EA044424B1 - CRYSTAL FORMS AND SALT FORMS OF KINASE INHIBITOR - Google Patents

Description

Cross reference to related applications

This application claims priority to US Provisional Application Serial No. 62/779430 and US Provisional Application Serial No. 62/856469, the entire contents of which are incorporated herein by reference.

Field of technology

The present invention relates to crystalline forms of the free base of the c-Met inhibitor, Compound 1. The present invention also relates to crystalline forms of salts of Compound 1. The present invention also relates to pharmaceutical compositions containing solid polymorphs of the free base and salts of Compound 1. The present invention also relates to methods of treating a disease, disorder or syndrome mediated, at least in part, by modulating protein kinase activity in vivo.

State of the art

Human Axl belongs to the Tyro3, Axl and Mer (TAM) subfamily of receptor tyrosine kinases, which includes Mer. TAM kinases are characterized by an extracellular ligand-binding domain consisting of two immunoglobulin-like domains and two type III fibronectin domains. Axl is overexpressed in a number of tumor cell types and was originally cloned from patients with chronic myelogenous leukemia. When overexpressed, Axl exhibits transformative potential. Axl signaling is thought to induce tumor growth through activation of proliferative and antiapoptotic signaling pathways. Axl has been associated with cancers such as lung cancer, myeloid leukemia, uterine cancer, ovarian cancer, gliomas, melanoma, thyroid cancer, renal cell carcinoma, osteosarcoma, gastric cancer, prostate cancer, and breast cancer. Overexpression of Axl leads to a poor prognosis for patients with these cancers.

Activation of Mer, like Axl, transmits information to downstream signaling pathways that induce tumor growth and tumor activation. Mer binds ligands such as the soluble Gas-6 protein. Binding of Gas-6 to Mer induces autophosphorylation of Mer on its intracellular domain, resulting in activation of a downstream signal. Overexpression of Mer in cancer cells leads to increased metastasis, most likely through the formation of a soluble Mer extracellular domain protein as a decoy receptor. Tumor cells secrete a soluble form of the extracellular receptor Mer, which reduces the ability of the soluble Gas-6 ligand to activate Mer on endothelial cells, leading to cancer progression.

Therefore, there is a need for compounds that inhibit TAM receptor tyrosine kinases, such as Axl and Mer, for the treatment of selected types of cancer.

The essence of the invention

The present invention provides crystalline forms of the free base and selected salts of compound 1, K-(4-fluorophenyl)-K-(4-((7-methoxy-6-(methylcarbamoyl)quinolin-4yl)oxy)phenyl)cyclopropane-1,1 -dicarboxamide having the structure

N

Connection 1

Compound 1 is disclosed in WO 2019/148044, the contents of which are incorporated herein by reference in their entirety.

Particular crystalline forms of an active pharmaceutical ingredient (API), such as Compound 1, may have several advantages over other crystalline or amorphous forms, such as increased stability during storage or processing, more favorable solubility, and increased bioavailability. Several stable crystalline forms of Compound 1 and certain salts of Compound 1 are reported herein.

In one aspect, the present invention provides a crystalline solid form of Compound 1 or a hydrate or solvate thereof.

In another aspect, the present invention relates to a crystalline salt form of the hydrochloric acid compound 1 of the compound

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HCl salt of compound 1 or its hydrate or solvate.

In one aspect, the present invention provides a crystalline fumaric acid salt form of Compound 1 having the general structure

Compound 1 hemifumarate or a hydrate or solvate thereof, wherein the crystalline salt form is Compound 1 hemifumarate - 0.5 fumaric acid, characterized as Compound 1 hemifumarate Form B.

In another aspect, the present invention provides a crystalline fumaric acid salt form of Compound 1 having the general structure

Compound 1 fumarate or a hydrate or solvate thereof, wherein the crystalline salt form is compound 1-fumaric acid fumarate.

In one aspect, the present invention provides a crystalline phosphoric acid salt form of Compound 1 having the general structure

Compound 1 phosphate form A or its hydrate or solvate characterized as compound 1 phosphate form A.

In yet another aspect, the present invention provides a method of treating a disease, disorder or syndrome mediated at least in part by modulating the activity of a protein kinase in vivo, comprising administering to a subject in need thereof a crystalline form or a crystalline salt form described herein, or pharmaceutical composition described herein.

In another aspect, the present invention provides a method for inhibiting a protein kinase, the method comprising contacting the protein kinase with a crystalline form or a crystalline salt form as described herein.

In yet another aspect, the present invention relates to a method for preparing Compound 1 hemifumarate Form B, comprising contacting Compound 1 with fumaric acid in an organic solvent to form a mixture and stirring the mixture.

Brief description of graphic materials

In fig. Figure 1 shows the XRPD pattern of Form A of compound 1.

In fig. Figure 2 shows a DSC thermogram of Form A of compound 1.

In fig. Figure 3 shows a TGA thermogram of Form A of compound 1.

In fig. 4 is a plot of the DVS Form A isotherm of Compound 1 from 5% RH to 95% RH.

In fig. 5A-5D are high temperature micrographs showing Form A of Compound 1 at (A) 28.6°C with no change in crystalline form, (B) 210.0°C with no change in crystalline form, (C) 230.0°C with some melting and decomposition and (D) 231.3°C with complete melting and decomposition.

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In fig. Figure 6 shows the Form B XRPD pattern of compound 1.

In fig. Figure 7 shows a TGA thermogram of Form B of compound 1.

In fig. Figure 8 shows the XRPD pattern C of compound 1.

In fig. Figure 9 shows a TGA form C thermogram of compound 1.

In fig. Figure 10 shows the Form D XRPD pattern of compound 1.

In fig. Figure 11 shows a TGA form D thermogram of compound 1.

In fig. Figure 12 shows the Form E XRPD pattern of compound 1.

In fig. Figure 13 shows a TGA form E thermogram of compound 1.

In fig. Figure 14 shows the XRPD pattern of Form F of compound 1.

In fig. Figure 15 shows a TGA thermogram of Form F of compound 1.

In fig. Figure 16 shows the XRPD pattern of Form G of compound 1.

In fig. Figure 17 shows a TGA thermogram of Form G of compound 1.

In fig. Figure 18 shows the XRPD pattern of Form H of Compound 1.

In fig. Figure 19 shows the XRPD pattern of Form K of compound 1.

In fig. Figure 20 shows a DSC thermogram of Form K of compound 1.

In fig. Figure 21 shows a TGA thermogram of Form K of compound 1.

In fig. Figure 22 shows the Form O XRPD pattern of compound 1.

In fig. Figure 23 shows the XRPD pattern of Form P of compound 1.

In fig. Figure 24 shows a DSC form P thermogram of compound 1.

In fig. Figure 25 shows the Q form XRPD pattern of compound 1.

In fig. Figure 26 shows the DSC form Q thermogram of compound 1.

In fig. Figure 27 shows a TGA form Q thermogram of compound 1.

In fig. 28 shows the XRPD pattern of compound 1 fumarate form A + compound 1 form A (free base form).

In fig. 29 shows the XRPD pattern of Compound 1 hemifumarate Form B.

In fig. 30 shows a DSC thermogram of Compound 1 hemifumarate Form B.

In fig. 31 shows a TGA thermogram of Compound 1 hemifumarate Form B.

In fig. 32 is a plot of the DVS isotherm of Compound 1 hemifumarate Form B.

In fig. 33A-33D are high temperature micrographs showing Compound 1 Hemifumarate Form B at (A) 26.4°C with no change in crystalline form, (B) 209.6°C with no change in crystalline form, (C) 222.1°C with some melting and (D) 223.1 °C with complete melting and some darkening indicating decomposition.

In fig. 34 shows the XRPD pattern of HCl form A of compound 1.

In fig. 35 shows the XRPD pattern of HCl form B of compound 1.

In fig. 36 shows the XRPD pattern of HCl form C of compound 1.

In fig. 37 shows the XRPD pattern of HCl form D of compound 1.

In fig. 38 shows the results of indexing Form A of Compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 39 shows the results of indexing Form B of Compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 40 depicts the results of indexing Form D of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 41 depicts the results of indexing the H form of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 42 depicts the results of indexing the O form of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 43 depicts the results of indexing the P form of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 44 depicts the results of indexing the Q form of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 45 depicts the results of indexing Form B of the hemifumarate of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

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In fig. 46 depicts the results of indexing HCl Form A of Compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived values.

In fig. 47 depicts the results of indexing the HCl form B of compound 1, including the tabulated space group consistent with the specified extinction symbol, unit cell parameters, and derived quantities.

In fig. 48 shows the XRPD pattern of compound 1 phosphate form A.

In fig. 49 shows the Form I XRPD pattern of compound 1.

In fig. 50 shows the XRPD pattern of Form J of compound 1.

In fig. Figure 51 shows the Form L XRPD pattern of compound 1.

In fig. 52 shows the XRPD pattern of Form M of compound 1.

In fig. 53 shows the XRPD pattern of Form N of compound 1.

In fig. 54 shows the 1H NMR spectrum of Compound 1 hemifumarate Form B in DMSO-d ₆ .

In fig. 55 shows the 1H NMR spectrum of Form A of compound 1 in DMSO-d ₆ .

In fig. 56 shows the 1H NMR spectrum of Form K of compound 1 in DMSO-d ₆ .

Detailed description of the invention

Definitions, abbreviations and acronyms.

Analytical techniques

Abbreviations/Acronyms Full title/description DSC Differential scanning calorimetry DVS Dynamic (aqueous) vapor sorption HSM High temperature microscopy NMR Nuclear magnetic resonance spectroscopy OM Optical microscopy PLM Polarized light microscopy TGA Thermogravimetry or thermogravimetric analysis XRPD X-ray powder diffractometry

Experimental techniques

Abbreviations/Acronyms Full title/description CC Emergency cooling C.P. Emergency deposition F.C. Fast cooling F.E. Fast evaporation R.C. Reaction-induced crystallization S.C. Slow cooling S.E. Slow evaporation VD Vapor diffusion VS Vapor voltage

other information

Abbreviations/Acronyms Full title/description ~ Approximately or approximately API Active Pharmaceutical Ingredient B/E Birefringence and extinction Endo/endo Endotherm or endothermic eq. Equivalent Exo/exo Exotherm or exothermic FB Free base FF Free form frz Freezer LIMS Laboratory Information Management System Max/max Maximum or maximums Observation Observation P.O. Preferred Orientation ppt Sediment or sedimentation ref Fridge RH Relative humidity RT Room temperature Sol./solution Solution vac. Vacuum wt.% Mass percentage

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Solvents

Abbreviations/Acronyms Full title/description ACN Acetonitrile AcOH Acetic acid DCM Dichloromethane DMSO Dimethyl sulfoxide EtOAc Ethyl acetate EtOH Ethanol HFIPA Hexafluoroisopropanol IPA Isopropyl alcohol, 2-propanol MEK Methyl ethyl ketone MeOH Methanol MTBE Methyl tert-butyl ether TFE 2,2,2-Trifluoroethanol THF Tetrahydrofuran

For the purposes of this document, the following definitions apply unless otherwise noted.

For the purposes of the present invention, chemical elements are identified in accordance with the periodic table of the elements, CAS version, Handbook of Chemistry and Physics, 95th ed. Additionally, general principles of organic chemistry are described in Organic Chemistry, 2nd ed., Thomas Sorrell, University Science Books, Sausalito: 2006, and March's Advanced Organic Chemistry, 7th ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2013, the entire contents of which are incorporated herein by reference.

As used herein, the term low/limited/significant hygroscopicity refers to a substance that exhibits <0.5/<2.0/>2.0 wt% water absorption within a specified RH range.

As used herein, the term stoichiometric hydrate refers to a crystalline substance with a specific water content over an extended RH range. Typical stoichiometric hydrates are hemihydrates, monohydrates, sesquihydrates, dihydrates, etc.

As used herein, the term variable hydrate refers to a crystalline substance with variable water content over an extended RH range, but without phase change.

As used herein, the chemical term referred to as form refers to a chemical compound or a salt thereof that consists of a single phase.

As used herein, the term low/limited/intermediate/good/high solubility refers to a substance having a solubility of <1/1-20/20-100/100-200/>200 mg/ml.

As used herein, the term crystalline refers to a substance that results in an XRPD pattern with sharp peaks (similar to the width of instrumental peaks) and weak diffuse scattering around the peaks.

As used herein, the term disordered crystalline refers to a substance that results in an XRPD pattern with broad peaks (relative to the width of the instrumental peaks) and/or significant diffuse scattering relative to the peaks. Disordered substances can be

1) microcrystalline;

2) crystalline with a high density of defects;

3) mixtures of crystalline and X-ray amorphous phases; or

4) a combination of the above.

As used herein, the term undersignal means that spectrographic analysis of a sample results in a spectrum or pattern (output) having insufficient signal above the expected background noise.

As used herein, the term single crystal phase refers to an XRPD pattern that is considered to contain evidence of a single crystalline form due to the fact that the Bragg peaks are indexed by a single unit cell. Indexing is the process of assigning Miller index labels to each peak in a diffraction pattern. During the indexing process, the size and shape of the crystalline unit cell is also determined.

As used herein, the term suspension refers to a suspension prepared by adding a sufficient amount of solids to a specified solvent under ambient conditions such that undissolved solids are present. A typical suspension involves stirring (usually by stirring or oscillating), an action also called suspending, in a stoppered vial at a specific temperature in

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As used herein, the term X-ray amorphous or amorphous refers to a substance characterized by diffuse scattering but lacking evidence of Bragg peaks in the XRPD diagram.

As used herein, the term crystalline refers to compounds in a solid state that have a periodic and repeating three-dimensional internal arrangement of atoms, ions, or molecules, such as crystals arranged in fixed geometric patterns or lattices that have rigid long-range order. The term crystalline does not necessarily mean that the compound exists in the form of crystals, but that it has a crystal-like internal structure.

As used herein, the term essentially crystalline refers to a solid that is predominantly arranged in fixed geometric patterns or lattices that have strong long-range order. For example, substantially crystalline materials have a crystallinity greater than about 85% (eg, a crystallinity greater than about 90%, a crystallinity greater than about 95%, or a crystallinity greater than about 99%). It should also be noted that the term essentially crystalline includes the attribute crystalline, which is defined in the previous paragraph.

The term patient for the purposes of the present invention includes humans and any other animals, in particular mammals, and other organisms. Thus, these methods are applicable to both human and veterinary applications. In a preferred embodiment, the patient is a mammal, and in a most preferred embodiment, the patient is a human. Examples of preferred mammals include mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses and primates.

Kinase-dependent diseases or conditions refer to pathological conditions that depend on the activity of one or more kinases. Kinases are directly or indirectly involved in signal transduction pathways for various cellular activities, including proliferation, adhesion, migration, differentiation and invasion. Diseases associated with kinase activity include tumor growth, pathological neovascularization that supports solid tumor growth, and are associated with other diseases in which there is excessive local vascularization, such as eye diseases (diabetic retinopathy, age-related macular degeneration, etc. .) and inflammation (psoriasis, rheumatoid arthritis, etc.).

A therapeutically effective amount is an amount of a crystalline form or crystalline salt form of the present invention that, when administered to a patient, relieves a symptom of the disease. The amount of the crystalline form or crystalline salt form of the present invention that constitutes a therapeutically effective amount will vary depending on the compound, the pathological condition and its severity, the age of the patient being treated, and the like. A therapeutically effective amount can be determined in the usual manner by one skilled in the art in the light of his or her own knowledge and in the light of the present disclosure.

The phrase pharmaceutically acceptable is used herein to refer to those compounds, substances, compositions and/or dosage forms that, within the bounds of sound medical judgment, are suitable for use in contact with tissues of humans and animals without excessive toxicity, irritation, allergic reaction, immunogenicity or other problem or complication, commensurate with a reasonable balance of benefit and risk.

As used herein, the phrase pharmaceutically acceptable excipient refers to a pharmaceutically acceptable substance, composition or carrier, such as a liquid or solid excipient, diluent, solvent or encapsulating agent. Excipients are generally safe, non-toxic and are not biologically or otherwise objectionable and include excipients acceptable for veterinary use as well as for pharmaceutical use in humans. In one embodiment, each component is pharmaceutically acceptable as defined herein. See, for example, Remington: The Science and Practice of Pharmacy, 21st ed.; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 6th ed.; Rowe et al, eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed.; Ash and Ash eds.; Gower Publishing Company: 2007; Pharmaceutical Pref ormulation and Formulation, 2nd ed.; Gibson ed.; CRC Press LLC: Boca Raton, Fla., 2009.

Cancer refers to cell-proliferative pathological conditions, including, but not limited to, from the heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; from the head and neck: squamous cell carcinomas of the head and neck, laryngeal and hypopharyngeal cancer, nasal cavity cancer

-6044424 and paranasal sinuses, cancer of the nasopharynx, cancer of the salivary glands, cancer of the oral cavity and oropharynx; from the lungs: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma, non-small cell lung cancer), alveolar (bronchiolar) carcinoma, alveolar sarcoma, alveolar soft tissue sarcoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, meso thelioma; from the colon: colorectal cancer, adenocarcinoma, stromal tumors of the gastrointestinal tract, lymphoma, carcinoids, Turco syndrome; from the gastrointestinal tract: stomach cancer, adenocarcinoma of the gastroesophageal junction, from the esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), from the stomach (carcinoma, lymphoma, leiomyosarcoma), from the pancreas (ductal adenocarcinoma, insulinoma, glucagonoma , gastrinoma, carcinoid tumors, vipoma), from the small intestine (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), from the large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); from the breast: metastatic breast cancer, ductal carcinoma in situ, invasive ductal carcinoma, tubular carcinoma, medullary carcinoma, mucinous carcinoma, lobular carcinoma in situ, triple negative breast cancer; from the genitourinary tract: from the kidney (adenocarcinoma, Wilms tumor [nephroblastoma], lymphoma, leukemia, renal cell carcinoma, metastatic renal cell carcinoma), from the bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma, urothelial cancer), prostate (adenocarcinoma, sarcoma, castrate-resistant prostate cancer, bone metastases, bone metastases associated with castrate-resistant prostate cancer), testicular (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma , fibroma, adenomatoid tumors, lipoma), clear cell carcinoma, papillary carcinoma, penile cancer, squamous cell carcinoma of the penis; from the liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; from the bones: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulocellular sarcoma), multiple myeloma, malignant giant cell chordoma, osteochondroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma , chondromyxofibroma, osteoid osteoma and giant cell tumors; from the thyroid gland: medullary thyroid cancer, differentiated thyroid cancer, papillary thyroid cancer, follicular thyroid cancer, Hürthle cell cancer and anaplastic thyroid cancer; from the nervous system: from the skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), from the meninges (meningioma, meningiosarcoma, gliomatosis), from the brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma [pinealoma] , glioblastoma multiforme, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma), NF1, neurofibromatosis, plexiform neurofibromas; gynecological: from the uterus (endometrial cancer), from the cervix (cervical carcinoma, precancerous cervical dysplasia), from the ovaries (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosathecal cell tumors, cell tumors Sertoli-Leydig, dysgerminoma, malignant teratoma), vulvar (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vaginal (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma [embryonic rhabdomyosarcoma], fallopian tubes (carcinoma ); hematological: blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), myelofibrosis, polycythemia vera, essential thrombocythemia, Hodgkin's disease [malignant lymphoma]; from the skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevi/moles, lipoma, angioma, dermatofibroma, keloids, psoriasis; and from the adrenal glands: neuroblastoma. Thus, the term cancer cell as used herein includes a cell affected by any of the above conditions. In some embodiments, a compound or combination disclosed herein can be used to treat diseases including HIV, sickle cell disease, graft-versus-host disease, acute graft-versus-host disease, chronic graft-versus-host disease, and sickle cell disease.

In general, the nomenclature used in this application is based on naming conventions adopted by the International Union of Pure and Applied Chemistry (IUPAC). The chemical structures presented herein were obtained using CHEMDRAW®. Any open valency exhibited on a carbon, oxygen, or nitrogen atom in the structures given herein indicates the presence of a hydrogen atom.

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which is 1-M'-(4-fluorophenyl)-1-M-[4-[7-methoxy-6-(methylcarbamoyl)quinolin4-yl]oxyphenyl]cyclopropane-1,1-dicarboxamide or K'-(4 -fluorophenyl)-C-[4-[7-methoxy-6(methylcarbamoyl)quinolin-4-yl]oxyphenyl]cyclopropane-1,1-dicarboxamide or its salt, solvate or hydrate.

In some embodiments of this aspect, the salt is an inorganic salt, an organic salt, a pharmaceutically acceptable salt, or a chiral salt.

In one aspect, the present invention provides a crystalline solid form of Compound 1

Compound 1 which is 1-M'-(4-fluorophenyl)-1-M-[4-[7-methoxy-6-(methylcarbamoyl)quinolin4-yl]oxyphenyl]cyclopropane-1,1-dicarboxamide or K'- (4-fluorophenyl)-C-[4-[7-methoxy-6(methylcarbamoyl)quinolin-4-yl]oxyphenyl]cyclopropane-1,1-dicarboxamide or its hydrate or solvate.

In one embodiment of this aspect, the crystalline solid form of Compound 1 is characterized as Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form K, Form O, or Form Q.

In one embodiment, the crystalline solid form is characterized as Form A of Compound 1.

In yet another further embodiment, Form A of Compound 1 is characterized by one or more of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 5.48, 9.93, 10.83, 10 .98, 11.36, 11.79, 12.04, 12.25, 12.62, 14.33, 14.67, 15.33, 16.02, 16.51, 16.77, 18.07 , 19.09, 19.34, 19.60, 20.00, 20.46, 20.85, 21.45, 21.55, 21.76, 22.16, 22.35, 22.58, 22 .87, 23.79, 24.11, 24.29, 24.35, 24.87, 25.42, 25.81, 26.09, 26.72, 27.04, 27.44, 27.77 , 27.98, 28.19 and 28.56.

In another embodiment, Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 5.48, 9.93, 11.36, 11.79, 12 .04, 12.62, 14.33, 14.67, 15.33, 16.51, 18.07, 19.09, 20.00, 21.55, 22.35, 22.58, 24.29 , 24.35, 24.87, 28.19 and 28.56.

In another embodiment, Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 10.83, 10.98, 11.36, 11.79, 12 .04, 14.33, 18.07, 19.09, 20.00, 22.58, 24.87 and 28.19.

In a further embodiment, Form A of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where the peaks are 10.83, 10.98, 11.36, 11.79, 12.04, 14.33, 18.07, 19.09, 20.00, 22.58, 24.87 and 28.19.

In yet another further embodiment, Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where the peaks are 5.48, 9.93, 11.36, 11.79, 12. 04, 12.62, 14.33, 14.67, 15.33, 16.51, 18.07, 19.09, 20.00, 21.55, 22.35, 22.58, 24.29, 24.35, 24.87, 28.19 and 28.56.

In yet another further embodiment, Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where the peaks are 5.48, 9.93, 10.83, 10.98, 11. 36, 11.79, 12.04, 12.25, 12.62, 14.33, 14.67, 15.33, 16.02, 16.51, 16.77, 18.07, 19.09, 19.34, 19.60, 20.00, 20.46, 20.85, 21.45, 21.55, 21.76, 22.16, 22.35, 22.58, 22.87, 23, 79, 24.11, 24.29, 24.35, 24.87, 25.42, 25.81, 26.09, 26.72, 27.04, 27.44, 27.77, 27.98, 28.19 and 28.56.

In yet another additional embodiment, Form A of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 1.

In one embodiment, Form A of Compound 1 exhibits an endotherm above 200° C. on the DSC thermogram. In one embodiment, Form A of Compound 1 is characterized by an endotherm with a decomposition onset temperature greater than 200° C. on the DSC thermogram.

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In another embodiment, Form A of Compound 1 exhibits weight loss above 200° C. on the TGA thermogram.

In another embodiment, Form A of Compound 1 exhibits a weight increase of about 0.8 to about 1.0 wt%, determined by DVS analysis, when removed from a 5% relative humidity environment and placed in a 95% relative humidity environment. relative humidity.

In one embodiment, the crystalline solid form is characterized as Form B of Compound 1.

In one embodiment, Form B of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 4.76, 9.58, 10.49, 10.97, 11 .27, 12.10, 13.26, 13.52, 14.52, 15.15, 15.42, 16.69, 17.29, 17.92, 18.34, 19.05, 19.25 , 19.48, 20.04, 20.59, 20.90, 21.39, 21.84, 22.25, 22.68, 22.84, 23.12, 23.32, 23.60, 24 .03, 24.79, 25.32, 25.65, 25.88, 26.50, 26.79, 27.25, 28.55 and 29.49.

In one embodiment, Form B of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 9.58, 10.49, 11.27, 12.10, 13 .26, 13.52, 15.15 and 16.69.

In another embodiment, Form B of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 10.49, 12.10, 13.26 and 13.52.

In another embodiment, Form B of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 10.49, 12.10, 13.26 and 13.52.

In yet another further embodiment, Form B of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 9.58, 10.49, 11.27, 12.10, 13. 26, 13.52, 15.15 and 16.69.

In yet another embodiment, Form B of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 4.76, 9.58, 10.49, 10.97, 11.27 , 12.10, 13.26, 13.52, 14.52, 15.15, 15.42, 16.69, 17.29, 17.92, 18.34, 19.05, 19.25, 19 .48, 20.04, 20.59, 20.90, 21.39, 21.84, 22.25, 22.68, 22.84, 23.12, 23.32, 23.60, 24.03 , 24.79, 25.32, 25.65, 25.88, 26.50, 26.79, 27.25, 28.55 and 29.49.

In yet another further embodiment, Form B of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 6.

In one embodiment, Form B of Compound 1 exhibits a first mass loss of ~0.3 wt% between temperatures of 38 and 92°C and a second mass loss of ~11.2 wt% between temperatures of 92 and 188°C on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form C of Compound 1.

In one embodiment, Form C of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 3.89, 4.63, 7.95, 9.31, 10 .54, 10.87, 11.14, 11.31, 11.49, 11.75, 12.22, 12.96, 13.59, 13.84, 14.01, 14.62, 14.79 , 15.46, 15.86, 16.07, 16.61, 16.73, 16.88, 17.64, 18.13, 18.73, 19.10, 19.42, 19.75, 20 .09, 20.47, 21.00, 21.65, 21.95, 22.47, 23.11, 23.46, 23.77, 24.84, 25.17, 26.14, 26.48 , 26.88, 27.72, 28.35, 28.70 and 28.96.

In one embodiment, Form C of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 3.89, 7.95, 9.31, 10.54, 12 .96, 16.61, 17.64 and 20.47.

In another embodiment, Form C of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 3.89, 7.95, 9.31 and 17.64.

In another embodiment, Form C of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 3.89, 7.95, 9.31 and 17.64.

In yet another further embodiment, Form C of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 3.89, 7.95, 9.31, 10.54, 12. 96, 16.61, 17.64 and 20.47.

In yet another embodiment, Form C of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 3.89, 4.63, 7.95, 9.31, 10.54 , 10.87, 11.14, 11.31, 11.49, 11.75, 12.22, 12.96, 13.59, 13.84, 14.01, 14.62, 14.79, 15 .46, 15.86, 16.07, 16.61, 16.73, 16.88, 17.64, 18.13, 18.73, 19.10, 19.42, 19.75, 20.09 , 20.47, 21.00, 21.65, 21.95, 22.47, 23.11, 23.46, 23.77, 24.84, 25.17, 26.14, 26.48, 26 .88, 27.72, 28.35, 28.70 and 28.96.

In yet another further embodiment, Form C of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 8.

In one embodiment, Form C of Compound 1 exhibits a first mass loss of ~0.4 wt% between temperatures of 40 and 75°C, a second mass loss of ~13.8 wt% between temperatures of 75 and 154°C, and a third mass loss of ~1 .9% between temperatures 190 and 220°C on the TGA thermogram.

- 9 044424

In one embodiment, the crystalline solid form is characterized as Form D of Compound 1.

In one embodiment, Form D of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.08, 5.43, 7.00, 9.62, 10 .21, 10.90, 12.31, 13.66, 14.06, 14.70, 15.35, 16.06, 16.39, 17.89, 18.17, 18.35, 18.53 , 18.80, 18.96, 19.15, 19.50, 20.09, 20.37, 20.58, 20.93, 21.31, 21.79, 21.97, 22.30, 22 .91, 23.12, 23.26, 23.62, 23.93, 24.37, 24.77, 24.99, 25.39, 25.96, 26.62, 27.10, 27.53 , 28.05, 28.38, 28.78, 29.09, 29.38 and 29.64.

In one embodiment, Form D of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.43, 7.00, 10.21, 18.96, 23 .62, 24.99, 26.62, 27.10 and 29.64.

In another embodiment, Form D of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.43, 7.00, 10.21 and 29.64.

In another embodiment, Form D of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.43, 7.00, 10.21 and 29.64.

In yet another further embodiment, Form D of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.43, 7.00, 10.21, 18.96, 23. 62, 24.99, 26.62, 27.10 and 29.64.

In yet another embodiment, Form D of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.08, 5.43, 7.00, 9.62, 10.21 , 10.90, 12.31, 13.66, 14.06, 14.70, 15.35, 16.06, 16.39, 17.89, 18.17, 18.35, 18.53, 18 .80, 18.96, 19.15, 19.50, 20.09, 20.37, 20.58, 20.93, 21.31, 21.79, 21.97, 22.30, 22.91 , 23.12, 23.26, 23.62, 23.93, 24.37, 24.77, 24.99, 25.39, 25.96, 26.62, 27.10, 27.53, 28 .05, 28.38, 28.78, 29.09, 29.38 and 29.64.

In yet another further embodiment, Form D of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 10.

In another embodiment, Form D of Compound 1 exhibits a weight loss of ~13.5 wt% between 38 and 130° C. on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form E of Compound 1.

In one embodiment, Form E of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.16, 6.13, 9.77, 10.37, 10 .82, 11.69, 13.73, 14.34, 14.79, 15.47, 15.79, 16.33, 16.64, 16.82, 17.60, 17.89, 18.16 , 18.72, 19.09, 19.59, 20.65, 21.73, 22.10, 22.72, 23.23, 23.54, 23.79, 24.78, 25.13, 26 .37, 26.91, 29.12 and 29.95.

In one embodiment, Form E of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.13, 9.77, 10.37, 13.73, 14 .79, 26.37, 29.12 and 29.95.

In another embodiment, Form E of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 10.37, 14.79, 26.37, and 29.95.

In a further embodiment, Form E of compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 10.37, 14.79, 26.37 and 29.95.

In yet another further embodiment, Form E of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.13, 9.77, 10.37, 13.73, 14. 79, 26.37, 29.12 and 29.95.

In yet another further embodiment, Form E of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.16, 6.13, 9.77, 10.37, 10. 82, 11.69, 13.73, 14.34, 14.79, 15.47, 15.79, 16.33, 16.64, 16.82, 17.60, 17.89, 18.16, 18.72, 19.09, 19.59, 20.65, 21.73, 22.10, 22.72, 23.23, 23.54, 23.79, 24.78, 25.13, 26, 37, 26.91, 29.12 and 29.95.

In yet another further embodiment, Form E of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 12.

In another embodiment, Form E of compound 1 exhibits a mass loss of ~8.2 wt.% between temperatures of 60 and 130°C on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form F of Compound 1.

In one embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.85, 7.44, 8.56, 10.95, 11 .75, 12.28, 13.65, 14.48, 14.94, 15.61, 16.27, 16.68, 17.84, 18.39, 19.25, 19.52, 20.30 , 21.62, 22.07, 22.83, 23.58, 24.33, 25.93, 26.20, 26.48, 27.79, 29.2 and 29.9.

In one embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.44, 8.56, 13.65, 16.27, 19 .25, 25.93, 29.2 and 29.9.

- 10 044424

In another embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.44, 8.56,

13.65 and 29.9.

In a further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.44, 8.56, 13.65 and 29.9.

In yet another further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.44, 8.56, 13.65, 16.27, 19. 25, 25.93, 29.2 and 29.9.

In yet another further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.85, 7.44, 8.56, 10.95, 11. 75, 12.28, 13.65, 14.48, 14.94, 15.61, 16.27, 16.68, 17.84, 18.39, 19.25, 19.52, 20.30, 21.62, 22.07, 22.83, 23.58, 24.33, 25.93, 26.20, 26.48, 27.79, 29.2 and 29.9.

In one embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.85, 7.44, 8.56, 10.95, 11 .75, 12.28, 13.65, 14.48, 14.94, 15.61, 16.27, 16.68, 17.84, 18.39, 19.25, 19.52, 20.30 , 21.62, 22.07, 22.83, 23.58, 24.33, 25.93, 26.20, 26.48 and 27.79.

In one embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.44, 8.56, 13.65, 16.27, 19 .25 and 25.93.

In another embodiment, Form F of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.44, 8.56 and 13.65.

In a further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.44, 8.56 and 13.65.

In yet another further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.44, 8.56, 13.65, 16.27, 19. 25 and 25.93.

In yet another further embodiment, Form F of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.85, 7.44, 8.56, 10.95, 11. 75, 12.28, 13.65, 14.48, 14.94, 15.61, 16.27, 16.68, 17.84, 18.39, 19.25, 19.52, 20.30, 21.62, 22.07, 22.83, 23.58, 24.33, 25.93, 26.20, 26.48 and 27.79.

In yet another further embodiment, Form F of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 14.

In another embodiment, Form F 1 exhibits a first mass loss of ~0.1 wt.% between temperatures of 38 and 77°C and a second mass loss of ~14.4 wt.% between temperatures of 77 and 178°C on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form G of Compound 1.

In one embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 4.72, 6.71, 9.47, 11.51, 11 .84, 13.04, 14.40, 15.12, 16.03, 16.28, 16.51, 17.04, 17.85, 18.04, 18.73, 19.29, 19.49 , 19.73, 20.72, 21.10, 22.61, 23.16, 24.10, 25.49, 26.47, 27.25, 27.84, 30.3 and 30.7.

In one embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.71, 9.47, 14.40, 17.04, 17 .85, 21.10, 30.3 and 30.7.

In another embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.71, 9.47, 30.3 and 30.7.

In a further embodiment, Form G of compound 1 is characterized by all of the following peaks in the XRPD pattern on a scale of 2-theta±0.2, where the peaks are 6.71, 9.47, 30.3 and 30.7.

In yet another additional embodiment, Form G of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.71, 9.47, 14.40, 17.04, 17. 85, 21.10, 30.3 and 30.7.

In yet another additional embodiment, Form G of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 4.72, 6.71, 9.47, 11.51, 11. 84, 13.04, 14.40, 15.12, 16.03, 16.28, 16.51, 17.04, 17.85, 18.04, 18.73, 19.29, 19.49, 19.73, 20.72, 21.10, 22.61, 23.16, 24.10, 25.49, 26.47, 27.25, 27.84, 30.3 and 30.7.

In one embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 4.72, 6.71,

- 11 044424

9.47, 11.51, 11.84, 13.04, 14.40, 15.12, 16.03, 16.28, 16.51, 17.04, 17.85, 18.04, 18, 73, 19.29, 19.49, 19.73,

20.72, 21.10, 22.61, 23.16, 24.10, 25.49, 26.47, 27.25 and 27.84.

In one embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.71, 9.47,

14.40, 17.04, 17.85 and 21.10.

In another embodiment, Form G of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.71 and 9.47.

In a further embodiment, Form G of compound 1 is characterized by all of the following peaks in the XRPD pattern on a scale of 2-theta±0.2, where the peaks are 6.71 and 9.47.

In yet another additional embodiment, Form G of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.71, 9.47, 14.40, 17.04, 17. 85 and 21.10.

In yet another additional embodiment, Form G of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 4.72, 6.71, 9.47, 11.51, 11. 84, 13.04, 14.40, 15.12, 16.03, 16.28, 16.51, 17.04, 17.85, 18.04, 18.73, 19.29, 19.49, 19.73, 20.72, 21.10, 22.61, 23.16, 24.10, 25.49, 26.47, 27.25 and 27.84.

In yet another further embodiment, Form G of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 16.

In one embodiment, Form G of compound 1 exhibits a mass loss of ~20.8 wt.% between temperatures of 40 and 165°C on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form H of Compound 1.

In another embodiment, Form H of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.30, 10.79, 11.42, 11.73, 12 .63, 14.01, 14.29, 14.67, 15.74, 16.41, 17.23, 17.52, 18.01, 18.31, 18.56, 19.04, 19.67 , 19.80, 20.32, 20.72, 21.53, 21.69, 21.95, 22.47, 23.14, 23.53, 24.33, 24.84, 25.13, 25 .38, 25.69, 26.75, 27.48, 28.19, 28.70, 29.09 and 29.60.

In another embodiment, Form H of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.30, 11.42, 11.73, 17.52, 18 .01, 18.56, 21.95 and 25.69.

In another embodiment, Form H of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.30, 17.52, 18.56 and 25.69.

In a further embodiment, Form H of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.30, 17.52, 18.56 and 25.69.

In yet another further embodiment, Form H of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.30, 11.42, 11.73, 17.52, 18. 01, 18.56, 21.95 and 25.69.

In yet another embodiment, Form H of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.30, 10.79, 11.42, 11.73, 12.63 , 14.01, 14.29, 14.67, 15.74, 16.41, 17.23, 17.52, 18.01, 18.31, 18.56, 19.04, 19.67, 19 .80, 20.32, 20.72, 21.53, 21.69, 21.95, 22.47, 23.14, 23.53, 24.33, 24.84, 25.13, 25.38 , 25.69, 26.75, 27.48, 28.19, 28.70, 29.09 and 29.60.

In yet another further embodiment, Form H of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 18.

In one embodiment, the crystalline solid form is characterized as Form I of Compound 1.

In a further embodiment, Form I of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 49.

In one embodiment, the crystalline solid form is characterized as Form J of Compound 1.

In a further embodiment, Form J of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 50.

In one embodiment, the crystalline solid form is characterized as Form K of Compound 1.

In one embodiment, Form K of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.73, 6.39, 8.10, 11.53, 11 .78, 12.83, 14.36, 15.56, 16.25, 17.42, 18.17, 19.07, 19.70, 19.89, 20.53, 21.11, 21.55 , 22.34, 22.50, 23.24, 23.76, 24.50, 25.94, 26.42, 27.76 and 28.28.

- 12 044424

In one embodiment, Form K of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.39, 8.10,

11.53, 19.89, 21.11, 22.34, 24.50 and 26.42.

In another embodiment, Form K of compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.39, 8.10, 22.34 and 24.50.

In a further embodiment, Form K of compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.39, 8.10, 22.34 and 24.50.

In yet another additional embodiment, Form K of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.39, 8.10, 11.53, 19.89, 21. 11, 22.34, 24.50 and 26.42.

In yet another additional embodiment, Form K of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.73, 6.39, 8.10, 11.53, 11. 78, 12.83, 14.36, 15.56, 16.25, 17.42, 18.17, 19.07, 19.70, 19.89, 20.53, 21.11, 21.55, 22.34, 22.50, 23.24, 23.76, 24.50, 25.94, 26.42, 27.76 and 28.28.

In yet another additional embodiment, Form K of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 19.

In another embodiment, Form K of Compound 1 exhibits an endotherm at approximately 226° C. on the DSC thermogram.

In another embodiment, Form K of Compound 1 exhibits a mass loss of ~0.2 wt.% between temperatures of 40 and 180°C on the TGA thermogram.

In one embodiment, the crystalline solid form is characterized as Form L of Compound 1.

In a further embodiment, Form L of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 51.

In one embodiment, the crystalline solid form is characterized as Form M of Compound 1.

In a further embodiment, Form M of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 52.

In one embodiment, the crystalline solid form is characterized as Form N of Compound 1.

In a further embodiment, Form N of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 53.

In one embodiment, the crystalline solid form is characterized as Form O of Compound 1.

In one embodiment, Form O of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.10, 9.01, 9.83, 10.68, 11 ,12, 11.33, 12.25, 12.99, 13.93, 14.51, 14.92, 15.55, 15.79, 17.14, 17.43, 17.58, 18.15 , 18.42, 19.35, 19.77, 20.24, 20.71, 20.90, 21.49, 21.68, 22.04, 22.36, 22.78, 23.37, 23 .96, 24.39, 24.92, 25.62, 26.20, 26.64, 26.93, 27.32, 27.68, 27.96, 28.26, 28.60 and 28.81 .

In one embodiment, Form O of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.10, 9.01, 14.92, 17.14, 17 .58, 23.96, 25.62 and 27.96.

In another embodiment, Form O of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.10, 14.92, 17.14, and 23.96.

In a further embodiment, Form O of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.10, 14.92, 17.14 and 23.96.

In yet another further embodiment, Form O of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.10, 9.01, 14.92, 17.14, 17. 58, 23.96, 25.62 and 27.96.

In yet another further embodiment, Form O of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.10, 9.01, 9.83, 10.68, 11. 12, 11.33, 12.25, 12.99, 13.93, 14.51, 14.92, 15.55, 15.79, 17.14, 17.43, 17.58, 18.15, 18.42, 19.35, 19.77, 20.24, 20.71, 20.90, 21.49, 21.68, 22.04, 22.36, 22.78, 23.37, 23, 96, 24.39, 24.92, 25.62, 26.20, 26.64, 26.93, 27.32, 27.68, 27.96, 28.26, 28.60 and 28.81.

In yet another further embodiment, Form O of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 22.

In one embodiment, the crystalline solid form is characterized as Form P of Compound 1.

- 13 044424

In one embodiment, Form P of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.99, 8.78,

9.40, 10.12, 11.99, 14.61, 14.87, 15.61, 15.98, 16.32, 16.62, 17.56, 17.62, 17.84, 18, 05, 18.43, 18.88, 19.22,

19.72, 19.85, 20.32, 20.91, 21.67, 22.04, 22.39, 22.93, 23.46, 23.71, 23.98, 24.11, 24, 43, 24.84, 25.74, 26.39,

26.64, 26.85, 27.77, 28.74, 29.26, 29.55 and 30.07.

In one embodiment, Form P of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 11.99, 14.61, 14.87, 20.91, 21 .67, 22.04, 22.93 and 26.85.

In yet another further embodiment, Form P of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 11.99, 14.61, 14.87, 20.91, 21. 67, 22.04, 22.93 and 26.85.

In yet another further embodiment, Form P of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.99, 8.78, 9.40, 10.12, 11. 99, 14.61, 14.87, 15.61, 15.98, 16.32, 16.62, 17.56, 17.62, 17.84, 18.05, 18.43, 18.88, 19.22, 19.72, 19.85, 20.32, 20.91, 21.67, 22.04, 22.39, 22.93, 23.46, 23.71, 23.98, 24, 11, 24.43, 24.84, 25.74, 26.39, 26.64, 26.85, 27.77, 28.74, 29.26, 29.55 and 30.07.

In a further embodiment, Form P of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 23.

In one embodiment, the crystalline solid form is characterized as Form Q of Compound 1.

In one embodiment, Form Q of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.11, 8.61, 9.06, 9.74, 10 .64, 10.89, 11.24, 11.33, 12.06, 12.24, 12.91, 13.82, 14.46, 14.83, 15.69, 15.76, 16.07 , 17.05, 17.31, 17.40, 17.78, 18.16, 18.42, 18.88, 19.08, 19.28, 19.56, 19.84, 20.07, 20 .70, 21.04, 21.38, 21.59, 21.91, 22.18, 22.30, 22.58, 22.78, 23.04, 23.23, 23.50, 23.81 , 24.01, 24.32, 24.86, 25.43, 25.80, 26.05, 26.20, 26.69, 27.02, 27.44, 27.63, 27.99, 28 .48, 28.75, 29.17 and 29.36.

In one embodiment, Form Q of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.11, 8.61, 9.06, 9.74, 15 .69, 16.07, 20.04 and 24.01.

In another embodiment, Form Q of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 8.61, 9.74, 16.07 and 20.04.

In a further embodiment, Form Q of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 8.61, 9.74, 16.07 and 20.04.

In yet another further embodiment, Form Q of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.11, 8.61, 9.06, 9.74, 15, 69, 16.07, 20.04 and 24.01.

In yet another further embodiment, Form Q of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.11, 8.61, 9.06, 9.74, 10, 64, 10.89, 11.24, 11.33, 12.06, 12.24, 12.91, 13.82, 14.46, 14.83, 15.69, 15.76, 16.07, 17.05, 17.31, 17.40, 17.78, 18.16, 18.42, 18.88, 19.08, 19.28, 19.56, 19.84, 20.07, 20, 70, 21.04, 21.38, 21.59, 21.91, 22.18, 22.30, 22.58, 22.78, 23.04, 23.23, 23.50, 23.81, 24.01, 24.32, 24.86, 25.43, 25.80, 26.05, 26.20, 26.69, 27.02, 27.44, 27.63, 27.99, 28, 48, 28.75, 29.17 and 29.36.

In yet another further embodiment, Form Q of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 25.

In another embodiment, Form Q of Compound 1 exhibits an endotherm at approximately 194-195° C. on the DSC thermogram. In another embodiment, Form Q of Compound 1 is characterized by an endotherm with a decomposition onset temperature of approximately 194-195° C. on the DSC thermogram.

In another embodiment, Form Q of Compound 1 exhibits a mass loss of ~11-12 wt.% between temperatures of 120 and 160° C. on the TGA thermogram.

In another aspect, the present invention relates to a crystalline salt form of the hydrochloric acid compound 1 of the compound

HCl salt of compound 1

- 14 044424 or its hydrate or solvate.

In one embodiment, a crystalline solid form characterized as

HCl Form A of Compound 1, HCl Form B of Compound 1, HCl Form C of Compound 1, or HCl Form D of Compound 1. In one embodiment, the crystalline solid form is characterized as

HCl form A of compound 1.

In one embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.19, 8.17, 9.84, 10.10 , 10.42, 11.07, 12.52, 12.76, 12.98, 13.49, 13.69, 13.89, 14.31, 14.84, 15.12, 15.68, 16 .34, 16.68, 17.08, 17.47, 17.96, 18.49, 19.23, 19.78, 20.31, 20.91, 21.16, 21.42, 22.10 , 22.81, 23.18, 23.89, 24.39, 25.20, 25.87, 26.34, 27.06, 27.59, 28.07, 28.4 and 30.0.

In one embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.19, 13.49, 13.69, 13.89 , 14.84, 15.12, 16.34, 16.68, 17.47, 18.49, 20.31, 23.18, 24.39, 25.87, 26.34, 27.06, 28 .07, 28.4 and 30.0.

In another embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 13.49, 17.47, 18.49, and 30.0 .

In a further embodiment, HCl Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 13.49, 17.47, 18.49 and 30.0.

In yet another additional embodiment, HCl Form A of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.19, 13.49, 13.69, 13.89, 14.84, 15.12, 16.34, 16.68, 17.47, 18.49, 20.31, 23.18, 24.39, 25.87, 26.34, 27.06, 28, 07, 28.4 and 30.0.

In yet another additional embodiment, HCl Form A of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.19, 8.17, 9.84, 10.10, 10.42, 11.07, 12.52, 12.76, 12.98, 13.49, 13.69, 13.89, 14.31, 14.84, 15.12, 15.68, 16, 34, 16.68, 17.08, 17.47, 17.96, 18.49, 19.23, 19.78, 20.31, 20.91, 21.16, 21.42, 22.10, 22.81, 23.18, 23.89, 24.39, 25.20, 25.87, 26.34, 27.06, 27.59, 28.07, 28.4 and 30.0.

In one embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.19, 8.17, 9.84, 10.10 , 10.42, 11.07, 12.52, 12.76, 12.98, 13.49, 13.69, 13.89, 14.31, 14.84, 15.12, 15.68, 16 .34, 16.68, 17.08, 17.47, 17.96, 18.49, 19.23, 19.78, 20.31, 20.91, 21.16, 21.42, 22.10 , 22.81, 23.18, 23.89, 24.39, 25.20, 25.87, 26.34, 27.06, 27.59 and 28.07.

In one embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 5.19, 13.49, 13.69, 13.89 , 14.84, 15.12, 16.34, 16.68, 17.47, 18.49, 20.31, 23.18, 24.39, 25.87, 26.34, 27.06 and 28 .07.

In another embodiment, HCl Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 13.49, 17.47 and 18.49.

In a further embodiment, HCl Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 13.49, 17.47 and 18.49.

In yet another additional embodiment, HCl Form A of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.19, 13.49, 13.69, 13.89, 14.84, 15.12, 16.34, 16.68, 17.47, 18.49, 20.31, 23.18, 24.39, 25.87, 26.34, 27.06 and 28, 07.

In yet another additional embodiment, HCl Form A of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 5.19, 8.17, 9.84, 10.10, 10.42, 11.07, 12.52, 12.76, 12.98, 13.49, 13.69, 13.89, 14.31, 14.84, 15.12, 15.68, 16, 34, 16.68, 17.08, 17.47, 17.96, 18.49, 19.23, 19.78, 20.31, 20.91, 21.16, 21.42, 22.10, 22.81, 23.18, 23.89, 24.39, 25.20, 25.87, 26.34, 27.06, 27.59 and 28.07.

In yet another further embodiment, HCl Form A of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 34.

In one embodiment, the crystalline solid form is characterized as HCl Form B of Compound 1.

In one embodiment, HCl Form B of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 8.20, 9.72, 9.81, 10.04 , 10.56, 12.52, 12.97, 13.32, 13.48, 13.81, 14.35, 14.95, 15.89, 16.63, 17.37, 17.83, 17 .99, 18.32, 19.15, 19.31, 19.51, 19.72, 20.17, 20.84, 21.04, 21.15, 21.30, 21.81, 22.02 , 22.65, 23.11, 23.40, 23.75, 24.76, 25.34, 25.74, 26.20, 26.90, 27.71, 27.98, 28.34 and 28 .98.

- 15 044424

In one embodiment, HCl Form B of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from

9.72, 15.89, 16.63, 17.37, 18.32, 19.51, 21.04, 21.30, 21.81, 23.40, 24.76, 26.20 and 27, 71.

In another embodiment, HCl Form B of Compound 1 is characterized by one or more peaks in the XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 9.72, 17.37, 18.32, and 19.51 .

In a further embodiment, HCl Form B of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 9.72, 17.37, 18.32 and 19.51.

In yet another further embodiment, HCl Form B of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 9.72, 15.89, 16.63, 17.37, 18.32, 19.51, 21.04, 21.30, 21.81, 23.40, 24.76, 26.20 and 27.71.

In yet another further embodiment, HCl Form B of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 8.20, 9.72, 9.81, 10.04, 10.56, 12.52, 12.97, 13.32, 13.48, 13.81, 14.35, 14.95, 15.89, 16.63, 17.37, 17.83, 17, 99, 18.32, 19.15, 19.31, 19.51, 19.72, 20.17, 20.84, 21.04, 21.15, 21.30, 21.81, 22.02, 22.65, 23.11, 23.40, 23.75, 24.76, 25.34, 25.74, 26.20, 26.90, 27.71, 27.98, 28.34 and 28, 98.

In yet another further embodiment, HCl Form B of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 35.

In one embodiment, the crystalline solid form is characterized as HCl Form C of Compound 1.

In yet another additional embodiment, HCl Form C of Compound 1 is characterized by one or more of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 2.5, 3.0, 4.3, 5, 1, 6.2, 6.8, 7.3, 7.8, 8.8, 10.6, 11.6, 12.5, 13.3, 13.8, 15.3, 15.7, 17.1, 17.8, 19.0, 19.4, 20.0, 20.5, 20.8, 21.5, 22.2, 22.6, 23.0, 23.5, 23, 9, 25.2, 26.2, 26.8, 27.2, 28.0, 28.9 and 29.5.

In one embodiment, HCl Form C of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 2.5, 3.0, 4.3, 6.2 , 7.3, 7.8 and 29.5.

In another embodiment, HCl Form C of Compound 1 is characterized by one or more peaks in the XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 2.5, 3.0, 4.3, 11.6 , 17.1, 19.0, 20.5, 26.8 and 29.5.

In one embodiment, HCl Form C of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 2.5, 3.0, 4.3, 6.2, 7, 3, 7.8 and 29.5.

In yet another further embodiment, HCl Form C of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 2.5, 3.0, 4.3, 6.2, 7.3, 7.8, 8.8, 11.6, 17.1, 19.0, 20.5, 26.8 and 29.5.

In yet another further embodiment, HCl Form C of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 2.5, 3.0, 4.3, 5.1, 6.2, 6.8, 7.3, 7.8, 8.8, 10.6, 11.6, 12.5, 13.3, 13.8, 15.3, 15.7, 17, 1, 17.8, 19.0, 19.4, 20.0, 20.5, 20.8, 21.5, 22.2, 22.6, 23.0, 23.5, 23.9, 25.2, 26.2, 26.8, 27.2, 28.0, 28.9 and 29.5.

In yet another further embodiment, HCl Form C of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 36.

In one embodiment, the crystalline solid form is characterized as HCl Form D of Compound 1.

In yet another additional embodiment, HCl Form D of Compound 1 is characterized by one or more of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 3.47, 5.27, 6.93, 8. 21, 8.97, 9.86, 10.16, 10.44, 10.69, 11.28, 12.26, 12.75, 13.27, 13.92, 14.23, 14.54, 14.95, 15.44, 15.58, 15.80, 16.08, 16.25, 17.84, 18.44, 18.65, 19.34, 19.75, 20.13, 20, 93, 21.29, 22.05, 22.69, 22.90, 23.69, 24.15, 24.39, 24.60, 24.91, 25.16, 26.27, 27.03, 27.61 and 28.37.

In one embodiment, HCl Form D of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 3.47, 5.27, 10.16, 10.69 , 12.26, 14.54, 14.95, 17.84, 20.93, 21.29, 22.05, 22.69, 22.90, 23.69, 24.91 and 25.16.

In a further embodiment, HCl Form D of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale; the peaks are 3.47, 5.27, 10.16, 10.69, 12.26, 14.54, 14.95, 17.84, 20.93, 21.29, 22.05, 22.69, 22.90, 23.69, 24.91 and 25.16.

In yet another further embodiment, HCl Form D of Compound 1 exhibits all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 3.47, 5.27, 6.93, 8.21, 8.97, 9.86, 10.16, 10.44, 10.69, 11.28, 12.26, 12.75, 13.27, 13.92, 14.23, 14.54, 14, 95, 15.44, 15.58, 15.80, 16.08, 16.25, 17.84, 18.44, 18.65, 19.34, 19.75, 20.13, 20.93, 21.29, 22.05, 22.69, 22.90, 23.69, 24.15, 24.39, 24.60, 24.91, 25.16, 26.27, 27.03, 27, 61 and 28.37.

- 16 044424

In a further embodiment, the Hcl form D of compound 1 is characterized by the pattern

XRPD, essentially identical to FIG. 37.

In one aspect, the present invention provides a crystalline fumaric acid salt form of Compound 1 having the general structure

Compound 1 hemifumarate or a hydrate or solvate thereof, wherein the crystalline salt form is Compound 1-0.5 fumaric acid hemifumarate.

In another aspect, the present invention provides a crystalline fumaric acid salt form of Compound 1 having the general structure

Compound 1 fumarate or a hydrate or solvate thereof, wherein the crystalline salt form is compound 1-fumaric acid fumarate.

In one embodiment, the crystalline solid form is characterized as Compound 1 fumarate Form A.

In a further embodiment, Fumarate Form A of Compound 1 exhibits an XRPD pattern substantially identical to FIG. 28.

In one aspect, the present invention includes a crystalline salt form of fumaric acid of Compound 1 having the structure

Compound 1 hemifumarate or its hydrate or solvate.

In one embodiment of this aspect, the fumaric acid salt crystalline form is characterized as Compound 1 hemifumarate Form B.

In one embodiment, the crystalline solid form is characterized as Compound 1 hemifumarate Form B.

In one embodiment, Compound 1 hemifumarate Form B is characterized by one or more of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.55, 7.92, 9.08, 9, 40, 10.81, 11.18, 13.24, 13.35, 14.47, 14.90, 15.14, 15.89, 16.64, 16.95, 17.14, 17.29, 17.44, 17.79, 18.24, 18.34, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22.29, 22, 48, 22.75, 23.82, 24.37, 24.97, 25.17, 25.69, 26.34, 26.75, 27.05, 27.35, 27.50 and 27.88.

In one embodiment, Compound 1 hemifumarate Form B is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.55, 9.08, 10.81, 13.24, 15.89, 16.95, 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21, 74, 22.29, 22.48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88.

In one embodiment, Compound 1 hemifumarate Form B is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 9.08, 10.81, 16.95, 17.44, 22.29, 22.48, 23.82, 24.37, 26.34 and 27.05.

In one embodiment, Compound 1 Hemifumarate Form B is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 9.08, 10.81, 16.95, 17.44, 22.29 , 22.48, 23.82, 24.37, 26.34 and 27.05.

In one embodiment, Hemifumarate Form B of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.55, 9.08, 10.81, 13.24, 15.89 , 16.95, 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22 .29, 22.48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88.

- 17 044424

In one embodiment, Compound 1 Hemifumarate Form B is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.55, 7.92, 9.08,

9.40, 10.81, 11.18, 13.24, 13.35, 14.47, 14.90, 15.14, 15.89, 16.64, 16.95, 17.14, 17, 29, 17.44, 17.79, 18.24,

18.34, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22.29, 22.48, 22.75, 23.82, 24, 37, 24.97, 25.17, 25.69,

26.34, 26.75, 27.05, 27.35, 27.50 and 27.88.

In yet another further embodiment, Compound 1 Hemifumarate Form B exhibits an XRPD pattern substantially identical to FIG. 29.

In one embodiment, Compound 1 hemifumarate Form B exhibits an endotherm at approximately 226° C. on the DSC thermogram. In one embodiment, Compound 1 hemifumarate Form B exhibits an endotherm with a decomposition onset temperature of approximately 226° C. on the DSC thermogram.

In one embodiment, Compound 1 hemifumarate Form B exhibits negligible weight loss at approximately 220° C. on the TGA thermogram.

In one embodiment, Compound 1 hemifumarate Form B exhibits a mass increase of approximately 0.2 wt%, as measured by DVS, in an environment when removed from 5% relative humidity and placed in 95% relative humidity.

In one aspect, the present invention provides a crystalline phosphoric acid salt form of Compound 1 having the general structure

Compound 1 phosphate form A or its hydrate or solvate characterized as compound 1 phosphate form A.

In one embodiment, Compound 1 Phosphate Form A is characterized by one or more of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.3, 6.8, 10.3, 10, 5, 11.4, 12.7, 13.8, 14.7, 15.7, 16.1, 17.3, 17.5, 18.1, 18.8, 19.4, 20.3, 20.9, 21.2, 22.1, 22.7, 23.2, 23.6, 24.7, 25.5, 27.4, 27.8, 28.5, 29.1 and 29, 3.

In one embodiment, Compound 1 Phosphate Form A is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.3, 6.8, 10.5, 12.7, 13.8, 16.1, 17.3, 18.1, 18.8, 19.4, 20.3, 20.9, 21.2, 22.1, 23.2, 24.7, 27, 4, 27.8 and 28.5.

In another embodiment, the phosphate form A of compound 1 is characterized by one or more peaks in the XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.3, 6.8, 13.8, 16.1, 19.4, 20.3, 23.2 and 24.7.

In a further embodiment, Compound 1 Phosphate Form A is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.3, 6.8, 13.8, 16.1, 19.4 , 20.3, 23.2 and 24.7.

In yet another further embodiment, Compound 1 Phosphate Form A is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.3, 6.8, 10.5, 12.7, 13 ,8, 16.1, 17.3, 18.1, 18.8, 19.4, 20.3, 20.9, 21.2, 22.1, 23.2, 24.7, 27.4 , 27.8 and 28.5.

In yet another further embodiment, Compound 1 Phosphate Form A is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.3, 6.8, 10.3, 10.5, 11 ,4, 12.7, 13.8, 14.7, 15.7, 16.1, 17.3, 17.5, 18.1, 18.8, 19.4, 20.3, 20.9 , 21.2, 22.1, 22.7, 23.2, 23.6, 24.7, 25.5, 27.4, 27.8, 28.5, 29.1 and 29.3.

In yet another further embodiment, Compound 1 Phosphate Form A exhibits an XRPD pattern substantially identical to FIG. 48.

In one aspect, the present invention includes a crystalline solid form of Compound 1

N

Compound 1 or a hydrate or solvate thereof, characterized as Form A of Compound 1 by at least one of the following:

(i) one or more peaks in the XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 5.48, 9.93, 10.83, 10.98, 11.36, 11.79 , 12.04, 12.25, 12.62, 14.33, 14.67, 15.33, 16.02,

- 18 044424

16.51, 16.77, 18.07, 19.09, 19.34, 19.60, 20.00, 20.46, 20.85, 21.45, 21.55, 21.76, 22, 16, 22.35, 22.58, 22.87,

23.79, 24.11, 24.29, 24.35, 24.87, 25.42, 25.81, 26.09, 26.72, 27.04, 27.44, 27.77, 27, 98, 28.19 and 28.56;

(ii) an endotherm with an initial decomposition temperature greater than 200°C in the DSC thermogram;

(iii) mass loss at temperatures above 200°C on the TGA thermogram;

(iv) increasing the weight by about 0.8 wt.% to about 1.0 wt.%, determined by DVS analysis, when removed from a 5% relative humidity environment and placed in a 95% relative humidity environment; and (v) a 1H NMR spectrum substantially identical to FIG. 55.

In one aspect, the present invention includes a crystalline solid form of Compound 1

Compound 1 or a hydrate or solvate thereof, characterized by at least one of the following:

(i) one or more peaks in the XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 5.48, 9.93, 10.83, 10.98, 11.36, 11.79 , 12.04, 12.25, 12.62, 14.33, 14.67, 15.33, 16.02, 16.51, 16.77, 18.07, 19.09, 19.34, 19 .60, 20.00, 20.46, 20.85, 21.45, 21.55, 21.76, 22.16, 22.35, 22.58, 22.87, 23.79, 24.11 , 24.29, 24.35, 24.87, 25.42, 25.81, 26.09, 26.72, 27.04, 27.44, 27.77, 27.98, 28.19 and 28 .56;

(ii) an endotherm with an initial decomposition temperature greater than 200°C in the DSC thermogram;

(iii) mass loss at temperatures above 200°C on the TGA thermogram;

(iv) increasing the weight by about 0.8 wt.% to about 1.0 wt.%, determined by DVS analysis, when removed from a 5% relative humidity environment and placed in a 95% relative humidity environment; and (v) a 1H NMR spectrum substantially identical to FIG. 55.

In one embodiment of this aspect, the crystalline solid form of Compound 1 is characterized as Form A.

In one embodiment, Form A of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 10.83, 10.98, 11.36, 11.79, 12 .04, 14.33, 18.07, 19.09, 20.00, 22.58, 24.87 and 28.19.

In another embodiment, Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where the peaks are 10.83, 10.98, 11.36, 11.79, 12.04, 14.33, 18.07, 19.09, 20.00, 22.58, 24.87 and 28.19.

In one embodiment, Form A of Compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.20 scale, where the peaks are 5.48, 9.93, 10.83, 10.98, 11.36, 11.79, 12.04, 12.25, 12.62, 14.33, 14.67, 15.33, 16.02, 16.51, 16.77, 18.07, 19.09, 19, 34, 19.60, 20.00, 20.46, 20.85, 21.45, 21.55, 21.76, 22.16, 22.35, 22.58, 22.87, 23.79, 24.11, 24.29, 24.35, 24.87, 25.42, 25.81, 26.09, 26.72, 27.04, 27.44, 27.77, 27.98, 28, 19 and 28.56.

In one embodiment, Form A of Compound 1 is characterized as Form A of Compound 1 by at least two of (i), (ii), (iii) and (iv).

In a further embodiment, Form A of Compound 1 is characterized as Form A of Compound 1 by at least three of (i), (ii), (iii) and (iv).

In yet another further embodiment, Form A of Compound 1 is characterized as Form A of Compound 1 by all of (i), (ii), (iii) and (iv).

In another aspect, the present invention includes a crystalline solid form of Compound 1

Compound 1 or a hydrate or solvate thereof, characterized as Form K of Compound 1 by at least one of the following:

(i) one or more peaks in the XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 6.39, 8.10, 11.53, 19.89, 21.11, 22.34 , 24.50 and 26.42;

(ii) an endotherm with an initial decomposition temperature of approximately 226°C in the DSC thermogram;

(iii) mass loss of ~0.2 wt.% between temperatures from 40 to 180°C in the TGA thermogram; And

- 19 044424 (v) 1H NMR spectrum, essentially identical to FIG. 56.

In another aspect, the present invention includes a crystalline solid form of Compound 1

Compound 1 or a hydrate or solvate thereof, characterized by at least one of the following:

(i) one or more peaks in the XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 6.39, 8.10, 11.53, 19.89, 21.11, 22.34 , 24.50 and 26.42;

(ii) an endotherm with an initial decomposition temperature of approximately 226°C in the DSC thermogram;

(iii) mass loss of ~0.2 wt.% between temperatures from 40 to 180°C in the TGA thermogram; and (v) a 1H NMR spectrum substantially identical to FIG. 56.

In one embodiment of this aspect, the crystalline solid form of Compound 1 is characterized as Form K.

In one embodiment, Form K of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 6.39, 8.10, 22.34, and 24.50.

In another embodiment, Form K of compound 1 has all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.39, 8.10, 22.34 and 24.50.

In another embodiment, Form K of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.39, 8.10, 11.53, 19.89, 21.11, 22.34, 24.50 and 26.42.

In another embodiment, Form K of Compound 1 is characterized as Form K of Compound 1 by at least two of (i), (ii) and (iii).

In a further embodiment, Form K of Compound 1 is characterized as Form K of Compound 1 by all of (i), (ii) and (iii).

In another aspect, the present invention includes a crystalline solid form of Compound 1

N

Compound 1 or a hydrate or solvate thereof, characterized as Form Q of Compound 1 by at least one of the following:

(i) one or more peaks in an XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 6.11, 8.61, 9.06, 9.74, 15.69, 16.07 , 20.04 and 24.01;

(ii) an endotherm with an initial decomposition temperature of approximately 194-195°C in the DSC thermogram; and (iii) mass loss of ~11-12 wt.% between temperatures from 120 to 160°C on the TGA thermogram.

In another aspect, the present invention includes a crystalline solid form of Compound 1

N

Compound 1 or a hydrate or solvate thereof, characterized by at least one of the following:

(i) one or more peaks in an XRPD pattern on a 2-theta±0.20 scale, where one or more peaks are selected from 6.11, 8.61, 9.06, 9.74, 15.69, 16.07 , 20.04 and 24.01;

(ii) an endotherm with an initial decomposition temperature of approximately 194-195°C in the DSC thermogram; and (iii) mass loss of ~11-12 wt.% between temperatures from 120 to 160°C on the TGA thermogram.

In one embodiment of this aspect, the crystalline solid form of Compound 1 is characterized as Form K.

- 20 044424

In one embodiment, Form Q of Compound 1 is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 8.61, 9.74,

16.07 and 20.04.

In another embodiment, Form Q of compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 8.61, 9.74, 16.07 and 20.04.

In another embodiment, Form Q of Compound 1 is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 6.11, 8.61, 9.06, 9.74, 15.69, 16.07, 20.04 and 24.01.

In one embodiment, Form Q of Compound 1 is characterized as Form Q of Compound 1 by at least two of (i), (ii) and (iii).

In a further embodiment, Form Q of Compound 1 is characterized as Form Q of Compound 1 by all of (i), (ii) and (iii).

In one aspect, the present invention includes a crystalline solid form of Compound 1

Compound 1 or a hydrate or solvate thereof, characterized as Compound 1 hemifumarate Form B by at least one of the following:

(i) one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.55, 9.08, 10.81, 13.24, 15.89, 16.95 , 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22.29, 22 .48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88;

(ii) an endotherm with an initial decomposition temperature of approximately 226°C in the DSC thermogram;

(iii) negligible mass loss at approximately 220°C on the TGA thermogram;

(iv) an increase in mass of approximately 0.2 wt.%, measured by DVS, in the environment when removed from 5% relative humidity and placed in 95% relative humidity; and (v) a 1H NMR spectrum substantially identical to FIG. 54.

In one aspect, the present invention includes a crystalline solid form of Compound 1

Compound 1 or a hydrate or solvate thereof, characterized by at least one of the following:

(i) one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 7.55, 9.08, 10.81, 13.24, 15.89, 16.95 , 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22.29, 22 .48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88;

(ii) an endotherm with an initial decomposition temperature of approximately 226°C in the DSC thermogram;

(iii) negligible mass loss at approximately 220°C on the TGA thermogram;

(iv) an increase in mass of approximately 0.2 wt.%, measured by DVS, in the environment when removed from 5% relative humidity and placed in 95% relative humidity; and (v) a 1H NMR spectrum substantially identical to FIG. 54.

In one embodiment of this aspect, the crystalline solid form of Compound 1 is characterized as Hemifumarate Form B.

In one embodiment, Compound 1 hemifumarate Form B is characterized by one or more peaks in an XRPD pattern on a 2-theta±0.2 scale, where one or more peaks are selected from 9.08, 10.81, 16.95, 17.44, 22.29, 22.48, 23.82, 24.37, 26.34 and 27.05.

In one embodiment, Compound 1 Hemifumarate Form B is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 9.08, 10.81, 16.95, 17.44, 22.29 , 22.48, 23.82, 24.37, 26.34 and 27.05.

In another embodiment, Compound 1 Hemifumarate Form B is characterized by all of the following peaks in the XRPD pattern on a 2-theta±0.2 scale, where the peaks are 7.55, 9.08, 10.81,

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13.24, 15.89, 16.95, 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21, 16, 21.74, 22.29, 22.48,

22.75, 23.82, 24.37, 26.34, 27.05 and 27.88.

In one embodiment, Compound 1 hemifumarate Form B is characterized as Form

The hemifumarate of compound 1 by at least two of (i), (ii), (iii) and (iv).

In a further embodiment, Compound 1 Hemifumarate Form B is characterized as Compound 1 Hemifumarate Form B by at least three of (i), (ii), (iii) and (iv).

In yet another further embodiment, Compound 1 Hemifumarate Form B is characterized as Compound 1 Hemifumarate Form B by all of (i), (ii), (iii) and (iv).

In another aspect, the present invention provides a pharmaceutical composition comprising a crystalline form or crystalline salt form as described herein and a pharmaceutically acceptable excipient.

In yet another aspect, the present invention provides a method of treating a disease, disorder or syndrome mediated at least in part by modulating the activity of a protein kinase in vivo, comprising administering to a subject in need thereof a crystalline form or a crystalline salt form as described herein, or a pharmaceutical composition described herein.

In one embodiment of this aspect, the disease, disorder or syndrome mediated, at least in part, by modulation of protein kinase activity in vivo is cancer.

In one aspect, the present invention provides a method for inhibiting a protein kinase, the method comprising contacting the protein kinase with a crystalline form or a crystalline salt form as described herein.

In one embodiment of this aspect, the protein kinase is Axl, Mer, c-Met, KDR, or a combination thereof.

In yet another aspect, the present invention relates to a method for preparing Compound 1 hemifumarate Form B, comprising contacting Compound 1 with fumaric acid in an organic solvent to form a mixture and stirring the mixture.

In one embodiment of this aspect, the organic solvent is acetone.

In another embodiment, the mixture is stirred at a temperature of approximately 50°C.

In another embodiment, the mixture is stirred for approximately 6 days.

Crystal forms of the present invention.

Form A of compound 1.

Form A of Compound 1 is likely the thermodynamically stable anhydrous/unsolvated form of Compound 1 at RT. Characterization of Form A of compound 1 is presented herein using XRPD, DSC, TGA, DVS and high temperature microscopy.

The XRPD pattern for Form A of Compound 1 is shown in FIG. 1, and the list of peaks of this pattern is presented in table. 1 below.

Table 1

Form A XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.48±0.20 16.106±0.587 7 9.93±0.20 8.901±0.179 4 10.83±0.20 8.165±0.150 22 10.98±0.20 8.052±0.146 31 11.36±0.20 7.785±0.137 85 11.79±0.20 7.502±0.127 thirty 12.04±0.20 7.346±0.122 33 12.25±0.20 7.222±0.118 4 12.62±0.20 7.009±0.111 17 14.33±0.20 6.175±0.086 29 14.67±0.20 6.035±0.082 18 15.33±0.20 5.777±0.075 20 16.02±0.20 5.529±0.069 5 16.51±0.20 5.365±0.065 13 16.77±0.20 5.283±0.063 9 18.07±0.20 4.904±0.054 25 19.09±0.20 4.645±0.048 29 19.34±0.20 4.586±0.047 16

- 22 044424

19.60±0.20 4.525±0.046 3 20.00±0.20 4.436±0.044 62 20.46±0.20 4.338±0.042 8 20.85±0.20 4.257±0.040 8 21.45±0.20 4.139±0.038 12 21.55±0.20 4.120±0.038 17 21.76±0.20 4.080±0.037 10 22.16±0.20 4.008±0.036 9 22.35±0.20 3.974±0.035 13 22.58±0.20 3.935±0.034 25 22.87±0.20 3.885±0.034 3 23.79±0.20 3.737±0.031 4 24.11±0.20 3.689±0.030 14 24.29±0.20 3.662±0.030 14 24.35±0.20 3.653±0.030 14 24.87±0.20 3.577±0.028 100 25.42±0.20 3.501±0.027 6 25.81±0.20 3.449±0.026 5 26.09±0.20 3.413±0.026 2 26.72±0.20 3.334±0.024 3 27.04±0.20 3.294±0.024 5 27.44±0.20 3.248±0.023 5 27.77±0.20 3.210±0.023 3 27.98±0.20 3.187±0.022 7 28.19±0.20 3.163±0.022 42 28.56±0.20 3.123±0.021 16

The XRPD pattern for Form A of compound 1 was successfully indexed, suggesting that the substance consists mainly or exclusively of a single crystalline phase. The unit cell volume corresponds to anhydrous/unsolvated compound 1.

Unit cell data for Form A of compound 1

Bravais type S-central monoclinic a [A] 35.918 b [A] 9,256 s [A] 17.368 <! (grad.) 90 β [deg.] 116.38 ⁷ [deg.] 90 Volume [AAcell] 5172.6 Chiral content? Undefined Symbol of extinction From 1 from 1 Spatial(s) group(s) Xie (9). C2/s (15)

DSC and TGA thermograms for Form A of Compound 1 are shown in FIG. 2 and 3 respectively. Negligible mass loss was noted by TGA up to 220°C, corresponding to anhydrous/unsolvated material. The sharp endotherm at ~230°C (onset of decomposition) in the DSC thermogram likely corresponds to simultaneous melting and decomposition. DSC thermograms for various Form A samples of compound 1 show inconsistency in the initial endotherm temperature. Variability in endotherm decomposition onset temperatures is likely due to concomitant decomposition. Due to the influence of decomposition, these endotherm decomposition start temperatures do not correspond to the actual melting points.

Form A of compound 1 was further analyzed by high temperature microscopy (Fig. 5A-5D). The heating observations are consistent with the DSC and TGA data described herein. The onset of melting with accompanying decomposition was observed at ~230°C, and color change was observed at ~231°C.

The limited hygroscopicity of Compound 1 Form A was observed by DVS (Figure 4). The substance stably absorbed ~0.91 wt.% water vapor at relative humidity from 5 to 95%. All this mass was lost by desorption with very little hysteresis. XRPD of the sample after DVS indicates no change in crystalline form.

- 23 044424

Determination of pKa and logP values for Form A of compound 1 was carried out using Pion Inc./Sirius

Analytical Instruments Ltd. Accordingly, Form A of compound 1 was determined to have a pKa

5.43±0.4, neutral logP 4.50±0.5 and cationic logP 1.79±0.8.

Form B of connection 1.

Form B of compound 1 is an acetic acid solvate obtained from a vapor diffusion experiment in acetic acid with diethyl ether.

The XRPD pattern for Form B of Compound 1 is shown in FIG. 6, and the list of peaks of this pattern is presented in table. 2 below.

table 2

XRPD peaks of form B of compound 1

2θ O Interatomic distance (A) Intensity(%) 4.76±0.20 18.536±0.778 52 9.58±0.20 9.227±0.192 51 10.49±0.20 8.430±0.160 21 10.97±0.20 8.060±0.147 13 11.27±0.20 7.847±0.139 94 12.10±0.20 7.308±0.120 63 13.26±0.20 6.671±0.100 27 13.52±0.20 6.546±0.096 21 14.52±0.20 6.096±0.084 61 15.15±0.20 5.845±0.077 37 15.42±0.20 5.743±0.074 13 16.69±0.20 5.306±0.063 28 17.29±0.20 5.124±0.059 23 17.92±0.20 4.946±0.055 13 18.34±0.20 4.834±0.052 15 19.05±0.20 4.654±0.048 76 19.25±0.20 4.608±0.047 32 19.48±0.20 4.554±0.046 25 20.04±0.20 4.426±0.044 49 20.59±0.20 4.309±0.041 18 20.90±0.20 4.247±0.040 17 21.39±0.20 4.151±0.038 31 21.84±0.20 4.066±0.037 18 22.25±0.20 3.991±0.035 8 22.68±0.20 3.917±0.034 17 22.84±0.20 3.890±0.034 25 23.12±0.20 3.844±0.033 22 23.32±0.20 3.811±0.032 46 23.60±0.20 3.767±0.031 24 24.03±0.20 3.701±0.030 17 24.79±0.20 3.589±0.029 eleven 25.32±0.20 3.515±0.027 100 25.65±0.20 3.471±0.027 13 25.88±0.20 3.441±0.026 10 26.50±0.20 3.361±0.025 7 26.79±0.20 3.326±0.024 10 27.25±0.20 3.270±0.024 14 28.55±0.20 3.124±0.021 52 29.49±0.20 3.026±0.020 22

The XRPD pattern was successfully indexed and the unit cell volume is large enough to accommodate solvated compound 1.

- 24 044424

Unit cell data for Form B of compound 1

Bravais type Prpmptpvnyjmonoklpn pcheskpy a [A] 19,400 b[A] 9,289 _C [A] 16,992 s (deg.) 90 β [deg.] 108.17 [deg.] 90 Volume [A^cell] 2909.4 Chiral content* ⁷ Achiral Symbol of extinction R 1 21/s 1 Space group(s) R21/S (14)

The proton NMR spectrum for Form B of compound 1 corresponds to the chemical structure of compound 1 with 1 mole of acetic acid per mole of API present.

The TGA thermogram for Form B of compound 1 indicates a mass loss of ~11.2% between 92°C and 188°C (Figure 7). Assuming that the mass loss corresponds solely to the loss of acetic acid, it would be approximately 1.1 mol/mol.

Based on the TGA data, an experiment was carried out to dry Form B of compound 1 by heating the substance at ~200°C for ~5 min. The solids were observed to turn black, indicating decomposition under these conditions.

Form C connection 1.

Form C of Compound 1 is a solvate of HFIPA obtained by antisolvent precipitation from HFIPA using MTBE.

The XRPD pattern for Form C of Compound 1 is shown in FIG. 8, and the list of peaks of this pattern is presented in table. 3 below.

Table 3

Form C XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 3.89±0.20 22.667±1.164 32 4.63±0.20 19.085±0.825 47 7.95±0.20 11.115±0.279 51 9.31±0.20 9.495±0.204 20 10.54±0.20 8.386±0.159 23 10.87±0.20 8.131±0.149 36 11.14±0.20 7.934±0.142 35 11.31±0.20 7.820±0.138 39 11.49±0.20 7.698±0.134 45 11.75±0.20 7.528±0.128 28 12.22±0.20 7.239±0.118 84 12.96±0.20 6.826±0.105 35 13.59±0.20 6.512±0.095 25 13.84±0.20 6.392±0.092 35 14.01±0.20 6.315±0.090 28 14.62±0.20 6.054±0.082 65 14.79±0.20 5.984±0.080 32 15.46±0.20 5.727±0.074 27 15.86±0.20 5.583±0.070 21 16.07±0.20 5.512±0.068 24 16.61±0.20 5.333±0.064 49 16.73±0.20 5.294±0.063 52 16.88±0.20 5.248±0.062 44 17.64±0.20 5.024±0.057 100 18.13±0.20 4.889±0.053 24 18.73±0.20 4.734±0.050 69 19.10±0.20 4.642±0.048 51 19.42±0.20 4.567±0.047 88

- 25 044424

19.75±0.20 4.490±0.045 93 20.09±0.20 4.417±0.044 54 20.47±0.20 4.335±0.042 44 21.00±0.20 4.227±0.040 38 21.65±0.20 4.102±0.037 51 21.95±0.20 4.045±0.036 48 22.47±0.20 3.953±0.035 36 23.11±0.20 3.846±0.033 41 23.46±0.20 3.789±0.032 53 23.77±0.20 3.740±0.031 52 24.84±0.20 3.581±0.028 31 25.17±0.20 3.535±0.028 73 26.14±0.20 3.406±0.026 21 26.48±0.20 3.364±0.025 19 26.88±0.20 3.314±0.024 17 27.72±0.20 3.216±0.023 23 28.35±0.20 3.145±0.022 18 28.70±0.20 3.108±0.021 49 28.96±0.20 3.081±0.021 34

The Form C proton NMR spectrum of compound 1 matches the chemical structure of compound 1 and indicates the presence of 0.6 mol HFIPA and 0.05 mol MTBE per mole API.

TGA analysis of Form C of Compound 1 shows a mass loss of ~13.8% between 75°C and 154°C (Figure 9). Assuming that the mass loss corresponds to the loss of HFIPA, this is equivalent to approximately 0.5 mol/mol. An additional stage of mass loss of ~1.9% is observed between 190°C and 220°C, possibly due to the onset of decomposition.

Form D connection 1.

Form D of Compound 1 is a MeOH solvate obtained from a flash cooling experiment in MeOH and as a minor component of a mixture with Form A of Compound 1 from suspension in MeOH at below ambient temperature.

The XRPD pattern for Form D of compound 1 is shown in FIG. 10, and the list of peaks of this pattern is presented in table. 4 below.

Table 4

Form D XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.08±0.20 17.369±0.683 4 5.43±0.20 16.269±0.599 5 7.00±0.20 12.617±0.360 10 9.62±0.20 9.183±0.190 7 10.21±0.20 8.659±0.169 100 10.90±0.20 8.111±0.148 4 12.31±0.20 7.183±0.116 7 13.66±0.20 6.478±0.094 5 14.06±0.20 6.295±0.089 10 14.70±0.20 6.022±0.082 2 15.35±0.20 5.768±0.075 19 16.06±0.20 5.514±0.068 2 16.39±0.20 5.403±0.065 4 17.89±0.20 4.954±0.055 5 18.17±0.20 4.878±0.053 10 18.35±0.20 4.831±0.052 23 18.53±0.20 4.785±0.051 9 18.80±0.20 4.716±0.050 5 18.96±0.20 4.678±0.049 4 19.15±0.20 4.631±0.048 3 19.50±0.20 4.549±0.046 5 20.09±0.20 4.416±0.044 4 20.37±0.20 4.357±0.042 5 20.58±0.20 4.312±0.041 9 20.93±0.20 4.241±0.040 thirty

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21.31±0.20 4.166±0.039 22 21.79±0.20 4.075±0.037 10 21.97±0.20 4.043±0.036 eleven 22.30±0.20 3.983±0.035 8 22.91±0.20 3.878±0.033 7 23.12±0.20 3.844±0.033 6 23.26±0.20 3.822±0.032 6 23.62±0.20 3.763±0.031 13 23.93±0.20 3.716±0.031 3 24.37±0.20 3.649±0.029 18 24.77±0.20 3.592±0.029 6 24.99±0.20 3.560±0.028 15 25.39±0.20 3.506±0.027 14 25.96±0.20 3.430±0.026 10 26.62±0.20 3.346±0.025 9 27.10±0.20 3.287±0.024 8 27.53±0.20 3.238±0.023 5 28.05±0.20 3.178±0.022 10 28.38±0.20 3.143±0.022 9 28.78±0.20 3.100±0.021 6 29.09±0.20 3.068±0.021 4 29.38±0.20 3.038±0.020 3 29.64±0.20 3.011±0.020 4

The XRPD pattern was successfully indexed and the unit cell volume could accommodate solvated compound 1 with up to 3 moles of MeOH.

Unit cell data for Form D of compound 1

Bravais type Elementary monoclinic a [A] 5,034 b[A] 18,352 s [A] 34,622 O. (deg/ 90 β [deg.] 92.14 7 [deg.] 90 Volume [A ³ /cell] 3196.3 Chiral content? Achiral Symbol of extinction R 1 21L1 1 Spatial(s) Р21/п(14) group(s)

The proton NMR spectrum for Form D of compound 1 corresponds to the chemical structure of compound 1 with 2 moles of MeOH present per mole of API.

TGA analysis of Form D of Compound 1 indicates that the compound readily desolvates upon heating, losing ~13.5 wt% between 38 and 130°C (Figure 11). This mass loss would be equivalent to ~2.6 moles of MeOH, greater than the 2 moles detected by proton NMR. Therefore, the mass loss determined by TGA may be due to the loss of MeOH and additional volatiles such as water.

Form D of compound 1 was dried in vacuum at ~80-81°C for 1 day, which resulted in transformation into an amorphous substance with several small peaks determined by XRPD.

Form E connection 1.

Form E of Compound 1 is a THF solvate obtained from a flash cooling experiment in THF and a flash precipitation experiment from a THF/water-heptane mixture. It should be noted that the solids collected in the flash-chill experiment were white, while almost all other forms of compound 1 were tan, brown, or rusty in color.

The XRPD pattern for Form E of compound 1 is shown in FIG. 12, and the list of peaks of this pattern is presented in table. 5 below.

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Table 5

Form E XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.16±0.20 17.123±0.664 9 6.13±0.20 14.403±0.469 12 9.77±0.20 9.043±0.185 7 10.37±0.20 8.520±0.164 6 10.82±0.20 8.172±0.151 17 11.69±0.20 7.564±0.129 5 13.73±0.20 6.443±0.093 5 14.34±0.20 6.173±0.086 4 14.79±0.20 5.986±0.081 5 15.47±0.20 5.725±0.074 8 15.79±0.20 5.607±0.071 6 16.33±0.20 5.423±0.066 8 16.64±0.20 5.322±0.064 13 16.82±0.20 5.265±0.062 18 17.60±0.20 5.034±0.057 5 17.89±0.20 4.955±0.055 8 18.16±0.20 4.882±0.053 7 18.72±0.20 4.736±0.050 37 19.09±0.20 4.645±0.048 13 19.59±0.20 4.529±0.046 5 20.65±0.20 4.298±0.041 6 21.73±0.20 4.087±0.037 100 22.10±0.20 4.019±0.036 14 22.72±0.20 3.910±0.034 7 23.23±0.20 3.825±0.032 5 23.54±0.20 3.777±0.032 6 23.79±0.20 3.737±0.031 6 24.78±0.20 3.591±0.029 5 25.13±0.20 3.541±0.028 4 26.37±0.20 3.377±0.025 3 26.91±0.20 3.310±0.024 4 29.12±0.20 3.064±0.021 6 29.95±0.20 2.981±0.019 9

The proton NMR spectrum corresponds to the chemical structure of compound 1 with 0.7 moles of THF present per mole of API.

The TGA thermogram shows a mass loss of ~8.2% between 60 and 130°C (Fig. 13). Assuming that THF is the only volatile species, this weight loss corresponds to ∼0.7 mol/mol, which is consistent with the proton NMR spectrum.

Based on these data, Form E of Compound 1 was dried in vacuo at ~77°C for 1 day, resulting in a new disordered species designated Form M of Compound 1.

Form F connection 1.

Form F of Compound 1 is a chloroform solvate (~0.7 mol chloroform) obtained by slow evaporation from chloroform and from a suspension at RT in chloroform (mixture with Form L of Compound 1).

The XRPD pattern for Form F of Compound 1 is shown in FIG. 14, and the list of peaks of this pattern is presented in table. 6 below.

Table 6

Form F XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.85±0.20 15.090±0.515 57 7.44±0.20 11.876±0.319 22 8.56±0.20 10.326±0.241 21 10.95±0.20 8.071±0.147 45 11.75±0.20 7.524±0.128 20 12.28±0.20 7.202±0.117 16

- 28 044424

13.65±0.20 6.480±0.094 36 14.48±0.20 6.112±0.084 21 14.94±0.20 5.924±0.079 17 15.61±0.20 5.673±0.072 40 16.27±0.20 5.443±0.066 21 16.68±0.20 5.310±0.063 72 17.84±0.20 4.968±0.055 61 18.39±0.20 4.820±0.052 39 19.25±0.20 4.606±0.047 45 19.52±0.20 4.544±0.046 19 20.30±0.20 4.371±0.043 31 21.62±0.20 4.106±0.038 24 22.07±0.20 4.024±0.036 72 22.83±0.20 3.892±0.034 46 23.58±0.20 3.770±0.032 100 24.33±0.20 3.655±0.030 16 25.93±0.20 3.434±0.026 31 26.20±0.20 3.399±0.025 14 26.48±0.20 3.364±0.025 15 27.79±0.20 3.208±0.023 81

The proton NMR spectrum for Form F of compound 1 corresponds to the chemical structure of compound 1 with 0.7 moles of chloroform per mole of API present.

The TGA thermogram for Form F of compound 1 shows a mass loss of ~14.4% between 77°C and 178°C, which is equivalent to a loss of 0.7 mol/mol chloroform if it is the only volatile species (Figure 15).

Based on the TGA data, a drying experiment was performed in which Compound 1 Form F was heated at ~175°C for ~13 min. The substance was completely converted to a new substance designated Form K of Compound 1. It should be noted that a color change to yellow and brown was observed during the heating experiment.

Form G connection 1.

Similar to Form F of Compound 1, Form G of Compound 1 is also a chloroform solvate (~1 mol chloroform) that is formed by precipitation from chloroform at below room temperature.

The XRPD pattern for Form G of compound 1 is shown in FIG. 16, and the list of peaks of this pattern is presented in table. 7 below.

Table 7

Form G XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 4.72±0.20 18.724±0.794 37 6.71±0.20 13.153±0.391 17 9.47±0.20 9.330±0.197 eleven 11.51±0.20 7.684±0.133 42 11.84±0.20 7.468±0.126 14 13.04±0.20 6.782±0.104 8 14.40±0.20 6.148±0.085 9 15.12±0.20 5.853±0.077 9 16.03±0.20 5.526±0.069 19 16.28±0.20 5.439±0.066 31 16.51±0.20 5.365±0.065 27 17.04±0.20 5.199±0.061 32 17.85±0.20 4.966±0.055 31 18.04±0.20 4.914±0.054 38 18.73±0.20 4.733±0.050 15 19.29±0.20 4.598±0.047 25 19.49±0.20 4.551±0.046 26 19.73±0.20 4.495±0.045 38 20.72±0.20 4.284±0.041 55 21.10±0.20 4.207±0.039 15 22.61±0.20 3.930±0.034 100

- 29 044424

23.16±0.20 3.838±0.033 20 24.10±0.20 3.689±0.030 19 25.49±0.20 3.492±0.027 13 26.47±0.20 3.365±0.025 48 27.25±0.20 3.270±0.024 18 27.84±0.20 3.202±0.023 12

The proton NMR spectrum for form F of compound 1 corresponds to the chemical structure of compound 1 with 0.9 mol/mol chloroform present.

TGA thermogram for Form G of Compound 1 as shown in FIG. 17 shows a mass loss of ~20.8% between 40 and 165°C. Assuming chloroform is the only volatile species, this is equivalent to ~1.2 mol, slightly higher than the 0.9 mol found by proton NMR. This may indicate a small amount of additional volatiles, such as water, or partial drying of the material before NMR analysis. It should be noted that the weight loss determined by TGA for Form G of Compound 1 begins at a lower temperature than the weight loss for Form F of Compound 1 (40 and 77°C), indicating that partial drying at ambient conditions is possible.

A sample of Form G of Compound 1 was heated at ~175°C for ~10 min similar to the drying conditions for Form F of Compound 1. This experiment resulted in conversion to the same form, Form K of Compound 1.

Form H of compound 1.

Form H of compound 1 is a probable solvate of DCM that was formed by steam exposure of amorphous compound 1 using DCM.

The XRPD pattern for Form H of Compound 1 is shown in FIG. 18, and the list of peaks of this pattern is presented in table. 8 below.

Table 8

Form H XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 6.30±0.20 14.027±0.445 23 10.79±0.20 8.196±0.152 44 11.42±0.20 7.743±0.135 37 11.73±0.20 7.536±0.128 15 12.63±0.20 7.005±0.110 19 14.01±0.20 6.316±0.090 18 14.29±0.20 6.193±0.086 eleven 14.67±0.20 6.034±0.082 14 15.74±0.20 5.625±0.071 20 16.41±0.20 5.396±0.065 22 17.23±0.20 5.143±0.059 23 17.52±0.20 5.057±0.057 34 18.01±0.20 4.921±0.054 49 18.31±0.20 4.841±0.052 20 18.56±0.20 4.777±0.051 22 19.04±0.20 4.658±0.048 100 19.67±0.20 4.510±0.045 71 19.80±0.20 4.480±0.045 85 20.32±0.20 4.367±0.043 26 20.72±0.20 4.283±0.041 34 21.53±0.20 4.123±0.038 27 21.69±0.20 4.095±0.037 41 21.95±0.20 4.047±0.036 52 22.47±0.20 3.954±0.035 thirty 23.14±0.20 3.841±0.033 64 23.53±0.20 3.777±0.032 96 24.33±0.20 3.656±0.030 53 24.84±0.20 3.581±0.028 25 25.13±0.20 3.541±0.028 48 25.38±0.20 3.506±0.027 68 25.69±0.20 3.464±0.027 24 26.75±0.20 3.330±0.024 22

- 30 044424

27.48±0.20 3.243±0.023 65 28.19±0.20 3.163±0.022 34 28.70±0.20 3.108±0.021 33 29.09±0.20 3.068±0.021 23 29.60±0.20 3.015±0.020 36

The XRPD pattern was successfully indexed and the unit cell volume could accommodate compound 1 with up to 1 mol of DCM.

Unit cell data for Form H of Compound 1

Bravais type Elementary monoclinic a [A] 5.718 b [A] 32,737 s [A] 15,491 n. (deg.) 90 β [deg.] 91.46 Ϊ [deg.] 90 Volume [A ³ /cell] 2898.8 Chiral content? Achiral Symbol of extinction R 1 21L1 1 Spatial(s) P21 41(14) group(s)

The sample was initially analyzed using solvent-wet XRPD. The solids were air-dried at ambient conditions for ∼2 h to remove residual solvent that may interfere with characterization; however, the substance partially desolvated, becoming a disordered substance with peaks similar to a mixture of Form H of Compound 1 and Form A of Compound 1. Therefore, Form H of Compound 1 exhibits poor physical stability under ambient conditions and has not been further characterized.

Form K connection 1.

Form K of compound 1 consists of anhydrous/unsolvated compound 1 and was prepared by drying two different chloroform solvates, forms F and G of compound 1, at ~175°C.

The XRPD pattern for Form K of compound 1 is shown in FIG. 19, and the list of peaks of this pattern is presented in table. 9 below.

Table 9

Form K XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.73±0.20 15.398±0.537 35 6.39±0.20 13.811±0.431 32 8.10±0.20 10.912±0.269 38 11.53±0.20 7.668±0.133 100 11.78±0.20 7.506±0.127 31 12.83±0.20 6.896±0.107 28 14.36±0.20 6.164±0.085 50 15.56±0.20 5.689±0.073 53 16.25±0.20 5.449±0.067 38 17.42±0.20 5.086±0.058 82 18.17±0.20 4.878±0.053 36 19.07±0.20 4.649±0.048 28 19.70±0.20 4.502±0.045 71 19.89±0.20 4.460±0.044 95 20.53±0.20 4.322±0.042 66 21.11±0.20 4.205±0.039 32 21.55±0.20 4.121±0.038 34 22.34±0.20 3.977±0.035 24 22.50±0.20 3.948±0.035 24 23.24±0.20 3.825±0.032 42 23.76±0.20 3.742±0.031 24 24.50±0.20 3.630±0.029 42 25.94±0.20 3.432±0.026 37

- 31 044424

26.42±0.20 3.371±0.025 43 27.76±0.20 3.211±0.023 29 28.28±0.20 3.153±0.022 34

The proton NMR spectrum for Form K of compound 1 matches the chemical structure of compound 1 and shows no signs of decomposition (minor amounts of chloroform detected).

The DSC and TGA thermograms for Form K of Compound 1 are shown in FIG. 20 and 21 respectively. Negligible mass loss was noted by TGA up to 180°C, corresponding to anhydrous/unsolvated material. The endotherm at ~220°C (onset of decomposition) determined by DSC likely corresponds to simultaneous melting and decomposition.

Form O of compound 1.

Form O of Compound 1 is a probable TFE solvate of Compound 1 that was obtained from one or more salt filter experiments in solvent systems containing TFE.

The XRPD pattern for Form O of Compound 1 is shown in FIG. 22, and the list of peaks of this pattern is presented in table. 10 below.

Table 10

Form O XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 6.10±0.20 14.467±0.474 24 9.01±0.20 9.809±0.217 16 9.83±0.20 8.991±0.182 8 10.68±0.20 8.277±0.155 20 11.12±0.20 7.953±0.143 20 11.33±0.20 7.806±0.137 37 12.25±0.20 7.221±0.117 33 12.99±0.20 6.810±0.104 18 13.93±0.20 6.351±0.091 38 14.51±0.20 6.099±0.084 44 14.92±0.20 5.932±0.079 41 15.55±0.20 5.694±0.073 8 15.79±0.20 5.607±0.071 10 17.14±0.20 5.170±0.060 12 17.43±0.20 5.083±0.058 74 17.58±0.20 5.042±0.057 41 18.15±0.20 4.885±0.053 eleven 18.42±0.20 4.812±0.052 7 19.35±0.20 4.583±0.047 100 19.77±0.20 4.486±0.045 34 20.24±0.20 4.385±0.043 57 20.71±0.20 4.286±0.041 22 20.90±0.20 4.246±0.040 25 21.49±0.20 4.131±0.038 25 21.68±0.20 4.096±0.037 25 22.04±0.20 4.030±0.036 17 22.36±0.20 3.973±0.035 56 22.78±0.20 3.900±0.034 23 23.37±0.20 3.803±0.032 27 23.96±0.20 3.711±0.031 92 24.39±0.20 3.647±0.029 54 24.92±0.20 3.570±0.028 eleven 25.62±0.20 3.474±0.027 9 26.20±0.20 3.398±0.025 16 26.64±0.20 3.343±0.025 8 26.93±0.20 3.308±0.024 6 27.32±0.20 3.262±0.023 9 27.68±0.20 3.221±0.023 35 27.96±0.20 3.189±0.022 12 28.26±0.20 3.156±0.022 eleven

- 32 044424

28.60±0.20 3.118±0.021 22 28.81±0.20 3.096±0.021 eleven

The XRPD pattern was successfully indexed and the unit cell volume could accommodate compound 1 with up to 1 mol of TFE. The substance was not further characterized.

Unit cell data for Form O of compound 1

Bravais type Triclinical a [A] 10,079 b [A] 10,592 s [A] 14,589 o (deg.) 98.17 β [deg.] 90.51 y [deg.] 103.21 Volume [A ³ /cell] 1,499.5 Chiral content? Undefined Symbol of extinction R Space group(s) Pl(1). RG(2) Form P of connection 1. Form P of compound 1 is a probable hydrate of compound 1, which was observed only in as a mixture with a trace amount of Form A of Compound 1. The XRPD pattern for Form P of Compound 1 is shown in FIG. 22, and the list of peaks of this pattern

presented in table. 11 below.

Table 11

Form P XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.99±0.20 14.745±0.492 8 8.78±0.20 10.063±0.229 5 9.40±0.20 9.404±0.200 5 10.12±0.20 8.738±0.172 8 11.99±0.20 7.376±0.123 64 14.61±0.20 6.058±0.082 thirty 14.87±0.20 5.953±0.080 41 15.61±0.20 5.674±0.072 7 15.98±0.20 5.542±0.069 10 16.32±0.20 5.428±0.066 22 16.62±0.20 5.330±0.064 4 17.56±0.20 5.047±0.057 10 17.62±0.20 5.030±0.057 10 17.84±0.20 4.967±0.055 4 18.05±0.20 4.910±0.054 4 18.43±0.20 4.810±0.052 10 18.88±0.20 4.695±0.049 12 19.22±0.20 4.614±0.048 12 19.72±0.20 4.499±0.045 15 19.85±0.20 4.469±0.045 22 20.32±0.20 4.367±0.043 6 20.91±0.20 4.246±0.040 100 21.67±0.20 4.097±0.037 thirty 22.04±0.20 4.029±0.036 47 22.39±0.20 3.967±0.035 eleven 22.93±0.20 3.875±0.033 38 23.46±0.20 3.789±0.032 8 23.71±0.20 3.749±0.031 8 23.98±0.20 3.708±0.030 7 24.11±0.20 3.688±0.030 10 24.43±0.20 3.640±0.029 15 24.84±0.20 3.581±0.028 6 25.74±0.20 3.459±0.026 6 26.39±0.20 3.374±0.025 15

- 33 044424

26.64±0.20 3.344±0.025 18 26.85±0.20 3.318±0.024 39 27.77±0.20 3.210±0.023 17 28.74±0.20 3.104±0.021 5 29.26±0.20 3.049±0.020 7 29.55±0.20 3.020±0.020 15 30.07±0.20 2.970±0.019 17

Despite existing as a mixture, the XRPD pattern for the P form of compound 1 was successfully indexed. The peaks at 11.36, 14.35 and 28.18° are inconsistent with the indexing solution and are associated with Form A of Compound 1. The unit cell volume per molecule is larger than that of Form A of Compound 1 and can potentially accommodate up to 2 moles more /mol of water.

Unit cell data for Form P of compound 1

Bravais type Primitive monocl] ΐΗΐ easier than skpy a [A] 6,152 b[A] 23.805 s [A] 18.961 O. (grad.) 90 β [deg.] 97.82 '1 [deg.] 90 Volume [A ³ /cell] 2 751.0 Chiral Achiral content? Symbol of extinction R 1 21 /p 1 Spatial(s) P2i/n( 14) group(s)

The proton NMR spectrum of the mixture is consistent with the chemical structure of compound 1 with a minor presence of THF.

The DSC thermogram is shown in Fig. 24. Small overlapping broad endotherms are observed at 90 and 109°C, consistent with dehydration. No other thermal events such as recrystallization or melting are observed after these events, indicating a loss of crystallinity after dehydration.

Form Q of compound 1.

Form Q of compound 1 was characterized by XPRD, DSC, TGA and SEM (not shown in the figures).

The XRPD pattern for Form Q of Compound 1 is shown in FIG. 25, and the list of peaks of this pattern is presented in table. 12 below.

Table 12

Form Q XRPD peaks of compound 1

2θ O Interatomic distance (A) Intensity(%) 6.11±0.20 14.454±0.473 18 8.61±0.20 10.266±0.238 15 9.06±0.20 9.755±0.215 48 9.74±0.20 9.071±0.186 21 10.64±0.20 8.305±0.156 45 10.89±0.20 8.117±0.149 15 11.24±0.20 7.869±0.140 25 11.33±0.20 7.803±0.137 25 12.06±0.20 7.331±0.121 8 12.24±0.20 7.223±0.118 10 12.91±0.20 6.852±0.106 12 13.82±0.20 6.401±0.092 21 14.46±0.20 6.120±0.084 42 14.83±0.20 5.969±0.080 23 15.69±0.20 5.645±0.072 8 15.76±0.20 5.619±0.071 8 16.07±0.20 5.510±0.068 7 17.05±0.20 5.195±0.060 10

- 34 044424

17.31±0.20 5.118±0.059 62 17.40±0.20 5.092±0.058 69 17.78±0.20 4.985±0.056 8 18.16±0.20 4.881±0.053 18 18.42±0.20 4.813±0.052 6 18.88±0.20 4.697±0.049 8 19.08±0.20 4.647±0.048 20 19.28±0.20 4.601±0.047 100 19.56±0.20 4.535±0.046 69 19.84±0.20 4.471±0.045 8 20.07±0.20 4.420±0.044 58 20.70±0.20 4.287±0.041 10 21.04±0.20 4.220±0.040 14 21.38±0.20 4.153±0.038 thirty 21.59±0.20 4.112±0.038 8 21.91±0.20 4.054±0.037 29 22.18±0.20 4.004±0.036 17 22.30±0.20 3.984±0.035 20 22.58±0.20 3.934±0.034 50 22.78±0.20 3.900±0.034 eleven 23.04±0.20 3.856±0.033 eleven 23.23±0.20 3.826±0.032 34 23.50±0.20 3.783±0.032 7 23.81±0.20 3.734±0.031 55 24.01±0.20 3.703±0.030 13 24.32±0.20 3.657±0.030 87 24.86±0.20 3.579±0.028 10 25.43±0.20 3.500±0.027 8 25.80±0.20 3.450±0.026 5 26.05±0.20 3.417±0.026 10 26.20±0.20 3.398±0.025 10 26.69±0.20 3.337±0.025 16 27.02±0.20 3.297±0.024 8 27.44±0.20 3.247±0.023 10 27.63±0.20 3.225±0.023 38 27.99±0.20 3.185±0.022 22 28.48±0.20 3.131±0.022 14 28.75±0.20 3.103±0.021 16 29.17±0.20 3.059±0.021 17 29.36±0.20 3.040±0.020 20

The XRPD pattern for the Q form of compound 1 was successfully indexed using a unit cell that is isostructural with the 2,2,2-trifluoroethanol (TFE) solvate, the O form of compound 1.

Unit cell data for Q form of compound 1

Bravais type a [A] b [A] C [A] 0. (deg.) β [deg.] 7 [deg.] Volume [A’/cell] Chiral content? Extinction symbol Spatial group(s)

Triclinical

10.044

10.691

14.626

98.58

91.18

103.65

1,506.5

Undefined

RR1 (1).RG(2)

DSC (Fig. 26) showed a small, shallow peak, followed by a broad peak that coincided with mass loss in thermogravimetric analysis, and a final sharp endotherm with an onset of ~194-195°C.

- 35 044424

The TGA mass loss that corresponded to the broad endotherm in DSC was approximately ~11-12% (Figure 27).

Scanning electron microscopy images were obtained for two samples of form Q of compound 1 at 100-5000* magnification. The first sample contained agglomerates larger than 200 µm, which consisted of flocs smaller than 20 µm. The second sample consisted of blades no less than 50 µm long and no more than 5 µm wide.

Form A of compound 1 fumarate.

Compound 1 fumarate Form A was prepared from an elevated temperature suspension consisting of acetone, fumaric acid and the free base of Compound 1. The elevated temperature suspension was stirred for 4 days and then the room temperature suspension was added for an additional day.

The XRPD pattern for fumarate form A of compound 1 is shown in FIG. 28.

Compound 1 hemifumarate form B.

Hemifumarate Form B of Compound 1 is the anhydrous salt of Compound 1. The form is physically stable at 40°C and 75% RH for 15 days, and disproportionation was not noticeable in either acetone or water. In addition, the form is not hygroscopic according to DVS analysis. By DSC and high temperature microscopy, Compound 1 hemifumarate Form B exhibits onset of melting at approximately 225°C, which is higher compared to other crystalline salts identified in this study.

Compound 1 hemifumarate Form B was prepared using the method described below.

Compound 1 (199.0 mg) was combined with 2 molar equivalents of fumaric acid (88.8 mg). The mixture was suspended in 10 ml of acetone at ~50°C. A scoop filled with seeds of Compound 1 hemifumarate Form B was added to the suspension. After 6 days at ~50°C, the pale pink solids were collected by vacuum filtration and dried on filter paper in air under reduced pressure for approximately 5 min.

The XRPD pattern for Hemifumarate Form B of Compound 1 is shown in FIG. 29, and the list of peaks of this pattern is presented in table. 13 below.

Table 13

XRPD peaks of compound 1 hemifumarate form B

2θ O Interatomic distance (A) Intensity(%) 7.55±0.20 11.702±0.310 14 7.92±0.20 11.158±0.281 5 9.08±0.20 9.732±0.214 84 9.40±0.20 9.397±0.199 eleven 10.81±0.20 8.179±0.151 38 11.18±0.20 7.909±0.141 8 13.24±0.20 6.681±0.100 12 13.35±0.20 6.627±0.099 8 14.47±0.20 6.114±0.084 4 14.90±0.20 5.942±0.079 5 15.14±0.20 5.846±0.077 6 15.89±0.20 5.573±0.070 13 16.64±0.20 5.325±0.064 4 16.95±0.20 5.227±0.061 63 17.14±0.20 5.168±0.060 20 17.29±0.20 5.124±0.059 21 17.44±0.20 5.080±0.058 35 17.79±0.20 4.982±0.056 5 18.24±0.20 4.860±0.053 20 18.34±0.20 4.834±0.052 17 19.16±0.20 4.629±0.048 74 19.91±0.20 4.456±0.044 10 20.19±0.20 4.395±0.043 eleven 20.42±0.20 4.346±0.042 9 20.70±0.20 4.287±0.041 20 21.16±0.20 4.196±0.039 8 21.74±0.20 4.085±0.037 10 22.29±0.20 3.986±0.035 28 22.48±0.20 3.952±0.035 26

- 36 044424

22.75±0.20 3.905±0.034 16 23.82±0.20 3.733±0.031 25 24.37±0.20 3.650±0.030 48 24.97±0.20 3.563±0.028 4 25.17±0.20 3.535±0.028 7 25.69±0.20 3.465±0.027 6 26.34±0.20 3.380±0.025 100 26.75±0.20 3.330±0.024 7 27.05±0.20 3.294±0.024 49 27.35±0.20 3.258±0.023 8 27.50±0.20 3.241±0.023 6 27.88±0.20 3.198±0.022 16

The XRPD pattern of compound 1 hemifumarate Form B was successfully indexed, indicating that the substance consists of a single crystalline phase. It has a triclinic unit cell containing two molecules of compound 1 and one molecule of fumaric acid. The formula unit volume calculated from the indexing solution corresponds to the anhydrous form.

Unit cell data for compound 1 hemifumurate form B

Bravais type Triclinical a [A] 10,711 b[A] 11,401 s [A] 12,626 O. (grad.) 86.78 β [deg.] 67.95 [deg.] 77.89 Volume [A ³ /cell] 1,396.8 Chiral content? Undefined Symbol of extinction R- Spatial(s) R1 (1), RG (2) group(s)

The 1H NMR spectra are consistent with the chemical structure of compound 1. Based on peak integration, approximately 0.5 mol of fumaric acid per mole of compound 1 is present, corresponding to the hemifumarate salt of compound 1. A negligible amount of residual acetone is also noticeable.

The TGA thermogram curve (Fig. 31) depicts negligible mass loss before decomposition. The DSC thermogram curve (Fig. 30) shows one endotherm with an onset of ~225°C (132.6 J/g).

High temperature micrographs (FIGS. 33A-33D) for Hemifumarate Form B of Compound 1 confirm that this phenomenon occurs as melting with simultaneous decomposition.

The DVS isotherm (Fig. 32) indicates that the form is not hygroscopic. Compound 1 hemifumarate Form B gains/loses less than 0.2% mass during the sorption/desorption experiment without hysteresis.

The substance recovered from the experiment remained unchanged and was identified as Form B hemifumarate of compound 1 by XRPD.

The physical stability of hemifumarate Form B was investigated. Compound 1 hemifumarate Form B was suspended in water at ambient conditions for approximately 24 hours. In a separate experiment, the substance was washed repeatedly with acetone. Both extracts were identified by XRPD analysis as Compound 1 hemifumarate Form B, indicating that these conditions did not cause salt disproportionation. In addition, the substance exposed to 75% RH/40°C for approximately 2 weeks did not change as determined by XRPD.

Hemifumarate Form B of Compound 1 was successfully reproduced on a laboratory scale of 200 and 1 g. This suggests that Hemifumarate Form B of Compound 1 can be produced relatively easily and reproducibly.

HCl form A of compound 1.

HCl Form A of Compound 1 was prepared by suspending Compound 1 and HCl in TGF at room temperature.

The XRPD pattern for HCl Form A of Compound 1 is shown in FIG. 34, and the list of peaks of this pattern is presented in table. 14 below.

- 37 044424

Table 14

XRPD peaks of Form A of HCl salt of compound 1

2θ O Interatomic distance (A) Intensity(%) 5.19±0.20 17.007±0.655 55 8.17±0.20 10.809±0.264 26 9.84±0.20 8.980±0.182 27 10.10±0.20 8.749±0.173 8 10.42±0.20 8.480±0.162 22 11.07±0.20 7.986±0.144 9 12.52±0.20 7.064±0.112 12 12.76±0.20 6.934±0.108 17 12.98±0.20 6.817±0.105 25 13.49±0.20 6.560±0.097 10 13.69±0.20 6.463±0.094 13 13.89±0.20 6.372±0.091 21 14.31±0.20 6.185±0.086 25 14.84±0.20 5.966±0.080 10 15.12±0.20 5.856±0.077 6 15.68±0.20 5.647±0.072 eleven 16.34±0.20 5.420±0.066 7 16.68±0.20 5.310±0.063 10 17.08±0.20 5.188±0.060 5 17.47±0.20 5.072±0.058 24 17.96±0.20 4.936±0.055 26 18.49±0.20 4.794±0.051 10 19.23±0.20 4.613±0.048 31 19.78±0.20 4.484±0.045 28 20.31±0.20 4.369±0.043 14 20.91±0.20 4.244±0.040 18 21.16±0.20 4.196±0.039 22 21.42±0.20 4.145±0.038 44 22.10±0.20 4.019±0.036 45 22.81±0.20 3.896±0.034 100 23.18±0.20 3.835±0.033 38 23.89±0.20 3.722±0.031 eleven 24.39±0.20 3.646±0.029 36 25.20±0.20 3.532±0.028 7 25.87±0.20 3.441±0.026 8 26.34±0.20 3.380±0.025 48 27.06±0.20 3.293±0.024 14 27.59±0.20 3.230±0.023 9 28.07±0.20 3.176±0.022 10

The XRPD pattern of Form A of compound 1 HCl was successfully indexed.

Unit cell data for Form A of compound 1

Tim Brave Elementary monoclinic a [A] 10.803 b [A] 9.043 s [A] 33.901 o (deg.) 90 r [deg.] 91.90 y [deg.] 90 Volume [A ³ /cell] 3,310.0 Chiral content? Achiral Symbol of extinction R 1 21/s 1 Spatial) R21/s (14) group(s)

- 38 044424

HCl form B of compound 1.

HCl Form B of Compound 1 was prepared by suspending Compound 1 and HCl in chloroform at elevated temperature.

The XRPD pattern for HCl form B of compound 1 is shown in FIG. 35, and the list of peaks of this pattern is presented in table. 15 below.

Table 15

XRPD peaks of the B salt HCl form of compound 1

2θ O Interatomic distance (A) Intensity(%) 8.20±0.20 10.772±0.262 9 9.72±0.20 9.094±0.187 15 9.81±0.20 9.006±0.183 18 10.04±0.20 8.804±0.175 5 10.56±0.20 8.374±0.158 9 12.52±0.20 7.065±0.112 9 12.97±0.20 6.823±0.105 20 13.32±0.20 6.640±0.099 eleven 13.48±0.20 6.565±0.097 6 13.81±0.20 6.408±0.092 42 14.35±0.20 6.166±0.085 5 14.95±0.20 5.922±0.079 10 15.89±0.20 5.573±0.070 5 16.63±0.20 5.328±0.064 13 17.37±0.20 5.101±0.058 5 17.83±0.20 4.970±0.055 15 17.99±0.20 4.926±0.054 20 18.32±0.20 4.839±0.052 eleven 19.15±0.20 4.630±0.048 13 19.31±0.20 4.592±0.047 15 19.51±0.20 4.546±0.046 17 19.72±0.20 4.498±0.045 18 20.17±0.20 4.399±0.043 17 20.84±0.20 4.260±0.040 9 21.04±0.20 4.219±0.040 19 21.15±0.20 4.198±0.039 17 21.30±0.20 4.168±0.039 26 21.81±0.20 4.071±0.037 28 22.02±0.20 4.034±0.036 25 22.65±0.20 3.922±0.034 100 23.11±0.20 3.846±0.033 8 23.40±0.20 3.799±0.032 24 23.75±0.20 3.744±0.031 16 24.76±0.20 3.593±0.029 26 25.34±0.20 3.511±0.027 5 25.74±0.20 3.458±0.026 13 26.20±0.20 3.399±0.025 56 26.90±0.20 3.311±0.024 21 27.71±0.20 3.217±0.023 6 27.98±0.20 3.186±0.022 8 28.34±0.20 3.146±0.022 5 28.98±0.20 3.079±0.021 10

HCl form C of compound 1.

The HCl form C of compound 1 was prepared by a method involving crystallization from methanol with MTBE as an antisolvent.

The XRPD pattern for HCl form C of compound 1 is shown in FIG. 36.

The XRPD pattern of HCl form C of compound 1 includes the following degree peaks on a 2-theta±0.2 scale: 2.5, 3.0, 4.3, 5.1, 6.2, 6.8, 7.3 , 7.8, 8.8, 10.6, 11.6, 12.5, 13.3, 13.8, 15.3, 15.7, 17.1, 17.8, 19.0, 19 ,4, 20.0, 20.5, 20.8, 21.5, 22.2, 22.6, 23.0, 23.5, 23.9, 25.2, 26.2, 26.8 , 27.2, 28.0, 28.9 and 29.5.

- 39 044424

HCl form D of compound 1.

HCl Form D of Compound 1 was first prepared by suspending Compound 1 in acetone at ~50°C, adding 2 molar equivalents of the acid, and stirring the resulting acidic suspension at ~50°C for 5 days. The product was collected by positive pressure filtration.

The XRPD pattern for HCl form D of compound 1 is shown in FIG. 37, and the list of peaks of this pattern is presented in table. 16 below.

Table 16

XRPD peaks of form D HCl salt of compound 1

2θ O Interatomic distance (A) Intensity(%) 3.47±0.20 25.471±1.469 100 5.27±0.20 16.748±0.635 40 6.93±0.20 12.745±0.367 eleven 8.21±0.20 10.758±0.262 thirty 8.97±0.20 9.856±0.219 24 9.86±0.20 8.964±0.181 18 10.16±0.20 8.700±0.171 87 10.44±0.20 8.469±0.162 31 10.69±0.20 8.271±0.154 65 11.28±0.20 7.838±0.139 18 12.26±0.20 7.215±0.117 46 12.75±0.20 6.939±0.108 22 13.27±0.20 6.668±0.100 19 13.92±0.20 6.357±0.091 17 14.23±0.20 6.220±0.087 31 14.54±0.20 6.085±0.083 52 14.95±0.20 5.921±0.079 65 15.44±0.20 5.734±0.074 eleven 15.58±0.20 5.682±0.072 eleven 15.80±0.20 5.603±0.070 12 16.08±0.20 5.508±0.068 16 16.25±0.20 5.451±0.067 13 17.84±0.20 4.967±0.055 45 18.44±0.20 4.807±0.052 20 18.65±0.20 4.755±0.051 33 19.34±0.20 4.586±0.047 19 19.75±0.20 4.492±0.045 23 20.13±0.20 4.408±0.043 23 20.93±0.20 4.241±0.040 75 21.29±0.20 4.171±0.039 45 22.05±0.20 4.028±0.036 52 22.69±0.20 3.916±0.034 51 22.90±0.20 3.880±0.033 49 23.69±0.20 3.753±0.031 63 24.15±0.20 3.682±0.030 21 24.39±0.20 3.647±0.029 33 24.60±0.20 3.616±0.029 38 24.91±0.20 3.571±0.028 67 25.16±0.20 3.536±0.028 52 26.27±0.20 3.389±0.025 39 27.03±0.20 3.296±0.024 47 27.61±0.20 3.228±0.023 36 28.37±0.20 3.144±0.022 28

Compound 1 phosphate form A.

Compound 1 phosphate Form A was prepared by adding 1 molar equivalent of phosphoric acid to a suspension of Compound 1 in chloroform and suspending at elevated temperature for 3 days.

The XRPD pattern for phosphate form A of compound 1 is shown in FIG. 48, and a list of peaks of this pattern is presented below.

- 40 044424

The XRPD pattern of Compound 1 Phosphate Form A includes the following degree scale peaks

2-theta ±0.2: 6.3, 6.8, 10.3, 10.5, 11.4, 12.7, 13.8, 14.7, 15.7, 16.1, 17, 3, 17.5, 18.1, 18.8, 19.4, 20.3, 20.9, 21.2,

22.1, 22.7, 23.2, 23.6, 24.7, 25.5, 27.4, 27.8, 28.5, 29.1 and 29.3.

General principles of introduction.

Administration of the crystalline forms or crystalline salt forms of the present invention in pure form or in an appropriate pharmaceutical composition can be accomplished by any of the accepted routes of administration or means for similar uses. Thus, administration may be, for example, orally, nasally, parenterally (intravenous, intramuscular or subcutaneous), topical, transdermal, intravaginal, intravesical, intracystemal or rectal, in the form of a solid, semi-solid, lyophilized powder or liquid dosage form such as, for example, tablets, suppositories, pills, soft and hard gelatin capsules, powders, solutions, suspensions, aerosols and the like, preferably in unit dosage forms suitable for easy administration of precise dosages.

The compositions will include a conventional pharmaceutical excipient and a crystalline form or crystalline salt form of the present invention as the active agent, and may further include other drugs, pharmaceuticals, excipients, adjuvants, etc. The compositions of the present invention can be used in combination with anticancer or other agents that are typically administered to a patient being treated for cancer. Adjuvants include preservatives, wetting agents, suspending agents, sweetening agents, flavoring agents, flavoring agents, emulsifying agents and dosing agents. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. It may also be desirable to include isotonic agents such as sugars, sodium chloride and the like. In addition, prolonged absorption of the injectable pharmaceutical form can be achieved by the use of absorption delaying agents, such as aluminum monostearate and gelatin.

If desired, the pharmaceutical composition of the present invention may also contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents and antioxidants and the like, such as, for example, citric acid, sorbitan, monolaurate, triethanol aminooleate, butylated hydroxytoluene etc.

Compositions suitable for parenteral injection may contain physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous excipients, diluents, solvents or carriers include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerin, etc.), suitable mixtures thereof, vegetable oils (such as olive oil) and organic esters for injection , such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

One preferred route of administration is oral, using a convenient dosage regimen that can be adjusted depending on the severity of the condition being treated.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In solid dosage forms, the active compound is mixed with at least one inert conventional excipient such as sodium citrate or dicalcium phosphate, or (a) excipients or diluents such as, for example, starches, lactose, sucrose, glucose, mannitol and silicic acid;

(b) binders such as cellulose derivatives, starch, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia;

(c) humectants, such as glycerin;

(d) disintegrants such as, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, croscarmellose sodium, complex silicates and sodium carbonate;

(e) dissolution retarders such as, for example, paraffin;

(f) absorption accelerators, such as, for example, quaternary ammonium compounds;

(g) wetting agents such as, for example, cetyl alcohol, and glycerol monostearate, magnesium stearate; and the like (h) adsorbents such as, for example, kaolin and bentonite; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or mixtures thereof.

- 41 044424

In the case of capsules, tablets and pills, the dosage forms may also contain buffering agents.

The solid dosage forms described above can be prepared using coatings and shells, such as enteric coatings and others well known in the art. They may contain sedatives and may also be formulated to release the active compound or compounds in a specific part of the intestinal tract in a delayed manner. Examples of embedded compositions that can be used are polymeric substances and waxes. The active compounds may also be in microencapsulated form, if desired, with one or more of the excipients mentioned above.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. Such dosage forms are prepared, for example, by dissolving, dispersing, etc. crystalline forms or crystalline salt forms of Compound 1 and optional pharmaceutical adjuvants in an excipient such as, for example, water, saline, aqueous dextrose, glycerin, ethanol and the like; solubilizing agents and emulsifiers such as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol and dimethylformamide; oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters; or mixtures of these substances, etc. to form a solution or suspension.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol, and sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, etc.

Compositions for rectal administration are, for example, suppositories, which can be prepared by mixing crystalline forms or crystalline salt forms of Compound 1, for example, with suitable non-irritating excipients such as cocoa butter, polyethylene glycol or suppository wax that is solid at normal temperatures, but is liquid at body temperature and therefore melts in a suitable body cavity and releases the active component therein.

Dosage forms for topical administration of a compound of the present invention include ointments, powders, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable excipient and any preservatives, buffers or propellants that may be required. Ophthalmic compositions, ophthalmic ointments, powders and solutions are also considered to be within the scope of the present invention.

Typically, depending on the intended route of administration, pharmaceutically acceptable compositions will contain from about 1% to about 99% by weight of the crystalline form or crystalline salt form of Compound 1 and from 99 to 1% by weight of a suitable pharmaceutical excipient. In one example, the composition will be from about 5% to about 75% by weight of the crystalline form or crystalline salt form of Compound 1, with the remainder being suitable pharmaceutical excipients.

Actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; for example, see Pharmaceutical Sciences, 21st ed., (Lippincott, Williams and Wilkins Philadelphia, PA, 2006). The composition administered will in any case contain a therapeutically effective amount of a crystalline form or crystalline salt form of Compound 1 or a pharmaceutically acceptable salt thereof for treating a pathological condition in accordance with the teachings of the present invention.

Crystalline forms or crystalline salt forms of Compound 1 are administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of Compound 1, metabolic stability and duration of action of Compound 1, age, body weight, general health, sex, diet, regimen and time of administration, rate of elimination, combination of drugs, severity of specific pathological conditions and host to be treated. Crystalline forms or crystalline salt forms of Compound 1 can be administered to a patient at dosage levels ranging from about 0.1 mg to about 1000 mg per day. For a normal adult having a body weight of approximately 70 kg, an example is a dosage ranging from about 0.01 mg to about 100 mg per kilogram of body weight per day. However, the specific dosage used may vary. For example, the dosage may depend on a number of factors, including the needs of the patient, the severity of the condition being treated, and the pharmacological activity of the compound used. Determining optimal dosages for a particular patient is well known to one of ordinary skill in the art.

- 42 044424

Combination therapy.

The crystalline form or crystalline salt form of Compound 1 disclosed herein may be administered as a single therapy or in combination (co-administered) with one or more additional therapies to treat a disease or disorder, such as a hyperproliferation-associated disease or disorder such like cancer. Treatment methods that can be used in combination with a compound disclosed herein include (i) surgery;

(ii) radiation therapy (eg, gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes);

(iii) endocrine therapy;

(iv) adjuvant therapy, immunotherapy, CAR T-cell therapy; and (v) other chemotherapeutic agents.

The term co-administration refers to either the simultaneous administration or any method of separate sequential administration of the crystalline form or crystalline salt form of Compound 1 described herein and an additional active pharmaceutical ingredient or ingredients, including cytotoxic agents and radiation therapy. If the administration is not simultaneous, the compounds are administered in close temporal proximity to each other. Moreover, it does not matter whether the compounds are administered in the same dosage form, for example, one compound may be administered topically and another compound may be administered orally.

Generally, any agent that has activity against the disease or condition being treated can be co-administered. Examples of such cancer treatments can be found, for example, at https://www.cancer.gov/about-cancer/treatment/drugs (last accessed: January 22, 2019) and in publicly available sources such as Cancer Principles and Practice of Oncology by V.T. Devita and S. Hellman (editors), 11th edition (2018), Lippincott Williams & Wilkins Publishers. One skilled in the art will be able to discern which combinations of agents will be suitable based on the specific characteristics of the drugs and the disease involved.

In one embodiment, the method of treatment includes co-administration of a crystalline form or crystalline salt form of Compound 1 described herein and at least one immunotherapy. Immunotherapy (also called biological response modifier therapy, biologic therapy, biotherapy, immunotherapy, or biologic therapy) is a treatment that uses parts of the immune system to fight a disease. Immunotherapy can help the immune system recognize cancer cells or enhance the response against cancer cells. The different types of immunotherapy include active and passive types of immunotherapy. Active immunotherapies stimulate the body's own immune system, while passive immunotherapies typically use components of the immune system created outside the body.

Examples of active immunotherapy include, but are not limited to, vaccines, including cancer vaccines, tumor cell vaccines (autologous or allogeneic), dendritic cell vaccines, antigen vaccines, anti-idiotypic vaccines, DNA vaccines, viral vaccines or lymphocyte-based vaccines, tumor infiltrating (TIL), using interleukin-2 (IL-2) or lymphokine-activated killer (LAK) cell-based therapy.

Examples of passive immunotherapies include, but are not limited to, monoclonal antibodies and targeted therapies containing toxins. Monoclonal antibodies include naked antibodies and conjugated monoclonal antibodies (also called tagged, tagged or loaded antibodies). Naked monoclonal antibodies do not have a drug or radioactive substance attached, whereas conjugated monoclonal antibodies are attached, for example, to a chemotherapy drug (chemolabeled), a radioactive particle (radiolabeled), or a toxin (immunotoxin). Examples of these naked monoclonal antibody drugs include, but are not limited to, rituximab (Rituxan), an antibody to the CD20 antigen used to treat, for example, B-cell non-Hodgkin's lymphoma; trastuzumab (Herceptin), an antibody against the HER2 protein used, for example, to treat advanced breast cancer; alemtuzumab (Campath), an antibody against the CD52 antigen used to treat, for example, B-cell chronic lymphocytic leukemia (B-CLL); cetuximab (Erbitux), an antibody to the EGFR protein, used, for example, in combination with irinotecan to treat, for example, advanced colorectal and head and neck cancer; and bevacizumab (Avastin), which is an anti-angiogenesis therapy that targets the VEGF protein and is used, for example, in combination with chemotherapy to treat, for example, metastatic colorectal cancer. Examples of conjugated monoclonal antibodies include, but are not limited to, the radiolabeled antibody ibritumomab

- 43 044424 tiuxetan (Zevalin), which delivers radioactivity directly to malignant B lymphocytes and is used to treat, for example, B-cell non-Hodgkin lymphoma; the radiolabeled antibody tositumomab (Bexxar), which is used to treat, for example, some types of non-Hodgkin's lymphoma; and the immunotoxin gemtuzumab ozogamicin (Mylotarg), which contains calicheamicin and is used to treat, for example, acute myelogenous leukemia (AML). BL22 is a conjugated monoclonal antibody for the treatment of, for example, hairy cell leukemia, immunotoxins for the treatment of, for example, leukemias, lymphomas and brain tumors, and radiolabeled antibodies such as OncoScint, for example, for the treatment of colorectal and ovarian cancer and ProstaScint, for example , for the treatment of prostate cancer.

Additional examples of therapeutic antibodies that may be used include, but are not limited to, HERCEPTIN™ (trastuzumab) (Genentech, CA), which is a humanized anti-HER2 monoclonal antibody for the treatment of patients with metastatic breast cancer; REOPRO.RTM. (abciximab) (Centocor), which is a receptor against glycoprotein IIb/IIIa on platelets to prevent clot formation; ZENAPAX™ (daclizumab) (Roche Pharmaceuticals, Switzerland), which is an immunosuppressive humanized anti-CD25 monoclonal antibody for the prevention of acute renal allograft rejection; PANOREX™, which is a mouse IgG2a antibody to cell surface antigen 17-IA (Glaxo Wellcome/Centocor); BEC2, which is a mouse anti-idiotypic (GD3 epitope) IgG antibody (ImClone System); IMC-C225, which is a chimeric anti-EGFR IgG antibody (ImClone system); VITAXIN™, which is a humanized anti-integrin alpha V beta 3 antibody (Applied Molecular Evolution/Medlmmune); Campath 1H/LDP-03, which is a humanized anti-CD52 IgG1 antibody (Leukosite); Smart M195, which is a humanized anti-CD33 IgG antibody (Protein Design Lab/Kanebo); RITUXAN™, which is a chimeric anti-CD20 IgG1 antibody (IDEC Pharm/Genentech, Roche/Zettyaku); LYMPHOCIDE™, which is a humanized anti-CD22 IgG antibody (Immunomedics); LYMPHOCIDE™ Y-90 (Immunomedics); lymphoscan (Tc-99m labeled; radioimaging; Immunomedics); Nuvion (anti-CD3; Protein Design Labs); CM3 is a humanized anti-ICAM3 antibody (ICOS Pharm); IDEC-114 is a primatized anti-CD80 antibody (IDEC Pharm/Mitsubishi); ZEVALIN™ is a radiolabeled mouse anti-CD20 antibody (IDEC/Schering AG); IDEC-131 is a humanized anti-CD40L antibody (IDEC/Eisai); IDEC-151 is a primatized anti-CD4 antibody (IDEC); IDEC-152 is a primatized anti-CD23 antibody (IDEC/Seikagaku); anti-CD3 SMART is a humanized anti-CD3 IgG antibody (Protein Design Lab); 5G1.1 is a humanized anti-complement factor 5 (C5) antibody (Alexion Pharm); D2E7 is a humanized anti-TNF-alpha antibody (CAT/BASF); CDP870 is a humanized anti-TNF-alpha antibody. Fab fragment (Celltech); IDEC-151 is a primatized anti-CD4 IgG1 antibody (IDEC Pharm/SmithKline Beecham); MDX-CD4 is a human anti-CD4 IgG antibody (Medarex/Eisai/Genmab); CD20-streptavidin (+biotin-yttrium 90; NeoRx); CDP571 is a humanized anti-TNF-alpha antibody. IgG4 antibody (Celltech); LDP-02 is a humanized anti-alpha4 beta7 antibody (LeukoSite/Genentech); OrthoClone OKT4A is a humanized anti-CD4 IgG antibody (Ortho Biotech); ANTOVA™ is a humanized anti-CD40L IgG antibody (Biogen); ANTEGREN™ is a humanized anti-VLA-4 IgG antibody (Elan); and CAT-152 is a human anti-TGF-beta ₂ antibody (Cambridge Ab Tech). Others are presented in the following paragraphs.

Immunotherapies that can be used in combination with the crystalline form or crystalline salt form of Compound 1 described herein include adjuvant immunotherapies. Examples include cytokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), macrophage inflammatory protein (MIP)-1-alpha, interleukins (including IL-1, IL-2, IL-4, IL-6, IL-7, IL-12, IL-15, IL-18, IL-21 and IL-27), tumor necrosis factors (including TNF-alpha) and interferons (including IFN-alpha, IFN-beta and IFN-gamma); aluminum hydroxide (alum); bacillus Calmette-Guerin (BCG); snail limpet hemocyanin (KLH); incomplete Freund's adjuvant (IFA); QS-21; DETOX; levamisole; and dinitrophenyl (DNP) and combinations thereof, such as combinations of interleukins, such as IL-2, with other cytokines, such as IFN-alpha.

In various embodiments, the crystalline form or crystalline salt form of Compound 1 can be combined with an immunological therapy and/or an immunological therapeutic agent. In various embodiments, the immunotherapy and/or immunotherapy may include one or more of the following: adoptive cell transfer, angiogenesis inhibitor, Bacillus Calmette-Guerin therapy, biochemotherapy, cancer vaccine, chimeric antigen T-cell therapy

- 44 044424 receptor (CAR), cytokine therapy, gene therapy, immune checkpoint modulator, immunoconjugate, radioconjugate, oncolytic virus therapy or targeted drug therapy. An immunotherapy or an immunotherapeutic agent is collectively referred to herein as an immunotherapy agent.

The present disclosure provides a method of preventing, treating, reducing, inhibiting or controlling neoplasia, tumor or cancer in a subject in need thereof, comprising administering a therapeutically effective amount of a crystalline form or crystalline salt form of Compound 1 in combination with an immunotherapeutic agent. In one non-limiting embodiment, the method includes administering a therapeutically effective amount of a combination comprising a crystalline form or crystalline salt form of Compound 1 in combination with an immunotherapeutic agent. In various embodiments, the combination provides a cooperative effect, an additive effect, or a synergistic effect of reducing the number of cancer cells when treated with the combination compared to each treatment alone. In some embodiments, administration of a therapeutically effective amount of a combination comprising a crystalline form or crystalline salt form of Compound 1 and an immunotherapeutic agent results in synergistic antitumor activity and/or antitumor activity that is greater than the additive effect of administering the crystalline form or crystalline salt form alone forms of Compound 1 or an immunotherapeutic agent.

Various human cancers are characterized by numerous genetic and epigenetic changes that generate neoantigens potentially recognized by the immune system (Sjoblom et al. (2006), Science, 314: 268-74). The adaptive immune system, composed of T and B lymphocytes, has potent anticancer potential with broad capacity and exceptional specificity to respond to various tumor antigens. In addition, the immune system exhibits significant plasticity and a memory component. Successfully harnessing all of these characteristics of the adaptive immune system would make immunotherapy unique among all cancer treatments.

The present disclosure provides a combination of a crystalline form or crystalline salt form of Compound 1 and an immunotherapeutic agent. These exemplary combinations can be used to treat a subject suffering from cancer. In various embodiments, immunotherapies that find use in the compositions, formulations and methods of the present invention may include one or more agents or treatments, including adoptive cell transfer, angiogenesis inhibitor, bacillus Calmette-Guerin therapy, biochemotherapy, cancer vaccine , Chimeric antigen receptor (CAR) T-cell therapy, cytokine therapy, gene therapy, immune checkpoint modulator such as an immune checkpoint inhibitor, immunoconjugate, radioconjugate, oncolytic virus therapy, or targeted drug therapy.

In certain embodiments of the present disclosure, the therapeutically effective combination comprises a crystalline form or a crystalline salt form of Compound 1 and an immunotherapeutic agent. In various related embodiments, the crystalline form or crystalline salt form of Compound 1 enhances the activity of the immunotherapeutic agent.

In certain embodiments of each of the above aspects, as well as other aspects and embodiments described elsewhere herein, the immunotherapeutic agent enhances the activity of the crystalline form or crystalline salt form of Compound 1 of the present invention.

In certain embodiments of each of the above aspects, as well as other aspects and embodiments described elsewhere herein, the crystalline form or crystalline salt form of Compound 1 and the immunotherapeutic agent act synergistically. In various embodiments described herein, an exemplary immunotherapeutic agent is an immune cell modulator (e.g., T cells, dendritic cells, natural killer cells, etc.) selected from an agonist or a co-stimulatory molecule activator, wherein the modulator is a monoclonal antibody, a bispecific antibody containing one or more antigen-binding immune checkpoint groups, a trispecific antibody, or a multivalent antibody/fusion protein/construct that interacts with immune cells known in the art. In some embodiments, the immunotherapy agent may be an antibody that modulates a costimulatory molecule that binds to an antigen on the surface of an immune cell or cancer cell. In each of these various embodiments, the antibody modulator may be a monoclonal antibody, a polyclonal antibody, a bispecific antibody, an antibody in a trispecific or multispecific format, a fusion protein, or a fragment thereof, e.g., a diabody, a single chain (sc) diabody (scFv)2, a miniantibody, minibody, barnase-barstar, scFv-Fc, sc(Fab)2, trimeric construct

- 45 044424 antibodies, antibody-tribody construct, antibody-tribody construct, antibody-tribody construct, antibody-collatel construct, (scFv-TNFa)3 or antibody construct F(ab)3/DNL.

In certain embodiments of each of the above aspects, as well as other aspects and embodiments described in other sections herein, the immunotherapeutic agent is an agent that modulates immune responses, such as a checkpoint inhibitor or a checkpoint agonist. In some embodiments, the immunotherapeutic agent is an agent that enhances antitumor immune responses. In some embodiments, the immunotherapeutic agent is an agent that enhances cellular immunity. In some embodiments, the immunotherapeutic agent is an agent that enhances T cell activity. In some embodiments, the immunotherapeutic agent is an agent that enhances cytolytic T cell (CTL) activity.

In some embodiments, the treatment methods of the present invention may include administering a crystalline form or crystalline salt form of Compound 1 in combination with a molecule, such as a binding agent, such as an antibody or functional fragment thereof, that modulates (activates or inhibits) a checkpoint protein. A checkpoint inhibitor can be any molecule, agent, treatment and/or method of inhibiting an immune checkpoint and/or stimulating an immune checkpoint inhibitor, for example, by stimulating an intrinsic immune checkpoint inhibitor; inhibiting a transcription factor involved in immune checkpoint expression; and/or joint action with some additional external factor. For example, a checkpoint inhibitor may include a treatment that inhibits transcription factors involved in the expression of immune checkpoint genes or promotes the expression of transcription factors for tumor suppressor genes, such as BACH2 (Luan et al., (2016), Transcription Factors and Checkpoint Inhibitor Expression with Age: Markers of Immunosenescence, Blood, 128(22), 5983). Moreover, a checkpoint inhibitor can suppress the transcription of immune checkpoint genes; modification and/or processing of immune checkpoint mRNA; translation of immune checkpoint proteins; and/or molecules involved in immunity or immune checkpoint pathways, for example, PD-1 transcription factors such as HIF-1, STAT3, NF-κΒ and AP-1, or activation of common oncogenic pathways such as JAK/STAT, RAS/ERK, or PI3K/AKT/mTOR (Zerdes et al., Genetic, transcriptional and post-translational regulation of the programmed death protein ligand 1 in cancer: biology and clinical correlations, Oncogene, vol. 37, p. 4639-4661 (2018), the disclosure of which is incorporated herein by reference in its entirety).

Checkpoint inhibitors may include treatments, molecules, agents and/or methods that regulate immune checkpoints at the transcriptional level, for example, using co-suppression of the RNA interference pathway and/or post-transcriptional gene silencing (PTGS) (eg, microRNA , siRNA; silencing RNA, small interfering RNA or short interfering RNA (siRNA). Transcriptional regulation of checkpoint molecules has been shown to involve mir-16, which has been shown to target the 3'UTR of checkpoint mRNAs CD80, CD274 ( PD-L1) and CD40 (Leibowitz et al., Post-transcriptional regulation of immune checkpoint genes by mir-16 in melanoma, Annals of Oncology (2017) 28; v428-v448). Mir-33a has also been shown to be involved in regulation of PD-1 expression in lung adenocarcinoma (Boldini et al., Role of microRNA-33a in regulating the expression of PD-1 in lung adenocarcinoma, Cancer Cell Int. 2017, 17: 105, the disclosure of which is incorporated herein by reference in full).

T cell-specific aptamer-siRNA chimeras have been proposed as a highly specific way to inhibit molecules in the immune checkpoint pathway (Hossain et al., The aptamer-siRNA conjugates: reprogramming T cells for cancer therapy, Ther. Deliv., 2015 Jan, 6 (1): 1-4, the disclosure of which is incorporated herein by reference in its entirety).

Alternatively, members of the immune checkpoint pathway can be inhibited by treatments that affect associated pathways, such as metabolism. For example, excess glycolysis intermediate pyruvate in mitochondria from CAD macrophages promoted PD-L1 expression through induction of the bone morphogenetic protein 4/phosphorylated SMAD1/5/IFN regulatory factor 1 (BMP4/pSMAD1/5/IRF1) signaling pathway. Accordingly, the use of treatments that modulate the metabolic pathway may result in subsequent modulation of the PD-1/PD-L1 checkpoint immune inhibitory pathway (Watanabe et al., Pyruvate controls the checkpoint inhibitor PD-L1 and suppresses T cell immunity, J. Clin Invest ., 2017 Jun 30, 127(7): 2725-2738).

Checkpoint immunity can be regulated by oncolytic viruses that selectively replicate in tumor cells and induce acute immune responses in the tumor microenvironment, i.e. acting as genetic vectors carrying specific agents (for example, antibodies, miRNA, siRNA, etc.) to cancer cells and influencing their oncolysis and

- 46 044424 secretion of cytokines and chemokines to synergize with immune checkpoint inhibition (Shi et al., Cancer Immunotherapy: A Focus on the Regulation of Immune Checkpoints, Int J. Mol. Sci., 2018 May, 19(5): 1389) . Clinical trials are currently underway using the following viruses as checkpoint inhibitors: poliovirus, measles virus, adenoviruses, poxviruses, herpes simplex virus (HSV), coxsackieviruses, reoviruses, Newcastle disease virus (NDV), T-VEC ( herpes virus encoded by GM-CSF (granulocyte-macrophage colony-stimulating factor) and H101 (Shi et al., supra).

Checkpoint inhibitors may function at the translational level of checkpoint immunity. Translation of mRNA into protein is a major event in the regulation of gene expression, so inhibition of immune checkpoint translation represents a means by which the immune checkpoint pathway can be inhibited.

Inhibition of the immune checkpoint pathway can occur at any stage of the immune checkpoint translation process. For example, drugs, molecules, agents, treatments and/or methods can inhibit the initiation process (wherein the 40S ribosomal subunit is recruited to the 5' end of the mRNA and scans the 5UTR of the mRNA towards its 3' end. Inhibition can occur by targeting to the anticodon of the start RNA to transfer methionyl (tRNA) (Met-tRNAi), base-pair it with the start codon, or recruit the 60S subunit to initiate elongation and sequentially add amino acids to the translation of immune checkpoint-specific genes. Alternatively, a checkpoint inhibitor can inhibit translational checkpoints, preventing the formation of the ternary complex (TC), i.e. eukaryotic initiation factor (eIF)2 (or one or more of its α, β and γ subunits); GTP; and Met-tRNAi.

Checkpoint inhibition may result from destabilization of eIF2α by preventing its phosphorylation by protein kinase R (PKR), PERK, GCN2, or HRI, or by preventing the association of TC with the 40S ribosomal subunit and/or other initiation factors, thereby preventing the formation of preinitiation complex (PIC); inhibition of the eIF4F complex and/or its cap-binding protein eIF4E, scaffold protein eIF4G or helicase eIF4A. Methods that discuss translational control of cancer are described in Truitt et al., New frontiers in translational control of the cancer genome, Nat. Rev. Cancer, 2016 Apr 26, 16(5): 288-304, the disclosure of which is incorporated herein by reference in its entirety.

Checkpoint inhibitors may also include treatments, molecules, agents and/or methods that regulate immune checkpoints at the cellular and/or protein level, for example, by inhibiting the immune checkpoint receptor. Checkpoint inhibition may result from the use of antibodies, antibody fragments, antigen binding fragments, small molecules and/or other drugs, agents, treatments and/or methods.

Immune checkpoints refer to inhibitory pathways in the immune system that are responsible for maintaining self-tolerance and modulating the extent of the immune system response to minimize damage to peripheral tissues. However, tumor cells can also activate immune checkpoints to reduce the effectiveness of the immune response (block the immune response) against tumor tissues. Unlike most anticancer agents, checkpoint inhibitors do not directly target tumor cells, but rather target lymphocyte receptors or their ligands in order to enhance the endogenous antitumor activity of the immune system. (Pardoll, 2012, Nature Reviews Cancer, 12: 252-264).

In some embodiments, the immunotherapy agent is a PD-1 activity modulator, a PD-L1 activity modulator, a PD-L2 activity modulator, a CTLA-4 activity modulator, a CD28 activity modulator, a CD80 activity modulator, a CD86 activity modulator, a 4-1BB activity modulator, OX40 activity modulator, KIR activity modulator, Tim-3 activity modulator, LAG3 activity modulator, CD27 activity modulator, CD40 activity modulator, GITR activity modulator, TIGIT activity modulator, CD20 activity modulator, CD96 activity modulator, IDO1 activity modulator, cytokine, chemokine, interferon, interleukin, lymphokine, a member of the tumor necrosis factor (TNF) family, or an immunostimulating oligonucleotide. In some embodiments, an immune checkpoint modulator, i.e. inhibitor or antagonist, or activator or agonist, is, for example, CD28 modulator, 4-1BB modulator, OX40 modulator, CD27 modulator, CD80 modulator, CD86 modulator, CD40 modulator or GITR modulator, Lag-3 modulator, 41BB modulator, LIGHT modulator , CD40 modulator, GITR modulator, TGF-beta modulator, TIM-3 modulator, SIRP-alpha modulator, TIGIT modulator, VSIG8 modulator, BTLA modulator, SIGLEC7 modulator, SIGLEC9 modulator, ICOS modulator, B7H3 modulator, B7H4 modulator, FAS modulator and/ or BTNL2 modulator. In some embodiments, the immunotherapy agent is an immune checkpoint modulator as described above (e.g., an immune checkpoint modulator antibody that can

- 47 044424 be presented in the form of a monoclonal antibody, a bispecific antibody containing one or more antigen-binding immune checkpoint fragments, a trispecific antibody or a multivalent antibody/fusion protein/construct that interacts with immune cells known in the art).

In some embodiments, the immunotherapeutic agent is an agent that inhibits PD-1 activity. In some embodiments, the immunotherapeutic agent is an agent that inhibits the activity of PD-L1 and/or PD-L2. In some embodiments, the immunotherapeutic agent is an agent that inhibits CTLA-4 activity. In some embodiments, the immunotherapeutic agent is an agent that inhibits the activity of CD80 and/or CD86. In some embodiments, the immunotherapeutic agent is an agent that inhibits TIGIT activity. In some embodiments, the immunotherapeutic agent is an agent that inhibits KIR activity. In some embodiments, the immunotherapeutic agent is an agent that enhances or stimulates the activity of activating immune checkpoint receptors.

PD-1 (also known as programmed cell death protein 1, CD279, PDCD1) is a cell surface receptor that plays a critical role in regulating the balance between stimulatory and inhibitory signals in the immune system and maintaining peripheral tolerance (Ishida, Y. et al., 1992, EMBO J., 11, 3887; Kier, Mary E. et al., 2008, Annu Rev. Immunol., 26, 677-704; Okazaki, Taku et al., 2007. International Immunology, 19, 813-824 ). PD-1 is an inhibitory member of the immunoglobulin superfamily homologous to CD28. Structure PD-1 is a monomeric type 1 transmembrane protein consisting of a single extracellular immunoglobulin variable region-like domain and a cytoplasmic domain containing an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switching motif (ITSM). PD-1 expression is induced on T cells, B cells, natural killer (NK) cells and monocytes, for example upon activation of lymphocytes through T cell receptor (TCR) or B cell receptor (BCR) signaling (Kier, Mary E. et al., 2008, Annu Rev. Immunol., 26, 677-704; Agata, Y. et al., 1996, Int Immunol., 8, 765-72). PD-1 is a receptor for the ligands CD80, CD86, PD-L1 (B7-H1, CD274) and PD-L2 (B7-DC, CD273), which are members of the B7 family expressed on the cell surface (Freeman, Gordon et al. , 2000, J. Exp. Med., 192, 1027; Latchman, Y. et al., 2001, Nat. Immunol., 2: 261) When interacting with a ligand, PD-1 recruits phosphatases such as SHP-1 and SHP -2 to its intracellular tyrosine motifs, which subsequently dephosphorylate effector molecules activated by TCR or BCR signaling (Chemnitz, J. et al., 2004, J. Immunol., 173: 945-954; Riley, James L. , 2009, Immunological Reviews, 229: 114-125). Thus, PD-1 transmits inhibitory signals to T and B cells only when it is engaged simultaneously with the TCR or BCR.

PD-1 has been demonstrated to suppress effector T cell responses through both intrinsic and extrinsic functional cellular mechanisms. Inhibitory signaling through PD-1 causes a state of T cell unresponsiveness, whereby cells are unable to proliferate clonally or produce optimal levels of effector cytokines. PD-1 can also induce apoptosis in T cells through its ability to inhibit survival signals through costimulation, resulting in decreased expression of key antiapoptotic molecules such as Bcl-XL (Kier, Mary E. et al., 2008, Annu Rev. Immunol., 26: 677-704). In addition to these direct effects, recent publications indicate that PD-1 is involved in the suppression of effector cells by promoting the induction and maintenance of T regulatory cells (TREG). For example, PD-L1 expressed on dendritic cells has been shown to act in synergy with TGF-β to promote the induction of CD4+FoxP3+TREG with enhanced suppressive function (Francisco, Loise M. et al., 2009, J. Exp. Med., 206: 3015-3029).

TIM-3 (also known as T-cell immunoglobulin domain and mucin domain 3, TIM-3, hepatitis A virus cell receptor 2, HAVCR2, HAVcr-2, KIM-3, TIMD-3, TIMD3, Tim-3, and CD366 ) is a single-pass type I membrane protein with a mass of ~33.4 kDa, involved in immune responses (Sanchez-Fueyo et al., Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance, Nat. Immunol., 4: 1093-1101 (2003)).

TIM-3 is selectively expressed on Th1 cells and phagocytic cells (eg, macrophages and dendritic cells). Application of siRNA or blocking antibody to reduce human TIM-3 expression resulted in increased interferon γ (IFN-γ) secretion by CD4-positive T cells, indicating an inhibitory role of TIM-3 in human T cells. Analysis of clinical samples obtained from patients with autoimmune disease showed no expression of TIM-3 in CD4 positive cells. In particular, TIM-3 expression levels are lower and IFN-γ secretion is higher in clones

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T cells derived from the cerebrospinal fluid of patients with multiple sclerosis than in clones obtained from normal healthy individuals (Koguchi K. et al., J. Exp. Med., 203: 1413-8. (2006)).

TIM-3 is a galectin-9 ligand receptor, which is a member of the galectin family, molecules ubiquitously expressed on various cell types, and which binds β-galactoside; phosphatidylserine (PtdSer) (DeKryff et al., T cell/transmembrane, Ig, and mucin-3 allelic variants differentially recognize phosphatidylserine and mediate phagocytosis of apoptotic cells, J. Immunol., 2010 Feb 15, 184(4): 1918-30 ); high mobility group protein 1 (also known as HMGB1, HMG1, HMG3, SBP-1, HMG-1 and high mobility group box-1) (Chiba et al., Tumorinfiltrating DCs suppress nucleic acid-mediated innate immune responses through interactions between the receptor TIM-3 and the alarmin HMGB1, Nat. Immunol., 2012 Sep, 13(9): 832-42); and carcinoembryonic antigen-associated cell adhesion molecule 1 (also known as CEACAM1, BGP, BGP1, BGPI, carcinoembryonic antigen-associated cell adhesion molecule 1) (Huang et al., CEACAM1 regulates TIM-3-mediated tolerance and exhaustion, Nature, 2015 Jan 15, 517(7534): 386-90).

BTLA (also known as B- and T-lymphocyte attenuator, BTLA1, CD272 and B- and T-lymphocyte associated) is a type I single-pass membrane protein of ~27.3 kDa involved in the inhibition of lymphocytes during the immune response. BTLA is constitutively expressed in both B and T cells. BTLA interacts with HVEM (herpes virus entry mediator), a member of the tumor necrosis factor receptor (TNFR) family (Gonzalez et al., Proc. Natl. Acad. Sci., USA, 2005, 102: 1116-21). The interaction of BTLA, which belongs to the CD28 family of the immunoglobulin superfamily, and HVEM, the tumor necrosis factor (TNF) costimulatory receptor (TNFR), is unique in that it defines crosstalk between these two receptor families. BTLA contains a membrane-proximal immunoreceptor tyrosine-based inhibitory motif (ITIM) and a membrane-distal immunoreceptor tyrosine-based switch motif (ITSM). Disruption of ITIM or ITSM abolished the ability of BTLA to recruit SHP1 or SHP2, suggesting that BTLA recruits SHP1 and SHP2 in a manner distinct from PD-1, and both tyrosine-based motifs are required to block T cell activation. The cytoplasmic tail of BTLA also contains a third conserved tyrosine-containing motif in the cytoplasmic domain, similar in sequence to the Grb-2 recruitment site (YXN). Additionally, a phosphorylated peptide containing this N-terminal tyrosine-based BTLA motif can interact with GRB2 and the p85 subunit of PI3K in vitro, although the functional effects of this interaction remain unexplored in vivo (Gavrieli et al., Bioochem. Biophysi. Res. Commun ., 2003, 312, 1236-43). BTLA is a receptor for PTPN6/SHP-1 ligands; PTPN11/SHP-2; TNFRSF14/HVEM; and B7H4.

VISTA (also known as V-domain suppressor of Ig T cell activation VSIR, B7-H5, B7H5, GI24, PP2135, SISP1, DD1alpha, VISTA, C10orf54, chromosome 10 open reading frame 54, PD-1H and V-complex immunoregulatory receptor ) is a ~33.9 kDa type I single-pass membrane protein involved in the inhibitory response of T cells, embryonic stem cell differentiation through inhibition of BMP4 signaling and MMP14-mediated activation of MMP2 (Yoon et al., Control of signaling -mediated clearance of apoptotic cells by the tumor suppressor p53, Science, 2015 Jul 31, 349(6247): 1261669). VISTA interacts with the VSIG-3 ligand (Wang et al., VSIG-3 as a ligand of VISTA inhibits human T-cell function, Immunology, 2019 Jan, 156(1): 74-85).

LAG-3 (also known as lymphocyte activation gene 3, LAG3, CD223, and lymphocyte-activating gene 3) is a type I single-pass membrane protein of ~57.4 kDa involved in lymphocyte activation that also binds to HLA class II antigens. LAG-3 is a member of the immunoglobulin supergene family and is expressed on activated T cells (Huard et al., 1994, Immunogenetics, 39: 213), NK cells (Triebel et al., 1990, J. Exp. Med., 171: 1393-1405), regulatory T cells (Huang et al., 2004, Immunity, 21: 503-513; Camisaschi et al., 2010, J. Immunol., 184: 6545-6551; Gagliani et al., 2013, Nat. Med., 19: 739-746) and plasmacytoid dendritic cells (DC) (Workman et al., 2009, J. Immunol, 182: 1885-1891). LAG-3 is a membrane protein encoded by a gene located on chromosome 12 and is structurally and genetically related to CD4. Similar to CD4, LAG-3 can interact with MHC class II molecules on the cell surface (Baixeras et al., 1992, J. Exp. Med., 176: 327-337; Huard et al., 1996, Eur. J. Immunol. , 26: 1180-1186). It has been suggested that direct binding of LAG-3 to MHC class II plays a role in suppressing antigen-dependent stimulation of CD4+ T cells (Huard et al., 1994, Eur. J. Immunol., 24: 3216-3221), and blockade of LAG- 3 has also been shown to activate CD8+ lymphocytes in both tumor and autoantigen (Gross et al., 2007, J. Clin. Invest., 117: 3383-3392) and viral models (Blackburn et al., 2009, Nat. Immunol., 10: 29-37). In addition, the intracytoplasmic region of LAG-3 can interact with LAP (LAG-3-associated protein), which is a signal transduction molecule involved in suppressing the CD3/TCR activation pathway (Iouzalen et al., 2001, Eur. J. Immunol. , 31: 2885-2891). Moreover, CD4+ CD25+ regulatory T cells (Treg) have been shown to express LAG-3 upon activation, which promotes Treg cell suppressive activity

- 49 044424 (Huang, C. et al., 2004, Immunity, 21: 503-513). LAG-3 may also negatively regulate

T cell homeostasis by Treg cells in both T cell-dependent and -independent mechanisms (Workman, C.J. and Vignali, D.A., 2005, J. Immunol., 174: 688-695).

LAG-3 has been shown to interact with MHC class II molecules (Huard et al., CD4/major histocompatibility complex class II interaction analyzed with CD4- and lymphocyte activation gene-3 (LAG-3)Ig fusion proteins, Eur. J. Immunol., 1995 Sep, 25(9): 2718-21).

In addition, several kinases are known to be checkpoint inhibitors, such as CHEK-1, CHEK-2, and A2aR.

CHEK-1 (also known as CHK 1 kinase, CHK1 and checkpoint kinase 1) is a ~54.4 kDa serine/threonine protein kinase that is involved in checkpoint-mediated cell cycle arrest and activation of DNA repair in response to DNA damage and/or unreplicated DNA.

CHEK-2 (also known as CHK2 kinase, CDS1, CHK2, HuCds1, LFS2, PP1425, RAD53, hCds1 and checkpoint kinase 2) is a ~60.9 kDa serine/threonine protein kinase involved in checkpoint-mediated cell cycle arrest, activation of DNA repair and double-strand break-mediated apoptosis.

A2aR (also known as adenosine receptor A2A, ADORA2A, adenosine receptor A2a, A2aR, ADORA2, and RDC8) is a multipass membrane receptor with ~44.7 kDa of adenosine and other ligands.

In some embodiments, exemplary immunotherapies may include one or more antibody modulators targeting PD-1, PD-L1, PD-L2, CEACAM (e.g., CEACAM-1, -3, and/or -5), CTLA-4, TIM-3, LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, TGF-beta, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS and/or BTNL2, among others known in the art. In some embodiments, the immunotherapeutic agent is an agent that enhances natural killer (NK) cell activity. In some embodiments, the immunotherapeutic agent is an agent that inhibits the suppression of an immune response. In some embodiments, the immunotherapeutic agent is an agent that inhibits the activity of suppressor cells or the activity of suppressor cells. In some embodiments, the immunotherapy agent is an agent or therapy that inhibits Treg activity. In some embodiments, the immunotherapeutic agent is an agent that inhibits the activity of inhibitory immune checkpoint receptors.

In some embodiments, the combination of the present disclosure comprises a crystalline form or crystalline salt form of Compound 1 and an immunotherapeutic agent, wherein the immunotherapeutic agent comprises a T cell modulator selected from an agonist or a co-stimulatory molecule activator. In one embodiment, the agonist of the co-stimulatory molecule is selected from an agonist (e.g., an agonist antibody or antigen binding fragment or soluble fusion protein thereof) ligand of GITR, OX40, SLAM (e.g., SLAMF7), HVEM, LIGHT, CD2, CD27, CD28, CDS, ICAM- 1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40, BAFFR, CD7, NKG2C, NKp80, CD160, B7-H3 or CD83. In other embodiments, the combination of effector cells includes a T cell-attracting bispecific activator (e.g., a bispecific antibody molecule that binds to CD3 and a tumor antigen (e.g., EGFR, PSCA, PSMA, EpCAM, HER2, among others).

In some embodiments, the immunotherapy agent is a PD-1 activity modulator, a PD-L1 activity modulator, a PD-L2 activity modulator, a CTLA-4 activity modulator, a CD28 activity modulator, a CD80 activity modulator, a CD86 activity modulator, a 4-1BB activity modulator, OX40 activity modulator, KIR activity modulator, Tim-3 activity modulator, LAG3 activity modulator, CD27 activity modulator, CD40 activity modulator, GITR activity modulator, TIGIT activity modulator, CD20 activity modulator, CD96 activity modulator, IDO1 activity modulator, SIRP activity modulator alpha, TIGIT activity modulator, VSIG8 activity modulator, BTLA activity modulator, SIGLEC7 activity modulator, SIGLEC9 activity modulator, ICOS activity modulator, B7H3 activity modulator, B7H4 activity modulator, FAS activity modulator, BTNL2 activity modulator, cytokine, chemokine, interferon, interleukin, lymphokine, a member of the tumor necrosis factor (TNF) family, or an immunostimulatory oligonucleotide.

In some embodiments, the immunotherapy agent is an immune checkpoint modulator (e.g., an immune checkpoint inhibitor, e.g., a PD-1 activity inhibitor, a PD-L1 activity modulator, a PD-L2 activity modulator, a CTLA-4 modulator, or a CD40 agonist (e.g., anti-CD40 antibody molecule), (xi) an OX40 agonist (e.g., an anti-OX40 antibody molecule), or (xii) a CD27 agonist (e.g., an anti-CD27 antibody molecule).In one embodiment, the immunotherapeutic agent is an inhibitor of: PD-1,

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PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (for example, CEACAM-1, -3 and/or -5), VISTA, BTLA,

TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta, galectin 9, CD69, galectin-1, CD113, GPR56, CD48,

GARP, PD1H, LAIR1, TIM-1 and TIM-4. In one embodiment, the immune checkpoint molecule inhibitor inhibits PD-1, PD-L1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and/or

-5), CTLA-4 or any combination thereof.

In one embodiment, the immunotherapy agent is an agonist of a protein that stimulates T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L , GITR, GITRL, CD70, CD27, CD40, DR3 and CD28H.

In some embodiments, the immunotherapeutic agent used in the combinations disclosed herein (eg, in combination with a crystalline form or crystalline salt form of Compound 1 of the present invention) is an activator or agonist of a costimulatory molecule. In one embodiment, the costimulatory molecule agonist is selected from an agonist (e.g., an agonist antibody or antigen binding fragment or soluble fusion protein thereof) of a CD2 ligand, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4 -1BB (CD137), GITR, CD30, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3 or CD83.

Inhibition of an inhibitory molecule can be performed at the DNA, RNA or protein level. In embodiments, an inhibitory nucleic acid (eg, dsRNA, siRNA, or shRNA) can be used to inhibit expression of the inhibitory molecule. In other embodiments, the inhibitory signal inhibitor is a polypeptide, e.g., a soluble ligand (e.g., PD-1-Ig or CTLA-4 Ig), or an antibody thereof, or an antigen-binding fragment thereof, e.g., a monoclonal antibody, a bispecific antibody, comprising one or more antigen-binding immune checkpoint fragments, trispecific antibody or multivalent immune cell reactive antibody/fusion protein/construct known in the art that bind to an inhibitory molecule; for example, an antibody or fragment thereof (also referred to herein as an antibody molecule) that binds to PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, - 3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta, galectin 9, CD69, galectin-1, CD113, GPR56, CD48, GARP, PD1H, LAIR1, TIM-1 , TIM-4 or a combination thereof.

In some embodiments, the combination includes a crystalline form or a crystalline salt form of Compound 1 and an immunotherapeutic agent, wherein the immunotherapeutic agent is a monoclonal antibody or a bispecific antibody. For example, a monoclonal or bispecific antibody may specifically bind to a member of the c-Met pathway and/or an immune checkpoint modulator (e.g., a bispecific antibody binds to both the hepatocyte growth factor receptor (HGFR) and the immune checkpoint modulator described herein , such as an antibody that binds PD-1, PD-L1, PD-L2 or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, TIGIT , VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2 or CD27). In specific embodiments, the bispecific antibody specifically binds the human HGFR protein and one of PD-1, PD-L1, and CTLA-4.

In some embodiments of the methods described herein, the immunotherapy agent is a PD-1 antagonist, PD-L1 antagonist, PD-L2 antagonist, CTLA-4 antagonist, CD80 antagonist, CD86 antagonist, KIR antagonist, Tim-3 antagonist, antagonist LAG3, TIGIT antagonist, CD20 antagonist, CD96 antagonist or IDO1 antagonist.

In some embodiments, the PD-1 antagonist is an antibody that specifically binds PD-1. In some embodiments, the antibody that binds PD-1 is pembrolizumab (KEYTRUDA®, MK-3475; Merck), pidilizumab (CT-011; Curetech Ltd.), nivolumab (OPDIVO®, BMS-936558, MDX-1106; Bristol Myer Squibb), MEDI0680 (AMP-514; AstraZenenca/MedImmune), REGN2810 (Regeneron Pharmaceuticals), BGB-A317 (BeiGene Ltd.), PDR-001 (Novartis), or STI-A1110 (Sorrento Therapeutics). In some embodiments, an antibody that binds PD-1 is described in PCT publication WO 2014/179664, for example an antibody identified as APE2058, APE1922, APE1923, APE1924, APE 1950 or APE1963 (Anaptysbio), or an antibody comprising CDR regions of any of these antibodies. In other embodiments, the PD-1 antagonist is a fusion protein that contains the extracellular domain of PD-L1 or PD-L2, for example, AMP-224 (AstraZeneca-MedImmune). In other embodiments, the PD-1 antagonist is a peptide inhibitor, such as AUNP-12 (Aurigene).

In some embodiments, a PD-L1 antagonist is an antibody that specifically binds PD-L1. In some embodiments, the antibody that binds PD-L1 is atezolizumab (RG7446, MPDL3280A; Genentech), MEDI4736 (AstraZeneca/MedImmune), BMS-936559 (MDX-1105; Bristol Myers Squibb), avelumab (MSB0010718C; Merck KGaA) , KD033 (Kadmon), part of the KD033 antibody, or STI-A1014 (Sorrento Therapeutics). In some embodiments, an antibody that binds PD-L1 is described in a PCT publication

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WO 2014/055897, for example Ab-14, Ab-16, Ab-30, Ab-31, Ab-42, Ab-50, Ab-52 or Ab-55, or is an antibody that contains CDR regions of any of these antibodies, the description of which is incorporated herein by reference in its entirety.

In some embodiments, the CTLA-4 antagonist is an antibody that specifically binds CTLA-4. In some embodiments, the antibody that binds CTLA-4 is ipilimumab (YERVOY®; Bristol Myer Squibb) or tremelimumab (CP-675206; Pfizer). In some embodiments, the CTLA-4 antagonist is a CTLA-4 fusion protein or a soluble CTLA-4 receptor, such as KARR-102 (Kahr Medical Ltd.).

In some embodiments, the LAG3 antagonist is an antibody that specifically binds LAG3. In some embodiments, the antibody that binds LAG3 is IMP701 (Prima BioMed), IMP731 (Prima BioMed/GlaxoSmithKline), BMS-986016 (Bristol Myer Squibb), LAG525 (Novartis), and GSK2831781 (GlaxoSmithKline). In some embodiments, the LAG3 antagonist includes a soluble LAG3 receptor, such as IMP321 (Prima BioMed).

In some embodiments, a KIR antagonist is an antibody that specifically binds KIR. In some embodiments, the antibody that binds KIR is lirilumab (Bristol Myer Squibb/Innate Pharma).

In some embodiments, the immunotherapeutic agent is a cytokine, such as a chemokine, interferon, interleukin, lymphokine, or a member of the tumor necrosis factor family. In some embodiments, the cytokine is IL-2, IL15, or interferon gamma.

In some embodiments of any of the above aspects or aspects described in other sections herein, the cancer is selected from the group consisting of lung cancer (eg, non-small cell lung cancer (NSCLC)), kidney cancer (eg, urothelial renal carcinoma), cancer bladder (eg, urothelial (transitional cell) carcinoma of the bladder), breast cancer, colorectal cancer (eg, adenocarcinoma of the colon), ovarian cancer, pancreatic cancer, gastric carcinoma, esophageal cancer, mesothelioma, melanoma (eg, melanoma skin), head and neck cancer (eg, head and neck squamous cell carcinoma (HNSCC)), thyroid cancer, sarcoma (eg, soft tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, osteogenic sarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma , lymphangioendothelial sarcoma, leiomyosarcoma or rhabdomyosarcoma), prostate cancer, glioblastoma, cervical cancer, thymic carcinoma, leukemia (eg, acute lymphocytic leukemia (ALL), acute myelocytic leukemia (AML), chronic myelocytic leukemia (CML), chronic eosinophilic leukemia or chronic lymphocytic leukemia (CLL), lymphoma (eg, Hodgkin's lymphoma or non-Hodgkin's lymphoma (NHL)), myeloma (eg, multiple myeloma (MM)), mycosis fungoides, Merkel cell carcinoma, hematologic malignancy, hematologic tissue cancer, B-cell cancer, bronchial cancer, stomach cancer, brain or central nervous system cancer, peripheral nervous system cancer, uterine or endometrial cancer, oral or pharyngeal cancer, liver cancer, testicular cancer, biliary tract cancer, small intestine or appendix cancer, salivary cancer glands, adrenal cancer, adrenal cortical carcinoma, adenocarcinoma, inflammatory myofibroblastic tumor, gastrointestinal stromal tumor (GIST), colon cancer, myelodysplastic syndrome (MDS), myeloproliferative disease (MPD), polycythemia vera, chordoma, synovioma, Ewing tumor , squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous carcinoma, papillary carcinoma, papillary adenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma , embryonal carcinoma, Wilms tumor, bladder carcinoma, epithelial carcinoma, glioma, anaplastic astrocytoma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, diffuse lymphoblastoma, retinoblastoma s, follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, hepatocellular carcinoma, thyroid cancer, small cell carcinoma, essential thrombocythemia, agnogenic myeloid metaplasia, hypereosinophilic syndrome, systemic mastocytosis, familial hypereosinophilia, neuroendocrine cancer or carcinoid tumor.

In some embodiments, any of the above aspects or aspects described in other sections herein, the subject's cancer or tumor does not respond to immune checkpoint inhibition (e.g., any immune checkpoint inhibitor described herein, such as a PD-antagonist 1 or PD-L1 antagonist) or the subject's cancer or tumor has progressed after an initial response to immune checkpoint inhibition (eg, to any immune checkpoint inhibitor described herein, such as a PD-1 antagonist or PD-L1 antagonist).

- 52 044424

In various embodiments, the immunotherapeutic agent may comprise an antibody or an antigen-binding fragment thereof. For the purposes of this definition, immune checkpoint inhibitors include bispecific antibodies and multivalent antibodies/fusion protein/immune cell reactive constructs known in the art. In some embodiments, immunotherapies that contain bispecific antibodies may include bispecific antibodies that are bivalent and bind either the same epitope of an immune checkpoint molecule, two different epitopes of the same immune checkpoint molecule, or different epitopes of two different immune checkpoints.

Those skilled in the art can make several bispecific antibody formats known in the art to target one or more of CTLA4, PD1, PD-L1, TIM-3, LAG-3, various B-7 ligands, B7H3, B7H4 , CHK 1- and CHK 2-kinases, BTLA, A2aR, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, SIRP-alpha, TIGIT, VSIG8, SIGLEC7, SIGLEC9, ICOS, FAS, BTNL2 and others for use in combinations described herein.

In various embodiments, the immunotherapy agent may include a multivalent antibody/fusion protein/construct that interacts with immune cells.

In one embodiment of the present disclosure, a checkpoint inhibitor in combination with a crystalline form or crystalline salt form of Compound 1 is used to reduce or inhibit the metastasis of a primary tumor or cancer to other sites, or the formation or survival of metastatic tumors or cancers at other sites distant from the primary one. tumor or cancer, thereby suppressing or reducing the recurrence of the tumor or cancer or the progression of the tumor or cancer.

In a further embodiment of the present disclosure, provided herein is a combination therapy for the treatment of cancer that includes a crystalline form or crystalline salt form of Compound 1 and a checkpoint inhibitor with the ability to induce potent and durable immune responses with increased therapeutic effect and more controlled toxicity.

In a further embodiment of the present disclosure, provided herein is a combination therapy for the treatment of cancer that includes a crystalline form or crystalline salt form of Compound 1 and an immune checkpoint inhibitor. In one embodiment of the present disclosure, there is provided a method of treating cancer and/or preventing the establishment of metastases by using a crystalline form or crystalline salt form of Compound 1 of the present invention that acts synergistically with a checkpoint inhibitor.

In additional embodiments, the present disclosure provides methods for one or more of the following:

1) reducing or inhibiting the growth, proliferation, motility or invasiveness of tumor or cancer cells that potentially or actually lead to the development of metastases;

2) reducing or inhibiting the formation or survival of metastases arising from the primary tumor or cancer in one or more other sites, positions or areas other than the primary tumor or cancer;

3) reducing or inhibiting the growth or proliferation of metastases in one or more other sites, positions or areas other than the primary tumor or cancer after the metastasis has formed or taken root;

4) reducing or inhibiting the formation or survival of additional metastases after the formation or survival of metastases;

5) long-term overall survival;

6) long-term progression-free survival; or

7) stabilization of the disease.

The methods involve administering to a subject in need thereof a crystalline form or a crystalline salt form of Compound 1 of the present invention in combination with a checkpoint inhibitor disclosed herein.

In one embodiment of the present disclosure, administration of a crystalline form or crystalline salt form of Compound 1 in combination with an immunotherapeutic agent provides a detectable or measurable improvement in the condition of a particular subject, such as alleviation or normalization of one or more adverse (physical) symptoms or consequences associated with the presence of a cell proliferative or a cellular hyperproliferative disorder, neoplasia, tumor or cancer or metastasis, i.e. therapeutic benefit or positive effect.

Therapeutic benefit or beneficial effect is any objective or subjective, transient, temporary or long-term improvement in a condition or pathology, or

- 53 044424 delayed onset, reduction in the severity, duration or frequency of adverse symptoms associated with or caused by cell proliferation or a cellular hyperproliferative disorder such as neoplasia, tumor or cancer or metastasis. They may lead to increased survival. A satisfactory clinical endpoint of a treatment method according to the present disclosure is achieved, for example, when there is a gradual or partial reduction in the severity, duration or frequency of one or more comorbidities, adverse symptoms or complications, or inhibition or abolition of one or more physiological, biochemical or cellular manifestations or characteristics of cell proliferation or a cellular hyperproliferative disorder such as neoplasia, tumor or cancer or metastasis. Thus, therapeutic benefit or improvement may include, but is not limited to, destruction of target proliferating cells (eg, neoplasia, tumor or cancer or metastasis) or elimination of one or more, most or all pathologies, adverse symptoms or complications associated with proliferation cells or a cellular hyperproliferative disorder such as or related to neoplasia, tumor or cancer or metastasis. However, the therapeutic effect or improvement need not be the cure or complete destruction of all target proliferating cells (eg, neoplasia, tumor or cancer or metastasis) or the elimination of all pathologies, adverse symptoms or complications associated with cell proliferation or a cellular hyperproliferative disorder, such as neoplasia, tumor or cancer or metastasis, or related thereto. For example, partial destruction of a tumor or cancer cell mass or stabilization of a tumor or cancer mass, size, or number of cells by inhibiting the progression or exacerbation of a tumor or cancer may reduce mortality and prolong life expectancy by at least a few days, weeks, or months, even if part or the bulk of the tumor or cancer, size or number of cells is preserved.

Specific non-limiting examples of therapeutic effect include reduction of neoplasia, tumor or cancer or metastatic volume (cell size or mass) or number of cells; inhibiting or preventing the increase in the volume of neoplasia, tumor or cancer (eg, stabilization); slowing or inhibiting the progression, exacerbation or metastasis of neoplasia, tumor or cancer; or inhibiting the proliferation, growth or metastasis of a neoplasia, tumor or cancer.

In one embodiment of the present disclosure, administration of an immunotherapeutic agent in combination therapy with a crystalline form or crystalline salt form of Compound 1 provides a detectable or measurable improvement or overall response according to irRC (derived from time point response estimates and based on tumor load), including one of several of the following:

(i) complete resolution according to irCR of all lesions, measurable or non-measurable, and no new lesions (confirmed by repeat, serial assessment at least 4 weeks after the date of first documentation);

(ii) a >50% reduction in irPR tumor burden compared with baseline (confirmed by serial assessment at least 4 weeks after first documentation).

Optionally, any method described herein may not be immediately effective. For example, treatment may be followed by an increase in the number or mass of neoplasia, tumor or cancer cells, but over time there may be a possible stabilization or decrease in tumor cell mass, size or number of cells in a particular subject.

Additional adverse symptoms and complications associated with neoplasia, tumor, cancer and metastasis that can be suppressed, attenuated, reduced, delayed or prevented include, for example, nausea, anorexia, lethargy, pain and discomfort. Thus, partial or complete reduction or reduction in the severity, duration or frequency of adverse symptoms or complications associated with or caused by a cellular hyperproliferative disorder, improvement in the quality of life and/or well-being of the subject, for example, all of increased energy, appetite, psychological well-being constitute specific non-limiting examples of therapeutic benefit.

Thus, therapeutic benefit or improvement may also include a subjective improvement in the quality of life of the subject being treated. In a further embodiment, the method prolongs or increases the lifespan (survival) of a subject. In another embodiment, the method improves the subject's quality of life.

In one embodiment, administration of the immunotherapeutic agent in combination therapy with the crystalline form or crystalline salt form of Compound 1 results in a clinically significant improvement in one or more markers of disease status and progression selected from one or more of the following:

(i) overall survival;

- 54 044424 (ii) progression-free survival;

(iii) overall response speed;

(iv) reducing metastatic disease;

(v) levels of circulating tumor antigens such as carbohydrate antigen 19.9 (CA19.9) and carcinoembryonic antigen (CEA) or others depending on the tumor;

(vii) nutritional status (weight, appetite, serum albumin);

(viii) pain control or analgesic use; and (ix) CRP/albumin ratios.

Treatment with the crystalline form or crystalline salt form of Compound 1 in combination with an immunotherapeutic agent induces more complex immunity, including not only the development of innate immunity and type 1 immunity, but also immunoregulation, which more effectively restores the corresponding immune functions.

In various exemplary methods, a checkpoint inhibitor antibody (monoclonal or polyclonal, bispecific, trispecific or multivalent antibody/fusion protein/construct that interacts with immune cells) directed to a checkpoint molecule of interest (eg, PD-1) can be sequenced and then the polynucleotide sequence can be cloned into an expression or propagation vector. The sequence encoding the antibody or antigen binding fragment thereof of interest can be stored in the vector in a host cell, and the host cell can then be expanded and frozen for later use. The production of recombinant monoclonal antibodies in cell culture can be accomplished by cloning antibody genes from B cells using methods known in the art. See, for example, Tiller et al., 2008, J. Immunol. Methods, 329: 112; US Patent No. 7314622.

Pharmaceutical compositions containing a crystalline form or crystalline salt form of Compound 1 in accordance with the present disclosure will contain an effective amount of a crystalline form or crystalline salt form of Compound 1, an immunotherapeutic agent, and/or both, typically dispersed in a pharmaceutically acceptable excipient. The phrases pharmaceutically or pharmacologically acceptable refer to molecular entities and compositions that do not cause adverse, allergic or other undesirable reactions when administered to an animal, such as, for example, a human, as appropriate. The preparation of a pharmaceutical composition that contains a crystalline form or a crystalline salt form of Compound 1 will be known to those skilled in the art in light of the present disclosure illustrated in Remington's Pharmaceutical Sciences, 21st ed., (Lippincott, Williams and Wilkins Philadelphia, PA, 2006). In addition, it should be understood that for administration to animals (for example, humans), drugs must meet standards of sterility, pyrogenicity, general safety and purity. A specific example of a pharmacologically acceptable excipient for a combination composition containing a crystalline form or crystalline salt form of Compound 1 in admixture with an immunotherapeutic agent described herein is borate buffer or sterile saline (0.9% NaCl).

Formulations of an immunotherapeutic agent, e.g., an immune checkpoint modulator antibody, used in accordance with the present disclosure may be prepared for storage by admixing the antibody having the required purity with optional pharmaceutically acceptable excipients or stabilizers as described and illustrated in detail in Remington's Pharmaceutical Sciences 21st ed. (Lippincott, Williams and Wilkins Philadelphia, PA, 2006), in the form of lyophilized formulations or aqueous solutions and/or suspensions. Acceptable excipients, buffers or stabilizers are non-toxic to recipients at the dosages and concentrations used and include acceptable aqueous and/or non-aqueous excipients that can be used in the pharmaceutical compositions of the present disclosure, for example water, ethanol, polyols (such as glycerin, propylene glycol, polyethylene glycol etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants, buffers such as phosphate, citrate and other organic acids. Antioxidants may include, for example, (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like;

(2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alphatocopherol, and the like; And

- 55 044424 (3) metal ion chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.;

(4) preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkylparabens such as methyl or ethylparaben, pyrocatechin, resorcinol, cyclohexanol, 3-pentanol and m-cresol);

(5) low molecular weight substances (less than approximately 10 residues).

Other examples of pharmaceutically acceptable excipients may include polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium, metal ion complexes (eg Zn-protein complexes) and/or nonionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

In one illustrative embodiment, the pharmaceutical compositions may optionally contain pharmaceutically acceptable excipients necessary to provide exemplary physiological conditions, such as pH adjusting, buffering, and tonicity adjusting agents, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, and sodium lactate. In some embodiments, the anti-checkpoint inhibitor antibodies or antigen-binding fragments thereof of the present disclosure are formulated and may be lyophilized prior to use according to prior art lyophilization and dissolution techniques, in a suitable storage and reconstitution excipient. In one exemplary pharmaceutical composition containing one or more anti-checkpoint inhibitor antibodies or an antigen-binding fragment thereof, the composition is formulated as a sterile, preservative-free solution of one or more anti-checkpoint inhibitor antibodies or antigen-binding fragments thereof for intravenous or subcutaneous administration. The composition may be provided either as a prefilled pen syringe for single use, or as a prefilled glass syringe containing 1 ml for single use, for example, or as an institutional bottle for single use. Preferably, the pharmaceutical composition containing the anti-checkpoint inhibitor antibody or antigen-binding fragment thereof is clear and colorless, with a pH of about 6.9-5.0, preferably a pH of 6.5-5.0, and even more preferably a pH of range from about 6.0 to about 5.0. In various embodiments, the formulations containing the pharmaceutical compositions, when dissolved and administered to a subject, may contain from about 500 mg to about 10 mg, or from about 400 mg to about 20 mg, or from about 300 mg to about 30 mg, or from about 200 mg up to approximately 50 mg of checkpoint antibody or antigen-binding fragment thereof per ml of solution. Exemplary excipients for injection or infusion may include mannitol, citric acid monohydrate, dibasic sodium phosphate dihydrate, monobasic sodium phosphate dihydrate, polysorbate 80, sodium chloride, sodium citrate, and water for parenteral administration, such as intravenous, intramuscular, intraperitoneal, or subcutaneous administration.

In another illustrative embodiment, one or more immunotherapeutic agents or an antigen binding fragment thereof is formulated for intravenous or subcutaneous administration as a sterile aqueous solution containing 1-75 mg/ml, or more preferably about 5-60 mg/ml, or even more preferably about 10 -50 mg/ml or even more preferably about 10-40 mg/ml antibody with sodium acetate, polysorbate 80 and sodium chloride at a pH in the range of about 5 to 6. Preferably, the formulation for intravenous or subcutaneous administration is a sterile aqueous solution, containing 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mg/ml of an immunotherapeutic agent, for example, an antibody to an immune checkpoint inhibitor or an antigen-binding fragment thereof with 20 mM sodium acetate, 0.2 mg/ml polysorbate 80 and 140 mM sodium chloride at pH 5.5. Additionally, the solution containing the anti-checkpoint inhibitor antibody or antigen-binding fragment thereof may contain, among many other compounds, histidine, mannitol, sucrose, trehalose, glycine, polyethylene glycol, EDTA, methionine and any combination thereof, and many other compounds known in the art. areas.

In one embodiment, the pharmaceutical composition of the present disclosure contains the following components: 5-500 mg of an immunotherapeutic agent or an antigen binding fragment thereof of the present disclosure, 10 mM histidine, 5% sucrose and 0.01% polysorbate 80 at pH 5.8, with a crystalline form or the crystalline salt form of Compound 1. This composition may be presented in the form of a lyophilized powder. When the powder is dissolved in full, the composition retains the same composition. Alternatively, the powder may be diluted to half its volume, in which case the composition contains 10-500 mg

- 56 044424 immunotherapeutic agent or antigen binding fragment thereof according to the present disclosure, mM histidine, 10% sucrose and 0.02% polysorbate 80 at pH 5.8.

In one embodiment, a portion of the dose is administered via an intravenous bolus and the remainder via an infusion of the immunotherapy formulation. For example, from about 0.001 mg/kg to about 200 mg/kg, such as from about 0.001 mg/kg to about 100 mg/kg, or from about 0.001 mg/kg to about 50 mg/kg, or from about 0.001 mg/kg up to about 10 mg/kg intravenous injection of the immunotherapeutic agent or antigen binding fragment thereof may be administered as a bolus, and the remaining dose of antibody may be administered by intravenous injection. A predetermined dose of the immunotherapeutic agent or antigen binding fragment thereof can be administered, for example, over a period of 1 to 2 or up to 5 hours.

In a further embodiment, a portion of the dose is administered by subcutaneous injection and/or bolus infusion and the remainder by infusion of the immunotherapy formulation. At some exemplary dosages, the immunotherapy formulation may be administered subcutaneously at a dose of from about 0.001 mg/kg to about 200 mg/kg, such as from about 0.001 mg/kg to about 100 mg/kg, or from about 0.001 mg/kg to about 50 mg /kg, or from about 0.001 mg/kg to about 10 mg/kg intravenous injection of an immunotherapeutic agent or an antigen-binding fragment thereof. In some embodiments, the dose may be administered as a bolus and the remainder of the immunotherapy agent may be administered by subcutaneous or intravenous injection. A predetermined dose of the immunotherapeutic agent or antigen binding fragment thereof can be administered, for example, over a period of 1 to 2 or up to 5 hours.

The formulation herein may also contain more than one active compound, as appropriate, for the particular indication being treated, preferably compounds with additional activities that do not adversely affect each other. For example, it may be necessary to provide one or more immunotherapies with different specificities. Alternatively or in addition, the composition may contain an anti-inflammatory agent, a chemotherapeutic agent, a cytotoxic agent, a cytokine, a cell growth inhibitory agent, and/or a small molecule antagonist. Such molecules are present in the appropriate combination in amounts that are effective for the intended use.

Formulations intended for in vivo administration must be sterile or substantially sterile. This condition is easily achieved by filtration through sterile filtration membranes.

In various embodiments, exemplary formulations of the pharmaceutical compositions described herein can be prepared using methods generally known in the art of preparing pharmaceutical compositions. Typically, such preparation methods may include the step of bringing the active ingredient into association with an excipient or one or more other excipients, and then, if necessary, packaging the product in the desired single-dose or multi-dose dosage unit.

In some embodiments, the composition containing the crystalline form or crystalline salt form of Compound 1 may also be delivered in a vesicle, and the immunotherapeutic agent may be delivered in the same liposomal formulation or in a separate formulation compatible with the liposomal formulation containing the crystalline form or crystalline salt form of compound 1. In some illustrative examples, a liposome containing one or more liposomal surface moieties, e.g., polyethylene glycol, antibodies, and antibody fragments that target a desired tumor surface antigen, receptor, growth factor, glycoprotein, glycolipid, or neoantigen that are selectively transported to certain cells or organs thus enhance targeted drug delivery.

In another embodiment, the crystalline form or crystalline salt form of Compound 1 can be delivered in a vesicle, particularly a liposome (see Langer, Science, 249: 1527-1533 (1990); Treat et al., in liposomes in the therapy of infectious disease and cancer, LopezBerestein and Fidler (eds.), Liss, N.Y., pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid.).

In yet another embodiment, a crystalline form, or a crystalline salt form of Compound 1, or a composition containing a combination, or a composition containing an immunotherapeutic agent, can be delivered in a controlled release system. In one embodiment, a pump can be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng., 14: 201 (1987); Buchwald et al., Surgery, 88: 507 (1980); Saudek et al. ., N. Engl. J. Med., 321: 574 (1989)). In another embodiment, the controlled release system of the crystalline form or crystalline salt form of Compound 1 may comprise

- 57 044424 polymeric substances to provide sustained, intermediate, pulsatile or alternative release (see Medical applications of controlled release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled drug bioavailability, Drug product design and performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem., 23: 61 (1983); see also Levy et al., Science, 228: 190 (1985); During et al., Ann. Neurol., 25: 351 (1989); Howard et al., J. Neurosurg., 71: 105 (1989)). Other controlled release systems discussed in the review by Langer (Science, 249: 1527-1533 (1990)) can be used.

The optimal concentration of active ingredient(s) in the selected vehicle can be determined empirically, in accordance with procedures well known to the skilled artisan, and will depend on the required primary pharmaceutical formulation and the mode of administration adopted.

The present disclosure also provides a pharmaceutical package or kit containing one or more containers filled with one or more ingredients of the pharmaceutical compositions of the present disclosure, which at a minimum will contain a crystalline form or crystalline salt form of Compound 1 and one or more anti-checkpoint inhibitor antibodies or an antigen binding fragment thereof described herein. In other embodiments, the kit may contain one or more additional containers providing a pharmaceutically acceptable excipient, such as a diluent. In one embodiment, the kit may contain at least one container, wherein the container may contain a crystalline form or a crystalline salt form of Compound 1, an anti-checkpoint inhibitor antibody, or an antigen binding fragment thereof of the present disclosure. The kit may also contain a number of instructions for preparing and administering the final pharmaceutical composition to a subject in need thereof to treat a disease or disorder mediated by the checkpoint molecule.

In some embodiments of the present disclosure, the immunotherapeutic agent is a population of immune cells that can be administered in combination with a crystalline form or crystalline salt form of Compound 1 to treat a subject suffering from cancer. In some embodiments, the immunotherapeutic agent is a population of immune cells, such as white blood cells (nuclear white blood cells), containing (eg, expressing) a receptor that binds to an antigen of interest. A leukocyte according to the present disclosure may be, for example, a neutrophil, eosinophil, basophil, lymphocyte or monocyte. In some embodiments, the leukocyte is a lymphocyte. Examples of lymphocytes include T cells, B cells, natural killer (NK) cells, or NKT cells. In some embodiments, the T cell is a CD4+ Th (T helper) cell, a CD8+ cytotoxic T cell, a γδT cell, or a regulatory (suppressor) T cell. In some embodiments, the immune cell is a dendritic cell.

The immune cells of the present disclosure are, in some embodiments, genetically engineered to express an antigen binding receptor. A cell is considered engineered if it contains an engineered (exogenous) nucleic acid. The engineered nucleic acids of the present disclosure can be introduced into a cell using any known (eg, standard) method. For example, the engineered nucleic acid can be introduced into the cell by electroporation (see, for example, Heiser W.C., Transcription Factor Protocols: Methods in Molecular Biology™, 2000, 130: 117-134), chemical (for example, using calcium phosphate or lipid) transfection (see, for example, Lewis W. H. et al., Somatic Cell Genet. 1980 May; 6(3): 333-47; Chen C. et al., Mol. Cell Biol., 1987 August, 7(8 ): 2745-2752), fusions with bacterial protoplasts containing recombinant plasmids (see, for example, Schaffner W., Proc. Natl. Acad. Sci., USA, 1980 April, 77(4): 2163-7), microinjection purified DNA directly into the cell nucleus (see, for example, Capecchi M.R., Cell, 1980 November, 22 (2, pt 2): 479-88) or retroviral transduction.

Some aspects of the present disclosure provide an adaptive cell-based approach that includes isolating immune cells (e.g., T cells) from a subject suffering from cancer, genetically engineering the immune cells (e.g., to express an antigen binding receptor, such as a chimeric antigen receptor), expanding cells ex vivo and then reintroducing the immune cells to the subject. This method results in a greater number of engineered immune cells in a subject compared to what can be achieved using conventional gene delivery and vaccination methods. In some embodiments, immune cells are isolated from a subject, expanded ex vivo without genetic modification, and then reintroduced to the subject.

Immune cells of the present disclosure contain receptors that bind to antigens, such as an antigen encoded by an exogenously delivered nucleic acid, provided herein. In some embodiments, the leukocyte

- 58 044424 is modified (eg genetically modified) to express a receptor that binds to an antigen. The receptor may be, in some embodiments, a naturally occurring antigen receptor (typically expressed on an immune cell), a recombinant antigen receptor (typically not expressed on an immune cell), or a chimeric antigen receptor (CAR). Naturally occurring and recombinant antigen receptors covered by this disclosure include T cell receptors, B cell receptors, NK cell receptors, NKT cell receptors, and dendritic cell receptors. Chimeric antigen receptor refers to an artificial immune cell receptor that is engineered to recognize and bind antigen expressed by tumor cells. Typically, a CAR is engineered for a T cell and is a chimera of the T cell receptor (TcR) complex signaling domain and an antigen recognition domain (eg, a single chain fragment (scFv) of an antibody) (Enblad et al., Human Gene Therapy, 2015, 26 (8): 498-505), the disclosure of which is incorporated herein by reference in its entirety.

In some embodiments, the antigen binding receptor is a chimeric antigen receptor (CAR). A T cell that expresses a CAR is called a CAR T cell. The CAR T cell receptor, in some embodiments, comprises a T cell receptor (TcR) complex signaling domain and an antigen recognition domain (e.g., a single chain fragment (scFv) of an antibody) (Enblad et al., Human Gene Therapy, 2015, 26(8): 498-505), the disclosure of which is incorporated herein by reference in its entirety.

There are four generations of CAR, each containing different components. First-generation CARs attach antibody-derived scFv to the CD3-zeta (zeta or z) intracellular signaling domain of the T cell receptor via hinge and transmembrane domains. Second-generation CARs contain an additional domain, such as CD28, 4-1BB (41BB), or ICOS, to provide a co-stimulatory signal. Third-generation CARs contain two co-stimulatory domains fused to the CD3-zeta chain of the TcR. Third generation costimulatory domains may comprise, for example, a combination of CD3z, CD27, CD28, 4-1BB, ICOS or OX40. CARs in some embodiments comprise an ectodomain (e.g., CD3), typically derived from a single chain variable fragment (scFv), a hinge, a transmembrane domain, and an endodomain with one (first generation), two (second generation), or three (third generation) signaling domains, derived from CD3Z and/or costimulatory molecules (Maude et al., Blood, 2015, 125(26): 4017-4023; Kakarla and Gottschalk, Cancer J., 2014, 20(2): 151-155), the disclosure of which is included incorporated herein by reference in its entirety.

In some embodiments, the chimeric antigen receptor (CAR) is a T cell redirected for universal cytokine killing (TRUCK), also known as a fourth generation CAR. TRUCKs are CAR-redirected T cells used as carriers to produce and release a transgenic cytokine that accumulates in target tissue, such as target tumor tissue. The transgenic cytokine is released upon exposure of the CAR target. TRUCK cells can deposit various therapeutic cytokines into targets. This can result in therapeutic concentrations at the target site and eliminate systemic toxicity.

CARs typically differ in their functional properties. The CD3-zeta T-cell receptor signaling domain, when engaged, activates and induces T-cell proliferation, but can also lead to anergy (lack of response of the body's defense mechanisms, which leads to direct induction of tolerance in peripheral lymphocytes). Lymphocytes are considered anergic if they do not respond to a specific antigen. The addition of a costimulatory domain to second-generation CARs improved the replicative capacity and persistence of the engineered T cells. Similar antitumor effects are observed in vitro with CD28 or 4-1BB CARs, however, preclinical in vivo studies indicate that 4-1BB CARs may induce higher proliferation and/or resistance. Clinical trials indicate that both of these second-generation CARs are able to induce significant T cell proliferation in vivo, but CARs containing the 4-1BB co-stimulatory domain appear to persist longer. To increase the effectiveness of third-generation CARs, several signaling domains (costimulatory) are combined. Fourth-generation CARs are further modified with a constitutive or inducible transgenic cytokine expression cassette that is released by the CAR T cell to modulate the T cell response. See, for example, Enblad et al., Human Gene Therapy, 2015, 26(8): 498-505; Chmielewski and Hinrich, Expert Opinion on Biological Therapy, 2015, 15(8): 1145-1154, the disclosures of which are incorporated herein by reference in their entirety.

In some embodiments, the exemplary immunotherapeutic agent is a first generation chimeric antigen receptor CAR. In some embodiments, the chimeric antigen receptor is a second generation CAR. In some embodiments, the chimeric antigen receptor is a third generation CAR. In some embodiments, the chimeric antigen receptor is

- 59 044424

Fourth generation CAR or T cell redirected for universal cytokine killing (TRUCK).

In some embodiments, the chimeric antigen receptor (CAR) comprises an extracellular domain comprising an antigen binding domain, a transmembrane domain, and a cytoplasmic domain. In some embodiments, the CAR is entirely human. In some embodiments, the antigen binding domain of the CAR is specific for one or more antigens. In some embodiments, the spacer domain or hinge domain is located between the extracellular domain (containing the antigen binding domain) and the transmembrane domain of the CAR or between the cytoplasmic domain and the transmembrane domain of the CAR. A spacer domain refers to any oligopeptide or polypeptide that has the function of linking a transmembrane domain to an extracellular domain and/or a cytoplasmic domain in a polypeptide chain. A hinge domain refers to any oligopeptide or polypeptide that functions to provide flexibility to a CAR or its domains or to prevent steric hindrance to a CAR or its domains. In some embodiments, the spacer domain or hinge domain may comprise up to 300 amino acids (eg, 10 to 100 amino acids or 5 to 20 amino acids). In some embodiments, one or more spacer domains may be included in other regions of the CAR.

In some embodiments, the CAR of the present disclosure comprises an antigen binding domain, such as a single chain Fv (scFv), specific for a tumor antigen. The choice of binding domain depends on the type and number of ligands that define the target cell surface. For example, the antigen binding domain may be selected to recognize a ligand that acts as a cell surface marker on target cells associated with a particular pathological condition, such as cancer or an autoimmune disease. Thus, examples of cell surface markers that can act as ligands for the antigen binding domain in a CAR of the present disclosure include markers associated with cancer cells and/or other forms of pathological cells. In some embodiments, a CAR is designed to target a tumor antigen of interest by designing a required antigen binding domain that specifically binds to an antigen on a tumor cell encoded by an engineered nucleic acid provided herein.

An antigen binding domain (eg, scFv) that specifically binds to a target or epitope is a term understood in the art, and methods for determining such specific binding are also known in the art. A molecule is said to exhibit specific binding if it reacts or associates more frequently, more rapidly, more persistently, and/or with greater affinity with a particular target antigen than with alternative targets. An antigen binding domain (eg, scFv) that specifically binds to a first target antigen may or may not specifically bind to a second target antigen. Thus, specific binding does not necessarily require exclusive binding (even though it may involve it).

In some embodiments, CAR-expressing immune cells are genetically modified to recognize multiple targets or antigens, thereby recognizing unique target or antigen expression profiles on tumor cells. Examples of CARs that can bind to multiple targets include split-signal CARs that limit full activation of immune cells against tumors expressing multiple antigens; tandem CARs (TanCARs), which contain extracellular domains with two scFvs; and generic extracellular domain CARs that introduce avidin or fluorescein isothiocyanate (FITC)-specific scFv to recognize tumor cells that have been incubated with labeled monoclonal antibodies (Mab).

A CAR is considered bispecific if it recognizes two separate antigens (has two distinct antigen recognition domains). In some embodiments, the bispecific CAR includes two separate domains to recognize antigens present in tandem on a single transgenic receptor (referred to as TanCAR; see, e.g., Grada Z. et al., Molecular Therapy Nucleic Acids, 2013; 2:e105, included in this document is incorporated by reference in its entirety). Thus, in some embodiments, the methods include delivering to a tumor a combination comprising a crystalline form or a crystalline salt form of Compound 1 and an immunotherapeutic agent, wherein the immunotherapeutic agent is an engineered nucleic acid that encodes an antigen, or delivering to a tumor an engineered nucleic acid that causes expression of a self-antigen, and delivery to the tumor of an immune cell expressing a bispecific CAR that binds to two antigens, one of which is encoded by an engineered nucleic acid.

In some embodiments, the CAR is an antigen-specific inhibitory CAR (iCAR), which can be used, for example, to avoid extratumor

- 60 044424 toxicity (Fedorov, V.D. et al., Sci. Transl. Med., published online December 11, 2013, incorporated herein by reference in its entirety). iCARs contain an antigen-specific inhibitory receptor, for example, to block nonspecific immunosuppression that may result from overexpression of the target tumor. iCARs can be based on, for example, CTLA-4 or PD-1 inhibitory molecules. In some embodiments, these iCARs block T cell responses from T cells activated by either their endogenous T cell receptor or an activating CAR. In some embodiments, this inhibitory effect is temporary.

In some embodiments, the CAR can be used in adoptive cell transfer, wherein the immune cells are removed from a subject and modified so that they express receptors specific for an antigen, for example, a tumor-specific antigen. Modified immune cells, which can then recognize and destroy cancer cells, are reintroduced to the subject (Pule et al., Cytotherapy, 2003, 5(3): 211-226; Maude et al., Blood, 2015, 125(26): 4017 -4023, each of which is incorporated herein by reference in its entirety).

In accordance with other aspects of this disclosure, the tumor antigenic component of the vaccine of the present invention is any natural or synthetic tumor-associated protein or peptide, or a combination of tumor-associated proteins and/or peptides, or glycoproteins or glycopeptides. In other aspects, the antigenic component may be patient specific or common to many or most patients with a particular type of cancer. In one aspect, the antigenic component consists of a cell lysate obtained from tumor tissue removed from the patient being treated. In another aspect, the lysate can be engineered or synthesized from an exosome obtained from tumor tissue. In another aspect, the antigenic component consists of a cell lysate derived from tumor tissue extracted from one or more unrelated individuals or tumor cell lines.

In various embodiments, an exemplary immunotherapy agent comprises one or more cancer vaccines for use in combination with a crystalline form or crystalline salt form of Compound 1. The tumor associated antigen component of the vaccine can be produced by any of various well known techniques. For individual protein components, the antigenic protein is isolated from tumor tissue or tumor cell line by standard chromatographic techniques such as high performance liquid chromatography or affinity chromatography or, alternatively, it is synthesized by standard recombinant DNA technology in a suitable expression system such as E. coli , yeast or plants. The tumor associated antigenic protein is then purified from the expression system using standard chromatographic techniques. In the case of peptide antigenic components, they are usually obtained through standard automated synthesis. Proteins and peptides can be modified by the addition of amino acids, lipids and other means to improve their delivery into a vaccine delivery system (eg, multilayer liposome). For a tumor-associated antigenic component obtained from the patient's own tumor, or tumors of other individuals, or cell lines, the tumor tissue or a single cell suspension obtained from the tumor tissue is typically homogenized in a suitable buffer. The homogenate may also be fractionated, for example by centrifugation, to isolate specific cellular components, such as cell membranes or soluble material. The tumor material can be applied directly, or tumor-associated antigens can be extracted for inclusion in a vaccine using a buffer containing a low concentration of a suitable agent, such as a detergent. An example of a suitable detergent for the extraction of antigenic proteins from tumor tissue, tumor cells and tumor cell membranes is diheptanoyl phosphatidylcholine. Exosomes derived from tumor tissue or tumor cells, whether autologous or heterologous to the patient, can be used as an antigenic component for inclusion in a vaccine or as starting material for the extraction of tumor-associated antigens.

In some embodiments of this disclosure, the combination therapy includes a crystalline form or crystalline salt form of Compound 1 in combination with an immunotherapeutic agent that is a cancer vaccine. In various examples, the cancer vaccine includes at least one tumor-associated antigen, at least one immunostimulant, and optionally at least one cellular immunotherapy agent. In some embodiments, the immunostimulatory component in a cancer vaccine of the present disclosure is a biological response modifier (BRM) capable of enhancing the therapeutic efficacy of the cancer vaccine to induce humoral and cellular immune responses against cancer cells in a patient. In one aspect, the immunostimulant is a cytokine

- 61 044424 or a combination of cytokines. Examples of such cytokines include interferons such as IFN-gamma, interleukins such as IL-2, IL-15 and IL-23, colony stimulating factors such as M-CSF and GM-CSF, and tumor necrosis factors. In another aspect, the immunostimulatory component of a disclosed cancer vaccine includes one or more adjuvant-type immunostimulatory agents, such as APC toll-like receptor agonists or costimulatory/membrane cell adhesion proteins, with or without immunostimulatory cytokines. Examples of toll-like receptor agonists include lipid A and CpG and co-stimulatory/adhesion proteins such as CD80, CD86 and ICAM-1.

In some embodiments, the immunostimulant is selected from the group consisting of IFN-gamma (IFN-γ), IL-2, IL-15, IL-23, M-CSF, GM-CSF, tumor necrosis factor, lipid A, CpG, CD80, CD86 and ICAM-1, or combinations thereof. In other aspects, the cellular immunotherapy agent is selected from the group consisting of dendritic cells, tumor-infiltrating T lymphocytes, chimeric antigen receptor-modified effector T cells directed to the patient's tumor type, B lymphocytes, natural killer cells, bone marrow cells, and any other cells of the patient's immune system or combinations thereof. In one aspect, the cancer vaccine immunostimulator includes one or more cytokines such as interleukin 2 (IL-2), GM-CSF, M-CSF and interferon-gamma (IFN-γ), one or more toll-like receptor agonists and/or adjuvants, such as monophosphoryl lipid A, lipid A, muramyl dipeptide (MDP) lipid conjugate and double-stranded RNA, or one or more costimulatory membrane proteins and/or cell adhesion proteins such as CD80, CD86 and ICAM-1, or any combination of the above. In one aspect, the cancer vaccine includes an immunostimulant that is a cytokine selected from the group consisting of interleukin 2 (IL-2), GM-CSF, M-CSF, and interferon-gamma (IFN-γ). In another aspect, the cancer vaccine includes an immunostimulant that is a toll-like receptor agonist and/or an adjuvant selected from the group consisting of monophosphoryl lipid A, lipid A and muramyl dipeptide (MDP) lipid conjugate, and double-stranded RNA. In another aspect, the cancer vaccine includes an immunostimulant that is a costimulatory membrane protein and/or a cell adhesion protein selected from the group consisting of CD80, CD86 and ICAM-1.

In various embodiments, the immunotherapy agent may include a cancer vaccine, wherein the cancer vaccine administers any tumor antigen that can potentially be used for the fusion protein construct of the present invention and, in particular, the following:

(a) cancer-testicular antigens, including NY-ESO-1, SSX2, SCP1, as well as RAGE, BAGE, GAGE and MAGE family polypeptides, for example GAGE-1, GAGE-2, MAGE-1 MAGE-2, MAGE -3, MAGE-4, MAGE-5, MAGE-6 and MAGE-12, which can be used, for example, to target melanoma, lung, head and neck, NSCLC, breast, gastrointestinal and bladder tumors;

(b) mutated antigens, including p53, associated with various solid tumors, for example, colorectal, lung, head and neck cancer; p21/Ras, associated with, for example, melanoma, pancreatic cancer and colorectal cancer; CDK4, associated with melanoma, for example; MUM1, associated with melanoma, for example; caspase-8, associated with, for example, head and neck cancer; CIA 0205, associated with, for example, bladder cancer; HLA-A2-R1701, beta-catenin, associated with melanoma, for example; TCR associated with, for example, T-cell non-Hodgkin's lymphoma; BCR-abl, associated, for example, with chronic myelogenous leukemia; triosephosphate isomerase; KIA 0205; CDC-27 and LDLR-FUT;

(c) overexpressed antigens, including galectin 4, associated with, for example, colorectal cancer; galectin 9, associated with, for example, Hodgkin's disease; proteinase 3, associated with, for example, chronic myelogenous leukemia; WT 1, associated, for example, with various leukemias; carbonic anhydrase, associated, for example, with kidney cancer; aldolase A, associated with, for example, lung cancer; PRAME, associated with melanoma, for example; HER-2/neu, associated with, for example, breast, colon, lung and ovarian cancer; mammaglobin, an alpha-fetoprotein associated with hepatoma, for example; KSA associated with, for example, colorectal cancer; gastrin, associated, for example, with pancreatic and esophageal cancer; telomerase catalytic protein, MUC-1, associated with, for example, breast and ovarian cancer; G-250, associated with, for example, renal cell carcinoma; p53 associated, for example, with breast and colon cancer; and carcinoembryonic antigen associated with, for example, breast cancer, lung cancer, and gastrointestinal cancers such as colorectal cancer;

(d) common antigens, including melanoma-melanocytic differentiation antigens such as MART-1/Melan A; gpl00; MC1R; melanocyte-stimulating hormone receptor; tyrosinase; tyrosinase-related protein-1/TRP1 and tyrosinase-related protein-2/TRP2, associated with, for example, melanoma;

- 62 044424 (e) prostate-associated antigens, including PAP, PSA, PSMA, PSH-P1,

PSM-P1, PSM-P2, associated, for example, with prostate cancer;

(f) immunoglobulin idiotypes associated with myeloma and B-cell lymphomas.

In some embodiments, one or more TAAs may be selected from pi 5, Hom/Mel-40, H-Ras, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein-Barr virus antigens, EBNA antigens human papilomavirus (HPV), including E6 and E7, virus antigens B and C, lymphotropic T-cell virus antigens, TSP-180, pl85erbB2, pl 80erbB-3, c-met, mn-23H1, TAG-72-4 , CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, pi 6, TAGE, PSCA, CT7, 43-9F, 5T4, 791 Tgp72, beta-HCG, BCA225, BTAA, CA 125, CA 15-3 (CA 27.29/BCAA), CA 195, CA 242, CA-50, CAM43, CD68/KP1, CO-029, FGF-5, Ga733 (EpCAM), HTgp-175, M344, MA-50, MG7 -Ag, MOV18, NB/70K, NY-CO-1, RCAS1, SDCCAG16, TA-90 (Mac-2 binding protein/cyclophilin C associated protein), TAAL6, TAG72, TLP, TPS or any combination thereof.

In some embodiments, the present disclosure provides a crystalline form or crystalline salt form of Compound 1 for use in combination with a cancer vaccine, which may include a tumor antigen comprising all, a portion of, or specific immunogenic epitopes of a human protein.

In various embodiments, an exemplary immunotherapeutic agent may include mRNA suitable for encoding any one or more of the above cancer antigens useful for the synthesis of a cancer vaccine. In some illustrative embodiments, an mRNA cancer vaccine may have one or more of the following properties:

a) the mRNA encoding each cancer antigen is interspersed with cleavage-sensitive sites;

b) the mRNA encoding each cancer antigen is linked directly to each other without a linker;

c) the mRNA encoding each cancer antigen is linked to each other by a single nucleotide linker;

d) each cancer antigen includes 20-40 amino acids and a centrally located SNP mutation;

e) at least 40% of cancer antigens have the highest affinity for MHC class I molecules from the subject;

f) at least 40% of the cancer antigens have the highest affinity for MHC class II molecules from the subject;

g) at least 40% of cancer antigens have a predicted binding affinity IC >500 nM for HLA-A, HLA-B and/or DRB1;

h) mRNA encodes 1 to 15 cancer antigens;

i) 10-60% of cancer antigens have MHC class I binding affinity and 10-60% of cancer antigens have MHC class II binding affinity; and/or

j) The mRNA encoding cancer antigens is designed so that the cancer antigens are ordered to minimize pseudoepitopes.

In various embodiments, a combination comprising a crystalline form or crystalline salt form of Compound 1 and a cancer vaccine immunotherapeutic agent disclosed herein can be used to antagonize an immune response in a subject against a cancer antigen. The method includes administering to a subject an RNA vaccine containing at least one RNA polynucleotide having an open reading frame encoding at least one antigenic polypeptide or an immunogenic fragment thereof, thereby inducing in the subject an immune response specific to the antigenic polypeptide or an immunogenic fragment thereof, in combination with administration of a crystalline form or a crystalline salt form of Compound 1, either in the same composition or in a separate composition, administered simultaneously or sequentially, wherein the antiantigen polypeptide antibody titer in the subject increases after vaccination compared to the antiantigen polypeptide antibody titer in the subject vaccinated prophylactically effective dose of traditional cancer vaccine. An antigenic polypeptide antibody is a serum antibody that specifically binds to an antigenic polypeptide.

A prophylactically effective dose is a therapeutically effective dose that prevents the development of cancer at a clinically acceptable level. In some embodiments, the therapeutically effective dose is the dose specified in the vaccine package insert. The conventional vaccine used herein refers to a vaccine different from the mRNA vaccines of the present invention. For example, a conventional vaccine includes, but is not limited to, live microbial vaccines, killed microbial vaccines, subunit vaccines, protein-antigen vaccines, DNA vaccines, and the like. In illustrative embodiments, a conventional vaccine is a vaccine that has received regulatory approval and/or is registered by a national regulatory authority

- 63 044424 medicinal products, for example by the Food and Drug Administration (FDA) in the USA or the European Medicines Agency (EMA).

In some embodiments, the antibody titer against the antigenic polypeptide in a subject increases from 1 log to 10 log after vaccination compared to the antibody titer against the antigenic polypeptide in a subject vaccinated with a prophylactically effective dose of a conventional cancer vaccine. In some embodiments, the antibody titer against the antigenic polypeptide in a subject is increased by up to 1 log after vaccination compared to the antibody titer against the antigenic polypeptide in a subject vaccinated with a prophylactically effective dose of a conventional cancer vaccine. In some embodiments, the antibody titer against the antigenic polypeptide in a subject is increased by up to 2 logs following vaccination compared to the antibody titer against the antigenic polypeptide in a subject vaccinated with a prophylactically effective dose of a conventional cancer vaccine.

Aspects of the present invention provide nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotide is present in a formulation for in vivo administration to a host that provides an antibody titer that exceeds the seroprotection criterion for the first antigen for an acceptable percentage of people. In some embodiments, the antibody titer produced by the mRNA vaccines of the present invention is a neutralizing antibody titer. In some embodiments, the neutralizing antibody titer is greater than that of the protein vaccine. In other embodiments, the titer of neutralizing antibodies produced by the mRNA vaccines of the present invention is greater than that of a protein-adjuvated vaccine. In other embodiments, the titer of neutralizing antibodies produced by the mRNA vaccines of the present invention is 1000-10000, 1200-10000, 1400-10000, 1500-10000, 1000-5000, 1000-4000, 1800-10000, 2000-10000, 2000 -5000, 2000-3000, 2000-4000, 3000-5000, 3000-4000 or 2000-2500. The neutralization titer is usually expressed as the maximum serum dilution required to achieve a 50% reduction in plaque burden.

In preferred aspects, the RNA vaccine immunotherapeutics of the present disclosure (eg, mRNA vaccines) produce prophylactically and/or therapeutically effective levels, concentrations and/or titers of antigen-specific antibodies in the blood or serum of the vaccinated subject. As defined herein, the term antibody titer refers to the amount of antigen-specific antibody produced in a subject, such as a human. In illustrative embodiments, the antibody titer is expressed as the reciprocal of the highest dilution (in serial dilution) that still provides a positive result. In illustrative embodiments, the antibody titer is determined or measured by enzyme-linked immunosorbent assay (ELISA). In illustrative embodiments, the antibody titer is determined or measured by a neutralization assay, such as a microneutralization assay. In some aspects, the antibody titer measurement is expressed as a ratio, such as 1:40, 1:100, and the like.

In illustrative embodiments of the present invention, the effective vaccine provides an antibody titer greater than 1:40, greater than 1:100, greater than 1:400, greater than 1:1000, greater than 1:2000, greater than 1:3000, greater than 1:4000, greater than 1:500 , more than 1:6000, more than 1:7500, more than 1:10000. In illustrative embodiments, the antibody titer is produced or achieved 10 days after vaccination, 20 days after vaccination, 30 days after vaccination, 40 days after vaccination, or 50 days or more after vaccination. In illustrative embodiments, a titer is produced or achieved after administration of a single dose of the vaccine to a subject. In other embodiments, the titer is established or achieved after multiple doses, for example, after a first and second dose (eg, a booster dose). In illustrative aspects of the invention, antigen-specific antibodies are measured in units of μg/ml or measured in units of IU/L (international units per liter) or mIU/ml (international milliunits per ml). In illustrative embodiments of the invention, the effective vaccine provides greater than 0.5 μg/ml, greater than 0.1 μg/ml, greater than 0.2 μg/ml, greater than 0.35 μg/ml, greater than 0.5 μg/ml, greater than 1 µg/ml, more than 2 µg/ml, more than 5 µg/ml or more than 10 µg/ml. In illustrative embodiments of the invention, the effective vaccine provides greater than 10 mIU/ml, greater than 20 mIU/ml, greater than 50 mIU/ml, greater than 100 mIU/ml, greater than 200 mIU/ml, greater than 500 mIU/ml, or greater than 1000 mIU/ml. In illustrative embodiments, the level or concentration of antibodies is produced or achieved 10 days after vaccination, 20 days after vaccination, 30 days after vaccination, 40 days after vaccination, or 50 or more days after vaccination. In illustrative embodiments, the level or concentration is produced or achieved after administration of one dose of the vaccine. In other embodiments, the level or concentration is produced or achieved after multiple doses, such as after a first and second dose (eg, a booster dose). In illustrative embodiments, the antibody level or concentration is detected or measured by an enzyme-linked immunosorbent

- 64 044424 studies (ELISA). In illustrative embodiments, the antibody level or concentration is determined or measured by a neutralization assay, such as a microneutralization assay. Also provided are nucleic acid vaccines containing one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide or a concatemeric polypeptide, wherein the RNA polynucleotide is present in the composition for administration in vivo to a host to produce a longer-lasting high antibody titer than the antibody titer caused by an mRNA vaccine containing a stabilizing element or formulated with an adjuvant and encoding a first antigenic polypeptide. In some embodiments, the RNA polynucleotide is formulated to produce neutralizing antibodies within one week of a single administration. In some embodiments, the adjuvant is selected from a cationic peptide and an immunostimulatory nucleic acid. In some embodiments, the cationic peptide is protamine.

Immunotherapeutic agents containing a nucleic acid-based vaccine containing one or more RNA polynucleotides with an open reading frame containing at least one chemical modification or optionally not containing a nucleotide modification, an open reading frame encoding a first antigenic polypeptide or a concatemeric polypeptide, wherein the RNA polynucleotide is present in a formulation for in vivo administration to a host such that the level of antigen expression in the host is significantly greater than the level of antigen expression produced by an mRNA vaccine containing a stabilizing element or containing an adjuvant and encoding the first antigenic polypeptide.

In other aspects, nucleic acid vaccines are provided comprising one or more RNA polynucleotides having an open reading frame containing at least one chemical modification, or optionally not containing a nucleotide modification, an open reading frame encoding a first antigenic polypeptide or a concatemer polypeptide, wherein the vaccine has less than 10 times less RNA polynucleotide than is required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotide is present at a dosage of 25-100 μg.

Aspects of the present invention also provide a vaccine administration unit comprising from 10 to 400 μg of one or more RNA polynucleotides having an open reading frame containing at least one chemical modification or optionally no nucleotide modification, wherein the open reading frame encodes a first antigenic polypeptide or concatemer a polypeptide and pharmaceutically acceptable excipient formulated for delivery to a human subject. In some embodiments, the vaccine further comprises a cationic lipid nanoparticle.

Aspects of the present invention relate to methods of creating, maintaining or restoring antigenic memory to a tumor in an individual or population of individuals, comprising administering to said individual or population an antigenic vaccine with a memory enhancing nucleic acid comprising (a) at least one RNA polynucleotide containing at least at least one chemical modification, or optionally free of nucleotide modifications, and two or more codon optimized open reading frames, wherein said open reading frames encode a set of reference antigenic polypeptides; and (b) optionally a pharmaceutically acceptable excipient.

In some embodiments, the vaccine is administered to an individual via a route selected from the group including intramuscular administration, intradermal administration, and subcutaneous administration. In some embodiments, the administration step includes contacting muscle tissue of the subject with a device suitable for injecting the composition. In some embodiments, the administration step includes contacting muscle tissue of the subject with a device suitable for injecting the composition in combination with electroporation.

Aspects of the present invention provide methods of vaccinating a subject comprising administering to the subject a single dose of 25 to 400 μg/kg of a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide or concatemer polypeptide in an effective amount to vaccination of the subject.

In other aspects, nucleic acid-based vaccines are provided comprising one or more RNA polynucleotides having an open reading frame containing at least one chemical modification, an open reading frame encoding a first antigenic polypeptide or a concatemer polypeptide, wherein the vaccine has less than 10-fold less RNA polynucleotide than is required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotide is present at a dosage of 25-100 μg.

- 65 044424

In some embodiments, the crystalline form or crystalline salt form of Compound 1 can be used in combination with a bispecific antibody immunotherapy agent. The bispecific antibody may include a protein construct having a first antigen binding moiety and a second antigen binding site that binds to a cytotoxic immune cell. The first antigen binding site can bind to a tumor antigen that is specifically treated with the combination of the present invention. For example, the first antigen binding fragment may bind to a non-limiting example of tumor antigens selected from EGFR, HGFR, Her2, Ep-CAM, CD20, CD30, CD33, CD47, CD52, CD133, CEA, gpA33, mucins, TAG-72, CIX, PSMA , folate binding protein, GD2, GD3, GM2, VEGF. Included among others are VEGFR, αVβ3 integrin, α5β1 integrin, MUC1, ERBB2, ERBB3, MET, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, and tenascin. In some embodiments, the first antigen-binding moiety has specificity for a protein or peptide that is overexpressed on a tumor cell relative to a corresponding non-tumor cell. In some embodiments, the first antigen binding fragment has specificity for a protein that is overexpressed on a tumor cell relative to a corresponding non-tumor cell. As used herein, the term corresponding non-tumor cell refers to a non-tumor cell that belongs to the same cell type as the original tumor cell. It is noted that such proteins do not necessarily differ from tumor antigens. Non-limiting examples include carcinoembryonic antigen (CEA), which is overexpressed in most carcinomas of the colon, rectum, breast, lung, pancreas and gastrointestinal tract; heregulin receptors (HER-2, neu or c-erbB-2), which are often overexpressed in breast, ovarian, colon, lung, prostate and cervical cancers; epidermal growth factor receptor (EGFR), which is highly expressed in a number of solid tumors, including breast, head and neck, non-small cell lung and prostate tumors; asialoglycoprotein receptor; transferrin receptor; serpin enzyme complex receptor, which is expressed on hepatocytes; fibroblast growth factor receptor (FGFR), which is overexpressed on pancreatic ductal adenocarcinoma cells; vascular endothelial growth factor receptor (VEGFR), for gene therapy against angiogenesis; folate receptor, which is selectively overexpressed in 90% of nonmucosal ovarian carcinomas; cell surface glycocalyx; carbohydrate receptors; and polymeric immunoglobulin receptor.

The second antigen binding fragment is any molecule that specifically binds to an antigen, protein or polypeptide expressed on the surface of a cytotoxic immune cell (CIK cell). Non-limiting examples of antigens expressed on the surface of cytotoxic immune cells suitable for use with the present disclosure may include CD2, CD3, CD4, CD5, CD8, CD11a, CD11 b, CD14, CD16a, CD27, CD28, CD45, CD45RA, CD56, CD62L , Fc receptor, LFA, LFA-1, TCRαβ, CCR7, macrophage inflammatory protein 1a, perforin, PD-1, PD-L1, PD-L2 or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR , TGF-beta, TIM-3, SIRP-alpha, TIGIT, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2, CD27 and Fas ligand. In some embodiments, the second antigen binding fragment binds to the CD3 of a cytotoxic immune cell, such as a CIK cell. In some embodiments, the second antigen-binding fragment binds to CD56 of a cytotoxic immune cell. In some embodiments, the second antigen-binding fragment binds to the Fc receptor of a cytotoxic immune cell. In some embodiments, the Fc region of the bispecific antibody binds to the Fc receptor of a cytotoxic immune cell. In some embodiments, the second antigen-binding moiety is any molecule that specifically binds to an antigen expressed on the surface of a cytotoxic immune cell (eg, a CIK cell). The second antigen-binding moiety is specific for an antigen on a cytotoxic immune cell. Examples of cytotoxic immune cells include, but are not limited to, CIK cells, T cells, CD8+ T cells, activated T cells, monocytes, natural killer (NK) cells, NK T cells, lymphokine-activated killer (LAK) cells, macrophages, and dendritic cells. The second antigen-binding moiety specifically binds to an antigen expressed on the surface of a cytotoxic immune cell. Non-limiting examples of antigens expressed on the surface of cytotoxic immune cells suitable for modulation with the present disclosure may include CD2, CD3, CD4, CD5, CD8, CD11a, CD11 b, CD14, CD16a, CD27, CD28, CD45, CD45RA, CD56, CD62L , Fc receptor, LFA, LFA-1, TCRαβ, CCR7, macrophage inflammatory protein 1a, perforin, PD-1, PD-L1, PD-L2 or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR , TGF-beta, TIM-3, SIRP-alpha, TIGIT, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2, CD27 and Fas ligand. In other embodiments, the bispecific antibody modulator is a costimulatory molecule activator (eg, an OX40 agonist). In one embodiment, the OX40 agonist is a bispecific antibody molecule to OX40 and another tumor antigen or costimulatory antigen.

- 66 044424

An OX40 agonist can be administered alone or in combination with other immunomodulators, for example, in combination with an inhibitor (antibody construct) of PD-1, PD-L1, CTLA-4, CEACAM (e.g. CEACAM-1, -3 and/or -5) , TIM-3 or LAG-3. In some embodiments, the anti-OX40 antibody molecule is a bispecific antibody that binds to GITR and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3 and/or -5), TIM-3 or LAG-3. In one illustrative embodiment, an OX40 antibody molecule is administered in combination with an anti-PD-1 antibody molecule (eg, an anti-PD-1 antibody molecule described herein). The OX40 antibody molecule and the anti-PD-1 antibody molecule may be in the form of a separate antibody composition or as a bispecific antibody molecule. In other embodiments, an OX40 agonist may be administered in combination with another co-stimulatory molecule, e.g., a GITR agonist, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB ( CD137), CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3 or CD83 ligand. In some embodiments, the second antigen binding moiety binds to an Fc receptor on a cytotoxic immune cell, such as a CIK cell.

In some embodiments, the bispecific antibody immunotherapy agent has tumor antigen and CIK cell specificity that brings the tumor cell expressing the tumor antigen closer to the CIK cell, resulting in elimination of the tumor cell due to the antitumor cytotoxicity of the CIK cell. In some embodiments, the bispecific antibody is specific for a tumor antigen but not specific for a CIK cell, however, the Fc region of the bispecific antibody can bind to the Fc receptor of the CIK cell, which in turn results in the formation of a tumor cell in close proximity to CIK cells, which leads to the destruction of the tumor cell due to the antitumor cytotoxicity of the CIK cell. In some embodiments, the bispecific antibody is specific for a CIK cell but not specific for a tumor cell, however, the Fc region of the bispecific antibody can bind to the Fc receptor of the tumor cell, which in turn brings the tumor cell closer to the CIK cell, resulting in the destruction of the tumor cell. cells due to the antitumor cytotoxicity of the CIK cell.

In some embodiments, a crystalline form or crystalline salt form of Compound 1 can be used in combination with a multivalent anti-immune cell antibody/fusion protein/engineered immunotherapy agent. In various embodiments, an exemplary immunotherapy agent may include a multivalent antibody/fusion protein/immune cell interacting construct that may comprise a recombinant structure, such as all engineered antibodies, that do not mimic the original IgG structure. In this case, different strategies for multimerization of antibody fragments are used. For example, shortening the peptide linker between the V domains results in the self-association of scFv into a dimer (diabody; 55 kDa). Bispecific diabodies are formed as a result of non-covalent association of two fragments VHA-VLB and VHB-VLA expressed in the same cell. This results in the formation of heterodimers with two different binding sites. Single-chain diabodies (sc-diabodies) are bispecific molecules in which the VHA-VLB and VHB-VLA fragments are linked by an additional third linker. Tandem diabodies (Tandab) are tetravalent bispecific antibodies produced by two sc diabodies.

Also included are di-diabodies known in the art. This 130 kDa molecule is formed by fusion of the diabody to the N-terminus of the CH3 domain of IgG, resulting in an IgG-like structure. Other derivatives of diabodies are triatera and tetrabodies, which fold into trimeric and tetrameric fragments by shortening the linker to less than 5 or 0-2 residues. Also included are (scFv) ₂ constructs known as bispecific T cell capture (BITE). BITEs are bispecific single chain antibodies consisting of two scFv antibody fragments connected by a flexible linker that are directed against surface antigen on target cells and CD3 on T cells. Examples of divalent (Fab)2 and trivalent (Fab)3 antibody formats are also provided. Minibodies and trimeric bodies derived from scFv are also presented. Exemplary constructs useful for targeting tumor antigens may include one or more of the following: diabody, single chain (sc)-diabody (scFv)2, minibody, minibody, barnazabarstar, scFv-Fc, sc(Fab)2, trimeric antibody constructs, tribody antibody constructs, trimerbody antibody constructs, tribody antibody constructs, collatebody antibody constructs, (scFv-TNFa)3, F(ab)3/DNL. Examples of cytotoxic immune cells include, but are not limited to, CIK cells, T cells, CD8+ T cells, activated T cells, monocytes, natural killer (NK) cells, NK T cells, lymphokine-activated killer (LAK) cells, macrophages, and dendritic cells.

In some embodiments, the crystalline form or crystalline salt form of Compound 1 can be used in combination with a radioconjugate immunotherapy agent.

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In various embodiments, a radioconjugate is a small molecule or large molecule (herein referred to as a cell-targeting agent), such as a polypeptide, antibody, or antibody fragment thereof, that is bound or otherwise attached to a radionuclide, or multiple radionuclides, such that binding of the radioconjugate with its target (protein or molecule on or in a cancer cell) will result in the death or morbidity of said cancer cell. In various embodiments, the radioconjugate may be a cell-targeting agent labeled with a radionuclide, or the cell-targeting agent may be bound or otherwise attached to a particle or microparticle or nanoparticle containing a plurality of radionuclides, wherein the radionuclides are the same or different. Methods for synthesizing radioconjugates are known in the art and may involve a class of immunoglobulins or antigen binding portions thereof that are conjugated to a toxic radionuclide.

In some embodiments, a molecule that binds to a cancer cell may be known as a cell-targeting agent. An exemplary cell-targeting agent as used herein may allow drug-containing nanoparticles or radionuclide to target specific types of cells of interest. Examples of cell-targeting agents include, but are not limited to, small molecules (eg, folate, adenosine, purine) and large molecules (eg, peptide or antibody) that bind to or target a tumor-associated antigen. Examples of tumor-associated antigens include, but are not limited to, adenosine receptors, alpha v beta 3, aminopeptidase P, alpha-fetoprotein, cancer antigen 125, carcinoembryonic antigen, cCaveolin1, chemokine receptors, clusterin, oncofetal antigens, CD20, epithelial tumor antigen associated melanoma antigen, Ras, p53, Her2/Neu, ErbB2, ErbB3, ErbB4, folic acid receptor, prostate-specific membrane antigen, prostate-specific antigen, purine receptors, radiation-induced cell surface receptor, serpin B3, serpin B4, squamous cell carcinoma antigen, thrombospondin, tumor antigen 4, tumor associated glycoprotein 72, tyrosinases and tyrosine kinases. In some embodiments, the cell-targeting agent is folate or a folic acid derivative that specifically binds to folate receptors (FRs). In some embodiments, the cell-targeting agent is an antibody, bispecific antibody, trispecific antibody, or an antigen-binding construct thereof that specifically binds to a cancer antigen selected from EGFR, HGFR, Her2, Ep-CAM, CD20, CD30, CD33, CD47 , CD52, CD133, CEA, gpA33, mucins, TAG-72, CIX, PSMA, folate binding protein, GD2, GD3, GM2, VEGF. Included among others are VEGFR, αVβ3 integrin, α5β1 integrin, MUC1, ERBB2, ERBB3, MET, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, and tenascin.

The use of folate as a targeting agent in the radioconjugate also allows for targeting of both tumor cells and regulatory T (Treg) cells for destruction. It is well known that large numbers of Treg cells suppress tumor immunity. In particular, Treg cells suppress (foreign and self-reactive) T cells without killing them through contact-dependent secretion or secretion of cytokines (eg, IL-10, TGF-beta, etc.). FR4 is selectively activated on Treg cells. The antibody was shown to block FR4-depleted Treg cells and provoke tumor immunity in tumor-bearing mice. Thus, folate-coated PBM nanoparticles carrying a cytotoxic agent will recruit FR-expressing cells for their destruction, both directly (i.e., BrCa cells) and indirectly (i.e., breast tumor-associated and peripheral Tregs -cells) inhibit tumor progression.

In another further embodiment, the targeting agent is an antibody or peptide or multivalent antibody/fusion protein/construct involving immune cells capable of binding tumor associated antigens, consisting of, but not limited to, the following: adenosine receptors, alpha v beta 3, aminopeptidase P , alpha fetoprotein, cancer antigen 125, carcinoembryonic antigen, caveolin-1, chemokine receptors, clusterin, oncofetal antigens, CD20, human growth factor receptor (HGFR), epithelial tumor antigen, melanoma-associated antigen, MUC1, Ras, p53, Her2/Neu, ErbB2, ErbB3, ErbB4, folate receptor, prostate-specific membrane antigen, prostate-specific antigen, purine receptors, radiation-induced cell surface receptor, serpin B3, serpin B4, squamous cell carcinoma antigen, thrombospondin, tumor associated antigen 4 glycoprotein 72, tyrosinase, tyrosine kinases, etc.

In some embodiments, the crystalline form or crystalline salt form of Compound 1 described herein can be used in combination with a vaccination protocol for the treatment of cancer. In some embodiments, the crystalline form or crystalline salt form of Compound 1 described herein can be used in combination with an immunotherapeutic agent, such as a vaccine. In various variants

- 68 044424 examples of vaccines include vaccines used to stimulate an immune response to cancer antigens.

An amount of both the crystalline form and the crystalline salt form of Compound 1 described herein and one or more additional therapeutic agents (in those compositions that contain the additional therapeutic agent as described above) that can be combined with auxiliary materials to obtain standard dosage form will vary depending on the patient being treated and the specific route of administration. In some embodiments, the compositions of the present invention are formulated so that a dose of 0.01 to 100 mg/kg body weight/day of the present invention can be administered.

The additional therapeutic agent and the crystalline form or crystalline salt form of Compound 1 disclosed herein may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions may be less than that required in monotherapy using that therapeutic agent alone, or there may be fewer side effects to the patient when using a lower dose. In some embodiments, a dosage of from 0.01 to 10,000 μg/kg body weight/day of the additional therapeutic agent may be administered in such compositions.

In some embodiments, crystalline forms or crystalline salt forms of Compound 1 disclosed herein can be combined with one or more inhibitors of the following kinases for the treatment of a disease disclosed herein, such as cancer: Aktl, Akt2, AIZ, TGF-3R, PKA, PKG, PKC, CaM kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, 1NS-R, IGF-1R, IR-R, PDGFaR, PDGFp/R, CSFIR, KIT , FLK-II, KDR/FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR/Flt2, Flt4, EphAl, EphA2, EphA3 , EphB2, EphB4, Tie2, Src, Fyn, Lek, Fgr, Btk, Fak, SYR, FRK, JAK, ABL, ALK, CDK7, CDK12, CDK13, KRAS and B-Raf. In some embodiments, the crystalline forms or salt forms of Compound 1 disclosed herein can be combined with one or more inhibitors of the CD47 and MALT1 proteins for the treatment of cancer.

In some embodiments, crystalline forms or crystalline salt forms of Compound 1 disclosed herein can be used in combination with one or more poly[ADP-ribose] polymerase (PARP) inhibitors to treat a disease disclosed herein, such as cancer. Exemplary PARP inhibitors include, but are not limited to, olaparib (Lynparza®), rucaprib (Rubraca®), niraparib (Zejula®), talzoparib (Talzenna®), and TPST-1120.

In some embodiments, the crystalline forms or crystalline salt forms of Compound 1 disclosed herein can be used in combination therapy with any of the kinase inhibitors disclosed herein to treat diseases such as cancer. Exemplary kinase inhibitors include imatinib, baricitinib, gefitinib, erlotinib, sorafenib, dasatinib, sunitinib, lapatinib, nilotinib, pirfenidone, zanubrutinib, upadacitinib, fedratinib, entrectinib, alpelisib, pazopanib, crizotinib, vemurafenib, vandetani b, ruxolitinib, axitinib, bosutinib, regorafenib, tofacitinib, cabozantinib, ponatinib, trametinib, dabrafenib, afatinib, ibrutinib, ceritinib, idelalisib, nintedanib, palbociclib, lenvatinib, cobimetinib, abemaciclib, acalabrutinib, alectinib, binimetinib, brigatinib, encorafenib, er dafitinib, everolimus, fostamatinib, gilter, larotrectinib, lorlatinib, netarsudil, osimertinib, pexidartinib, ribociclib, temsirolimus, XL-147, XL-765, XL-499 and XL-880. In some embodiments, a compound described herein can be used in combination with an HSP90 inhibitor (e.g., XL888), liver X receptor (LXR) modulators, retinol-related orphan receptor gamma (RORy) modulators, a CK1 inhibitor, a CK1-a inhibitor, a a Wnt pathway (eg, SST-215) or a mineralocorticoid receptor inhibitor (eg, esaxeronone or XL-550) for treating a disease disclosed herein, such as cancer.

In some embodiments, crystalline forms or crystalline salt forms of Compound 1 disclosed herein can be used in combination with polatuzumab vedotin to treat a disease disclosed herein, such as cancer.

Labeled compounds and methods of analysis.

Another aspect relates to the labeled crystalline form or crystalline salt form of the present invention (labeled with a radioactive isotope, fluorescent label, etc.), useful not only in imaging methods, but also in assays, both in vitro and in vivo, for localization and quantification of TAM kinases in tissue samples, including human ones, and to identify TAM kinase ligands by inhibiting the binding of a labeled compound. Accordingly, the present invention includes TAM kinase assays that contain such labeled compounds.

The present invention also includes an isotopically labeled crystalline form or a crystalline salt form of the present invention. Isotopically or radioactively labeled

-69044424 compound is a compound of the present invention in which one or more atoms are replaced or replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (ie, naturally occurring). Suitable radionuclides that may be included in the crystalline form or crystalline salt form of the present invention include, but are not limited to, the following: ² H (also denoted D as deuterium), ³ H (also denoted T as tritium), ⁿ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N ^{, 15} O, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³⁵ S, ³⁶ Cl, 82 Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I and ¹³¹ I. The radionuclide that is included in a given radiolabeled compound will depend on the particular application of the radiolabeled compound. For example, for metalloprotease labeling and in vitro competition assays, compounds that include ³ H, ¹⁴ C, ⁸² Br, ¹²⁵ I, ¹³¹ I, or ³⁵ S will generally be most suitable. will be ⁿ C, ¹⁸ F, ¹²⁵ I, ¹²³ I, ¹²⁴ I, ¹³¹ I, ⁷⁵ Br, ⁷⁶ Br or ⁷⁷ Br. In some embodiments, crystalline forms or crystalline salt forms described herein, in which one or more hydrogens are replaced by deuterium, for example, hydrogen bonded to a carbon atom. Such compounds exhibit increased resistance to metabolism and are thus useful for increasing the half-life of any compound when administered to a mammal, especially a human.

It is understood that a radiolabeled or labeled compound is a compound that contains at least one radionuclide. In some embodiments, the radionuclide is selected from the group consisting of ³ H, ¹⁴ C, ¹²⁵ 1, ³⁵ S, and ⁸² Br.

The present invention may further include synthetic methods for incorporating radioisotopes into the crystalline form or crystalline salt form of the present invention. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and those skilled in the art will readily recognize the methods applicable to the compounds of the present invention.

The labeled compound of the present invention can be used in a screening assay for identifying/evaluating compounds. For example, a newly synthesized or identified compound (ie, a test compound) that is labeled can be assessed for its ability to bind TAMs by monitoring the change in its concentration upon contact with TAM kinases by tracking the label. For example, a test compound (labeled) can be assessed for its ability to reduce the binding of another compound known to bind to TAM kinase (ie, a reference compound). Accordingly, the ability of a test compound to compete with a standard compound for binding to TAM kinase directly correlates with its binding affinity. Conversely, in some other screening assays the standard compound is labeled but the test compounds are unlabeled. Accordingly, the concentration of the labeled standard compound is monitored to assess the competition between the standard compound and the test compound and thereby determine the relative binding capacity of the test compound.

Preparations and examples

General experimental techniques.

Experiments with aqueous suspension. Salts of compound 1, which were determined to have a water solubility of less than 1 mg/ml, were suspended in 20 ml of water at ambient temperature for 1 day. The solids were then collected by vacuum filtration and analyzed by XRPD.

Extreme Cooling (CC). Concentrated solutions of compound 1 and various counterions were prepared in MeOH at elevated temperature with stirring. Closed vials containing hot solutions were transferred to a freezer (-20°C) and quickly cooled. The solids formed were collected. If solids were not present, additional crystallization techniques were used.

Emergency precipitation (CP). Pure solutions of compound 1 and coformer were prepared in various solvents at RT. Aliquots of the various antisolvents were added to the solution, slowly, with gentle stirring until the solids precipitated out of solution. The mixtures were left to stir for the indicated time. The solids formed were collected by pressurized filtration.

Fast cooling (FC). Concentrated solutions of compound 1 and various counterions were prepared in acetone or MeOH at elevated temperature with stirring. Closed vials containing hot solutions were transferred to a work surface at ambient temperature. The solids formed were collected. If solids were not present, additional crystallization techniques were used.

Fast evaporation (FE). Pure solutions of compound 1 and the coformer were prepared in various solvents. The vials were left uncapped and the solvent was evaporated under ambient conditions.

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Suspension for interconversion. A Form A suspension of Compound 1 was prepared by adding sufficient solids to the given solvent system at ambient conditions such that undissolved solids were present. The mixture was then stirred for additional time to ensure saturation. Solids of the forms of interest were then added to an aliquot of the saturated solution (filtered through a 0.2-μm nylon filter) so that undissolved solids were present. The mixture was then stirred at ambient temperature for additional time and solids were isolated.

Isolation techniques. Typically, isolation was performed quickly after removal of samples from appropriate temperature control devices to minimize equilibration with ambient temperature before isolation of solids.

Liquid phase decanting. Some solids isolated from solution-based crystallization techniques were collected by centrifuging the suspension (if necessary) and removing the liquid phase, leaving wet solids. Solids were rapidly dried (eg, air-dried or nitrogen-dried) unless the moisture content analyzed was specified herein.

High pressure filtration. Solids were collected onto 0.2-μm nylon or PTFE filters by forcing the suspension through a syringe and Swinnex filter holder assembly. Typically, solids were quickly dried by blowing air through a filter with a 20 mL syringe. When referred to herein as analyzed moisture, the solids remained wet with the mother liquor. Prior to analysis, some samples were additionally quickly dried under a gentle stream of nitrogen gas.

Vacuum filtration. Solids were collected onto paper or nylon filters by vacuum filtration and quickly air-dried on the filters under reduced pressure before transferring to the vial.

Reaction crystallization (RC). A mixture of compound 1 and various coformers were combined into acetone-based suspensions at elevated temperature such that the molarity of the coformer was 2 times API. The solution was stirred for the indicated time. If clear solutions were observed, additional crystallization techniques were used.

Stability test. Various salts of compound 1 were placed in open vials in a chamber with a relative humidity of 75% (saturated sodium chloride solution). The relative humidity chamber was placed in an oven at 40°C for 15-16 days. Samples were analyzed by PLM and XRPD at the end of the period.

Slow cooling (SC). Concentrated solutions of compound 1 and various coformers were prepared in various solvents at elevated temperatures with stirring. The vials were sealed in the heated sampling block and the hot plate was turned off, allowing the vials to gradually cool to ambient temperature in the heated vial block. When cooled to ambient temperature, pure solutions were further cooled in a refrigerator (5-7°C) and/or freezer (-20°C). If solids were not present, additional crystallization techniques were used.

Slow evaporation. Solutions were prepared in various solvents with stirring and typically filtered through a 0.2-μm nylon or PTFE filter. Each solution was allowed to evaporate from a covered vial (eg, loosely sealed or covered with perforated aluminum foil) at ambient conditions unless otherwise noted. The solutions were left to evaporate to dryness, unless indicated as partial evaporation (solid present in a small amount of remaining solvent), in which case the solids were recovered as described herein.

Solubility assessment. Aliquots of various solvents were added to measured amounts of Compound 1 with stirring (usually ultrasonically) at the indicated temperatures until complete dissolution was achieved as judged by visual observation. If dissolution occurs after the first aliquot is added, the values are shown as >. If dissolution does not occur, values are indicated as <.

Assessment of solubility in water. Aliquots of water were added to measured amounts of various salts of compound 1 under sonication.

Experiments with suspension. Saturated solutions of compound 1 and various coformers were prepared in various solvents and solvent mixtures. The mixtures were stirred at ambient and elevated temperatures for the indicated times. Solids were collected using the above procedure and additional crystallization techniques were used as necessary.

Solvent removal in a vacuum oven. Salts of compound 1, which were determined to be solvates by various analytical methods, were subjected to solvent removal attempts. The samples were placed in a vacuum oven at temperatures ranging from

- 71 044424 environment up to 80°C for the specified time. Samples were analyzed by XRPD and/or TGA to determine the success of solvent removal.

Vapor diffusion. Concentrated solutions were prepared in various solvents and typically filtered through a 0.2-μm nylon or PTFE filter. The filtered solution was dispensed into a small vial, which was then placed inside a larger vial containing the antisolvent. The small vial was left uncovered and the larger vial was capped to allow vapor diffusion. Any solids present were isolated as described herein.

Steam voltage. The selected solids were transferred to a small vial, which was then placed inside a larger vial containing solvent. The small vial was left uncovered and the larger vial was capped to provide vapor tension at the specified temperature.

A coformer means one or more of the pharmaceutically acceptable bases and/or pharmaceutically acceptable acids disclosed herein in connection with Compound 1. Exemplary coformers as used herein include fumaric acid, HCl, and phosphoric acid.

Instrumental techniques.

Differential scanning calorimetry (DSC). DSC was performed using a Mettler-Toledo DSC3+ differential scanning calorimeter. Temperature calibration was carried out using adamantane, phenyl salicylate, indium, tin and zinc. The sample was placed in a sealed or open aluminum DSC tray and the weight was accurately recorded. A weighed aluminum pan, shaped like a sample tray, was placed on the reference side of the cell. Samples were analyzed at temperatures from -30 to 250°C at an increasing rate of 10°C/min. Although the thermograms are plotted using a reference temperature (x-axis), the results are presented in accordance with sample temperatures.

Dynamic vapor sorption (DVS).

a) VTI. Automatic vapor sorption (VS) data are collected on a VTI SGA-100 vapor sorption analyzer. NaCl and PVP were used as calibration standards. Samples were dried before analysis. Sorption and desorption data were collected at relative humidity ranging from 5% to 95% in 10% increments while purging with nitrogen. The equilibrium criterion used for analysis was a weight change of less than 0.0100% in 5 min with a maximum equilibration time of 3 h. The data were not corrected for the initial moisture content of the samples.

b) Intrinsic. Automatic vapor sorption (VS) data are collected on a Surface Measurement System DVS Intrinsic instrument. Samples were not dried before analysis. Sorption and desorption data were collected at relative humidity ranging from 5 to 95% in 10% increments while purging with nitrogen. The equilibrium criterion used for analysis was a weight change of less than 0.0100% in 5 min with a maximum equilibration time of 3 h. The data were not corrected for the initial moisture content of the samples.

High temperature microscopy (HSM). High temperature microscopy was performed using a Linkam heating stage (FTIR 600) mounted on a Leica DM LP microscope equipped with a SPOT Insight™ color digital camera. Temperature calibration was performed using USP melting point standards. The samples were placed on a coverslip and a second coverslip was placed on top of the sample. As the stage was heated, each sample was visually observed using a 20x objective using crossed polarizers and a first order red compensator. Images were acquired using SPOT software (version 4.5.9).

Optical microscopy. Samples were observed under a Motic or Wolfe optical microscope with crossed polarizers or under a Leica stereo microscope with a first order red compensator and crossed polarizers.

Determination of pKa and logP. Determination of pKa and logP was carried out by Pion Inc./Sirius Analytical Instruments Ltd. to East Sussex, United Kingdom.

Solution Proton Nuclear Magnetic Resonance (1H NMR) Spectroscopy: The 1H NMR spectrum of the solution was obtained through Spectral Data Services in Champaign, Illinois. Samples were prepared by dissolving approximately 5-10 mg of sample in DMSO-d ₆ . Data acquisition parameters appear on the first page of each spectrum in the Data section of this report.

Thermogravimetric analysis (TGA). Thermogravimetric analyzes were performed using a Mettler Toledo TGA/DSC3+ analyzer. Temperature calibration was carried out using phenyl salicylate, indium, tin and zinc. The sample was placed in an aluminum tray. The open tray was inserted into the TG oven. The furnace was heated in a nitrogen atmosphere. Each sample was heated from ambient temperature to 350°C at a rate of 2, 5, or 10°C/min. Although the thermograms are plotted using a reference temperature (x-axis), the results are presented in accordance with sample temperatures.

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X-ray powder diffraction (XRPD).

a) Display. XRPD patterns were collected by a PANalytical X'Pert PRO MPD diffractometer using an incident beam of Cu Kα radiation generated using a long, precisely focused source and a nickel filter at room temperature (298 Kelvin). The diffractometer was configured to use a symmetrical Bragg-Brentano geometry. Prior to analysis, a silicon sample (NIST SRM 640e) was analyzed to ensure that the observed Si 111 peak position corresponded to the NIST certified position. A sample sample was placed into the well. Anti-scatter slits (SS) were used to minimize the background generated by air. Soller slits for the incident and diffracted beams were used to minimize expansion due to axial divergence. X-ray diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located at a distance of 240 mm from the sample and Data Collector v software. 2.2b. Data acquisition parameters for each pattern are displayed above the image in the Data section of this report, including divergence slit (DS) and incident beam SS.

b) Transfer. XRPD patterns were collected by a PANalytical X'Pert PRO MPD diffractometer using an incident beam of Cu radiation obtained through a long, precisely focused Optix source at room temperature (298 Kelvin). An elliptically graded multilayer mirror was used to focus Cu Kα x-rays through the sample and onto the detector. Prior to analysis, a silicon sample (NIST SRM 640e) was analyzed to ensure that the observed Si 111 peak position corresponded to the NIST certified position. A sample sample was clamped between 3-μm-thick films and analyzed in transmission geometry. To minimize the background created by air, a beam limiter, a short scatter-preventing extension, and a scatter-preventing cutting edge were used. Soller slits for the incident and diffracted beams were used to minimize expansion due to axial divergence. X-ray diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located at a distance of 240 mm from the sample and Data Collector v software. 2.2b. Data acquisition parameters for each pattern are displayed above the image in the Data section of this report, including the divergence slit (DS) to the mirror.

XRPD indexing.

Indexing and structure refinement are computational studies. Within a figure for a given indexed XRPD pattern, the correspondence between the resolved peak positions indicated by the bands and the observed peaks indicate a consistent unit cell definition. Successful pattern indexing indicates that the sample consists primarily of a single crystalline phase unless otherwise noted. Groups of spaces corresponding to the assigned attenuation symbol, unit cell parameters and derived quantities are tabulated.

Examples.

Preparation Example 1: Synthesis of Compound 1.

Step 1: N-(4-fluorophenyl)-N-(4-hydroxyphenyl)cyclopropane-1,1-dicarboxamide (4).

EDCI

To a solution of compound 2 (10 g, 44.80 mmol, 1 eq.) and compound 3 (5.87 g, 53.8 mmol, 1.2 eq.) in dimethylacetamide (DMA) (60 ml) was added 3-(ethyliminomethyleneamino) -N,N-Dimethyl-propan-1-amine hydrochloride (EDCI) (10.31 g, 53.8 mmol, 1.2 eq.). The mixture was stirred vigorously at 20°C until the reaction was complete. The mixture was poured into aqueous (aq) saturated NaHCO ₃ (400 ml) and extracted using EtOAc (4x100 ml). The combined organic phases were washed with aqueous saturated NaCl (100 ml), dried over anhydrous (anhydrous) Na ₂ SO ₄ and concentrated. Compound 4 (21 g, crude) (50% purity) was obtained.

1H NMR (400 MHz, DMSO- _d6 ) δ 10.16 (br s, 1H), 9.72 (br s, 1H), 7.61 (dd, 2H), 7.34 (d, 2H), 7.13 (t, 2H), 6.68 (d, 2H), 1.42 (s, 4H);

MS (EI) for C ₁₇ H ₁₅ FN ₂ O ₃ , detected value 314.9 (MH+).

Step 2: Methyl 4-[4-[[1-[(4-fluorophenyl)carbamoyl]cyclopropane-carbonyl]amino]phenoxy]-7methoxyquinoline-6-carboxylate (6).

anisole

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A mixture of compound 4 (5.99 g, 9.5 mmol, 1.2 eq.), compound 5 (2 g, 8.0 mmol, 1.0 eq.), Pd(OAc) ₂ (89 mg, 397, 4 µmol, 0.05 eq.), rac-2-(di-tert-butylphosphino)-1,1'-binaphthyl (TrixiePhos, 316.71 mg, 794.7 µmol, 0.1 eq.) and K ₃ PO ₄ (2.53 g, 11.9 mmol, 1.5 eq.) in anisole (50 ml) was stirred at 110°C for 2 hours under nitrogen atmosphere. The mixture was filtered and the filtrate was concentrated. The residue was purified by flash chromatography on silica gel (1:1 pertolium ether:EtOAc to 20:1 EtOAc:MeOH). Compound 6 was obtained (2.6 g, 61.8% yield).

1H NMR (400 MHz, CDCl ₃ ) δ 9.38 (s, 1H), 8.80 (s, 1H), 8.63 (d, 2H), 7.64 (d, 2H), 7.54- 7.41 (m, 3H), 7.18 (d, 2H), 7.09-7.01 (m, 2H), 6.43 (d, 1H), 4.05 (s, 3H), 3 .97 (s, 3H), 1.78-1.72 (m, 2H), 1.69-1.63 (m, 2H);

MS (EI) for C ₂₉ H ₂₄ FN ₃ O ₆ detected value 530.0 (MH+).

Step 3: 4-[4-[[1-[(4-fluorophenyl)carbamoyl]cyclopropane-carbonyl]amino]phenoxy]-7methoxyquinoline-6-carboxylic acid (7).

To a solution of compound 6 (1.8 g, 3.4 mmol, 1 eq.) in tetrahydrofuran (THF) (15 mL) and MeOH (15 mL) was added 2 M aqueous NaOH (7 mL, 4.1 eq.). The mixture was stirred at 6-13°C for 4 hours. The mixture was adjusted to pH approximately 8 using 1 M aqueous HCl and concentrated to remove solvent. Water (50 ml) was added and the mixture was adjusted to pH approximately 6 using 1 M aqueous HCl. The resulting precipitate was filtered off, washed with water (2*10 ml) and dried in vacuum. Compound 7 was obtained (1.7 g, 97.0% yield).

1H NMR (400 MHz, DMSO-d6) δ 10.22 (s, 1H), 10.08 (s, 1H), 8.65 (d, 1H), 8.48 (s, 1H), 7.77 (d, 2H), 7.64 (dd, 2H), 7.47 (s, 1H), 7.25 (d, 2H), 7.15 (t, 2H), 6.45 (d, 1H) , 3.96 (s, 3H), 1.47 (s, 4H);

MS (EI) for C ₂₈ H ₂₂ FN ₃ O ₆ , detected value 516.1 (MH+).

Step 4: 1-N'-(4-fluorophenyl)-1-N-[4-[7-methoxy-6-(methylcarbamoyl)quinolin-4yl]oxyphenyl]cyclopropane-1,1-dicarboxamide (1).

A solution of compound 7 (300 mg, 582.0 µmol, 1 eq.), HATU (332 mg, 873.2 µmol, 1.5 eq.) and DIEA (301 mg, 2.3 mmol, 406 µl, 4 eq. ) in DMF (10 ml) was stirred at 6-10°C for 1 hour. Methamine hydrochloride (79 mg, 1.2 mmol, 2.0 eq.) was added and the mixture was stirred at 6-10°C for 17 hours The mixture was filtered and the resulting filtrate was purified by preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm, gradient: 33-63% acetonitrile in 10 mM aqueous NH4HCO3, flow rate: 25 ml/min). Compound 1 was obtained (105.4 mg, 34.3% yield).

1H NMR (400 MHz, DMSO- _d6 ) δ 10.20 (s, 1H), 10.06 (s, 1H), 8.65 (d, 1H), 8.61 (s, 1H), 8. 42-8.33 (m, 1H), 7.77 (d, 2H), 7.68-7.61 (m, 2H), 7.51 (s, 1H), 7.25 (d, 2H) , 7.19-7.11 (m, 2H), 6.46 (d, 1H), 4.02 (s, 3H), 2.84 (d, 3H) 1.47 (s, 4H);

MS (EI) for C ₂₉ H ₂₅ FN ₄ O ₅ , detected value 529.1 (MH+).

Example 1: Preparation of Compound 1 Fumarate Form A.

Fumaric acid (1 eq.) in acetone was added to the free base of compound 1 (1 eq.) and the resulting reddish suspension was stirred at approximately 50°C for 4 days. The SC suspension was then brought to RT and stirred for an additional 1 minute to obtain a pink suspension. The solids were then removed by high pressure filtration to obtain a mixture of fumarate form A and free base form A.

Example 2. Preparation of Hemifumarate Form B of Compound 1.

Fumaric acid (2 eq.) in acetone was added to the free base of compound 1 (1 eq.) and the resulting reddish suspension was stirred at approximately 50°C for 6 days, resulting in an off-white suspension. The solids were then removed by high pressure filtration of the hot solution to yield hemifumarate Form B.

Example 3. Preparation of HCl-form A of compound 1.

eq. HCl was added to the free base of compound 1 in THF and the resulting dark reddish suspension was stirred at RT for 3 days to obtain the resulting thick off-white suspension. The solids were then removed by high pressure filtration to obtain HCl Form A.

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Example 4. Preparation of HCl-form B of compound 1.

eq. HCl was added to the free base of Compound 1 in chloroform and the resulting reddish suspension was stirred at approximately 50°C for 3 days to result in a pale pink suspension. The solids were then removed by high pressure filtration to obtain HCl Form B.

Example 5. Preparation of HCl-form C of compound 1.

eq. HCl was added to the free base of Compound 1 in methanol at approximately 60°C to result in a yellowish suspension. The solution was then CC to approximately -20°C and kept cold for approximately 2 days to obtain a clear orange solution. Partial FE provided a clear red solution, and then four volumes of MTBE antisolvent were added and the solution was stirred for 1 day at RT to give an off-white solid, HCl Form C of Compound 1, which was separated by high pressure filtration.

Example 6. Preparation of HCl-form D of compound 1.

eq. HCl was added to the free base of compound 1 at approximately 50°C and the resulting pink suspension was stirred at 50°C for 5 days. The HCl Form D solid of Compound 1 was separated by high pressure filtration.

Example 7. Preparation of Form A of Compound 1.

It is likely that Form A of Compound 1 is the most thermodynamically stable crystalline form of the free base of Compound 1. Accordingly, several procedures lead to the formation of this form. A list of some possible procedures for obtaining Form A of compound 1 is given in Table. 17. This list is in table. 17 is not exhaustive, there are likely many more procedures that will provide this form.

Table 17

Selected procedures for preparing Form A of Compound 1

Solvent Conditions ACN/water 80:20 1) Suspension at 2-8°C for 14 days; or 2) suspension at room temperature for 14 days Chloroform Suspension at 57°C for 2 days DCM Suspension at room temperature for 14 days Ethyl acetate Suspension at 76°C for 3 days Ethanol 1) Suspension at room temperature for 14 days; or 2) suspension at 76°C for 3 days Ethanol/water 90:10 Suspension at room temperature for 14 days Isopropyl alcohol 1) Suspension at room temperature for 14 days; or 2) suspension at 76°C for 3 days Methanol 1) Suspension at room temperature for 14 days; 2) suspension at 57-58°C for 4 days; or 3) fast evaporation Methanol/ethyl acetate 3:2 Suspension at room temperature for 14 days 2,2,2-Trifluoroethanol 1) Slow evaporation; 2) fast evaporation; or 3) flash precipitation using diethyl ether as an antisolvent, then suspension for 1 day Tetrahydrofuran 1) Suspension at room temperature for 14 days; or 2) suspension at 57-58°C for 4 days Tetrahydrofuran/water 50:50 Suspension at room temperature for 14 days

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Example 8. Preparation of Form B of Compound 1.

Compound 1 was dissolved in AcOH and crystallized by VD using diethyl ether as an antisolvent.

Example 9 Preparation of Form C of Compound 1.

Compound 1 was dissolved in HFIPA and crystallized by CP using MTBE as antisolvent.

Example 10 Preparation of Form D of Compound 1.

Compound 1 was dissolved in methanol and crystallized by CC. The mixture was then suspended at 2-8°C to obtain Form D.

Example 11 Preparation of Form E of Compound 1.

Method A: Compound 1 was dissolved in THF and crystallized by CC.

Method B: Compound 1 was dissolved in 90:10 THF:water and precipitated by CP.

Example 12. Preparation of Form F of Compound 1.

Method A: Compound 1 was dissolved in chloroform and crystallized by SE.

Method B: Compound 1 was suspended in chloroform.

Example 13 Preparation of Form G of Compound 1.

Compound 1 was dissolved in chloroform and crystallized by placing the mixture in a freezer.

Example 14 Preparation of Form H of Compound 1.

Form H was prepared by VS amorphous compound 1 with DCM.

Example 15 Preparation of Form K fumarate of compound 1.

Form K of compound 1 was prepared by removing the solvent from Form F or Form G, which are solvates of chloroform.

Example 16 Preparation of Form O of Compound 1.

The O form of compound 1 was discovered during attempts to apply the salt with various counterions in solvent systems containing TFE and is likely to be a solvate of TFE.

Example 17 Preparation of Compound 1 Phosphate Form A.

One molar equivalent of phosphoric acid was added to a suspension of compound 1 in chloroform, and the resulting mixture was suspended for 3 days at approximately ~50°C. The product was isolated by high pressure filtration.

Example 18 Preparation of Form I of Compound 1.

Compound 1 in a 90:10 THF/water mixture was flash-precipitated with heptane and then stirred at freezing temperatures for 7 days.

Example 19 Preparation of Form J of Compound 1.

Compound 1 was suspended in acetone for 14 days.

Example 20 Preparation of Form L of Compound 1.

Compound 1 was suspended in chloroform for 14 days.

Example 21 Preparation of Form M of Compound 1.

Dehydrate Form E of Compound 1 in a vacuum oven at ~77°C for 1 day.

Example 22. Preparation of Form N of Compound 1.

Compound 1 was suspended in a 70:30 TFE/MTBE mixture for 7 days at room temperature.

Other embodiments

The foregoing description is described in more detail by way of illustrations and examples for the purposes of clarity and understanding. The invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many changes and modifications may be made within the spirit and scope of the invention. It will be apparent to one skilled in the art that changes and modifications may be made within the scope of the appended claims. Therefore, it should be understood that the above description is intended to be illustrative and not limiting.

Therefore, the scope of the invention should not be determined by reference to the foregoing description, but instead should be determined by reference to the following appended claims, together with the full scope of equivalents to which such claims are entitled.

Claims (11)

CLAIM 1. Crystalline form of the fumaric acid salt of compound 1, having the structure Compound 1 hemifumarate or a hydrate or solvate thereof, characterized as Compound 1 Hemifumarate Form B, wherein Compound 1 Hemifumarate Form B is characterized by three or more peaks in an XRPD pattern on a 2-theta±0.2 scale, wherein three or more peaks are selected from 7, 55, 9.08, 10.81, 13.24, 15.89, 16.95, 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21.74, 22.29, 22.48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88. 2. Hemifumarate Form B of Compound 1 according to claim 1, wherein three or more peaks are selected from 9.08, 10.81, 16.95, 17.44, 22.29, 22.48, 23.82, 24.37 , 26.34 and 27.05. 3. Form B hemifumarate of compound 1 according to claim 1 or 2, characterized by all of the following peaks in the XRPD pattern on a scale of 2-theta ± 0.2, where the peaks are 9.08, 10.81, 16.95, 17, 44, 22.29, 22.48, 23.82, 24.37, 26.34 and 27.05. 4. Form B hemifumarate of compound 1 according to claim 1, characterized by all of the following peaks in the XRPD pattern on a scale of 2-theta ± 0.2, where the peaks are 7.55, 9.08, 10.81, 13.24, 15.89, 16.95, 17.14, 17.29, 17.44, 18.24, 19.16, 19.91, 20.19, 20.42, 20.70, 21.16, 21, 74, 22.29, 22.48, 22.75, 23.82, 24.37, 26.34, 27.05 and 27.88. 5. Form B of the hemifumarate of compound 1 according to any one of claims 1 to 4, further characterized by an endotherm with an onset temperature of approximately 226° C. on the DSC thermogram. 6. Form B of the hemifumarate of compound 1 according to any one of claims 1 to 5, further characterized by negligible weight loss at a temperature of approximately 220° C. on the TGA thermogram. 7. Hemifumarate Form B of Compound 1 according to any one of claims 1 to 6, further characterized by a weight increase of approximately 0.2 wt.%, as measured by DVS, in an environment when removed from 5% relative humidity and placed at 95 % relative humidity. 8. A pharmaceutical composition containing a crystalline form or a crystalline salt form according to any one of claims 1 to 7 and a pharmaceutically acceptable excipient. 9. A method of treating a disease, disorder or syndrome mediated at least in part by modulating the activity of a protein kinase in vivo, comprising administering to a subject in need thereof a crystalline form or a crystalline salt form according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 1 .8. 10. The method of claim 9, wherein said disease, disorder or syndrome mediated at least in part by modulating protein kinase activity in vivo is cancer. 11. A method of inhibiting a protein kinase, the method comprising contacting the protein kinase with a crystalline form or a crystalline salt form according to any one of claims 1 to 7 or with a pharmaceutical composition according to claim 8, wherein said protein kinase is Axl, Mer, c-Met , KDR or a combination thereof.