WO2020051230A1 - Amorphous pharmaceutical compositions and uses thereof - Google Patents

Abstract

The present invention features pharmaceutical compositions including an autotax in (ATX) inhibitor and methods for the treatment of ATX-related disorders (e.g., cancer, inflammation, fibrosis, and pain) with such compositions.

Description

AMORPHOUS PHARMACEUTICAL COMPOSITIONS AND USES THEREOF

BACKGROUND OF THE INVENTION

Autotaxin (ATX) is a secreted enzyme of the ectonucleotide pyro-phosphatase/

phosphodiesterase family and is also known as Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP-2 or NPP2). ATX plays a role in driving pathological conditions, including fibrosis, arthritic inflammation, neurodegeneration, neuropathic pain, and cancer. ATX is the fundamental regulator of the conversion of Lysophosphatidylcholine (LPC) to Lysophosphatidic Acid (LPA). LPA is a bioactive lipid that affects migration, proliferation, and survival of various cell types.

Inhibition of ATX has been shown to reduce LPA levels in pathological settings. Reduction of LPA may provide therapeutic benefits in diseases with unmet medical need, including cancer, lymphocyte homing, chronic inflammation, neuropathic pain, fibrotic diseases such as Idiopathic Pulmonary Fibrosis (IPF), thrombosis, and cholestatic pruritus which are caused, mediated, and/or propagated by increased LPA levels and/or activation of ATX.

Fibrotic diseases are chronic, debilitating and often lethal pathologies driven by a dysregulated response to tissue or organ injury. Fibrosis can develop in the liver, kidney, lung, dermis, vasculature, gut, and other sites. Fibrosis develops due to action of pathways including growth factors, cytokines, integrin, and lipids.

ATX, LPA, and LPA Receptor (LPAR) pathways have been implicated in fibrotic disease. For example, profiling studies show increased levels of ATX, LPA, and LPARs in various rodent models of fibrosis and in human patient fluids and biopsy tissue. LPA can induce proliferative, survival, and chemotactic responses in transformed cell lines, indicating that LPA may exert pro-inflammatory and pro- fibrotic responses in cells known to be critical in fibrotic disease, including: fibroblasts, smooth muscle cells, macrophages, epithelial and endothelial cells, and leukocytes. Gene-targeted mouse models have implicated LPARs in fibrosis pathogenesis. Inhibitors of LPARs indicate that antagonism of receptors within this pathway blocked or reversed fibrosis in the lung, liver, kidney, and skin in rodents. Cell type- specific gene targeting studies have showed that ATX plays a role in the development of lung fibrosis and inflammatory arthritis.

ATX and LPA have also been implicated in tumor progression and metastasis. ATX may be responsible for increased LPA levels in ascites and plasma of ovarian cancer patients since ATX converts LPC to LPA. Increased levels of LPA, altered receptor expression, and altered responses to LPA may contribute to initiation, progression, or outcome of ovarian cancer. LPA has also been linked to prostate, breast, melanoma, head and neck, bowel, brain, and thyroid cancers.

LPA has been shown to promote tumor cell survival, proliferation, invasion, and migration into neighboring tissues, which can result in the formation of metastases. Additionally, LPA promotes cytoskeletal remodeling that may enhance migratory and invasive properties of cells, which may contribute to cancer metastasis. These biological and pathobiological processes of LPA are initiated through the activation of G-protein coupled receptors.

Transcriptome analyses of more than 350 normal tissues and more than 1700 malignant tissues demonstrate that ATX is expressed in a variety of carcinomas and sarcomas, underscoring the potential contribution of LPA to metastatic disease. Accordingly, when treating patients with diseases, such as cancer, lymphocyte homing, chronic inflammation, neuropathic pain, fibrotic diseases, thrombosis, and cholestatic pruritus, it is desirable to lower LPA levels. This can be accomplished through inhibition of enzymes involved in LPA biosynthesis, such as ATX.

Since ATX is expressed in tumors and affects tumor cell proliferation and invasion into neighboring tissues both of which can lead to the formation of metastases, ATX is a target for anti-tumor therapy. Moreover, in angiogenesis, ATX, taken with other anti-angiogenetic factors, brings about blood vessel formation. Angiogenesis supplies tumors with nutrients during tumor growth. Therefore, inhibition of angiogenesis is a target for anti-tumor therapy, leading to starvation of a tumor.

ATX has also been implicated in nerve injury-induced neuropathic pain. LPA biosynthesis, through ATX, is the source of LPA for LPA1 receptor-mediated neuropathic pain. Therefore, targeted inhibition of ATX-mediated LPA biosynthesis may represent a novel treatment to prevent nerve injury- induced neuropathic pain.

As described in International Patent Publication No. WO2015/154023, Compound I, shown below, has been found to modulate the activity of ATX:

I

Accordingly, pharmaceutical compositions of compound I, or pharmaceutically acceptable salts thereof, are useful for the treatment of ATX-related disorders such as cancer, lymphocyte homing, chronic inflammation, neuropathic pain, fibrotic diseases, thrombosis, or cholestatic pruritus.

SUMMARY OF THE INVENTION

The present invention features compositions (e.g., pharmaceutical compositions) including compound I, methods of producing such compositions, and methods for the treatment of ATX-related disorders with such compositions.

Accordingly, in an aspect, the invention features a composition (e.g., a pharmaceutical composition) including amorphous compound I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In some embodiments, the pharmaceutical composition includes at least 90% by weight of amorphous compound I (e.g., the pharmaceutical composition is substantially free of crystalline compound I).

In some embodiments, the pharmaceutical composition further includes at least 0.5% by weight of water (e.g., at least 0.6%, at least 0.7%, at least 0.8%, at least 0.9%, at least 1 .0%, at least 1 .1 %, at least 1 .2%, at least 1 .3%, at least 1 .4%, at least 1 .5%, at least 1 .6%, at least 1 .7%, at least 1 .8%, at least 1 .9%, at least 2.0%, at least 2.1 %, at least 2.2%, at least 2.3%, at least 2.4%, at least 2.5%). In some embodiments, the pharmaceutical composition further includes between 0.5% and 2.5% by weight of water (e.g., between 0.5% and 1 %, between 0.7% and 1 .5%, between 1 % and 2%, between 1 .5% and 2.5%, between 0.9 and 2.2%). The amount of water in the composition may be measured by any method known in the art, e.g., by Karl Fischer analysis.

In some embodiments, the pharmaceutical composition further includes at least 0.1 % (at least 0.2%, at least 0.3%, at least 0.4%, at least 0.5%, at least 0.6%, at least 0.7%, at least 0.8%, at least 0.9%, or at least 1 .0%, or 0.1 -0.3%, 0.2-0.5%, 0.3-0.6%, 0.4%-0.8%, 0.5-0.9%, or 0.7-1 .0%) by weight of solvent (e.g., acetone or ethanol). The amount of solvent in the composition may be measured by any method known in the art, e.g., by gas chromatography analysis.

In some embodiments, the pharmaceutical composition is substantially free of amorphous state stabilizing agents (e.g., polymers such as povidone, crospovidone, poloxamer, hydroxypropyl methylceullulose acetate succinate (HPMCAS), hydroxypropyl methylcellulose phthalate (HP-MCP), hydroxypropyl methylcellulose (HPMC), or a polymethacrylate).

In some embodiments, the amorphous compound I has an endothermic onset at about 125±1 °C (e.g., about 125.04 or 125.34) in differential scanning calorimetry (DSC) profile.

In some embodiments, the amorphous compound I has a loss of weight from 30 Ό to 150 °C of less than 3.5% (e.g., less than 3.4%, less than 3.3%, less than 3.2%, less than 3.1 %, less than 3.0%, less than 2.9%, less than 2.8%, less than 2.7%, less than 2.6%, less than 2.5%, less than 2.4%, less than 2.3%, less than 2.2%, less than 2.1 %, less than 2.0%, less than 1 .9%, less than 1 .8%, less than 1 .7%, less than 1 .6%, less than 1 .5%, less than 1 .4%, less than 1 .3%, less than 1 .2%, less than 1 .1 %, less than 1 .0%) as measured by thermal gravimetric analysis. In some embodiments, the amorphous compound I has a loss of weight from 30 Ό to 1 50 °C of less than 2.0%.

In some embodiments, the amorphous compound I comprises spray-dried compound I. In some embodiments, the amorphous compound I is spray-dried from acetone. In some embodiments, the amorphous compound I is spray-dried from acetone and water (e.g., acetone and water in a 17:3 mixture).

In some embodiments, the amorphous compound I includes particles having a shriveled sphere morphology.

In some embodiments, the purity of the pharmaceutical composition decreases by less than 1 % (e.g., less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1 %) upon storage (e.g., at room temperature and/or exposure to light) for at least 5 days (e.g., at least 1 week, at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 1 1 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 16 months, at least 17 months, at least 18 months, at least 19 months, at least 20 months, at least 21 months, at least 22 months, at least 23 months, or at least 24 months).

In some embodiments, the composition comprises at least 5 mg (e.g., at least 10 mg, at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least 100 mg, at least 1 50 mg, at least 200 mg, at least 250 mg, at least 300 mg, at least 350 mg, at least 400 mg, at least 450 mg, at least 500 mg) of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof. In some embodiments, the composition comprises between 5 mg and 1000 mg (e.g., 5-25 mg, 15-50 mg, 25-100 mg, 50-200 mg, 100-250 mg, 200-500 mg, 300-600 mg, 500-750 mg, 600-1000 mg) of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof. In some embodiments, the composition comprises about 5 mg (e.g., about 1 0 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 100 mg, about 1 50 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg) of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preparing any of the foregoing compositions (e.g., pharmaceutical compositions). This method includes: (a) combining compound I and a solvent (e.g., dichloromethane, methanol, acetone, tetrahydrofuran, water, or combinations thereof); and (b) evaporating (e.g., spray-drying) the solvent of the mixture of step (a). In some embodiments, the method is conducted on large scale (e.g., at least 1 .0 kg of compound I is used in step (a)).

In another aspect, the invention features a method of preparing any of the foregoing compositions (e.g., pharmaceutical compositions). This method includes: (a) combining compound I and one or more solvents (e.g., ethanol); and (b) precipitating compound I from the mixture of step (a) by addition of an additional solvent (e.g., water).

In some embodiments, the solvent is acetone or a mixture of acetone and water (e.g., a 17:3 mixture of acetone and water).

In some embodiments, the method further includes drying the product of step (b) in an oven for at least 24 hours (e.g., at least 36 hours, at least 48 hours, or at least 72 hours). In some embodiments, the purity of the pharmaceutical composition decreases by less than 1 % (e.g., less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1 %) upon storage (e.g., at room temperature and/or exposure to light) for at least 5 days (e.g., at least 1 week, at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 1 1 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 16 months, at least 17 months, at least 18 months, at least 19 months, at least 20 months, at least 21 months, at least 22 months, at least 23 months, or at least 24 months).

In another aspect, the invention features a method of decreasing the level or activity of autotaxin (ATX) in a cell (e.g., a human cell) or subject (e.g., a human) in need thereof. This method includes administering to the subject or cell an effective amount of any of the foregoing compositions (e.g., pharmaceutical compositions).

In another aspect, the invention features a method of treating an ATX-related disorder in a subject in need thereof. This method includes administering an effective amount of any of the foregoing compositions (e.g., pharmaceutical compositions).

In some embodiments, the effective amount includes an amount which results in a reduction (e.g., at least 5% reduction, at least 1 0% reduction, at least 20%, reduction, at least 50% reduction, at least 75% reduction, at least 100% reduction) in LPA levels in the subject.

In some embodiments, the ATX-related disorder is cancer, lymphocyte homing, inflammation, pain, fibrotic diseases, thrombosis, or cholestatic pruritus

In some embodiments, the inflammation is chronic inflammation.

In some embodiments, the pain is neuropathic pain. In some embodiments, the fibrotic disease is lung fibrosis.

In some embodiments, the ATX-related disorder is lung fibrosis, asthma, chronic obstructive pulmonary disease (COPD), renal fibrosis, acute kidney injury, chronic kidney disease, liver fibrosis, skin fibrosis, fibrosis of the gut, breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, bone cancer, colon cancer, bowel cancer, head and neck cancer, melanoma, multiple myeloma, chronic lymphocytic leukemia, B cell lymphoma, T cell lymphoma, cancer pain, tumor metastasis, transplant organ rejection, scleroderma, ocular fibrosis, age related macular degeneration (AMD), diabetic retinopathy, collagen vascular disease, atherosclerosis, Raynaud's phenomenon, rheumatoid arthritis, osteoarthritis or neuropathic pain.

In some embodiments, the ATX-related disorder is cancer, e.g., breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, bone cancer, colon cancer, bowel cancer, head and neck cancer, melanoma, multiple myeloma, chronic lymphocytic leukemia, B cell lymphoma, T cell lymphoma, cancer pain, tumor metastasis, or nonalcoholic fatty liver disease. In some embodiments, the ATX- related disorder is fibrosis, e.g., lung fibrosis, renal fibrosis, liver fibrosis, skin fibrosis, ocular fibrosis, or fibrosis of the gut.

In some embodiments, the method includes administering one or more additional therapies to the subject. In some embodiments, the one or more addition therapies includes a corticosteroid,

immunosuppressant, analgesic, anti-cancer agent, anti-inflammatory agent, non-steroidal anti inflammatory agent, dual cyclooxygenase-1 and -2 inhibitor, cyclooxygenase-2 selective inhibitor, TNFa blocker, kinase inhibitor, chemokine receptor antagonist, bronchodilator, leukotriene receptor antagonist, leukotriene formation inhibitor, prostaglandin receptor antagonist, prostaglandin formation inhibitor, monoacylglycerol kinase inhibitor, phospholipase A1 inhibitor, phospholipase A2 inhibitor,

lysophospholipase D (lysoPLD) inhibitor, autotaxin inhibitor, or LPA receptor antagonist..

In some embodiments, the one or more addition therapies includes a corticosteroid,

immunosuppressant, analgesic, anti-inflammatory agent, non-steroidal anti-inflammatory agent, dual cyclooxygenase-1 and -2 inhibitor, cyclooxygenase-2 selective inhibitor, TNFa blocker, chemokine receptor antagonist, bronchodilator, leukotriene receptor antagonist, leukotriene formation inhibitor, prostaglandin receptor antagonist, prostaglandin formation inhibitor, monoacylglycerol kinase inhibitor, phospholipase A1 inhibitor, phospholipase A2 inhibitor, lysophospholipase D (lysoPLD) inhibitor, autotaxin inhibitor, or LPA receptor antagonist.

Definitions

In this application, unless otherwise clear from context, (i) the term“a” may be understood to mean“at least one”; (ii) the term“or” may be understood to mean“and/or”; (iii) the terms“comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms“about” and “approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.

As used herein, the term“activity of a protein,” refers to an activity related to the protein, or a related downstream effect. Activity of ATX includes, for example, lysophospholipase D activity that converts lysophosphatidylcholine into LPA. As used herein, the term“administration” refers to the administration of a composition to a subject or cell. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, or vitreal.

As used herein, the term“amorphous,” refers to a disordered state of a compound that lacks the long-range order which is characteristic of a crystal.

As used herein, the term“amorphous state stabilizing agent,” refers to pharmaceutically acceptable excipients which inhibit crystallization of a compound. For example, polymeric matrices have been shown to stabilize the amorphous state of compounds by stabilizing the amorphous state of the compound below the glass transition temperature. Amorphous state stabilizing agents include, but are not limited to, povidone, crospovidone, poloxamer, hydroxypropyl methylceullulose acetate succinate (HPMCAS), hydroxypropyl methylcellulose phthalate (HP-MCP), hydroxypropyl methylcellulose (HPMC), and polymethacrylates.

As used herein, the term“ATX-related disorder,” refers to any disease or disorder that may derive a therapeutic benefit from modulation (e.g., decrease) of the level or activity of ATX, e.g., lung fibrosis, asthma, chronic obstructive pulmonary disease (COPD), renal fibrosis, acute kidney injury, chronic kidney disease, liver fibrosis, skin fibrosis, fibrosis of the gut, breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, bone cancer, colon cancer, bowel cancer, head and neck cancer, melanoma, multiple myeloma, chronic lymphocytic leukemia, B cell lymphoma, T cell lymphoma, cancer pain, tumor metastasis, transplant organ rejection, scleroderma, ocular fibrosis, age related macular degeneration (AMD), diabetic retinopathy, collagen vascular disease, atherosclerosis, Raynaud's phenomenon, rheumatoid arthritis, osteoarthritis or neuropathic pain.

The term“decreasing the activity of autotaxin” as used herein, refers to an observable change in the level of a marker of autotaxin activity (e.g., a decrease in the level of iysophosphatidic acid) in a subject and/or cell, as measured using techniques and methods known in the art for the measurement of the marker. Decreasing the marker level in a subject and/or cell may result in a decrease of at least 1 % relative to prior to administration (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 98% or more relative to prior to

administration; e.g., up to 100% relative to prior to administration).

By“decreasing the level of autotaxin” is meant decreasing the level of autotaxin in a subject and/or cell (e.g., human). The level of autotaxin is determined by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample) to obtain the physical entity or value. “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third-party laboratory that directly acquired the physical entity or value). Methods to measure autotaxin level generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescent polarization, phosphorescence, immunohistochemical analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS), and flow cytometry, as well as assays based on a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners. In some embodiments, the level of autotaxin is decreased by about 5%, about 1 0%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 1 00%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; decreased by more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; or decreased by more than about 1 .2- fold, about 1 .4-fold, about 1 .5-fold, about 1 .8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, 25 about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more).

By“determining the level of a compound” is meant the detection of a compound, by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample or“analyzing a sample” as that term is defined herein) to obtain the physical entity or value. “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third-party laboratory that directly acquired the physical entity or value). Methods to measure compound levels generally include, but are not limited to, liquid chromatography (LC)-mass spectrometry.

By“determining the level of a protein” is meant the detection of a protein, or an mRNA encoding the protein, by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample or“analyzing a sample” as that term is defined herein) to obtain the physical entity or value. “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third-party laboratory that directly acquired the physical entity or value). Methods to measure protein level generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescent polarization, phosphorescence, immunohistochemical analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS), and flow cytometry, as well as assays based on a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners. Methods to measure mRNA levels are known in the art.

An“effective amount” of a compound may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit the desired response. A therapeutically effective amount encompasses an amount in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects. A therapeutically effective amount also encompasses an amount sufficient to confer benefit, e.g., clinical benefit.

By“level” is meant the amount of a compound or protein, or its activity. Typically, this level is compared to a reference. The reference can be any useful reference, as defined herein. By a “decreased level” or an“increased level” of a compound, protein, or activity of a protein is meant a decrease or increase in level, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an increase of more than about 1 0%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by less than about 0.01 - fold, about 0.02-fold, about 0.1 -fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less; or an increase by more than about 1 .2-fold, about 1 .4-fold, about 1 .5-fold, about 1 .8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more). A level of a compound or protein may, in some cases, be expressed in mass/vol (e.g., g/dL, mg/mL, pg/mL, ng/mL) or as a percentage relative to the total compound in a sample.

As used herein, the term“marker of autotaxin activity,” refers to a compound or protein which is implicated in the autotaxin pathway. For example, as is known in the art, autotaxin has

lysophospholipase D activity that converts lysophosphatidylcholine into lysophosphatidic acid.

Accordingly, either lysophosphatidylcholine or lysophosphatidic acid could be utilized as markers of autotaxin activity. In some embodiments, an increase in lysophosphatidylcholine as compared to a reference is a marker of a decrease in autotaxin activity. In some embodiments, a decrease in lysophosphatidic acid as compared to a reference is a marker of a decrease in autotaxin activity.

The term“pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient. A pharmaceutical composition may be one manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gel cap, suspension, solution, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

As used herein, the term“pharmaceutically acceptable salt” means any pharmaceutically acceptable salt of compound I. For example, pharmaceutically acceptable salts of compound I include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: in Remington (Remington: The Science and Practice of Pharmacy, (22nd ed.) ed. L.V. Allen, Jr., 2013, Pharmaceutical Press, Philadelphia, PA). The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable organic acid.

Compound I may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids. Suitable pharmaceutically acceptable acids and methods for preparation of the appropriate salts are well-known in the art. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, gentisate,

glucoheptonate, glycerophosphate, glycolate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate salts.

A“pharmaceutically acceptable excipient,” as used herein, refers any inactive ingredient (for example, a vehicle capable of suspending or dissolving the active compound) having the properties of being nontoxic and non-inflammatory in a subject. Typical excipients include, for example:

antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: butylated optionally substituted hydroxyltoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxylpropyl cellulose, optionally substituted hydroxylpropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those of ordinary skill in the art are familiar with a variety of agents and materials useful as excipients.

By a“reference” is meant any useful reference used to compare compound levels or activity. The reference can be any sample, standard, standard curve, or level or activity that is used for comparison purposes. The reference can be a normal reference sample or a reference standard or level. A “reference sample” can be, for example, a control, e.g., a predetermined negative control value such as a “normal control” or a prior sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (e.g., a cell or tissue) from a subject not having a disease; a sample from a subject that is diagnosed with a disease, but not yet treated with a compound of the invention; a sample from a subject that has been treated by a compound of the invention; or a sample of a purified compound (e.g., any described herein) at a known normal concentration. By“reference standard or level” is meant a value or number derived from a reference sample. A“normal control value” is a pre-determined value indicative of non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range (“between X and Y”), a high threshold (“no higher than X”), or a low threshold (“no lower than X”). A subject having a measured value within the normal control value for a particular biomarker is typically referred to as“within normal limits” for that biomarker. A normal reference standard or level can be a value or number derived from a normal subject not having a disease or disorder (e.g., cancer); a subject that has been treated with a compound of the invention. In preferred embodiments, the reference sample, standard, or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of levels of a purified compound, e.g., any described herein, within the normal reference range can also be used as a reference.

As used herein, the term“subject” or“patient” refers to any organism to which a composition in accordance with the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.

The term“substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological and/or chemical arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term“substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

As used herein, the terms“treat,”“treated,” or“treating” mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e. , not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present disclosure; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

DETAILED DESCRIPTION The present invention features pharmaceutical compositions including compound I or a pharmaceutically acceptable salt thereof and methods for the treatment of ATX-related disorders with such compositions.

Compound I

The pharmaceutical compositions of the invention comprise amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof. Compound I has the structure:

I

In some embodiments, a pharmaceutical composition of the invention includes 20-70% amorphous compound I or amorphous pharmaceutically acceptable salt thereof by total weight, e.g.,

20%, 30%, 40%, 50%, 60%, or 70%. In some embodiments, a pharmaceutical composition includes greater than 20% compound I or pharmaceutically acceptable salt thereof by total weight, e.g., greater than 25%, greater than 30%, greater than 40%, greater than 50%, greater than 60%, greater than 70%, greater than 80%, or greater than 90%.

Compositions

The present invention includes pharmaceutical compositions comprising amorphous compound I, or amorphous pharmaceutically acceptable salts thereof, which is formulated for a desired mode of administration with or without one or more pharmaceutically acceptable and useful carriers.

The amorphous compounds can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

The pharmaceutical compositions of the invention comprise amorphous compound I (or an amorphous pharmaceutically acceptable salt thereof) as an active ingredient, optional pharmaceutically acceptable carrier(s) and optional other therapeutic ingredients or adjuvants. The compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

Amorphous compound I can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the invention can be presented as discrete units suitable for oral administration such as capsules, cachets, or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder (e.g., powder in capsules with or without excipients), as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion, or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof, can also be administered by controlled release means and/or delivery devices. The compositions can be prepared by any of the methods of pharmacy.

In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

A tablet containing the composition of this invention can be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent.

Molded tablets can be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. Each tablet preferably contains from about 0.05 mg to about 5 g of the active ingredient and each cachet or capsule preferably containing from about 0.05 mg to about 5 g of the active ingredient.

A formulation intended for the oral administration to humans may contain from about 0.5 mg to about 5 g of active agent, compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition. Unit dosage forms will generally contain between from about 1 mg to about 2 g of the active ingredient, typically 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

Amorphous compounds of the invention can be provided for formulation at high purity, for example, at least about 90%, 95%, or 98% pure by weight.

Amorphous pharmaceutical compositions of the invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

Amorphous pharmaceutical compositions of the invention suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

Amorphous pharmaceutical compositions of the invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, or dusting powder. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations may be prepared, utilizing compound I, or a pharmaceutically acceptable salt thereof, via conventional processing methods. As an example, a cream or ointment is prepared by admixing hydrophilic material and water, together with about 5 wt% to about 10 wt% of the compound, to produce a cream or ointment having a desired consistency.

Amorphous pharmaceutical compositions of this invention can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

In addition to the aforementioned carrier ingredients, the amorphous pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, or preservatives (including anti-oxidants). Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient.

Compositions containing amorphous compound I, or amorphous pharmaceutically acceptable salts thereof, may also be prepared in powder or liquid concentrate form.

Uses

Amorphous compound I inhibits the activity of ATX in animals, including humans, and are useful in the treatment and/or prevention of various diseases and conditions such as cancer, lymphocyte homing and inflammation, neuropathic pain, fibrotic diseases, thrombosis, and cholestatic pruritus which are caused, mediated, and/or propagated by increased LPA levels and/or the activation of ATX. In particular, amorphous compound I, and compositions thereof, are inhibitors of ATX, and are useful in treating conditions modulated, at least in part, by ATX.

In some embodiments, the invention includes a method of treating cancer comprising

administering to a mammal in need thereof a therapeutically effective amount of a composition of the invention.

In some embodiments, the invention includes a method of treating a cancer mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for treating a cancer, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating lymphocyte homing and inflammation comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein. In some embodiments, the invention includes a method of treating lymphocyte homing and inflammation mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for treating lymphocyte homing and inflammation, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating neuropathic pain comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating neuropathic pain mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for treating neuropathic pain, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating fibrotic diseases comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating fibrotic diseases mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for treating a fibrotic disease, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating thrombosis comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating thrombosis mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for treating thrombosis, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating cholestatic pruritus comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein. In some embodiments, the invention includes a method of treating cholestatic pruritus mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

In some embodiments, the invention includes a method of treating or a method of manufacturing a medicament for cholestatic pruritus, such as those described herein, which is mediated at least in part by ATX, comprising administering to a mammal in need thereof a therapeutically effective amount of a composition described herein.

The compositions of the invention are useful in the treatment of a variety of cancers, including, but not limited to, solid tumors, sarcoma, fibrosarcoma, osteoma, melanoma, retinoblastoma, rhabdomyosarcoma, glioblastoma, neuroblastoma, teratocarcinoma, hematopoietic malignancy, and malignant ascites. More specifically, the cancers include, but not limited to, lung cancer, bladder cancer, pancreatic cancer, kidney cancer, gastric cancer, breast cancer, colon cancer, prostate cancer (including bone metastases), hepatocellular carcinoma, ovarian cancer, esophageal squamous cell carcinoma, melanoma, an anaplastic large cell lymphoma, an inflammatory myofibroblastic tumor, and a

glioblastoma.

In some embodiments, the above methods are used to treat one or more of bladder, colorectal, non-small cell lung, breast, or pancreatic cancer. In some embodiments, the above methods are used to treat one or more of ovarian, gastric, head and neck, prostate, hepatocellular, renal, glioma, or sarcoma cancer.

In some embodiments, the invention includes a method, including the above methods, wherein the compound is used to inhibit cellular epithelial to mesenchymal transition (EMT).

In some embodiments, the method further comprises administering at least on additional active agent. In some embodiments, the invention includes a method of treating cancer comprising

administering to a mammal in need thereof a therapeutically effective amount of a composition described herein, wherein at least one additional active anti-cancer agent is used as part of the method.

In some embodiments, the invention includes a method of treating the disease described herein mediated at least in part by ATX comprising administering to a mammal in need thereof a therapeutically effective regimen comprising a composition described herein and at least one additional active agent. Generally, dosage levels on the order of from about 0.01 mg/kg to about 150 mg/kg of body weight per day are useful in the treatment of the above-indicated conditions, or alternatively about 0.5 mg to about 7 g per patient per day. For example, inflammation, cancer, psoriasis, allergy/asthma, disease and conditions of the immune system, disease and conditions of the Central Nervous System (CNS), may be effectively treated by the administration of from about 0.01 to 50 mg of the compound per kilogram of body weight per day, or alternatively about 0.5 mg to about 3.5 g per patient per day.

It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

EXAMPLES

While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention. As such, the following examples are provided to teach various aspects of the present invention. These examples represent individual embodiments of the aspects of this invention and one skilled in the art will recognize that additional examples can be generated in order to equally teach the aspects of the present invention.

Example 1. Preparation of amorphous compound I by spray-drying

Method: The compound I spray dried intermediates were produced using a ProCept 4M8-TRIX. The active spray solution formulations used are outlined in Table 1 . The spray solution was prepared the day of spraying to limit the solution hold time between solution preparations. Processing conditions for each of the solvent systems where selected based on previous conditions which have shown to produce particles with a shriveled sphere morphology.

Table 1 . Active Spray Solution Formulations

Spray Solution Preparation Method:

The solvent was dispensed into an appropriately sized container. A stir bar was inserted, and the container was placed on a magnetic stir plate. Stirring was begun and produced a gentle vortex. The solution was allowed to mix for at least 15 minutes. Compound I was added slowly to the vortex. The start and end time for compound I to become completely soluble and solution appearance was noted.

Spray Dried Material Preparation Method:

To begin sample preparation, the main power to the ProCept control unit and nitrogen unit was turned on and the process parameters entered. Once the pressure and oxygen level requirements were met for the spray dryer, the heater was engaged. The spray dryer was warmed up for at least one hour. The solvent wash spray solution was placed on a top loader balance and tared. Once the outlet temperature exceeded the target temperature for the solvent system, the spraying of the wash solution was begun. The wash solution bottle was tared, and pump speed determined to achieve a spray rate of 10 g/min. The inlet temperature was adjusted to target an outlet near to or at a target temperature for the solvent system. The active solution was dispensed, and feed rate determined. The pump speed was adjusted to maintain 10 g/min. The inlet temperature was adjusted to target an outlet near to or at the target temperature for the solvent system. After a steady state was reached, the inlet and outlet temperatures were recorded. The process was monitored until all solution was sprayed. The inlet temperature was adjusted as needed to target the required outlet temperature. The material was collected, and the mass weighed.

The spray dried intermediate material was tested by powder x-ray diffraction (PXRD), modulated differential scanning calorimetry (mDSC), thermogravimetric analysis (TGA), scanning electron microscope (SEM), bright field microscope (BFM) and polarized light microscope (PLM). The results are summarized in Table 2. These results demonstrate that all solvent systems rendered an amorphous version of compound I upon spray drying. All material was stored at 2-8 °C under desiccated conditions.

Table 2. Physical Characterization Summary

Example 2. Stability analysis of amorphous compound I in open conditions

Based on the initial results, all spray dried intermediates were placed on 40°C/75%RH open conditions for 1 week. The spray dried intermediate material was tested by powder x-ray diffraction (PXRD), modulated differential scanning calorimetry (mDSC), thermogravimetric analysis (TGA), scanning electron microscope (SEM), bright field microscope (BFM) and polarized light microscope (PLM). The results are summarized in Table 3.

Table 3. Open Condition Stability Data

All spray dried intermediates surprisingly remained amorphous with no observed crystalline growth over 1 -week stability at 40°C/75% RH. There was an increase in the percent weight loss for the TGA results varying from 1 to 2% for each of the solvent systems as compared with the initial TGA results. This is most likely due to water uptake from the high relative humidity. This increase had no observed effect on the physical stability of any of the spray dried intermediates. These results demonstrate that all solvent systems are viable to produce a stable amorphous form of compound I through spray drying.

Example 3. Stability analysis of amorphous compound I in closed conditions

To further test compound stability, two spray dried intermediates were placed in 40 Ό/75%BH closed desiccated conditions for 1 week. The spray dried intermediate material was tested by powder x- ray diffraction (PXRD), modulated differential scanning calorimetry (mDSC), thermogravimetric analysis (TGA), scanning electron microscope (SEM), bright field microscope (BFM) and polarized light microscope (PLM). The results are summarized in Table 4.

Table 4. Closed Condition Stability Data

The physical characterization testing showed that surprisingly both spray dried intermediates remained amorphous at 40 Ό/75%BH under closed desiccated conditions. Both spray dried

intermediates exhibited single Tg’s, an amorphous halo on the PXRD diffractogram, and no birefringence on the PLM images, indicating that there was no secondary crystal growth. TGA results did not show any meaningful change in residual solvent/water content when compared with the initial TGA results. These data demonstrate that both spray dried intermediates are stable under closed conditions.

Example 4. Kilogram scale preparation of amorphous compound I

Large scale preparation of amorphous compound I was also carried out. The spray solution consisted of neat compound I API in 100% acetone at 10% solids content and prepped using the weights described in Table 5. The solution was prepared the day of spraying by first adding acetone into a 36L stainless steel mixing vessel. The compound I API was then added to the solvent system and required 45 minutes of mixing with a top down mixer at a medium vortex. The solution had a clear appearance with very fine fiber-like particulates observed. The solution was filtered through a 1 .0 pm filter prior to spraying. The viscosity of the spray solution was measured to be 0.98 cP. Table 5. Compound I Solution Preparation Weights

The Mobile Minor was set up per Table 6 and warmed up for approximately one hour prior to spraying. The compound I spray solution was sprayed at a nozzle pressure of 400 psi and an outlet temperature of 40°C was targeted. Once steady state was reached, a sample was taken, and spraying was paused. The sample had a bulk/tap density of 0.192 g/ml, a particle size distribution of d(0.1 ) = 9.5 pm, d(0.5) = 22.9 pm, and d(0.9) = 48.9 pm, and SEM images showed a mixture of spheres and shriveled raisins. Targeting these settings generated a spray-dried intermediate (SDI) with acceptable physical characteristics; therefore, the remaining solution was sprayed at the same settings as the sample (Table 6). The SDI was dried overnight (~16 hours) in a Shel Vacuum Oven at 40°C under a pressure of -25 Hg vacuum with a nitrogen purge at 10 scfh. GC samples were taken at the beginning and end of drying for residual solvent analysis. As shown in Table 7, the samples taken after 16 hours of drying contained solvents beyond the limits for acetone. Therefore, drying of the SDI was continued for another day. Although the samples were found to be acceptable following 41 .5 hours of drying, the residual solvent concentration was close to the limit. For assurance, the sample was dried for an additional two days.

The GC results are shown in Table 7.

Table 6. Mobile Minor Setup/Set Points

Table 7. Compound I Residual Solvent Results

The SDI was unloaded from the oven after 4.5 days of drying. Subsequent tests were performed to analyze the bulk and tapped densities (Table 8) and particle size distribution (Table 9). The SDI was also analyzed via DSC and PXRD to verify that an amorphous SDI was generated. The amorphous diffractogram, lack of crystalline melt, and single T_g at 125.7 °C suggested that the SDI is homogenous and amorphous. The amount of water content in the compound was found by Karl Fischer analysis to be 0.8%. The chiral purity of the compound was found to be 99.59%.

The large-scale batch of compound I SDI was analyzed using a Phenom Pure Plus scanning electron microscope. The SDI consisted mostly of shriveled raisins and some spheres.

The compound I SDI achieved a bulk density of 0.2427 g/ml and a particle size distribution of d(0.1 ) = 9.63 pm, d(0.5) = 19.5 pm, and d(0.9) = 32.7 pm by targeting an outlet temperature of 40°C and a nozzle pressure of 400 psi. 1234.69 g of compound I SDI was collected after drying, resulting in a yield of 78.14%. The SEM images indicated that the SDI consisted mostly of shriveled raisins and some spheres. The DSC and PXRD results suggested that the SDI generated was homogenous and amorphous. The GC samples barely passed after 41 .5 hours of drying.

Table 8. Compound I Dried SDI Bulk/Tap Density Results

Table 9. Compound I Dried SDI Particle Size Distribution Results

Example 5. Stability analysis of kilogram scale batch of amorphous compound I

For the spray solution stability study, three stock solutions of 10% w-w compound I loading in acetone were prepared and placed at three conditions: 5°C (light protected), ambient (light protected), and unprotected exposure at ambient conditions in a light box. Samples were pulled and assessed for Assay/Rs and Chiral purity at t=0, t=1 day, and t=3 days respectively using HPLC. Results can be found in Table 10 and Table 1 1 below.

Table 10. Assay/RS Results - Spray Solution Stability

Table 1 1 . Chiral Purity Results - Spray Solution Stability

Compound I remained chemically stable in acetone for up to three days. A minor degradant peak was present at 0.06% area in the light exposed sample after three days of constant, direct exposure.

These results indicate that scaled up manufacturing/spray-drying would not require any special accommodations if prolonged storage of the solution containing vessel is not light permissible.

For further stability analysis, ~1 g of spray dried compound I amorphous material from the Example 4 batch taken following only 41 .5 hours of drying. This sample was then stored at ambient laboratory conditions for five days prior to analysis to mimic holding of wet massed material during typical larger scale spray drying productions. Chemical purity results are shown in Table 12 and Table 13 below.

Table 12. RS Results - Wet Hold Study

Table 13. Chiral Purity Results - Wet Hold Study

In comparing the chemical and physical purity results of the t=5-day Wet Hold Sample to that of the release t=0, immediately secondary dried SDI, there was essentially no difference. The data above suggests that a wet mass of spray dried amorphous compound I material can be held at ambient conditions for up to 5 days during a production setting with little or no effect on for final product quality.

Example 6. Preparation of amorphous compound I by evaporation

A slurry of compound I in 10% H2O in absolute ethanol (7.0 vol) was heated to 50 to 55 °C and stirred for 1 to 3 hours. The mixture was checked for dissolution and clarified at 45 to 50 °C. The filtrates were concentrated under reduced pressure at < 55 °C, transferred to drying trays, and dried under vacuum at up to 65 °C until the ethanol content was < 1 % w/w. The dried compound was sieved through a 1 4mm aperture sieve. The compound was obtained in a yield of 75 to 100% theoretical yield and 75 to 100% w/w.

Example 7. Stability analysis of amorphous compound I in closed conditions

To further test compound stability, amorphous compound I wasplaced in 25Ό/60%BH closed desiccated conditions for up to 24 months. The material was tested by appearance, Karl Fischer analysis, x-ray diffraction (XRPD), and high-performance liquid chromatography (HPLC). The results are summarized in Tables 14, 15, and 16. Table 14. Closed Condition Stability Data

nd= not detected

Table 15. Closed Condition Stability Data

nd= not detected Table 16. Closed Condition Stability Data

nd= not detected

The physical characterization testing showed that surprisingly the amorphous compound I remained amorphous and stable at 25°C/60%RH under closed desiccated conditions for up to 24 months. These data demonstrate that both spray dried intermediates are stable under closed conditions.

Example 8. Stability analysis of amorphous compound I in closed conditions

To further test compound stability, amorphous compound I placed in 25Ό/60%RH or

40Ό/75%RH closed desiccated conditions for up to 9 months. The material was tested by appearance, Karl Fischer analysis, x-ray diffraction (XRPD), and high-performance liquid chromatography (HPLC). The results are summarized in Tables 17 and 18.

Table 17. Closed Condition Stability Data at 25 °C/60% RH

NT=not tested

Table 18. Closed Condition Stability Data at 40 °C/75% RH

NT=not tested

The physical characterization testing showed that surprisingly the amorphous compound I remained amorphous and stable at 25°C/60%RH for up to 9 months and at 40°C/75%RH for up to 6 months under closed desiccated conditions. These data demonstrate that both spray dried intermediates are stable under closed conditions. OTHER EMBODIMENTS

It is to be understood that while the present disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the present disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and alterations are within the scope of the following claims.

Claims

What is claimed: CLAIMS

1 . A pharmaceutical composition comprising amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, wherein compound I has the structure:

I

2. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition comprises at least 90% by weight of amorphous compound I.

3. The pharmaceutical composition of claim 1 or 2, wherein the pharmaceutical composition further comprises at least 0.2% by weight of water.

4. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition comprises between 0.4% and 3% by weight of water.

5. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition comprises between 0.5% and 1 % by weight of water.

6. The pharmaceutical composition of any one of claims 1 to 5, wherein the amorphous compound I has an endothermic onset at about 125 °C in differential scanning calorimetry (DSC) profile.

7. The pharmaceutical composition of any one of claims 1 to 6, wherein the amorphous compound I has a loss of weight from 30 Ό to 150 °C of less than 2% as measured by thermal gravimetric analysis.

8. The pharmaceutical composition of any one of claims 1 to 7, wherein substantially no bi refringence is observed when the amorphous compound is analyzed by polarized light microscope.

9. The pharmaceutical composition of any one of claims 1 to 8, wherein an amorphous halo is observed when the amorphous compound is analyzed by x-ray diffraction.

10. The pharmaceutical composition of any one of claims 1 to 9, wherein the pharmaceutical composition is substantially free of amorphous state stabilizing agents.

1 1 . The pharmaceutical composition of any one of claims 1 to 10, wherein the purity of the pharmaceutical composition decreases by less than 1 % upon storage for at least 5 days.

12. The pharmaceutical composition of claim 1 1 , wherein the purity of the pharmaceutical composition decreases by less than 1 % upon storage for at least 6 months.

13. The pharmaceutical composition of claim 1 1 or 12, wherein the purity of the pharmaceutical composition decreases by less than 1 % upon storage for at least 24 months.

14. The pharmaceutical composition of any one of claims 1 1 to 13, wherein the storage comprises room temperature.

15. The pharmaceutical composition of any one of claims 1 1 to 14, wherein the storage comprises exposure to light.

16. The pharmaceutical composition of any one of claims 1 to 15, wherein the composition comprises about 5 mg of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof.

17. The pharmaceutical composition of any one of claims 1 to 15, wherein the composition comprises about 50 mg of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof.

18. The pharmaceutical composition of any one of claims 1 to 15, wherein the composition comprises about 250 mg of amorphous compound I, or an amorphous pharmaceutically acceptable salt thereof.

19. A method of producing a pharmaceutical composition of any one of claims 1 to 18, the method comprising:

(a) combining compound I and one or more solvents; and

(b) precipitating compound I from the mixture of step (a) by addition of an additional solvent.

20. A method of producing a pharmaceutical composition of any one of claims 1 to 18, the method comprising:

(a) combining compound I and a solvent; and

(b) evaporating the solvent of the mixture of step (a).

21 . The method of claim 20, wherein the evaporating comprises spray-drying.

22. The method of claim 20 or 21 , wherein the solvent is methanol, a methanol :water mixture, a tetrahydrofura water mixture, acetone, or an acetone:water mixture.

23 The method of any one of claims 20 to 22, wherein the solvent is acetone or an acetone:water mixture.

24. The method of any one of claims 20 to 23, wherein the method is conducted on large scale.

25. The method of claim 24, wherein at least 1 .0 kg of compound I is used in step (a).

26. The method of any one of claims 19 to 25, wherein the method further comprises drying the product of step (b) in an oven for at least 24 hours.

27. The method of any one of claims 19 to 26, wherein the purity of the pharmaceutical composition decreases by less than 1 % upon storage for at least 5 days.

28. The method of claim 27, wherein the storage comprises room temperature.

29. The method of claim 27 or 28, wherein the storage comprises exposure to light.

30. A method of decreasing the level or activity of autotaxin (ATX) in a cell or subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical composition of any one of claims 1 to 18.

31 . The method of claim 30, wherein the cell is a human cell.

32. The method of claim 30, wherein the subject is human.

33. A method of treating an ATX-related disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical composition of any one of claims 1 to 18.

34. The method of claim 33, wherein the ATX-related disorder is lung fibrosis, asthma, chronic obstructive pulmonary disease (COPD), renal fibrosis, acute kidney injury, chronic kidney disease, liver fibrosis, skin fibrosis, fibrosis of the gut, breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, glioblastoma, bone cancer, colon cancer, bowel cancer, head and neck cancer, melanoma, multiple myeloma, chronic lymphocytic leukemia, B cell lymphoma, T cell lymphoma, cancer pain, tumor metastasis, transplant organ rejection, scleroderma, ocular fibrosis, age related macular degeneration (AMD), diabetic retinopathy, collagen vascular disease, atherosclerosis, Raynaud's phenomenon, rheumatoid arthritis, osteoarthritis or neuropathic pain.

35. The method of claim 33 or 34, wherein the subject is human.