.83 16.7829 3.73 18.1828 34.80 17.0690 9.84 19.0276 20.04 17.3057 9.38 19.5461 38.96 17.4929 9.78

- 12 042667

20.0004 50.44 17.,7730 16.33 20.8037 38.08 18.2136 15.19 21.1204 27.38 18.4160 12.24 21.7227 32.60 18.7137 6.65 22.0621 78.29 19,1,252 12.24 22.8416 33.03 19.6214 21.10 23.1627 26.64 20.0040 25.77 24.4157 100.00 20.8028 15.49 25.3391 39.04 21.1629 11.75 26.9321 23.79 21.4304 8.00 27.8349 14.56 21.,7505 13.89 28.4163 12.04 22.1114 47.85 29.2666 25.40 22.8531 9.68 30.3829 10.40 23.1912 10.23 34.2390 3.40 24.4503 62.,76 35.8360 4.32 25.0031 15.78 38.0065 6.35 25.4128 18.58 38.9170 1.78 26.6621 4.52 26.9692 14.35 27.4037 4.15 27.8432 9.39 28.5253 6.78 29.3252 17.35 29.6443 6.08

- 13 042667

30.4781 5.95 31.1600 1.72 32.5213 2.02 32.9371 2.57 34.1290 1.91 35.6680 3.36 36.0502 2.33 36.7594 2.46 37.1729 1.34 (see also Figure 1) (see also Figure 2)

In table. 1A shows the endothermic peaks of the PTSA salts obtained in examples 1 and 2. _____________________________________________Table 1A

Example 1 Example 2 °C °C 153.66 145.66

Example 3 Particle Size Analysis

Particle size analysis (PSA) of the PTSC of the TGR-1202 salts prepared in Examples 1 and 2 was performed as described below using a Malvern Master Sizer 2000 instrument with a Scirocco 2000 dry powder system. The following instrument settings were used:

refractive index (RI) of the particles: 1.50;

absorption rate: 0.1;

dimming range: 1.5%;

analysis model: universal;

sensitivity: normal;

air pressure: 2.0 bar;

feed rate: 30%;

sample measurement time: 6 s;

background measurement time: 6 s;

number of measurement cycles: 1.

Procedure: 5 g of TGR-1202 was transferred to a sample measuring cuvette (ie Scirocco 2000 system) and air was turned on. The laser intensity was maintained above 70%. The measurement was started and the vibration speed was increased to achieve the degree of obscuration within the specified limits. After reaching the degree of darkening, which is within the specified limits, recorded the results of measurements. The analysis was carried out with three parallel measurements and the results were presented as the average of the three experiments.

Particle size distribution (PSD) for the products obtained in examples 1 and 2 are presented in table. 2.

________________________________________________Table 2

PTSA salt TGR-1202 Example d(0.1) µm d(0.5) µm d(0.9) µm Example 1 8.01 147.41 510.73 Example 2 1.07 3.98 12.18

- 14 042667

Example 4 Tableting of PTSA salts of TGR-1202.

The actual tablets were prepared containing 200 mg (as free base form) of the TGR-1202 PTSA salts of Examples 1 and 2 to study their dissolution characteristics. These compositions are designated as examples 4a and 4b, respectively. Particle size distribution for

The PTSA salts of TGR-1202 prepared in Examples 1 and 2 are shown in Example 3.

Example 4a. Six different tablets (T1, T2, T3, T4, T5 and T6) were prepared containing the PTSA salt of TGR-1202 prepared in Example 1.

Example 4b. Six different tablets (T1, T2, T3, T4, T5 and T6) were prepared containing the PTSA salt of TGR-1202 prepared in Example 2.

Used Equipment: Used tableting equipment is listed in Table 1. 3.

Table 3

Stage Name of equipment Cooking Sieves 20 or 24, 40 and 60 mesh (#) (841, 750, 420 and 250 microns) High Speed Mixer Granulator (RMG) Mixer Vessels made of stainless steel (SS) Fluid Bed Dryer (FBD) Multi Mill with 1.0mm Stainless Steel (SS) Screen Mixer Conta Blender Pressing Pressing machine Cadpress 17.8 x 8.8 mm, oval, D-Tooling system Tablet dust collectors Metal detectors (optional) Application coatings Coating Machine Mixer Vessels made of stainless steel (SS) Package Thermal induction sealer

Preparation of tablets: tablet formulations are given in table. 4.

- 15 042667

Table 4

Formulation of the composition for a dose of 200 mg (in terms of the free base form) No. Ingredients Example 4a Example 4b 1 PTSA salt TGR-1202 260.20 (salt) 260.20 (salt) 2 Hydroxypropyl Betadex 270.00 270.00 3 Microcrystalline cellulose (Avicel PH101) 41.80 41.80 4 Croscarmellose sodium (Ac-Di-Sol) 20.00 20.00 5 Hydroxypropylcellulose (Klucel LF) 6.00 6.00 6 Purified water q.s. (of necessity) q.s. 7 Croscarmellose sodium (Ac-Di-Sol) 70.00 70.00 8 magnesium stearate 2.00 2.00 The weight of the tablet itself 670.00 670.00 Coating 9 Opadry II Brown 40L565004 - 20.10 10 Purified water - q.s. Coated tablet weight - 690.10

Preparation process: Tablets were prepared as follows.

1. Dose the amount of starting materials in accordance with the recipe.

2. Screen the active pharmaceutical ingredient (API) along with hydroxypropyl betadex, croscarmellose sodium and microcrystalline cellulose through a 40 mesh (420 µm) sieve and collect in an integrated bulk container or double polyethylene bags.

3. Dissolve a weighed amount of hydroxypropyl cellulose (Klucel LF) in purified water with stirring in a stainless steel container until a clear solution (binder solution) is obtained.

4. Load the mixture from step 2 into a high speed mixer granulator (RMG) and mix for 8 minutes at low impeller speed with the chopper off.

5. Granulate the mixture from step 4 with the binder solution from step 3. Add additional water, if necessary, to obtain granules of suitable consistency. Spend additional mixing, if necessary. Spend periodic scraping, if necessary, during granulation.

6. Dry the wet mass in a fluid bed dryer (FBD) with an inlet temperature not exceeding 65° C. until a loss on drying (LOD) value of less than 3.0% w/w is achieved. Periodic scraping is carried out, if necessary.

7. Pass the dried granules from step 6 through a 24 mesh (750 µm) sieve and collect the sifted granules in a separate container. Grind the granules that have not passed through a 24 mesh (750 µm) sieve in a Multi mill equipped with a 1.0 mm stainless steel sieve at high speed with knives extended. The ground granules are collected and sifted through a 24 mesh sieve mounted on a vibrating sieving machine. Collect the granules that have passed through a 24 mesh sieve into a separate polyethylene bag.

8. If necessary, repeat step 7 again for granules that did not pass through the 24 mesh sieve. Collected in a built-in bulk container or double polyethylene bags.

9. Screen the croscarmellose sodium through a 40 mesh (420 µm) sieve and mix with the granules from the stage in a Conta mixer at 12 rpm for 8 minutes.

10. Screen the magnesium stearate through a 60 mesh (250 µm) sieve and mix with the mixture from step 9 in a Conta mixer at 12 rpm for 3 minutes.

11. Unload the lubricated granules into double polyethylene bags. Weigh and record the weight of the lubricated pellets.

12. Press the lubricated mixture using punches 17.8x8.8 mm oval with medium

-16042667 neem weighing 670.00 mg/tablet.

The coating procedure described below was carried out only for the tablets of example 4b.

13. Preparation of coating solution: Place a weighed amount of purified water in a container equipped with a mechanical agitator. Disperse Opadry II Brown 40L565004 in purified water with continuous stirring and mix for 45 minutes. Filter the resulting slurry through a 100 mesh (149 µm) sieve or nylon mesh.

14. Pressed tablets are loaded into a coating pan and coating is carried out until the required weight gain (2.5-3.5%) is achieved.

Process control data (IPC) for tablets according to examples 4a and 4b are given in table. 5 and 6, respectively.

Table 5

Example 4a: Process Control Data (IPC) for a 200 mg dose Parameter Rated value Average value Range Weight [mg] 670.00 ± 5% 669.70 664.8 - 678.6 Thickness [mm] - 5.31 5.27 - 5.35 Hardness [kp] 12.0±4.0 12.2 11.2-13.1 Decay [min] only registered 14-15 min brittleness no more than 1% 0.018%

Table 6

Example 4b: Process control data (IPC) for a dose of 200 mg Parameter Rated value Average value Range Weight [mg] 670.00 ± 5% 671.10 650-683 Thickness [mm] - — 5.33 - 5.42 Hardness [kp] 12.0±4.0 — 12.53 - 14.83 Decay [min] only registered 16-17 min brittleness no more than 1% 0.04%

Oral tablets remain one of the most effective treatments available. The effectiveness of such dosage forms is based on the dissolution of the drug in the fluids of the gastrointestinal tract before absorption into the systemic circulation. The dissolution rate of a tablet is thus related to its characteristics.

The dissolution of the actual tablet of Examples 4a and 4b was carried out using the following parameters: 900 ml of 0.1N aqueous HCl with 1% sodium lauryl sulfate (SDS), paddle stirrer according to USP - Apparatus 1, stirring speed 100 rpm, and 6 parallel measurements (n=6). The criteria for dissolution in a medium with pH 1 are at least 75% (Q=75) in 120 minutes. The results of dissolution measurements are presented in table. 7 below.

- 17 042667

Table 7

Time (min) Example 4a Example 4b % drug release Relative Standard Deviation (RSD) % drug release Relative Standard Deviation (RSD) 10 24 10.1 7 11.7 15 39 8.3 15 12.4 thirty 70 10.3 44 8.9 45 78 12.3 72 5.6 60 79 13.3 91 2.3 90 81 12.8 98 2.4 120 82 12.1 100 2.6

Example 5 Pharmacokinetics.

The oral bioavailability of tablets (examples 4a and 4b) was evaluated in healthy volunteers. The protocol of pharmacokinetic studies is given below.

All healthy subjects were fast overnight (12 hours) before dosing and for 4.0 hours after administration of the study composition. The test compositions (tablets) were prepared as described in examples 4a and 4b. Blood samples were taken and placed in microcentrifuge tubes containing an anticoagulant. Blood samples were immediately centrifuged and the isolated plasma samples were frozen at temperatures below -80°C and stored until analysis. Plasma concentrations of test substances in all samples were analyzed by liquid chromatography with tandem mass spectrometry (LC/MS-MS). Pharmacokinetic parameters were determined (i.e. Cmax, AUC _0-t , ^T max, ^{and t} 1/2 ⁾ ·

The tablet composition containing the PTSA salt TGR-1202 prepared according to Example 2 exhibited a C _max value about 2.5 times greater and an area under the curve (AUC) about 1.9 times greater than a tablet composition containing PTSA-salt TGR-1202, prepared in accordance with example 1. The results are presented in table. 8 below.

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Table 8

Pharmacokinetics in humans Options Tablets according to Example 4a Tablets according to Example 4b Number of healthy volunteers 12 12 Dose (mg) 200 200 Median T _max (h) 2 3 Stax (ng/ml) 155.97 395.49 *Т1/ ₂ (h) 74.92 73.59 AUCo-t (µg x h/ml) 5.06 9.43 AUCo-mf (µg x h/ml) 7.28 13.92

Although the invention set forth herein is described with reference to specific embodiments, it should be understood that such embodiments are only an illustrative representation of the principles and applications of the present invention. Thus, it should be understood that numerous modifications may be made to the illustrative embodiments, and that other variations may be devised without departing from the spirit and scope of the present invention described above. It is intended that the appended claims define the scope of the invention, and that they thus cover the methods and structures falling within the scope of these claims and their equivalents.

All publications, patents and patent applications referred to in this specification are hereby incorporated by reference to the same extent as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated herein by reference. .

Claims (14)

1. Crystalline p-toluenesulfonic acid salt of the compound where the specified salt has a value of d(0.9) from 5 to 50 microns. 2. Crystalline salt according to claim 1, characterized in that the specified salt has a value of (i) d(0.9) from 5 to 25 microns; (ii) d(0.5) from 1 to 10 µm; and/or (iii) d(0,l) from 0.5 to 1.5 µm. 3. Crystalline salt according to claim 1 or 2, characterized in that the specified salt has a value of (i) d(0.9) from 5 to 15 microns; (ii) d(0.5) from 2 to 5 µm; and/or (iii) d(0,l) from 0.5 to 1.0 µm. 4. A crystalline salt according to any of the preceding claims, characterized in that said salt is a crystalline form A and exhibits a powder diffraction pattern having one or more peaks selected from 5.01, 10.09, 15.91, 16.13, 16.34, 20.00, 22.06 and 24.42±0.2°20 and/or exhibit a differential scanning calorimetry (DSC) curve with a characteristic endothermic peak at about 154°C. - 19 042667 5. A crystalline salt according to any one of the preceding claims, characterized in that the ratio of p-toluenesulfonic acid to compound is 1:1. 6. Crystal form A of the p-toluenesulfonic acid salt of the compound where said crystalline form exhibits an x-ray powder pattern having one or more peaks selected from 5.01, 10.09, 15.91, 16.13, 16.34, 20.00, 22.06, and 24.42±0, 2° 2θ. 7. A crystalline form according to claim 6, characterized in that said form exhibits an x-ray powder pattern substantially as shown in FIG. 1. 8. A crystalline form according to claim 6 or 7, characterized in that said form has a d(0.5) value from 1 to 10 µm or from 2 to 5 µm. 9. A crystalline form according to any one of claims 6 to 8, characterized in that said form has a d(0.9) value of 5 to 50 µm, or 5 to 25 µm, or 5 to 15 µm. 10. A crystalline form according to any one of claims 6 to 9, characterized in that said form has a d(0.1) value of 0.5 to 1.5 µm or 0.5 to 1.0 µm. 11. A crystalline form according to any one of claims 6 to 10, characterized in that said form exhibits a differential scanning calorimetry (DSC) curve with a characteristic endothermic peak at about 154°C. 12. A pharmaceutical composition for the treatment of a PI3K associated disease or disorder, comprising a crystalline salt according to any one of the preceding claims and a pharmaceutically acceptable excipient. 13. The use of a crystalline salt according to any one of claims 1 to 11 for the treatment of a PI3K associated disease or disorder selected from leukemias and lymphomas. 14. Use according to claim 13, characterized in that the PI3K-associated disease, disorder or condition is acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, Burkitt's lymphoma; hematopoietic tumors of myeloid origin, acute myelogenous leukemia, chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia; chronic lymphocytic leukemia or small lymphocyte lymphoma.