EA044352B1 - SELECTIVE INHIBITORS OF PROTEINARGININE METHYL TRANSFERASE 5 (PRMT5) - Google Patents

Description

Cross reference to related application

The invention claims priority under U.S. Provisional Patent Application No.

62/902,322, filed September 18, 2019, incorporated herein by reference in its entirety.

Field of technology

This invention relates to PRMT5 inhibitors and methods of using them.

State of the art

Protein arginine methylation is a common post-translational modification that regulates numerous cellular processes, including gene transcription, mRNA splicing, DNA repair, cellular protein localization, cell fate determination, and signal transduction. There are three types of methyl-arginine species: ω NG monomethylarginine (MMA), ω NG asymmetric dimethylarginine (ADMA) and ω NG, N'G symmetric dimethylarginine (SDMA). The formation of methylated arginines is catalyzed by the protein arginine methyl transferase (PRMT) family of methyltransferases. There are currently nine PRMTs annotated in the human genome. Most of these enzymes are type I enzymes (PRMT1, -2, -3, -4, 6, -8), which are capable of mono- and asymmetric dimethylation of arginine, using Sadenosylmethionine (SAM) as a methyl donor. PRMT-5, -7 and -9 are considered type II enzymes that catalyze the symmetrical dimethylation of arginines. Each PRMT species contains characteristic seven beta-strand methyltransferase motifs ( Katz et al., 2003 ), as well as additional double E and THW sequence motifs specific to the PRMT subfamily.

PRMT5 is a general transcriptional repressor that functions with numerous transcription factors and repressor complexes, including BRG1 and hBRM, Blimp 1 and Snail. This enzyme, after being recruited to the promoter, symmetrically dimethylates H3R8 and H4R3. It is important to note that the H4R3 site is a major target for PRMT1 (ADMA) methylation and is generally considered a transcriptional activation mark. Thus, both marks H4R3me2s (repressive; me2s indicates SDMA modification) and H4R3me2a (active; me2a indicates ADMA modification) are produced in vivo. The specificity of PRMT5 for H3R8 and H4R3 may be altered by its interaction with COPR5, and this may possibly play an important role in determining the corepressor status of PRMT5.

Role of PRMT in cancer.

Aberrant expression of PRMTs has been identified in human cancers, and PRMTs are considered therapeutic targets. A global analysis of histone modifications in prostate cancer showed that histone H4R3 dimethylation was positively correlated with increasing grade, and these changes predicted clinical outcome.

PRMT5 levels have been shown to be elevated in a panel of lymphoid cancer cell lines as well as clinical samples of mantle cell lymphoma. PRMT5 interacts with a number of substrates that are involved in various cellular processes, including RNA processing, signal transduction, and transcriptional regulation. PRMT5 can directly modify histone H3 and H4, resulting in suppression of gene expression. Overexpression of PRMT5 can stimulate cell growth and induce transformation by directly suppressing tumor suppressor genes. Pal et al., Mol. Cell. Biol. 2003, 7475; Pal et al., Mol. Cell. Biol. 2004, 9630; Wang et al., Mol. Cell. Biol. 2008, 6262; Chung et al., J Biol Chem 2013, 5534. In addition to its well-documented oncogenic functions in transcription and translation, the transcription factor MYC also ensures proper splicing of precursor mRNA as an important step in lymphomagenesis. Koh et al., Nature 2015, 523 7558; Hsu et al., Nature 2015 525, 384.

The discovery of anticancer addictions has the potential to inform therapeutic strategies and to identify putative drug targets. By combining data from comprehensive genomic profiling of cancer cell lines and functional characterization of cancer cell dependencies, loss of the enzyme methylthioadenosine phosphorylase (MTAP) was recently found to cause selective dependence on protein arginine methyltransferase 5 (PRMT5) and its binding partner WDR77. MTAP is often lost due to its proximity to the often deleted tumor suppressor gene, CDKN2A. Cells containing MTAP deletions have increased intracellular concentrations of methylthioadenosine (MTA, a metabolite cleaved by MTAP). In addition, MTA specifically inhibits the enzymatic activity of PRMT5. Administration of either MTA or a small molecule PRMT5 inhibitor shows preferential impairment of cell viability for MTAP-null cancer cell lines compared to isogenic MTAP-expressing counterparts. Together, these results indicate that PRMT5 is a potential vulnerability across multiple cancer lines complemented by a common passenger genomic alteration.

The role of PRMT5 in hemoglobinopathies.

The developmental switch in the human globin gene subtype from fetal to adult, which begins at birth, heralds the onset of hemoglobinopathy, b-thalassemia, and sickle cell disease (SCD). The observation that increased expression of the adult globin gene (in the setting of hereditary persistence of fetal hemoglobin [HPFH]) significantly reduces the clinical severity of thalassemia and SCD has led to the search for therapeutic strategies to reverse silencing of the gamma globin gene. Central to gamma gene silencing is DNA methylation, which targets critical CpG dinucleotides adjacent to the gene transcription start site in adult bone marrow erythroid cells. It has been shown that these marks are created due to the recruitment of the DNA methyltransferase, DNMT3 A, to the gamma promoter by the protein arginine methyltransferase PRMT5. Zhao et al., Nat Struct Mol Biol. 2009 16, 304. PRMT5-mediated histone H4R3 methylation recruits DNMT3 A, histone binding and DNA methylation in gene silencing.

PRMT5 induces the repressive histone mark H4R3me2s, which serves as a template for direct DNMT3 A binding and subsequent DNA methylation. Loss of PRMT5 binding or enzymatic activity results in demethylation of CpG dinucleotides and gene activation. In addition to the H4R3me2s mark and DNA methylation, PRMT5 binding to the gamma promoter and its enzymatic activity are required for the assembly of a multiprotein complex at the gamma promoter that induces a range of coordinated repressive epigenetic marks. Disruption of this complex leads to reactivation of gamma gene expression. These studies provide the basis for the development of PRMT5 inhibitors as targeted therapeutics for thalassemia and SCD.

Summary of the invention

The invention relates to pharmaceutically acceptable salts of a compound of formula I:

This invention also relates to the hydrochloride, phosphate and tartrate salts of formula I.

Also described are crystalline forms of such salts, as well as pharmaceutical compositions and methods of using such salts.

Brief description of graphic materials

In fig. 1 shows the XRPD of formula IA-Form I.

In fig. Figure 2 shows a DSC thermogram of formula IA-form I.

In fig. 3 shows a TGA of formula IA-form I.

In fig. 4 shows the XRPD of Formula IA-Form II.

In fig. 5 shows the TGA profile of Formula IA-Form II.

In fig. 6 shows the XRPD of formula IA-form IIa.

In fig. Figure 7 shows a DSC thermogram of formula IA-form IIa.

In fig. 8 shows the XRPD of formula IA-form III.

In fig. Figure 9 shows a DSC thermogram of formula IA-form III.

In fig. 10 shows the TGA profile of Formula IA-Form III.

In fig. 11 shows the XRPD of Formula IA-Form IV.

In fig. 12 shows a DSC thermogram of formula IA-form IV.

In fig. 13 shows the TGA profile of Formula IA-Form IV.

In fig. 14A shows XRPD of Formula III-Form I.

In fig. 14B shows an XRPD of Formula III-Form II.

In fig. 15A shows a DSC thermogram of Formula III-Form I.

In fig. 15B shows a DSC thermogram of Formula III-Form II.

In fig. 16A shows the TGA profile of Formula III-Form I.

In fig. 16B shows the TGA profile of Formula III-Form II.

In fig. 17 shows the XRPD of formula IC.

In fig. Figure 18 shows a DSC thermogram of formula IC.

In fig. 19 shows the TGA profile of formula IC.

In fig. Figure 20 shows the 1H NMR spectrum of formula IC.

In fig. 21 shows XRPD of the free base of formula I.

Detailed Description of Illustrative Embodiments

The invention may be more fully disclosed with reference to the following description, including the following definitions and examples. Certain features of the present compounds and methods that are described herein in the context of individual aspects may also be presented in combination in one aspect. Alternatively, various features of the present compounds and methods, which for brevity are described in the context of one aspect, may also be presented separately or in any subcombination.

Pharmaceutically acceptable means approved or approved by a regulatory agency of the federal or state government or appropriate authority in countries other than the United States, or that is listed in the United States Pharmacopoeia or other generally accepted pharmacopoeia for use in animals, such as humans.

Pharmaceutically acceptable salt refers to a salt of a compound of the present invention that is pharmaceutically acceptable and that has the required pharmacological activity of the parent compound. In particular, such non-toxic salts may be addition salts of inorganic or organic acids and addition salts of bases. In particular, such salts include: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or formed with organic acids such as acetic acid, propionic acid, caproic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalene sulfonic acid, 4-toluenesulfonic acid , camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, etc.; or (2) salts formed by replacing an acidic proton present in the parent compound with a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion; or is consistent with an organic base such as ethanolamine, diethanolamine, triethanolamine, Nmethylglucamine and the like. Salts further include, by way of example, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium and the like; and when the compound contains the basic functionality, salts of non-toxic organic or inorganic acids such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.

The term pharmaceutically acceptable excipient refers to a substance that is non-toxic, biologically tolerated and otherwise biologically acceptable for administration to a subject, such as an inert substance added to a pharmaceutical composition or otherwise used as a carrier medium, carrier or diluent to facilitate administration of the agent and compatible with him. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

The term solvate refers to the physical association of a compound of formula I with one or more solvent molecules.

The term subject includes people. The terms person, patient, and subject are used interchangeably throughout this document.

The term providing medical care or treating any disease or disorder in one embodiment refers to eliminating the disease or disorder (ie, stopping or reducing the progression of the disease or at least one of its specified clinical symptoms). In another embodiment, providing medical care or treatment refers to the elimination of at least one physical parameter that may not be noticeable to the subject. In yet another embodiment, providing medical care or treatment refers to reducing the intensity of a disease or disorder from a physical (eg, stabilization of a noticeable symptom) or physiological (eg, stabilization of a physical parameter) perspective, or both. In yet another embodiment, providing medical care or treatment relates to slowing the progression of a disease or disorder. In some embodiments, the provision of medical care or treatment refers to preventive treatment, i.e. preventing the onset of a disease or disorder.

The compounds of this invention and equivalent expressions are intended to cover pharmaceutically acceptable salts of a compound of Formula I as described herein, as well as subgenera thereof, which include stereoisomers (e.g., enantiomers, diastereomers) and structural isomers (e.g., tautomers), where applicable context.

As used herein, the term isotopic variant refers to a compound that contains proportions of isotopes in one or more of the atoms constituting such compound that exceed their natural abundance. For example, an isotopic version of a compound may be radiolabeled, i.e. contain one or more radioactive isotopes or may be labeled with non-radioactive isotopes, such as, for example, deuterium ( ^2H or D), carbon-13 ( ^13C ), nitrogen-15 ( ^15N ), or the like. It should be understood that in a compound where such an isotopic substitution has been made, the following

- 3 044352 atoms, if present, may vary so that, for example, any hydrogen atom may be ² H/D, any carbon atom may be ¹³ C, or any nitrogen atom may be ¹⁵ N and that the presence and placement of such atoms may determined by those skilled in the art.

It should also be understood that compounds having the same molecular formula but differing in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are called isomers. Isomers that differ in the arrangement of atoms in space are called stereoisomers, for example, diastereomers, enantiomers and atropoisomers. The compounds of this invention may contain one or more asymmetric centers; therefore, such compounds can be obtained as individual (R)- or (8)-stereoisomers at each asymmetric center or mixtures thereof. Unless otherwise indicated, the description or name of a particular compound in the specification and claims is intended to include all stereoisomers and mixtures thereof, racemic or otherwise. If a structure has one chiral center but no stereochemistry is specified for that center, then both enantiomers, individually or as a mixture of enantiomers, are included in the structure. If a structure has more than one chiral center but no stereochemistry is specified for the centers, then all enantiomers and diastereomers, individually or as a mixture, are included in the structure. Methods for determining stereochemistry and isolating stereoisomers are well known in the art.

In some aspects, the invention provides pharmaceutically acceptable salts of a compound of Formula I:

In some embodiments, the pharmaceutically acceptable salt is phosphoric acid, sulfuric acid, hydrochloric acid, ascorbic acid, L-tartaric acid, ethane-1,2-disulfonic acid or 1-hydroxy-2-naphthoic acid, and oxalic acids.

In some embodiments, the pharmaceutically acceptable salt of a compound of Formula I is a hydrochloride salt, i.e. formulas IA.

HCI(IA).

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is a phosphate salt, i.e. formula III.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is a tartrate salt, i.e. IC formulas.

In some embodiments, the tartrate is L-tartrate. In other embodiments, the tartrate is D-tartrate.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is a sulfate salt, i.e. formula III.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is an ascorbic acid salt, i.e. IE formulas.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is an ethane-1,2-disulfonic acid salt, i.e. IF formulas.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is the 1-hydroxy-2-naphthoate salt, i.e. IG formulas.

In other embodiments, the pharmaceutically acceptable salt of the compound of Formula I is an oxalate, i.e. formula III.

In some aspects, the present invention provides crystalline forms of pharmaceutically acceptable salts of Formula I.

In some embodiments, the invention provides crystalline forms of salts of Formula IA, Formula III, or Formula IC.

Crystalline forms of salts of formula ΙΑ, III or IC in accordance with the present invention may have useful properties including one or more of the following properties: chemical or polymorphic purity, fluidity, solubility, dissolution rate, bioavailability, morphology or crystal habit, stability, e.g. chemical stability, thermal stability and mechanical stability with respect to polymorphic transformation, stability at

-4044352 storage; hygroscopicity, low residual solvent content and advantageous handling and transport characteristics such as compression ratio or bulk density.

A crystalline form may be referred to herein as having graphical data as shown in the figure. Such data includes, for example, X-ray powder diffraction (XRPD) patterns, differential scanning calorimetry (DSC) thermograms, or thermogravimetric analysis (TGA) profiles. As is known in the art, graphical data potentially provides additional technical information to further determine the appropriate solid shape, which may not necessarily be described by reference to numerical values or peak positions alone. Thus, the term substantially as illustrated in reference to graphical data in a figure herein means a pattern that is not necessarily identical to those depicted herein, but is within the limits of experimental error or variation when considered by one skilled in the art. One skilled in the art can easily compare the graphical data in the figures shown here with graphical data obtained for an unknown crystalline form and confirm whether the two sets of graphical data represent the same crystalline form or two different crystalline forms.

The solid crystalline form may be referred to herein as being polymorphically pure or substantially free from any other form. As used in this context, the expression essentially free from any other forms will mean that the solid form contains about 20% or less, about 10 or less, about 5 or less, about 2 or less, about 1 or less, or 0% any other forms of the test compound, as measured, for example, by XRPD. For example, a solid form of Formula IA described herein as being substantially free from any other solid forms will have greater than about 80, greater than about 90, greater than about 95, greater than about 98, greater than about 99 wt. /wt.% or about 100% of the test solid form of formula IA. Accordingly, in some embodiments, the disclosed solid forms of Formula IA may contain from about 1% to about 20% w/w, from about 5% to about 20% w/w, or from about 5% to about 10% w/w ./wt.% one or more other solid forms of formula IA.

Herein, unless otherwise stated, the XRPD peaks described herein are measured using Cu Κα radiation, λ=1.54 A.

The modifier about should be considered as describing a range defined by the absolute values of the two final indicators. For example, the expression about 2 to about 4 also describes the range from 2 to 4. When used to modify a singular number, the term about refers to plus or minus 10% of the specified number and includes the specified number. For example, about 10% means a range of 9% to 11%, and about 1 means 0.9-1.1.

In some aspects, the present invention provides a crystalline form of the hydrochloride salt of Formula I, i.e. formulas IA. In some embodiments, the crystalline form of Formula IA is Form I (Formula IA-Form I). In some embodiments, Formula IA-Form I is substantially free from any other solid form of Formula IA.

In some embodiments, Formula IA-Form I exhibits substantially the same XRPD profile as shown in FIG. 1. XRPD profile of Formula IA-Form I shown in FIG. 1, contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), distances between lines (d-values) and relative intensities, as shown in Table. 1:

- 5 044352

Table 1

XRP data for the crystalline form of Formula IA-Form I shown in FIG. 1

Angle (2-theta degrees ± 0.2 2-theta degrees) d(A) value Relative intensity 7,999 11.0435 2.5 12.08 7.3205 5 12,822 6.8986 5.4 13.88 6.3747 5.8 16,541 5.3549 3.6 17.28 5.1275 6.7 18,601 4.7662 9.7 19.48 4.5532 4.9 21.18 4.1914 16.4 21.84 4.0662 6.4 22,781 3.9003 2.9 23.84 3.7293 100 26.1 3.4114 AND 26.98 3,302 26.2 27.68 3.2201 3.4 28.5 3.1292 4.6 28,979 3.0786 4.8 30.32 2.9454 3.1 31,162 2.8678 3.8 31.88 2.8048 1.6 32,499 2.7527 19.5 33.84 2.6467 3.7 35.02 2.5602 1.4 36.06 2.4886 4.4 36.8 2.4403 2 39,421 2.2839 2.3

In some embodiments of the present disclosure, Formula IA-Form I is characterized by an XRPD profile containing a peak at one of the angles listed in Table 1. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing more than one peak at one of the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing two peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing three peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing four peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing five peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing six peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile comprising seven peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing eight peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing nine peaks selected from the angles set forth in Table 1 above. 1. In other aspects, Formula IA Form I is characterized by an XRPD profile containing ten peaks selected from the angles listed in Table 1 above. 1. In other aspects, Formula IA-Form I is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 1.

In some embodiments, Formula IA-Form I is characterized by an XRPD profile containing a peak at 23.8 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA Form I has an XRPD profile containing peaks at 21.2 and 23.8 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form I has an XRPD profile containing peaks at 21.2, 23.8 and 27.0 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA Form I has an XRPD profile containing peaks at 21.2, 23.8, 27.0, and 32.5 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IA-Form I is characterized by an XRPD profile containing peaks at two or more of 21.2, 23.8, 27.0, and 32.5 degrees ± 0.2 degrees 2theta.

In some embodiments, Formula IA-Form I may be characterized by substantially the same DSC thermogram as shown in FIG. 2. As shown in FIG. 2, formula IA-form I forms an endothermic peak at 244.19°C, with a peak onset temperature of 234.71°C and an enthalpy of melting of 252.8 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula IA-Form I is characterized by a DSC thermogram containing an endothermic peak at about 244°C. In other embodiments of the present disclosure, Formula IA-Form I has a DSC enthalpy of fusion of about 253 J/g.

In some embodiments, Formula IA-Form I may have a TGA profile substantially as shown in FIG. 3 when heated at a rate of 20°C/min. As shown in FIG. 3, Formula IA-Form I lost about 18.4% of its mass when heated to about 300°C.

In some embodiments of the present disclosure, Formula IA-Form I is characterized by an XRPD profile containing peaks at one or more of 21.2, 23.8, 27.0, and 32.5 degrees ± 0.2 degrees 2-theta, and a DSC thermogram , containing an endothermic peak at about 244°C when heated at a rate of 10°C/min.

In some embodiments, the crystalline form of Formula IA is Form II (Formula IA-Form II). In some embodiments, Formula IA-Form II is substantially free from any other solid form of Formula IA.

In some embodiments, Formula IA-Form II exhibits substantially the same XRPD profile as shown in FIG. 4. XRPD profile of Formula IA-Form II shown in FIG. 4 contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), distances between lines (d-values) and relative intensities, as shown in Table. 2:

table 2

XRPD data for the crystalline form of the formula

IA-Form II shown in FIG. 4

Angle (2-theta degrees ± 0.2 2-theta degrees) d(A) value Relative intensity 3.94 22.4059 73.3 6,618 13.3443 20.7 12,199 7.2496 58.1 13,458 6,574 37.4 14,839 5.9649 51.1 17,462 5.0746 50.4 18.36 4.8281 55.2 19.84 4.4713 44.8 21,361 4.1562 31.1 22,341 3.9761 32.2 22,778 3.9007 45.2 23,877 3.7236 24.1 25.46 3.4955 100 28,019 3.1819 47.4 28.66 3.1122 45.2 29,858 2.99 19.6 31.62 2.8272 35.2

In some embodiments of the present disclosure, Formula IA-Form II is characterized by an XRPD profile containing a peak at one of the angles listed in Table 1. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing more than one peak at one of the angles specified in Table 1 above. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing two peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA Form II is characterized by an XRPD profile containing three peaks selected from the angles specified in Table 1 above. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing five peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing six peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing ten peaks selected from the angles listed above in the table. 2. In other aspects, Formula IA-Form II is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 2.

In some embodiments, Formula IA-Form II is characterized by an XRPD profile containing a peak at 25.5 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA form II is characterized by an XRPD profile containing peaks at 14.8, 17.5 and 25.5 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form II has an XRPD profile containing peaks at 14.8, 17.5, 18.4, 24.0 and 25.5 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form II is characterized by an XRPD profile containing peaks at 14.8, 17.5, 18.4,

24.0, 25.5, 28.0 and 28.7 degrees ± 0.2 degrees 2-theta. In other embodiments, formula IA-form

II is characterized by an XRPD profile containing peaks at 17.5, 18.4, 19.8, 24.0, 25.5, 28.0 and 28.7 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IA-Form II is characterized by an XRPD profile containing peaks at three or more of 17.5, 18.4, 19.8, 24.0, 25.5, 28.0, and 28.7 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form II is characterized by an XRPD profile containing peaks at four or more of 17.5, 18.4, 19.8, 24.0, 25.5, 28.0, and 28.7 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form II is characterized by an XRPD profile containing peaks at five or more of 17.5, 18.4, 19.8, 24.0, 25.5, 28.0, and 28.7 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form II is characterized by an XRPD profile containing peaks at six or more of 17.5, 18.4, 19.8, 24.0, 25.5, 28.0, and 28.7 degrees ±0.2 degrees 2-theta.

In some embodiments, Formula IA-Form II may have a TGA profile substantially as shown in FIG. 5 when heated at a rate of 20°C/min. As shown in FIG. 5, Formula IA-Form II lost about 3% of its mass when heated to about 225°C.

In some embodiments, the crystalline form of Formula IA is Form IIa (Formula IA-Form IIa). In some embodiments, Formula IA-Form IIa is substantially free from any other solid form of Formula IA.

In some embodiments, Formula IA-Form IIa exhibits substantially the same XRPD profile as shown in FIG. 6. XRPD profile of Formula IA-Form IIa shown in FIG. 6 contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), line spacing (d-values) and relative intensities, as shown in Table. 3:

Table 3

XRPD data for the crystalline form of Formula IA-Form IIa shown in FIG. 6

Angle (degrees 2-theta ±0.2 degrees 2-theta) d value (A) Relative intensity 3.96 22.2923 34.6 4.639 19.0309 42.1 12,519 7.0649 89.3 14.02 6.3117 79.3 14,879 5.9489 76.2 15.64 5.6613 22.5 16,597 5.337 21.5 17.76 4.9899 33.5 18.38 4.823 50.5 19.4 4.5717 17 20,418 4.3461 17 21.5 4.1296 41.9 22,257 3.9909 12.3 23,481 3.7856 24.3 24,938 3.5675 81.9 26.08 3.4139 100 26,962 3.3042 27.5 27,638 3.2248 32.2 28,339 3.1466 34.6 29.46 3.0294 29.3 30,683 2.9114 14.7 31.94 2.7997 27.2 34.122 2.6255 16.5

In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by an XRPD profile containing a peak at one of the angles listed in Table 1. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing more than one peak at one of the angles specified in Table 1 above. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing two peaks selected from the angles shown in Table 1 above. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing three peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA Form IIa is characterized by an XRPD profile containing five peaks selected from the angles specified in Table 1 above. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing

- 8 044352 six peaks selected from the angles indicated in the table above. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing ten peaks selected from the angles listed above in the table. 3. In other aspects, Formula IA-Form IIa is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 3.

In some embodiments, Formula IA-Form IIa is characterized by an XRPD profile containing a peak at 26.1 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA Form IIa has an XRPD profile containing peaks at 14.0, 14.9, and 26.1 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IIa has an XRPD profile containing peaks at 12.5, 14.0, 14.9, 18.4, and 26.1 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IIa has an XRPD profile containing peaks at 12.5, 14.0, 14.9, 18.4, 24.9, and 26.1 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IIa has an XRPD profile containing peaks at 12.5, 14.0, 14.9, 18.4, 24.9, 26.1 and 28.3 degrees ± 0.2 degrees 2 -theta.

In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by an XRPD profile containing peaks at three or more of 12.5, 14.0, 14.9, 18.4, 24.9, 26.1, and 28.3 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA Form IIa is characterized by an XRPD profile containing peaks at four or more of 12.5, 14.0, 14.9, 18.4, 24.9, 26.1, and 28.3 degrees ±0 ,2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by an XRPD profile containing peaks at five or more of 12.5, 14.0, 14.9, 18.4, 24.9, 26.1, and 28.3 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by an XRPD profile containing peaks at six or more of 12.5, 14.0, 14.9, 18.4, 24.9, 26.1, and 28.3 degrees ±0.2 degrees 2-theta.

In some embodiments, Formula IA-Form IIa may be characterized by substantially the same DSC thermogram as shown in FIG. 7. As shown in FIG. 7, formula IA-form IIa forms an endothermic peak at 199.44°C, with a peak onset temperature of 194.14°C and an enthalpy of fusion of 55.02 J/g; followed by an exothermic peak; followed by an endothermic peak at 244.53°C, with an onset temperature of 236.29°C and an enthalpy of fusion of 327.1 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by a DSC thermogram containing an endothermic peak at about 199°C. In other embodiments of the present disclosure, Formula IA-Form IIa has a DSC enthalpy of fusion of about 55 J/g.

In some embodiments of the present disclosure, Formula IA-Form IIa is characterized by an XRPD profile containing peaks at one or more of 12.5, 14.0, 14.9, 18.4, 24.9, 26.1, and 28.3 degrees ± 0.2 degrees 2-theta, and a DSC thermogram containing an endothermic peak at about 199°C when heated at a rate of 10°C/min.

In some embodiments, the crystalline form of Formula IA is Form III (Formula IA-Form III). In some embodiments, Formula IA-Form III is substantially free from any other solid form of Formula IA.

In some embodiments, Formula IA-Form III exhibits substantially the same XRPD profile as shown in FIG. 8. XRPD profile of Formula IA-Form III shown in FIG. 8, contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), distances between lines (d-values) and relative intensities, as shown in Table. 4:

- 9 044352

Table 4

XRPD data for the crystalline form of the formula

IA-Form III shown in FIG. 8

Angle (2-theta degrees ± 0.2 2-theta degrees) d value (A) Relative intensity 8,059 10.9618 100 12,499 7.0758 15.4 13.7 6.4584 13.5 14.54 6.0872 13.5 15.26 5.8014 15.5 16,219 5.4606 37.8 16,841 5.2602 12.4 18.82 4.7113 31.5 20,038 4.4276 5.5 20,659 4.2958 13.1 21,279 4,172 14.9 23.3 3.8146 54.4 23,774 3.7395 15.6 24.46 3.6362 19.1 26,699 3.3361 7.3 27,681 3.22 5.4 28.78 3.0995 12.2 29,641 3.0114 12.6 32.16 2,781 2.6 32,839 2.7251 7.1 34.2 2.6196 2.7 34,964 2.5641 2.8 38,018 2.3649 5.4 42,678 2.1168 2.6

In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing a peak at one of the angles listed in Table 1. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing more than one peak at one of the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing two peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA Form III is characterized by an XRPD profile containing three peaks selected from the angles listed in Table 1 above. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing five peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing six peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing ten peaks selected from the angles listed above in the table. 4. In other aspects, Formula IA-Form III is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 4.

In some embodiments, Formula IA-Form III is characterized by an XRPD profile containing a peak at 8.1 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA form III is characterized by an XRPD profile containing peaks at 8.1 and 23.3 degrees ± 0.2 degrees 2theta. In other embodiments, Formula IA-Form III is characterized by an XRPD profile containing peaks at 8.1, 12.5, 16.2 and 23.3 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form III has an XRPD profile containing peaks at 8.1, 12.5, 16.2, 18.8, 23.3, and 24.5 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form III is characterized by an XRPD profile containing peaks at 8.1, 12.5, 13.7, 14.5, 16.2, 18.8, 23.3, and 24.5 degrees ±0 ,2 degrees 2-theta. In other embodiments, Formula IA-Form III is characterized by an XRPD profile containing peaks at 8.1, 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23. 3 and 24.5 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at three or more of 8.1, 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at four or more of 8.1, 12.5,

- 10 044352

13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at five or more of 8.1, 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at six or more of 8.1, 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at seven or more of 8.1, 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta.

In some embodiments, Formula IA-Form III may be characterized by substantially the same DSC thermogram as shown in FIG. 9. As shown in FIG. 9, formula IA-form III forms an endothermic peak at 120.83°C, with a peak onset temperature of 113.61°C and an enthalpy of fusion of 187.5 J/g; followed by an exothermic peak at 163.21°C, with an onset temperature of 158.31°C and an enthalpy of fusion of 67.85 J/g; followed by an endothermic peak at 192.59°C, with an onset temperature of 190.01°C and an enthalpy of fusion of 66.36 J/g; followed by an endothermic peak at 233.74°C, with an onset temperature of 227.54°C and an enthalpy of fusion of 88.63 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula IA-Form III is characterized by a DSC thermogram containing an endothermic peak at about 121°C. In other embodiments of the present disclosure, Formula IA-Form III has a DSC enthalpy of fusion of about 187.5 J/g.

In some embodiments, Formula IA-Form III may have a TGA profile substantially as shown in FIG. 10 when heating at a rate of 20°C/min. As shown in FIG. 10, Formula IA-Form III lost about 4.1% of its mass when heated to about 125°C.

In some embodiments of the present disclosure, Formula IA-Form III is characterized by an XRPD profile containing peaks at one or more of 12.5, 13.7, 14.5, 15.3, 16.2, 18.8, 21.2, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta, and a DSC thermogram containing an endothermic peak at about 121°C when heated at a rate of 10°C/min.

In some embodiments, the crystalline form of Formula IA is Form IV (Formula IA-Form IV). In some embodiments, Formula IA-Form IV is substantially free from any other solid form of Formula IA.

In some embodiments, Formula IA-Form IV exhibits substantially the same XRPD profile as shown in FIG. 11. XRPD profile of Formula IA-Form IV shown in FIG. 11 contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), line spacing (d-values) and relative intensities, as shown in Table. 5:

Table 5

XRPD data for the crystalline form of the formula

IA-Form IV shown in FIG. eleven

Angle (degrees 2-theta ±0.2 degrees 2-theta) d(A) value Relative intensity 4.04 21.8507 100 6.58 13.4221 3 7,961 11.0963 6.8 12,359 7,156 4.3 13,441 6.5823 7 14.84 5.9648 8.9 18,418 4.8131 6.5 19.86 4.4668 7 21.26 4.1757 7.3 22,719 3.9107 14.1 23,698 3.7513 2.7 25,361 3,509 AND 26.32 3.3833 3.3 27,841 3.2019 11.6 28,657 3.1125 3.4 30,581 2.9209 13.8 31,177 2.8664 4.1 31.86 2.8065 3.9 34,599 2.5903 4.2 38,699 2.3248 2.8 42,799 2.1111 4.8

In some embodiments of the present disclosure, Formula IA-Form IV is characterized by

- 11 044352 XRPD profile containing a peak at one of the angles indicated in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing more than one peak at one of the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing two peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA Form IV is characterized by an XRPD profile containing three peaks selected from the angles listed in Table 1 above. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing five peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing six peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing ten peaks selected from the angles listed above in the table. 5. In other aspects, Formula IA-Form IV is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 5.

In some embodiments, Formula IA-Form IV is characterized by an XRPD profile containing a peak at 4.0 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA Form IV is characterized by an XRPD profile containing peaks at 4.0 and 22.7 degrees ± 0.2 degrees 2theta. In other embodiments, Formula IA-Form IV has an XRPD profile containing peaks at 4.0, 22.7, 27.8 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IV has an XRPD profile containing peaks at 22.7, 27.8, 30.6 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IV has an XRPD profile containing peaks at 14.8, 22.7, 27.8, and 30.6 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula IA-Form IV has an XRPD profile containing peaks at 4.0, 14.8, 22.7, 27.8, and 30.6 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IA-Form IV is characterized by an XRPD profile containing peaks at three or more of 4.0, 14.8, 22.7, 27.8, and 30.6 degrees ± 0.2 degrees 2-theta . In some embodiments of the present disclosure, Formula IA-Form IV is characterized by an XRPD profile containing peaks at four or more of 4.0, 14.8, 22.7, 27.8, and 30.6 degrees ± 0.2 degrees 2-theta .

In some embodiments, Formula IA-Form IV may be characterized by substantially the same DSC thermogram as shown in FIG. 12. As shown in FIG. 12, formula IA form IV forms an endothermic peak at 169.03°C with an onset temperature of 161.67°C, and an enthalpy of fusion of 25.82 J/g, followed by an exothermic peak at 192.83°C, followed by an endothermic peak at 243 .32°C with an onset peak temperature of 238.22°C and an enthalpy of 366.9 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula IA-Form IV is characterized by a DSC thermogram containing an endothermic peak at about 169°C. In other embodiments of the present disclosure, Formula IA-Form IV has a DSC enthalpy of fusion of about 26 J/g.

In some embodiments, Formula IA-Form IV may have a TGA profile substantially as shown in FIG. 13 when heated at a rate of 20°C/min. As shown in FIG. 13, formula IA-form IV lost about 12.4 (1.135% + 11.23% of its mass when heated to about 300°C.

In some embodiments of the present disclosure, Formula IA-Form IV is characterized by an XRPD profile comprising peaks at one or more of 4.0, 14.8, 22.7, 27.8, and 30.6 degrees ± 0.2 degrees 2-theta , and a DSC thermogram containing an endothermic peak at about 169°C when heated at a rate of 10°C/min.

In some aspects, the present invention provides a crystalline form of the phosphate salt of Formula I, i.e. formula III.

In some embodiments, the crystalline salt of Formula III is Formula III-Form I, which exhibits substantially the same XRPD profile as shown in FIG. 14A. The XRPD profile of Formula III-Form I shown in FIG. 14A, contains reflection angles (2-theta degrees ± 0.2 2-theta degrees), line spacing (d-values) and relative intensities, as shown in Table 14A. 6:

- 12 044352

Table 6

XRPD data for the crystalline form of formula III-form I shown in FIG. 14A

Angle (degrees 2-theta ±0.2 degrees 2-theta) d(A) value Relative intensity 9,298 9.5039 4.7 11.56 7.6487 7.5 13.08 6,763 10.9 13,679 6,468 3.9 14.74 6,005 7.5 15,539 5.6978 4.7 18.22 4.8651 32.7 18,819 4.7114 23.8 19.62 4.5209 55.9 20.36 4.3581 Yu, 1 22.6 3.9311 27.1 23.88 3.7232 28.8 24.86 3.5786 100 25.74 3.4582 27.6 27,001 3.2995 25.6 27.66 3.2223 41 29,239 3.0518 7.9 29.88 2.9878 8.9 30,699 2.9099 6.1 31.82 2.8099 AND 33.48 2.6743 8.8 36.138 2.4835 5.4 37,019 2.4264 5.2 38.101 2.3599 5 39.138 2.2997 6.7 39,999 2.2522 7.8 40.7 2.215 3.7 41,717 2.1634 5.4 43.8 2.0652 9

In some embodiments of the present disclosure, Formula IB-Form I is characterized by an XRPD profile containing a peak at one of the angles listed in Table 1. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing more than one peak at one of the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing two peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing three peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing five peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing six peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 6. In other aspects, Formula III Form I is characterized by an XRPD profile containing ten peaks selected from the angles listed in Table 1 above. 6. In other aspects, Formula III-Form I is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 6.

In some embodiments, Formula III-Form I is characterized by an XRPD profile containing a peak at 24.9 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III Form I is characterized by an XRPD profile containing peaks at 18.2, 19.6 and 24.9 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III-Form I has an XRPD profile containing peaks at 18.2, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III-Form I has an XRPD profile of peaks at 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IB-Form I is characterized by an XRPD profile containing peaks at three or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2

- 13 044352 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form I is characterized by an XRPD profile containing peaks at four or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form I is characterized by an XRPD profile containing peaks at five or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form I is characterized by an XRPD profile containing peaks at six or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form I is characterized by an XRPD profile containing peaks at seven or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta.

In some embodiments, Formula III-Form I may be characterized by substantially the same DSC thermogram as shown in FIG. 15A. As shown in FIG. 15 A, formula III form I forms an endothermic peak at 200.6°C with an onset temperature of 198.96°C and an enthalpy of fusion of 53.99 J/g followed by endothermic peaks at 224.96°C and 235.97°C at heating at a rate of 10°C/min. In some embodiments of the present disclosure, Formula III, Form I is characterized by a DSC thermogram containing an endothermic peak at about 201°C. In other embodiments of the present disclosure, Formula III-Form I has a DSC enthalpy of fusion of about 54 J/g. In some embodiments, Formula III-Form I may have a TGA profile substantially as shown in FIG. 16A when heating at a speed of 20°C/min. As shown in FIG. 16A, Formula III-Form I lost about 6% of its mass when heated to about 250°C.

In some embodiments of the present disclosure, Formula IB-Form I is characterized by an XRPD profile comprising peaks at one or more of 18.2, 18.8, 19.6, 24.9, 25.7, and 27.0 degrees ± 0.2 degrees 2-theta, and a DSC thermogram containing an endothermic peak at about 201°C when heating at a rate of 10°C/min.

In some embodiments, the crystalline salt of Formula III is Formula III-Form II, which exhibits substantially the same XRPD profile as shown in FIG. 14V. The Formula III-Form II XRPD profile shown in FIG. 14B, contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), line spacing (d-values) and relative intensities, as shown in Table. 6B:

Table 6B

XRPD data for the crystalline form of the formula

III-Form II shown in FIG. 14V

Angle (2-theta degrees ± 0.2 2-theta degrees) d(A) value Relative intensity 3.84 22.9927 6.4 7,322 12,063 2.1 9.02 9.7961 9.4 12.72 6.9537 14.4 13,439 6.5832 7.2 14.34 6.1715 8.6 15,138 5.8477 4.7 17.88 4.9568 33 18,699 4.7414 9.8 19.32 4.5904 41.7 21,159 4.1953 7.2 22.28 3.9868 24.3 23.6 3.7667 25.5 24.64 3.6101 100 25.56 3.4821 26.5 26.82 3.3214 AND 2 27.42 3.25 35.5 28,821 3.0951 3.7 29,641 3.0114 10.9 31,599 2.8291 8.4 33.1 2.7041 2.6 35,937 2.4969 3.3 37,979 2.3672 3.2 38,859 2.3156 3.9 39.72 2.2674 6.5 43.5 2.0787 4.3

In some embodiments of the present disclosure, Formula IB-Form II is characterized by

- 14 044352 XRPD profile containing a peak at one of the angles indicated in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing more than one peak at one of the angles specified in Table 1 above. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing two peaks selected from the angles specified in Table 1 above. 6B. In other aspects, Formula III Form II is characterized by an XRPD profile containing three peaks selected from the angles listed in Table 1 above. 6B. In other aspects, the III-Form II formula is characterized by an XRPD profile containing four peaks selected from the angles listed above in the table. 6B. In other aspects, the III-form II formula is characterized by an XRPD profile containing five peaks selected from the angles listed above in the table. 6B. In other aspects, the III-Form II formula is characterized by an XRPD profile containing six peaks selected from the angles listed above in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing seven peaks selected from the angles listed above in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing eight peaks selected from the angles listed above in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing nine peaks selected from the angles listed above in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing ten peaks selected from the angles listed above in the table. 6B. In other aspects, Formula III-Form II is characterized by an XRPD profile containing more than ten peaks selected from the angles listed above in the table. 6B.

In some embodiments, Formula III-Form II is characterized by an XRPD profile containing a peak at 24.6 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III Form II is characterized by an XRPD profile containing peaks at 19.3, 24.6 and 27.4 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III-Form II is characterized by an XRPD profile containing peaks at 19.3, 22.3, 23.6, 24.6 and 27.4 degrees ± 0.2 degrees 2-theta. In other embodiments, Formula III-Form II has an XRPD profile of peaks at 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, Formula IB-Form II is characterized by an XRPD profile containing peaks at three or more of 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form II is characterized by an XRPD profile comprising peaks at four or more of 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form II is characterized by an XRPD profile containing peaks at five or more of 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, Formula III-Form II is characterized by an XRPD profile containing peaks at six or more of 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta.

In some embodiments, Formula III-Form II may be characterized by substantially the same DSC thermogram as shown in FIG. 15V. As shown in FIG. 15B, Formula III Form II forms an endothermic peak at 228.56°C, with an onset temperature of 225.70°C and an enthalpy of fusion of 140.4 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula III-Form II is characterized by a DSC thermogram containing an endothermic peak at about 229°C. In other embodiments of the present disclosure, Formula III, Form II has a DSC enthalpy of fusion of about 140 J/g.

In some embodiments, Formula III-Form II may have a TGA profile substantially as shown in FIG. 16V when heating at a rate of 20°C/min. As shown in FIG. 16B, Formula III-Form II lost about 7.3% of its mass when heated to about 275°C.

In some embodiments of the present disclosure, Formula IB-Form II is characterized by an XRPD profile comprising peaks at one or more of 19.3, 22.3, 23.6, 24.6, 25.6, and 27.4 degrees ± 0.2 degrees 2-theta, and a DSC thermogram containing an endothermic peak at about 229°C when heated at a rate of 10°C/min.

In some aspects, the present invention provides a crystalline form of the tartrate salt of formula I, i.e. IC formulas.

In some embodiments, the IC formula exhibits an XRPD profile substantially as shown in FIG. 17. XRPD profile of formula IC shown in FIG. 17 contains reflection angles (degrees 2-theta ± 0.2 degrees 2-theta), line spacing (d-values) and relative intensities, as shown in Table. 7:

- 15 044352

Table 7

XRPD data for the crystalline form of formula IC shown in FIG. 17

Angle (2-theta degrees ± 0.2 2-theta degrees) d value (A) Relative intensity 5.5 16.0539 7.9 9,779 9.0371 14.3 11.14 7.9361 35.7 12,376 7.1462 6.6 13.48 6,563 7.7 14,479 6.1126 9.5 18.38 4.8231 100 19.44 4.5623 82.4 19.94 4.4491 93.9 20.84 4.2589 I.5 21,481 4.1333 62.4 22.78 3.9004 15.6 25.54 3.4849 28.4 26,339 3.3809 78.5 27,881 3.1973 19.8 29.24 3.0517 5.8 30.28 2.9492 21.7 31,779 2.8134 4.8 33.14 2,701 32.7 35.98 2,494 8.7 36,941 2.4313 13.2 37,919 2.3708 7.6 40.28 2.2371 9.3 41.28 2.1852 15.1 43,398 2.0834 7.8

In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing a peak at one of the angles specified in the table. 7. In other aspects, the IC formula is characterized by an XRPD profile containing more than one peak at one of the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing two peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing three peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing four peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing five peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing six peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing seven peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing eight peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing nine peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing ten peaks selected from the angles specified in the table above. 7. In other aspects, the IC formula is characterized by an XRPD profile containing more than ten peaks selected from the angles indicated in the table above. 7.

In some embodiments, the IC formula is characterized by an XRPD profile containing a peak at 18.4 degrees ± 0.2 degrees 2-theta. In other embodiments, the IC formula is characterized by an XRPD profile containing peaks at 18.4, 19.9 and 21.5 degrees ± 0.2 degrees 2-theta. In other embodiments, the IC formula is characterized by an XRPD profile containing peaks at 18.4, 19.4, 19.9, 21.5, and 26.3 degrees ± 0.2 degrees 2-theta. In other embodiments, the IC formula is characterized by an XRPD profile containing peaks at 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, and 30.2 degrees ± 0.2 degrees 2-theta. In other embodiments, the IC formula is characterized by an XRPD profile containing peaks at 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, 30.2, and 33.1 degrees ± 0.2 degrees 2-theta.

In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing peaks at three or more of 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, 30.2, and 33.1 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing peaks at four or more of 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, 30.2, and 33.1 degrees ±0.2 degrees 2-theta. In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing peaks at five or more of 11.4, 18.4, 19.4, 19.9,

- 16 044352

21.5, 26.3, 30.2 and 33.1 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing peaks at six or more of 11.4

18.4, 19.4, 19.9, 21.5, 26.3, 30.2 and 33.1 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile containing peaks at seven or more of 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, 30.2, and 33.1 degrees ±0.2 degrees 2-theta.

In some embodiments, the IC formula may be characterized by substantially the same DSC thermogram as shown in FIG. 18. As shown in FIG. 18, formula IC forms an endothermic peak at 190.04°C, with a peak onset temperature of 180.70°C and an enthalpy of fusion of 19.00 J/g when heated at a rate of 10°C/min. In some embodiments of the present disclosure, Formula IC is characterized by a DSC thermogram containing an endothermic peak at about 190°C. In other embodiments of the present disclosure, Formula IC has a DSC fusion enthalpy of about 19 J/g.

In some embodiments, the IC formula may have a TGA profile substantially as shown in FIG. 19 when heated at a rate of 20°C/min. As shown in FIG. 19, Formula IC lost about 9.3% of its mass when heated to about 275°C.

In some embodiments of the present disclosure, the IC formula is characterized by an XRPD profile comprising peaks at one or more of 11.4, 18.4, 19.4, 19.9, 21.5, 26.3, 30.2, and 33.1 degrees ± 0.2 degrees 2-theta, and a DSC thermogram containing an endothermic peak at about 190°C when heated at a rate of 10°C/min.

Pharmaceutical compositions and methods of administration.

Subject pharmaceutical compositions are generally formulated to provide a therapeutically effective amount of a compound of the present invention as an active ingredient or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Optionally, the pharmaceutical compositions contain a pharmaceutically acceptable salt and/or its coordination complex, as well as one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and excipients, diluents, including a sterile aqueous solution and various organic solvents, penetration enhancers, solubilizers and adjuvants. .

The subject pharmaceutical compositions may be administered alone or in combination with one or more other agents, which are typically also administered in the form of pharmaceutical compositions. If desired, one or more of the compounds of this invention and the other agent(s) may be mixed to form a formulation, or both components may be formulated as separate formulations for use in combination separately or simultaneously.

In some embodiments, the concentration of one or more compounds provided in the pharmaceutical compositions of the present invention is less than 100, 90, 80, 70, 60, 50, 40, 30, 20, 19, 18, 17, 16, 15, 14, 13 , 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0 ,3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008 , 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0,0002 or 0,0001 % (or a number in the range defined by and including any two numbers above) m/m, m/v or v/v.

In some embodiments, the concentration of one or more compounds of this invention is greater than 90, 80, 70, 60, 50, 40, 30, 20, 19.75, 19.50, 19.25, 19, 18.75, 18.50 , 18.25, 18, 17.75, 17.50, 17.25, 17, 16.75, 16.50, 16.25, 16, 15.75, 15.50, 15.25, 15, 14 .75, 14.50, 14.25, 14, 13.75, 13.50, 13.25, 13, 12.75, 12.50, 12.25, 12, 11.75, 11.50, 11 ,25, 11, 10.75, 10.50, 10.25, 10, 9.75, 9.50, 9.25, 9, 8.75, 8.50, 8.25, 8, 7.75 , 7.50, 7.25, 7, 6.75, 6.50, 6.25, 6, 5.75, 5.50, 5.25, 5, 4.75, 4.50, 4.25 , 4, 3.75, 3.50, 3.25, 3, 2.75, 2.50, 2.25, 2, 1.75, 1.50, 1.25, 1, 0.9, 0 .8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05 , 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006 ( or a number within the range defined by and including any two numbers above) m/m, m/rev or v/rev.

In some embodiments, the concentration of one or more compounds of this invention ranges from about 0.0001 to about 50, from about 0.001 to about 40, from about 0.01 to about 30, from about 0.02 to about 29, from about 0.03 to about 28, about 0.04 to about 27, about 0.05 to about 26, about 0.06 to about 25, about 0.07 to about 24, about 0.08 to about 23, from about 0.09 to about 22, from about 0.1 to about 21, from about 0.2 to about 20, from about 0.3 to about 19, from about 0.4 to about 18, from about 0 .5 to about 17, from about 0.6 to about 16, from about 0.7 to about 15, from about 0.8 to about 14, from about 0.9 to about 12, from about 1 to about 10% m /m, m/about or about/about.

In some embodiments, the concentration of one or more compounds of this invention ranges from about 0.001 to about 10, from about 0.01% to about 5, from about 0.02 to about 4.5, from about 0. 03 to about 4, about 0.04 to about 3.5, about 0.05 to about 3, about 0.06 to about 2.5, about 0.07 to about 2, about 0.08 to about 1.5, from about 0.09 to about 1, from about 0.1 to about 0.9%, m/m, m/v or v/v.

In some embodiments of this invention, the amount of one or more compounds of this invention is equal to or less than 10, 9.5, 9.0, 8.5, 8.0, 7.5, 7.0, 6.5, 6.0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0, 0.95, 0.9, 0, 85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0.0002 or 0 , 0001 g (or a number in the range defined by and including any two numbers above).

In some embodiments, the amount of one or more compounds of this invention is greater than 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001 , 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0 .0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085 , 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6 , 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4 .5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 g (or a number in the range defined by and including any two numbers above) .

In some embodiments, the amount of one or more compounds of this invention is in the range of 0.0001-10, 0.0005-9, 0.001-8, 0.005-7, 0.01-6, 0.05-5, 0.1- 4, 0.5-4 or 1-3 g.

The compounds of this invention may be effective over a wide range of dosages. For example, in the treatment of adults, examples of dosages that may be used include dosages of 0.01 to 1000, 0.5 to 100, 1 to 50 mg per day, and 5 to 40 mg. The approximate dosage is 10 to 30 mg per day. The exact dosage will depend on the route of administration, the form of administration of the compound, the subject being treated, the body weight of the subject being treated, and the preference and experience of the attending physician.

The pharmaceutical composition of the present invention generally contains the active ingredient (i.e., the compound of the present invention) of the present invention or a pharmaceutically acceptable salt and/or a coordination complex thereof, as well as one or more pharmaceutically acceptable excipients, carriers, including, without limitation, inert solid diluents and fillers, diluents, sterile aqueous solution and various organic solvents, penetration enhancers, solubilizers and adjuvants.

Described below are non-limiting examples of pharmaceutical compositions and methods for their preparation.

Pharmaceutical compositions for oral administration.

In some embodiments, a pharmaceutical composition for oral administration is provided comprising a compound of the present invention and a pharmaceutical excipient suitable for oral administration.

In some embodiments, the invention provides a solid pharmaceutical composition for oral administration containing: (i) an effective amount of a compound of the invention; optionally (ii) an effective amount of the second agent; and (iii) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further comprises: (iv) an effective amount of a third agent.

In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral consumption. The pharmaceutical compositions of the present invention suitable for oral administration may be presented in discrete dosage forms such as capsules, wafers or tablets, or liquids or aerosol dispensers, each containing a predetermined amount of the active ingredient in the form of a powder or granules, solution or a suspension in an aqueous or non-aqueous liquid liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion.

Such dosage forms can be prepared by any of the pharmaceutical methods, but all methods include the step of binding the active ingredient to a carrier that contains one or more of the desired ingredients. Typically, compositions are prepared by uniformly and thoroughly mixing the active ingredient with liquid carriers or finely divided solid carriers, or both, and then, if necessary, forming the product into the desired form. For example, a tablet may be made by compression or molding, optionally with one or more additional ingredients. Compressed tablets can be prepared by compressing in a suitable device the active ingredient in bulk form such as powder or granules, optionally mixed with an excipient such as, but not limited to, a binder, lubricant, inert diluent and/or surfactant or dispersant means. Molded tablets can be prepared by molding a mixture of

- 18 044352 powder compound, moistened with an inert liquid diluent.

The present invention further covers anhydrous pharmaceutical compositions and dosage forms containing the active ingredient, since water may promote the degradation of some compounds. For example, water may be added (eg, 5%) in the pharmaceutical field to simulate long-term storage to determine characteristics such as shelf life or stability of formulations over time. The anhydrous pharmaceutical compositions and dosage forms of this invention can be prepared using anhydrous or low-moisture ingredients under low-humidity conditions. The pharmaceutical compositions and dosage forms of the present invention that contain lactose may be made anhydrous if significant exposure to moisture and/or humidity is expected during manufacture, packaging and/or storage. The anhydrous pharmaceutical composition can be prepared and stored in such a way as to maintain its anhydrous nature. Accordingly, anhydrous compositions may be packaged using materials known to prevent exposure to water, allowing them to be included in appropriate regulatory kits. Examples of acceptable packaging include, but are not limited to, hermetically sealed foil, plastic, or the like, unit dose containers, blister packs, and contour packs.

The active ingredient may be combined with the pharmaceutical carrier by thorough mixing in accordance with conventional pharmaceutical formulation techniques. The carrier can take a variety of forms depending on the form of preparation desired for administration. In the preparation of oral dosage form compositions, the carrier may be any of the usual pharmaceutical vehicles, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like. in the case of oral liquid preparations (such as suspensions, solutions and elixirs) or aerosols; or in the case of oral solid preparations, in some lactose-free embodiments, carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders and tablet disintegrating agents may be used. For example, for solid oral preparations, acceptable carriers include powders, capsules and tablets. If necessary, tablets can be coated using standard aqueous or non-aqueous methods.

Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as gum acacia, sodium alginate, alginic acid, other alginates, tragacanth powder gum, guar gum, cellulose and its derivatives (for example, ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose, microcrystalline cellulose and mixtures thereof.

Examples of suitable excipients for use in the pharmaceutical compositions and dosage forms described herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dexrates, kaolin, mannitol, silicic acid, sorbitol , starch, pre-gelatinized starch and mixtures thereof.

Disintegration regulators can be used in the compositions of this invention to produce tablets that disintegrate when released into an aqueous environment. Too much disintegrating agent may produce tablets that may disintegrate in the vial. Too little of it may be insufficient to disintegrate, which may affect the rate and extent of release of the active ingredient(s) from the dosage form. Thus, a sufficient amount of disintegration agent that is neither too little nor too much to adversely affect the release of the active ingredient(s) can be used to prepare the dosage forms of the compounds described herein. The amount of disintegration agent used may vary depending on the type of formulation and route of administration and can be readily determined by one skilled in the art. The pharmaceutical composition may use from about 0.5 to about 15 wt.% disintegration agent, or from about 1 to about 5 wt.% disintegration agent. Disintegration regulators that can be used to prepare the pharmaceutical compositions and dosage forms of this invention include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch. , other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums or mixtures thereof.

Lubricants that can be used to form the pharmaceutical compositions and dosage forms of this invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other

- 19 044352 glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (for example, peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar or mixtures thereof. Additional lubricants include, for example, silica gel, coagulated synthetic silica aerosol, or mixtures thereof. The lubricant may optionally be added in an amount of less than about 1% by weight of the pharmaceutical composition.

If aqueous suspensions and/or elixirs are desired for oral administration, their active ingredient may be combined with various sweetening or flavoring agents, coloring or coloring agents, and, if necessary, emulsifiers and/or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin and various combinations thereof.

Tablets may be uncoated or may be coated using known techniques to slow disintegration and absorption in the gastrointestinal tract and provide sustained action over a longer period of time. For example, a material that retards the release of the active ingredient, such as glyceryl monostearate or glyceryl distearate, may be used. Oral formulations may also be presented as hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed with a water or oily vehicle. , such as peanut oil, liquid paraffin or olive oil.

Surfactants that can be used to form the pharmaceutical compositions and dosage forms of this invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. Thus, a mixture of hydrophilic surfactants, lipophilic surfactants, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be used.

A suitable hydrophilic surfactant may have a substantially HLB value of at least 10, while a suitable lipophilic surfactant may have a substantially HLB value of equal to or less than about 10. An empirical parameter used to describe the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds, serves as a hydrophilic-lipophilic balance (HLB value). Surfactants with lower HLB values are more lipophilic or hydrophobic, and have greater solubility in oils, while surfactants with higher HLB values are more hydrophilic and have greater solubility in aqueous solutions.

Hydrophilic surfactants are essentially considered to be compounds that have an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds to which the HLB scale is not substantially applicable. Likewise, lipophilic (i.e., hydrophobic) surfactants are compounds that have an HLB value equal to or less than about 10. However, a surfactant's HLB value is a fairly rough estimate commonly used for formulating industrial, pharmaceutical, and cosmetic products. emulsions.

Hydrophilic surfactants can be either ionic or non-ionic. Suitable ionic surfactants include, but are not limited to, alkyl ammonium salts; fusidic acid salts; derivatives of fatty acids, amino acids, oligopeptides and polypeptides; glyceride derivatives of amino acids, oligopeptides and polypeptides; lecithins and hydrogenated lecithins; lysolecithins and hydrogenated lysolecithins; phospholipids and their derivatives; lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfate salts; fatty acid salts; docusate sodium; acyl lactylates; mono- and diacetylated tartaric acid esters of mono- and diglycerides; succinylated mono- and diglycerides; citric acid esters of mono- and diglycerides; and mixtures thereof.

In the above group, ionic surfactants include, for example: lecithins, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; alkyl sulfate salts; fatty acid salts; docusate sodium; acylactylates; mono- and diacetylated tartaric acid esters of mono- and diglycerides; succinylated mono- and diglycerides; citric acid esters of mono- and diglycerides; and mixtures thereof.

Ionic surfactants may be ionized forms of lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP -phosphatidylethanolamine, lactate esters of fatty acids, stearoyl-2-lactate, stearoyl lactylate, succinylated monoglycerides, mono/diacetylated tartaric acid esters of mono/diglycerides, citric acid esters of mono/diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate, linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate, lauroylcarnitines, palmitoylcarnitines, myristoylcarnitines and salts and mixtures thereof.

Hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltosides; alkylthioglucosides; lauryl macrogol glycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; esters of fatty acids and polyethylene glycol glycerol; esters of fatty acids and polyglycerol; fatty acid esters of polyoxyalkylene sorbitan, such as fatty acid esters and polyethylene glycol sorbitan; hydrophilic transesterification products of a polyol with at least one member from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; polyoxyethylene sterols, their derivatives and analogues; polyoxyethylated vitamins and their derivatives; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitol fatty acid esters and hydrophilic polyol transesterification products with at least one member from the group consisting of triglycerides, vegetable oils and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol or a saccharide.

Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castor oil, PEG35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG -6 caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides, polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phytosterol, PEG-30 soy sterol, PEG-20 trioleate, PEG-40 sorbitol oleate, PEG-80 sorbitol laurate , polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl- 10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol series, PEG 15-100 octylphenol series and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fatty alcohols; esters of fatty acids and glycerol; acetylated esters of fatty acids and glycerol; esters of fatty acids and lower alcohols; esters of fatty acids and propylene glycol; esters of fatty acids and sorbitol; esters of fatty acids and polyethylene glycol sorbitan; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and diglycerides; hydrophobic transesterification products of a polyol with at least one member from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; fat-soluble vitamins/vitamin derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include fatty acid esters of glycerol, fatty acid esters of propylene glycol, and mixtures thereof, or are hydrophobic transesterification products of a polyol with at least one member from the group consisting of vegetable oils, hydrogenated vegetable oils and triglycerides.

In one embodiment, the composition may contain a solubilizer to ensure good solubilization and/or dissolution of the compound of the present invention and minimize precipitation of the compound of the present invention. This may be particularly important for compositions for non-oral use, for example, compositions for injection. A solubilizer may also be added to increase the solubility of the hydrophilic drug and/or other components, such as surfactants, or to maintain the composition as a stable or homogeneous solution or dispersion.

Examples of suitable solubilizers include, without limitation, the following: alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediols and their isomers, glycerin, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; polyethylene glycol ethers having an average molecular weight of from about 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether (glycofurol) or methoxy PEG; amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone, Nhydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolac tone and its isomers; and other solubilizers known in the art, such as dimethylacetamide, dimethylisosorbide, N-methylpyrrolidones, monooctanoin, diethylene glycol monoethyl ether and water.

Mixtures of solubilizers can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethyl acetamide, N-methyl pyrrolidone, N-hydroxyethyl pyrrolidone, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol, transcutol, propylene glycol and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerin, triacetin, ethyl alcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that may be included is not particularly limited. The amount of a given solubilizer may be limited to a bioacceptable amount that can be readily determined by one skilled in the art. In some cases, it may be advantageous to include solubilizers in amounts well above the bioacceptable amount, for example, to achieve maximum drug concentration, with the excess solubilizer removed prior to delivery of the composition to the subject using conventional techniques such as distillation or evaporation. Thus, if present, the solubilizer may have a weight ratio of 10, 25, 50, 100%, or up to about 200% by weight, based on the total weight of the drug and other excipients. If desired, very small amounts of solubilizer can also be used, for example 5%>, 2%>, 1%) or even less. Typically, the solubilizer may be present in an amount from about 1%> to about 100%, usually from about 5 to about 25%> wt.

The composition may further include one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, but are not limited to, viscosity reducing agents, antifoaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscosity modulators, tonicity regulators, flavoring agents, colorants, fragrances, opacifying agents, suspending agents, binders, fillers, plasticizers, lubricants and mixtures thereof.

In addition, an acid or base may be included in the composition to facilitate production, improve stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminosilicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine , triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), etc. Also suitable are bases which are salts of a pharmaceutically acceptable acid, such as acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carboxylic acid, citric acid, fatty acids acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, parabromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid acid, thioglycolic acid, toluenesulfonic acid, uric acid, etc. Salts of polyprotic acids such as sodium phosphate, sodium hydrogenphosphate and sodium dihydrogenphosphate can also be used. If the base is a salt, the cation may be any convenient and pharmaceutically acceptable cation such as ammonium, alkali metals, alkaline earth metals, and the like. Examples may include, but are not limited to, sodium, potassium, lithium, magnesium, calcium and ammonium.

Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid and the like. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carboxylic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, parabromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, etc.

In some embodiments, the pharmaceutical composition contains a compound of formula IA, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of formula IA, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of Formula IA-Form I, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of Formula IA-Form II, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of Formula IA-Form IIa, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of Formula IA-Form III, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of Formula IA-Form IV, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

In some embodiments, the pharmaceutical composition contains a compound of formula III, mannitol, microcrystalline cellulose, crospovidone, sodium lauryl sulfate and magnesium stearate.

Pharmaceutical compositions for injection.

In some embodiments, the invention provides a pharmaceutical composition for injection comprising a compound of the present invention and a pharmaceutical excipient suitable for injection. The components and amounts of agents in the compositions are as described herein.

Forms into which the novel compositions of the present invention may be formulated for administration by injection include aqueous or oily suspensions or emulsions using sesame oil, corn oil, cottonseed oil or peanut oil, as well as elixirs, mannitol, dextrose or sterile aqueous solution and similar pharmaceutical carriers.

Aqueous solutions in physiological saline are also commonly used for injection. Ethanol, glycerin, propylene glycol, liquid polyethylene glycol, etc. can also be used. (and acceptable mixtures thereof), cyclodextrin derivatives and vegetable oils. Proper fluidity can be maintained, for example, by using a coating such as lecithin to maintain the desired particle size in the case of a dispersion and by using surfactants. Prevention of exposure to microorganisms can be achieved by using various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.

Sterile injection solutions are prepared by incorporating the compound of the present invention in the required amount in a suitable diluent with various other ingredients listed above, followed by sterilization by filtration if necessary. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile carrier medium that contains the basic dispersion medium and other necessary ingredients listed above. In the case of sterile powders for the preparation of sterile injection solutions, certain desirable methods of preparation are vacuum drying and freeze drying to produce a powder containing the active ingredient plus any additional required ingredient from a pre-sterile filtered solution thereof.

Pharmaceutical compositions for topical (eg, transdermal) application.

In some embodiments, the invention provides a pharmaceutical composition for transdermal delivery comprising a compound of the present invention and a pharmaceutical excipient suitable for transdermal delivery.

The compositions of the present invention may be formulated into solid, semi-solid or liquid forms suitable for topical or superficial application, such as gels, water-soluble jellies, creams, lotions, suspensions, foams, powders, suspensions, ointments, solutions, oils, pastes , suppositories, sprays, emulsions, physiological saline solutions, solutions based on dimethyl sulfoxide (DMSO). Typically, carriers with higher densities are able to provide an area of prolonged exposure to the active ingredients. Conversely, a composition in the form of a solution can

- 23 044352 ensure faster action of the active ingredient on the selected area.

The pharmaceutical compositions may also contain suitable carriers or excipients in a solid or gel phase, which are compounds that enhance the penetration or facilitate the delivery of therapeutic molecules across the stratum corneum barrier of the skin. Many such penetration-enhancing molecules are known to those skilled in the art and are useful in formulating compositions for external use.

Examples of such carriers and excipients include, but are not limited to, humectants (eg, urea), glycols (eg, propylene glycol), alcohols (eg, ethanol), fatty acids (eg, oleic acid), surfactants (eg, isopropyl myristate and lauryl sulfate). sodium), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes (eg menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin and polymers such as polyethylene glycols.

Another exemplary formulation for use in the methods of the present invention uses transdermal delivery devices (patches). Such transdermal patches can be used to provide continuous or intermittent infusion of a compound of the present invention in controlled amounts, with or without another agent.

The design and use of transdermal patches for the delivery of pharmaceutical agents are well known to those skilled in the art. See, for example, US Pat. No. 5,023,252, 4,992,445 and 5,001,139. Such patches can be designed for continuous, intermittent, or on-demand delivery of pharmaceutical agents.

Pharmaceutical compositions for inhalation.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. Preferably, the compositions are administered into the respiratory tract orally or intranasally for local or systemic effect. The compositions in preferred pharmaceutically acceptable solvents can be nebulized using inert gases. Nebulized solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be connected to a face mask or intermittent positive pressure ventilation device. Solutions, suspensions or powder compositions can be administered preferably orally or nasally, from devices that deliver the composition in an appropriate manner.

Other pharmaceutical compositions.

Pharmaceutical compositions can also be prepared from the compositions described herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural, or intraspinal administration. Methods for preparing such pharmaceutical compositions are well known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition, McGraw Hill, 20037ybg; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, ^20th Ed., Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all of which are incorporated herein in their entirety by reference.

Administration of the compounds or pharmaceutical composition of the present invention can be accomplished by any of the methods that provide delivery of the compounds to the site of action. These routes include oral routes, intraduodenal routes, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), topical (eg, transdermal application), rectal administration, local delivery by catheter, stent, or inhalation. The compounds can also be administered intraadiposally or intrathecally.

The amount of compound administered will depend on the subject being treated, the severity of the disorder or condition, the rate of administration, the location of the compound, and the judgment of the attending physician. However, an effective dose ranges from about 0.001 to about 100 mg per kg of body weight per day, preferably from about 1 to about 35 mg/kg/day, once or in divided doses. For a 70 kg person, the dose will be from about 0.05 to 7 g/day, preferably from about 0.05 to about 2.5 g/day. In some cases, dosage levels below the lower end of the above range may be more justifiable, while in other cases large doses can be used without any harmful side effect, for example, by dividing such large doses into several small doses to be administered throughout the day.

In some embodiments, a compound of this invention is administered as a single dose

- 24 044352 doses.

Typically, such administration will be by injection, such as intravenous injection, to rapidly administer the agent. However, if necessary, other routes of administration can be used. A single dose of a compound of this invention can also be used to treat an acute condition.

In some embodiments, the compound is administered in multiple doses. Dosing may be approximately one, two, three times, four times, five times, six times, or more than six times per day. Dosing may be approximately once a month, once every two weeks, once a week, or once a day. In another embodiment, a compound of this invention and another agent are administered together from about once per day to about 6 times per day. In another embodiment, administration of the compound of this invention and the agent lasts less than about 7 days. In yet another embodiment, administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is carried out and maintained for as long as necessary.

Administration of the compounds of this invention may continue for as long as necessary. In some embodiments, a compound of this invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a compound of this invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a compound of this invention is administered regularly and over a long period of time, for example, to treat chronic effects.

An effective amount of a compound of this invention may be administered in single or multiple doses by any of the acceptable routes of administration of agents having similar beneficial properties, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenous, intraperitoneal, parenteral, intramuscular, subcutaneous, orally , topically or by inhalation.

The compositions of the invention can also be delivered by an impregnated or coated device, such as a stent, for example, or a cylindrical polymer inserted into an artery. This route of administration may, for example, help prevent or reduce restenosis after procedures such as balloon angioplasty. Without being limited by theory, the compounds of this invention can slow down or inhibit the migration and proliferation of smooth muscle cells in the arterial wall involved in restenosis. The compound of this invention can be administered, for example, by local delivery from stent struts, stent grafts, implants, or a stent covering or sheath. In some embodiments, a compound of this invention is admixed into a matrix. Such a matrix may be a polymer matrix and may serve to bind the compound to the stent. Polymer matrices suitable for such applications include, for example, lactone-based polyesters or ester copolymers such as polylactide, polycaprolactone glycolide, polyorthoesters, polyanhydrides, polyamino acids, polysaccharides, polyphosphazenes, poly(alkoxy acid ester) copolymers (e.g. PEO-PLLA); polydimethylsiloxane, poly(ethylene vinyl acetate), acrylate-based polymers or copolymers (eg polyhydroxyethyl methyl methacrylate, polyvinyl pyrrolidinone), fluorinated polymers such as polytetrafluoroethylene and cellulose esters. Suitable matrices may be non-degradable or may degrade over time, releasing the compound or compounds. The compounds of this invention can be applied to the surface of a stent by various methods, such as dip/spinning coating, spray coating, dip coating and/or brush coating. The compounds may be applied in a solvent, after which the solvent is allowed to evaporate, thereby forming a compound layer on the stent. In an alternative embodiment, the connection may be located in the body of the stent or implant, for example, in microchannels or micropores. Once implanted, the compound diffuses out of the stent body and contacts the arterial wall. Such stents can be prepared by immersing a stent made to accommodate such micropores or microchannels in a solution of a compound of the present invention in a suitable solvent, followed by evaporation of the solvent. Excess drug on the stent surface can be removed by additional brief flushing with solvent. In other embodiments, the compounds of this invention may be covalently linked to a stent or implant. A linker can be used for covalent attachment, which degrades in vivo, releasing the compound of the invention. For this purpose, any biolabile linkage, such as ester, amide or anhydride linkage, can be used. The compounds of this invention can optionally be administered intravascularly from a balloon used during angioplasty. To reduce restenosis, it is also possible to carry out extravascular administration of compounds through the pericardium or through adventitial application of the compositions of this invention.

Various stenting devices are available that can be used as described, for example, in the following references, all of which are incorporated herein by reference: US Pat. No. 5,451,233; US Patent No. 5040548; US Patent No. 5,061,273; US Patent No. 5496346; patent

- 25 044352

US No. 5292331; US Patent No. 5674278; US Patent No. 3657744; US Patent No. 4739762; patent

US No. 5195984; US Patent No. 5292331; US Patent No. 5674278; US Patent No. 5879382; patent

US No. 6344053.

The compounds of the invention can be administered in dosage doses. It is known in the art that, due to intersubject variability in the pharmacokinetics of a compound, it is necessary to individualize the dosage regimen for optimal therapy. Determination of the dose of the compound of this invention can be accomplished through routine experimentation in accordance with this description.

When a compound of the present invention is administered as part of a composition containing one or more agents, and the agent has a shorter half-life than the compound of the present invention, the unit dosage form of the agent and the compound of the present invention can be adjusted accordingly.

The pharmaceutical composition to be tested may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulation, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical and external for use as an ointment or cream, or for rectal administration as a suppository. The pharmaceutical composition may be in a unit dosage form suitable for single administration of precise doses. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient, as well as a compound according to the present invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.

Typical forms for parenteral administration include solutions or suspensions of the active compound in sterile aqueous solutions, such as aqueous solutions of propylene glycol or dextrose. If desired, such dosage forms can be suitably buffered.

Methods of application.

The method typically involves administering to a subject a therapeutically effective amount of a compound of this invention. The therapeutically effective amount of the test combination of compounds may vary depending on the intended use (in vitro or in vivo) or the subject and the extent of the disease being treated, for example, the weight and age of the subject, the severity of the disease, the route of administration, etc., and may be easily determined by one skilled in the art. The term also refers to a dose that induces a specific response in target cells, such as decreased proliferation or inhibition of target protein activity. The specific dose will vary depending on the specific compounds selected, the dosage regimen to be followed, whether it is administered in combination with other compounds, the time of administration, the tissue into which it is administered, and the physical delivery system in which it is carried. .

As used herein, the term IC ₅₀ refers to the half-maximal inhibition concentration of an inhibitor when inhibiting a biological or biochemical function. This score indicates how much of a particular inhibitor is needed to inhibit a given biological process (or component of a process, ie enzyme, cell, cell receptor or microorganism) by half. In other words, it is the half-maximal (50%) inhibition concentration (IC) of the substance (50% IC or IC ₅₀ ). EC ₅₀ refers to the plasma concentration required to obtain 50% of the maximum effect in vivo.

In some embodiments of the present invention, the methods described herein use a PRMT5 inhibitor with an IC ₅₀ value of about or less than a predetermined value, as determined in an in vitro assay. In some embodiments, the PRMIT5 inhibitor inhibits PRMT5 with an IC ₅₀ value of about 1 or less, 2 or less, 5 or less, 7 or less, 10 or less, 20 or less, 30 or less, 40 or less, 50 or less, 60 or less, 70 or less, 80 or less, 90 or less, 100 or less, 120 or less, 140 or less, 150 or less, 160 or less, 170 or less, 180 or less, 190 or less, 200 or less , 225 or less, 250 or less, 275 or less, 300 or less, 325 or less, 350 or less, 375 or less, 400 or less, 425 or less, 450 or less, 475 or less, 500 or less, 550 or less, 600 or less, 650 or less, 700 or less, 750 or less, 800 or less, 850 or less, 900 or less, 950 nM or less, 1 mM or less, 1.1 or less, 1.2 or less, 1.3 or less, 1.4 or less, 1.5 or less, 1.6 or less, 1.7 or less, 1.8 or less, 1.9 or less, 2 or less, 5 or less, 10 or less, 15 or less, 20 or less, 25 or less, 30 or less, 40 or less, 50 or less, 60 or less, 70 or less, 80 or less, 90 or less, 100 or less , 200 or less, 300 or less, 400 or less, or 500 mM or less (or a number in the range defined by and including any two numbers above).

In some embodiments, the PRMT5 inhibitor selectively inhibits PRMT5 with an IC50 value that is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45. 50, 100, or 1000 times less (or a number in the range defined by and including any two numbers above) than the IC ₅₀ value for one, two, or three other PRMT5.

In some embodiments, the PRMT5 inhibitor selectively inhibits PRMT5 with an IC ₅₀ value that is less than about 1, 2, 5, 7, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 7 50, 800, 850, 900, 950 nM, 1 µM, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 µM (or a number in the range defined by and including any two numbers above), and said IC ₅₀ value is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100 or 1000 times less (or number in the range defined by and including any two numbers above) than the IC ₅₀ value for one, two or three other PRMT5.

The present methods are used to treat PRMT5-related disease. Any disease that directly or indirectly results from abnormal activity or expression levels of PRMT5 may constitute a putative disease.

Various diseases associated with PRMT5 have been reported. PRMT5 is involved, for example, in various human cancers, as well as in a number of hemoglobinopathies.

The examples of such pathological states that are not restrictive include, without restrictions, acanthomic, acinocynocchiatine carcinum, acoustic neuroma, acarol lending melanoma, acrossing, acute eosinophilic leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, acute megakaric leukemia, acute monocytal leukem Acute myeloblastic leukemia maturing, acute myeloid leukemia, acute myelogenous leukemia, acute promyelocytic leukemia, adamantinoma, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenomatoid odontogenic tumor, adrenocortical carcinoma, adult T-cell leukemia, aggressive NK-cell leukemia, AIDS-related cancer , AIDS-related lymphoma, alveolar soft tissue sarcoma, ameloblastic fibroma, anal cancer, anaplastic large cell lymphoma, anaplastic thyroid cancer, angioimmunoblastic T-cell lymphoma, angiomyolipoma, angiosarcoma, appendix cancer, astrocytoma, atypical teratoid rhabdoid tumor, basal cell carcinoma, basal-like carcinoma, B-cell leukemia, B-cell lymphoma, Bellini duct carcinoma, biliary tract cancer, bladder cancer, blastoma, bone cancer, bone tumor, brain stem glioma, brain tumor, breast cancer, Brenner tumor, tumor bronchi, bronchioloalveolar carcinoma, brown tumor, Burkitt's lymphoma, cancer of unknown primary site, carcinoid tumor, carcinoma, carcinoma in situ, carcinoma of the penis, carcinoma of unknown primary site, carcinosarcoma, Castleman's disease, embryonal tumor of the central nervous system, cerebellar astrocytoma, cerebral astrocytoma brain, cervical cancer, cholangiocarcinoma, chondroma, chondrosarcoma, chordoma, choriocarcinoma, choroid plexus papilloma, chronic lymphocytic leukemia, chronic monocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disease, chronic neutrophilic leukemia, clear cell tumor, colon cancer, color ectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, Degos disease, dermatofibrosarcoma protuberans, dermoid cyst, desmoplastic small round cell tumor, diffuse large B-cell lymphoma, dysembryoplastic neuroepithelial tumor, embryonal carcinoma, endodermal sinus tumor, endometrial cancer, uterine endometrial cancer, endometrioid tumor, T -cell lymphoma associated with enteropathy, ependymoblastoma, ependymoma, epidermoid cancer, epithelioid sarcoma, erythroleukemia, esophageal cancer, esthesioneuroblastoma, Ewing family tumor, Ewing family sarcoma, Ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, extra mammary Paget's disease, fallopian tube cancer, fetus in fetus, fibroma, fibrosarcoma, follicular lymphoma, follicular thyroid cancer, gallbladder cancer, ganglioglioma, ganglioneuroma, stomach cancer, gastric lymphoma, gastrointestinal cancer, carcinoid tumor of the gastrointestinal tract, gastrointestinal stromal tumor, germ cell tumor, germinoma, gestational choriocarcinoma, gestational trophoblastic tumor, giant cell tumor of bone, glioblastoma multiforme, glioma, gliomatosis cerebri, glomus tumor, glucagonoma, gonadoblastoma, granulosa cell tumor, hairy cell leukemia, head and neck cancer , heart cancer, hemoglobinopathies such as b-thalassemia and sickle cell disease (SCD), hemangioblastoma, hemangiopericytoma, hemangiosarcoma, hematological malignancy, hepatocellular carcinoma, hepatosplenic T-cell lymphoma, hereditary breast and ovarian cancer syndrome, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic glioma, inflammatory breast cancer, intraocular melanoma, islet cell carcinoma, islet cell tumor, juvenile myelomonocytic leukemia, Kaposi's sarcoma, kidney cancer, Klatskin tumor, Krukenberg tumor, laryngeal cancer, lentigo melanoma, leukemia, cancer of the lips and oral cavity , liposarcoma, lung cancer, luteoma, lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoid leukemia, lymphoma, macroglobulinemia, malignant fibrous histiocytoma, malignant fibrous histiocytoma of bone, malignant glioma, malignant mesothelioma, malignant peripheral nerve sheath tumor, malignant rhabdoid tumor, malignant Newt tumor MALT lymphoma, mantle cell lymphoma, mast cell leukemia, mastocytosis, mediastinal germ cell tumor, mediastinal tumor,

- 27 044352 medullary thyroid cancer, medulloblastoma, medulloepithelioma, melanoma, meningioma, Merkel carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck with an occult primary tumor, metastatic urothelial carcinoma, mixed Mullerian tumor, monocytic leukemia, oral cancer, mucinous tumor, syndrome multiple endocrine neoplasia, multiple myeloma, mycosis fungoides, myelodysplasia, myelodysplasia syndromes, myeloid leukemia, myeloid sarcoma, myeloproliferative disease, myxoma, nasal cancer, nasopharyngeal cancer, neoplasm, neurinoma, neuroblastoma, neurofibroma, neuroma, nodular melanoma, non-Hodgkin's lymphoma, numb nomal cancer skin, non-small cell lung cancer, ocular oncology, oligoastrocytoma, oligodendroglioma, oncocytoma, optic nerve sheath meningioma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, epithelial ovarian cancer, ovarian germ cell tumor, low-grade ovarian tumor, Paget's disease breast, Pancoast tumor, pancreatic cancer, papillary thyroid cancer, papillomatosis, paraganglioma, paranasal sinus cancer, parathyroid cancer, penile cancer, perivascular epithelioid cell tumor, pharyngeal cancer, pheochromocytoma, parenchymal tumor of the pineal gland of intermediate differentiation, pineoblastoma, pituicytoma, pituitary adenoma, pituitary tumor, plasma cell tumor, pleuropulmonary blastoma, polyembryoma, precursor of T-lymphoblastic lymphoma, primary central nervous system lymphoma, primary effusion lymphoma, primary hepatocellular cancer, primary liver cancer, primary peritoneal cancer, primitive neuroectoderm al tumor, prostate cancer, pseudomyxoma peritonei, rectal cancer, renal cell carcinoma, respiratory tract carcinoma involving the NUT gene on chromosome 15, retinoblastoma, rhabdomyoma, rhabdomyosarcoma, Richter's transformation, sacrococcygeal teratoma, salivary gland cancer, sarcoma, Schwannomatosis, carcinoma sebaceous gland, secondary neoplasm, seminoma, serous tumor, Sertoli-Leydig cell tumor, sex cord stromal tumor, Sézary syndrome, signet ring cell carcinoma, skin cancer, small blue round cell tumor, small cell carcinoma, small cell lung cancer, small cell lymphoma, small bowel cancer , soft tissue sarcoma, somatostatinoma, sooty wart, spinal cord tumor, spinal tumor, marginal zone lymphoma of the spleen, squamous cell carcinoma, gastric cancer, superficial spreading melanoma, supratentorial primitive neuroectodermal tumor, superficial epithelial-stromal tumor, synovial sarcoma, T-cell acute lymphoblastic leukemia, T-cell large-grained lymphocytic leukemia, T-cell leukemia, T-cell lymphoma, T-cell prolymphocytic leukemia, teratoma, terminal lymphatic cancer, testicular cancer, tecoma, throat cancer, thymic carcinoma, thymoma, thyroid cancer, transitional cell renal cancer pelvis and ureter, transitional cell carcinoma, bladder cancer, urethral cancer, urogenital neoplasm, uterine sarcoma, uveal melanoma, vaginal cancer, Werner Morrison syndrome, verrucous carcinoma, optic pathway glioma, vulvar cancer, Waldenström's macroglobulinemia, Warthin's tumor, Wilms' tumor or any combination thereof.

In some embodiments, the method is for treating a disease selected from the group consisting of tumor angiogenesis, a chronic inflammatory disease such as rheumatoid arthritis, atherosclerosis, inflammatory bowel disease, skin diseases such as psoriasis, eczema and scleroderma, diabetes, diabetic retinopathy , retinopathy of prematurity, age-related macular degeneration, hemangioma, glioma, melanoma, Kaposi's sarcoma, as well as ovarian, breast, lung, pancreatic, prostate, colon and epidermoid cancer.

In other embodiments, the method is for treating a disease selected from breast cancer, lung cancer, pancreatic cancer, prostate cancer, colon cancer, ovarian cancer, uterine cancer, or cervical cancer.

In other embodiments, the method is for treating a disease selected from leukemia, such as acute myeloid leukemia (AML), acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorders, acute myelogenous leukemia (AML) , chronic myelogenous leukemia (CML), mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma (MM), myelodysplastic syndrome (MDS), epidermoid carcinoma or hemoglobinopathies such as b-thalassemia and sickle cell anemia (SCA).

In yet other embodiments, the methods are for treating a disease selected from cancers with a deletion of the CDKN2A gene; types of cancer with deletion of the 9P gene; types of cancer with deletion of the MTAP gene; cancers caused by splice gene mutations, glioblastoma, non-small cell lung cancer (NSCLC), head and neck cancer, bladder cancer, or hepatocellular carcinoma.

In other embodiments, the method is for treating a disease selected from the following: breast cancer, lung cancer, pancreatic cancer, prostate cancer, colon cancer, ovarian cancer, uterine cancer, cervical cancer, leukemia, such as acute

- 28 044352 myeloid leukemia (AML), chronic lymphocytic leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorders, acute myelocytic leukemia (AML), chronic myeloid leukemia goats (CML), mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma (MM), myelodysplastic syndrome (MDS), squamous cell carcinoma, hemoglobinopathies such as b-thalassemia and sickle cell anemia (SCA), cancers with deletion of the CDKN2A gene; types of cancer with deletion of the 9P gene; types of cancer with deletion of the MTAP gene; cancers caused by splice mutations, glioblastoma, non-small cell lung cancer (NSCLC), head and neck cancer, bladder cancer, hepatocellular carcinoma, adenoid cystic carcinoma (ACC), primary central nervous system lymphoma, fallopian tube cancer, or non-Hodgkin's lymphoma.

In other embodiments, the method is for treating a disease selected from the following: adenoid cystic carcinoma (ACC), primary central nervous system lymphoma, fallopian tube cancer, or non-Hodgkin's lymphoma.

The compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered for the treatment of any of the described diseases, alone or in combination with drug treatment. Drug treatments include, for example, surgery and radiotherapy (eg, gamma radiation, neutron radiation therapy, electron beam therapy, proton therapy, brachytherapy, radionuclide therapy).

In other aspects, the compounds described herein, as well as pharmaceutical compositions containing them, can be administered to treat any of the described diseases, alone or in combination with one or more other agents.

In other methods, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with nuclear receptor agonist agents.

In other methods, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with nuclear receptor antagonist agents.

In other methods, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with an antiproliferative agent.

In other aspects, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered for the treatment of any of the described diseases, alone or in combination with one or more other chemotherapeutic drugs. Examples of other chemotherapy drugs include, for example, abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, polytransretinoic acid, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bendamustine, bevacizumab, bexarotene, bleomycin, bortezombi, bortezomib, intravenous busulfan, busulfan orally, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubi cin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorourasil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, ida rubicin, ifosfamide, imatinib mesylate, interferon alpha 2a, irinotecan, lapatinib ditosylate, lenalidomib, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mecloretamine, megestrol acetate, melphalan, mercaptopurine, methotrexane, methoxalen, mitomycin C, mitotane, mitoxantrone, nandrolone fenpropio nat, nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronate, panobinostat, panitumumab, pegaspargas, pegfilgrastim, pemetrexed disodium, pentostatin, pipobromane, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide , teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinstat and zoledronate.

In other aspects, the other agent is a therapeutic agent that targets an epigenetic regulator. Examples of epigenetic regulator agents include, for example, bromodomain inhibitors, histone lysine methyltransferases, histone arginine methyltransferases, histone demethylases, histone deacetylases, histone acetylases and DNA methyltransferases, as well as any combination thereof. Histone deacetylase inhibitors are preferred in some aspects and include, for example, vorinostat.

In other methods in which the disease being treated is cancer or other proliferative disease, the compounds of this invention, as well as pharmaceutical compositions containing them, can be administered in combination with targeted therapy agents. Targeted therapies include, for example, JAK kinase inhibitors (eg, ruxolitinib), PI3 kinase inhibitors (including selective PI3K delta inhibitors and broad spectrum PI3K inhibitors), MEK inhibitors, cyclic dependent kinase inhibitors (eg, CDK4/6 inhibitors), BRAF, mTOR inhibitors, proteasome inhibitors (eg, bortezomib, carfilzomib), HDAC inhibitors (eg, panobinostat, vorinostat), DNA methyltransferase inhibitors, dexa- 29 044352 metazone, brominated and additional terminal family members, BTK inhibitors (eg, ibrutinib, acalabrutinib ), BCL2 inhibitors (eg, venetoclax), MCL1 inhibitors, PARP inhibitors, FLT3 inhibitors and LSD1 inhibitors, as well as any combinations thereof.

In other methods in which the disease being treated is cancer or other proliferative disease, the compounds of this invention, as well as pharmaceutical compositions containing them, can be administered in combination with immune checkpoint inhibitor agents. Immune checkpoint inhibitors include, for example, PD-1 inhibitors, such as an anti-PD-1 monoclonal antibody. Examples of PD-1 K monoclonal antibodies include, for example, nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR1210, PDR001 and AMP-224, as well as combinations thereof. In some aspects of the invention, the anti-PD1 antibody is nivolumab. In some aspects of the invention, the anti-PD1 antibody is pembrolizumab. In some aspects, the immune checkpoint inhibitor is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some aspects, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C, or any combination thereof. In some aspects, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736. In other aspects, the immune checkpoint inhibitor is a CTLA-4 inhibitor, such as an anti-CTLA-4 monoclonal antibody. In some aspects, the anti-CTLA-4 antibody is ipilimumab.

In other methods in which the disease to be treated is cancer or other proliferative disease, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with an alkylating agent (for example, cyclophosphamide (CY), melphalan (MEL) and bendamustine), a proteasome inhibitor agent (eg, carfilzomib), a corticosteroid agent (eg, dexamethasone (DEX)), or an immunomodulatory agent (eg, lenalidomide (LEN) or pomalidomide (POM)), or any combination thereof.

In some embodiments, the disease being treated is an autoimmune condition or an inflammatory condition. In these aspects, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with a corticosteroid agent, such as, for example, triamcinolone, dexamethasone, fluocinolone, cortisone, prednisolone or flumetholone, or any combination thereof.

In other methods in which the disease being treated is an autoimmune condition or an inflammatory condition, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with an immunosuppressive agent, such as, for example, fluocinolone acetonide (RETISERT™), rimexolone (AL-2178, VEXOL™, ALCO™) or cyclosporine (RESTASIS™) or any combination thereof.

In some embodiments, the disease being treated is beta thalassemia or sickle cell disease. In these aspects, the compounds of the present invention, as well as pharmaceutical compositions containing them, can be administered in combination with one or more agents, such as, for example, HYDREA™ (hydroxyurea).

In some aspects, the present invention relates to methods for preparing the pharmaceutically acceptable salts described herein. In some embodiments, methods for preparing pharmaceutically acceptable salts are described in the examples below.

The examples and preparations presented below further illustrate and explain the compounds of the present invention and methods for preparing such compounds. It should be understood that the scope of the present invention is in no way limited by the scope of the following examples and preparations.

Experimental procedures.

Example 1. Synthesis of (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl-7H-pyrrolo[2,3d]pyrimidin-7-yl )tetrahydrofuran-3,4-diol (formula I).

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Step 1. Synthesis of 2-(2-bromo-5-chlorophenyl)ethanol (22a).

To a solution of 2-(2-bromo-5-chlorophenyl)acetic acid (20.0 g, 80.16 mmol) in THF (200 ml) was added borane in THF (240.49 ml, 240.49 mmol) and the mixture was stirred at 40°C for 8 hours. The mixture was quenched

MeOH at 0°C, concentrated and extracted with EA (400 mlx2). The combined organic layers were dried, concentrated and purified by eluting with combined flash chromatography with CH ₃ N/H2O (neutral) 5/95 to 95/5 to give 22b (18.1 g, 76.854 mmol, 95.9% yield) in in the form of a colorless oil. LCMS [M-18]: 217.0/219.0.

Step 2. Synthesis of 2-(2-bromo-5-chlorophenyl)ethoxy-tert-butyldimethylsilane (22b).

To a solution of 22a (18.1 g, 76.85 mmol) in DMF (200 ml), imidazole (7.85 g, 115.28 mmol) and TBMSCl (13.9 g, 92.23 mmol) was added and the mixture was stirred at 25°C for 8 hours. EA (800 ml) was added and the mixture was washed with saline (400 ml x 2). The organic layer was concentrated and purified by flash column chromatography (PE) to give 22b (26.7 g, 76.34 mmol, 99.3% yield) as a colorless oil.

Step 3. Synthesis of [2-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-chlorophenyl]-[(3aR,4R,6S,6aS)-4-(4chloropyrrolo[2,3-d]pyrimidine -7-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6yl]methanone (22c).

To a solution of 22b (8.91 g, 25.6 mmol) in dry THF (50 ml) was added n-BuLi (12.8 ml, 20.48 mmol) at -78°C and the mixture was stirred for 10 min in atmosphere nitrogen. A solution of 1Ad (4.0 g, 10.24 mmol) in dry THF (20 ml) was added and the mixture was stirred for 5 min at -78°C. TLC (PE:EA=8:1) showed that the reaction was complete. The reaction mixture was poured into dilute HCl (pH 6; pH kept < 8 during quenching process) The mixture was extracted with EA (200 mlx2), the combined organic layers were dried, concentrated and purified by elution using combined flash chromatography with CH3CN/H2O (neutral) from 5 /95 to 95/5 to give 22c (5.1 g, 8.60 mmol, 84% yield) as a yellow solid.

Step 4. Synthesis of (R)-[(3aR,4R,6R,6aR)-4-(4-chloropyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl3a,4,6,6a -tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]-[2-[[tert-butyl (dimethyl)silyl]oxymethyl]-5chlorophenyl]methanol (22d).

To a solution of 22c (5.0 g, 8.44 mmol) in THF (30 ml) at -78°C, DIBAL-H (16.88 ml, 25.31 mmol) was added and the mixture was stirred at -78°C in for 30 minutes. TLC (PE/EA=8/1) showed that SM Rf=0.5 was completely consumed by the main product Rf=0.4. The reaction mixture was poured into dilute HCl (pH 6, 400 ml, maintaining pH < 8 during the quench process) The mixture was extracted with EA (300 ml x 2) and the combined organic layers were dried and concentrated to give crude 22d (5.0 g) as a solid yellow substances.

Step 5. Synthesis of 2-[5-chloro-2-[(R)-hydroxy-[(3aR,4R,6R,6aR)-4-(4-chloropyrrolo[2,3-d]pyrimidin-7yl)-2 ,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methyl]phenyl]ethanol (22e).

To a solution of 22d (3.0 g, 5.17 mmol) in THF (50 mL) was added tetrabutylammonium fluoride (5.17 mL, 5.17 mmol). The mixture was stirred at 25°C for 40 minutes. The reaction mixture was poured into aqueous NH ₄ Cl and extracted with EA (100 ml). The combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and the solvent was concentrated under reduced pressure. The crude product was purified by flash column chromatography (PE:EA=15:1-3:1) to give 22e (2 g, 4.08 mmol, 79% yield) as a white solid. LCMS [M+H]: 480.1.

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Step 6. Synthesis of 4-chloro-7-[(3aR,4R,6R,6aR)-2,2-dimethyl-6-[(1R)-6-chloroisochroman-1-yl]-3a,4,6,6atetrahydrofuro [3,4-d][1,3]dioxol-4-yl]pyrrolo[2,3-d]pyrimidine (22f).

To a solution of 22e (2.0 g, 4.16 mmol) in THF (100 ml) and added tributylphosphine (2.1 ml, 8.33 mmol), isopropyl (NE)-N-isopropoxycarbomatocarbamate (1.72 ml, 8 .74 mmol) and pyridine (0.34 ml,

4.16 mmol) and the reaction mixture was stirred at 25°C for 16 hours.

TLC (PE/EA=3/1, Rf=0.4) showed that the starting material was consumed. The solvent was removed in vacuo and the crude product was purified by silica gel column chromatography using petroleum ether/EtOAc (10:1-5:1) as eluent to give 22f (1.7 g, 3.68 mmol, 88% yield) in in the form of yellow oil. LCMS [M+H]: 462.1.

Step 7. Synthesis of 4-methyl-7-[(3aR,4R,6R,6aR)-2,2-dimethyl-6-[(1R-)-6-chloroisochroman-1-yl]3a,4,6,6a -tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]pyrrolo[2,3-d]pyrimidine (22g).

Methylmagnesium bromide (3.68 ml, 11.04 mmol) was added dropwise to a solution of ferric acetylacetonate (0.13 g, 0.37 mmol) and 22f (1.7 g, 3.68 mmol) in THF (100 ml) at 5°C in a nitrogen atmosphere. The reaction mixture was warmed to room temperature and stirred for 1 hour. TLC (EA:PE=1:1, Rf=0.3) showed that the reaction was complete. Saturated NH ₄ Cl was added dropwise to quench the reaction, which was extracted with EA (200 mlx2), then dried over Na ₂ SO ₄ and concentrated. The residue was purified by flash column chromatography (PE:EA=10:1-1:1) to give 22g (900 mg, 1.93 mmol, 52.6% yield) as a white solid.

Step 8. Synthesis of (2R,3R,4S,5S)-2-(4-methylpyrrolo[2,3-d]pyrimidin-7-yl)-5-[(1R-)-6chloroisochroman-1-yl]tetrahydrofuran- 3,4-diols (formula I).

To a solution of HCl (6.0 ml, 12 mmol) in methanol (10 ml) was added 22g (900 mg, 2.04 mmol) and the reaction mixture was stirred at 40°C for 2 hours. The reaction mixture was concentrated and the residue was stirred with EA (50 ml) and filtered. The solid was purified by preparative HPLC eluting with CH ₃ CN/H ₂ O (0.1% NH4OH) from 5/95 to 95/5. Product fractions were extracted with EA (100 ml x 2 ) and the extracts were concentrated to give Formula I (550 mg, 1.34 mmol, 66% yield) as a white solid. LCMS [M+H]: 402.3. 1H NMR (400 MHz, DMSO- _d6 ): δ 8.67 (s, 1H), 7.76 (d, J=4.0 Hz, 1H), 7.22-7.31 (m, 3H) , 6.81 (d, J=3.6 Hz, 1H), 6.31 (d, J=7.6 Hz, 1H), 5.26 (d, J=7.2 Hz, 1H), 5 .13 (d, J=4.0 Hz, 1H), 4.90 (d, J=3.6 Hz, 1H), 4.48-4.54 (m, 1H), 4.42-4, 43 (m, 1H), 4.23-4.27 (m, 1H), 3.84-3.86 (m, 1H), 3.66-3.72 (m, 1H), 2.91- 2.99 (m, 1H), 2.70-2.74 (m, 1H), 2.67 (s, 3H). 1H NMR (400 MHz, DMSOd ₆ +D ₂ O): δ 8.86 (s, 1H), 7.77 (d, J=4 Hz, 1H), 7.22-7.31 (m, 3H) , 6.82 (d, J=3.6 Hz, 1H), 6.31 (d, J=7.6 Hz, 1H), 4.90 (d, J=3.6 Hz, 1H), 4 .49-4.53 (m, 1H), 4.42-4.43 (m, 1H), 4.24-4.28 (m, 1H), 3.83-3.85 (m, 1H) , 3.66-3.72 (m, 1H), 2.91-2.99 (m, 1H), 2.70-2.75 (m, 1H), 2.69 (s, 3H).

Example 2. Synthesis of (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl-7H-pyrrolo[2,3d]pyrimidin-7-yl )tetrahydrofuran-3,4-diol (formula I).

Add 5.84 g of Formula IA to a 250 mL round bottom flask. 60 ml of deionized water was added and stirred for 5 min to obtain a suspension (pH 1.6). 2.5 ml of concentrated NH4OH (37%) was slowly added and stirred to obtain a suspension (pH 10). Stirred for 3 hours, pH 10. Filtered the batch, first washed with 200 ml of water, then added 200 ml of heptane. The product was filter dried in an oven under vacuum (temperature 40° C.) to obtain 5.20 g (98.0%) of formula I. HPLC purity was 99.7%. formula I was crystallized using XRPD.

In some embodiments, the free base of Formula I may have the following XRPD peaks (see FIG. 21).

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Angle (2-theta degrees ± 0.2 2-theta degrees) d(A) value Relative intensity 6,561 13.4599 3.8 8,079 10,935 26.6 10,415 8.4866 4.3 11,456 7.7179 4.7 12.44 7.1093 21.6 14.52 6.0955 12.8 15,221 5.8162 19.9 16,221 5.4599 13.3 16,698 5.3049 13.5 18.74 4.7311 30.5 20 4.4358 7.4 21,099 4.2071 12 23.02 3.8603 100 23,978 3.7083 3.8 25,521 3.4873 6 26,096 3.4119 3.8 27,459 3.2455 10.8 28.34 3.1466 34.1 28.98 3.0786 12.6 30.82 2.8988 3 32.18 2.7793 5.9 32,899 2.7202 5.1 34,719 2.5817 6.3

Example 3.

The solids can be prepared by treating the free base of formula I with phosphoric, sulfuric, hydrochloric, ascorbic, Z-tartaric acid, ethane-1,2-disulfonic acid and 1-hydroxy-2-naphthoic acid and oxalic acids.

Example 4. Synthesis of the hydrochloride salt of (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl7H-pyrrolo[2,3-d]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IA).

To 60 mg of the free base of formula I (0.150 mmol) were added 1.5 ml of dichloromethane (DCM) and 2.0 ml of acetonitrile and the mixture was stirred to obtain a clear solution. Hydrochloric acid (1 M solution in isopropanol; 0.165 ml, 0.165 mmol, 1.10 eq.) and the resulting mixture was stirred at room temperature for 40 minutes. DCM was removed at 40-45°C to obtain a suspension. The suspension was stirred at 65°C for 60 minutes and then cooled to room temperature and stirred for 2 hours. The mixture was filtered and washed with methyl tert-butyl ether (MTBE). The filter cake was dried at 45-48°C under vacuum overnight to obtain 48.0 mg of the hydrochloride salt (Formula IA).

The XRPD profile of the hydrochloride salt is shown in FIG. 1.

The DSC profile of the hydrochloride salt is shown in FIG. 2.

The TGA of the hydrochloride salt is shown in FIG. 3.

The crystalline HCl salt formed during this procedure is Formula IA-Form I.

Example 5. Synthesis of the hydrochloride salt of (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl7H-pyrrolo[2,3-th]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IA).

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To 100.3 mg of formula I (0.25 mmol, 1.0 eq.) was added 4.0 ml of acetone and stirred for 5 minutes. Add 265 μl of 1.0 M HCl in IPA (0.263 mmol, 1.06 eq). The resulting mixture was stirred to form a fine suspension and then stirred continuously overnight. The mixture was filtered to obtain a solid, which was dried at 40°C under vacuum overnight to obtain 98.8 mg (90.0%) of the salt (Formula IA). The purity of the salt was 99.2% by HPLC. The crystallinity of the salt was confirmed by XPRD. Comparison of the HPLC peak area of the salt and the free base showed that the ratio of free base to hydrochloric acid was approximately 1:1.

Example 6. Synthesis of the hydrochloride salt of (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl7H-pyrrolo[2,3-d]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IA).

To purify and dry the RBF, crude PRT1063, 3476.2 g, (containing 1829.0 g TPR1063 based on the theoretical yield of previous steps) and MeOH (18.3 L) were charged. After loading conc. HCl (1720 ml, 5.0 eq.) into the reactor, the resulting solution was stirred at 35-45°C for approximately 8 hours. MTBE (55.0 L) was added to the reaction mixture and the resulting suspension was stirred at room temperature for approximately 2 hours. The suspension was cooled to 0-5°C and stirred for approximately 1 hour. The suspension was filtered and the filter cake was transferred back to RBF-MTBE (9.1 L) loaded and the suspension was stirred at room temperature for 0.5 -1 hour. The suspension was filtered and the filter cake was washed with MTBE (3.7 L). The filter cake was then dried on a funnel under vacuum for NLT for 2 h.

Recrystallization of Formula IA: For purification and drying, RBF was charged with Formula IA (1500 g) and MeOH (15 L). The mixture was heated to 50-60°C and stirred to form a clear solution. The solution was cooled to 20-30°C and filtered through a baked glass funnel. The filtrate was transferred to a clean and dried reactor, and the volume of filtrate in the reactor was recorded. The funnel was washed with MeOH (3.0 L). This MeOH wash was used to wash the flask to receive the filtrate before adding it to the reactor. The distillation equipment was assembled into a reactor. The filtered solution in the reactor was heated to reflux and the distillate was collected. While maintaining distillation in the reactor, IPA (15 L) was gradually added to the reactor at a rate necessary to maintain the original recorded volume of solution. After adding IPA, heptane (22.5 L) was gradually added to the reactor until the internal volume reached the recorded volume. The heating was stopped and the suspension was cooled to 15-25°C. The suspension was stirred at 15-25°C for approximately 2 hours. The batch was filtered and the filter cake was washed with heptane (4.5 L). The product was dried on the NLT filter for 2 hours by forcing air through the filter cake. The filter cake was transferred to trays and dried in an oven at 50°C under vacuum to constant weight.

Example 7.

Polymorphic forms of Formula IA have been identified as described below. These polymorphs Formula IA-Form I, Formula IA-Form II, Formula IA-Form IIa, Formula IA-Form III, Formula IA-Form IV can be prepared using the methods described below. Solubility of HCl salt at 21 ± 1°C and at 50 ± 1°C.

Add 3 ml of test solvent to a 4 ml vial, Formula IA is added until a cloudy solution is obtained at 21 ± 1°C. About 30 mg of additional formula IA was added to the cloudy solution. The mixture was stirred at 21 ± 1°C over a weekend, during which the temperature was controlled using an IKA® ETS-D5 temperature controller and an IKA® RCT basic safety device. The mixture was filtered using a syringe filter (PTFE, 0.22 μl, 13 mm, Agela Technologies Inc.). The saturated solution was transferred to an HPLC vial, diluted with methanol and analyzed by HPLC.

Add 2 ml of test solvent to a 4 ml vial, Formula IA is added until a cloudy solution is obtained at 50 ± 1°C. About 30 mg of additional formula IA was added to the cloudy solution. The mixture was stirred at 50 ± 1°C for 24 h, during which time the temperature was controlled using an IKA® ETS-D5 temperature controller and an IKA® RCT basic safety device. The mixture was filtered using a syringe filter (PTFE, 0.22 μl, 13 mm, Agela Technologies Inc.). The saturated solution was transferred to an HPLC vial, diluted with methanol and analyzed using

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HPLC. The results are presented in table. 8.

Table 8

Solvent Solubility at 21 ± 1 °C (mg/ml) Solubility at 50 ± 1 °C (mg/ml) MeCN 0.69 1.05 Methanol >50 > 50 EtOH 7.64 10.80 EtOAc 0.12 0.16 IP Ac 0.16 0.24 IPA 0.96 1.99 Water 14.68 43.76

Phase equilibration at 25 ± 1°C and at 50 ± 1°C Phase equilibration studies were designed to obtain information about the predominant crystal form, Formula IA was equilibrated in solvents at 25 ± 1°C and at 50 ± 1°C. The temperature was controlled using an IKA® ETS-D5 temperature controller and an IKA® RCT basic safety device.

To about 3 ml of solvent was added Formula IA Form I until a cloudy solution was obtained, then about 20 mg of additional Formula IA Form I was added to the cloudy solution. The mixture was stirred at 25 ± 1°C and at 50 ± 1°C for 2.0 days . The solid was filtered and analyzed by XRPD.

In these experiments, polymorphic form II (formula IA-form II) was obtained from phase equilibration in ethanol at 25°C, and polymorphic form III (formula IA-form III) was obtained from water at 21°C (Table 2). As a result of phase equilibration at 50 ± 1°C (Table 3), polymorphic form III was obtained from water, and form I was obtained from other solvents. The results are presented in table. 9.

Table 9

Solvent Resulting polymorphic form 25 ± 1 °C 50 ± 1 °C acetonitrile I I methanol I I ethanol II I Ethyl acetate I I Isopropyl acetate I I isopropanol I I water III III

Evaporation studies.

Evaporation studies have been conducted to identify the predominant crystalline form during uncontrolled evaporation. Experiments that did not result in any particulate matter (ie, clear thin films and oils) were not further examined. XRPD was used to study the solid state morphology of the crystalline forms of the evaporation samples at 20 and 50°C. In these studies, the saturated solution obtained from Formula IA-Form I was evaporated. The results are presented in table. 10.

Table 10

Solvent Resulting polymorphic form 20 ± 1 °C 50 ± 1 °C acetonitrile n/a N/A methanol I I ethanol I I Ethyl acetate n/a N/A Isopropyl acetate n/a N/A isopropanol IV IV (see Fig. 11) water N/A N/A

Antisolvent addition experiments.

A saturated solution or nearly saturated solutions of Formula IA were prepared by adding Formula IA-Form I to the solvent. An antisolvent was added to induce precipitation. Hexane, heptane, methyl tert-butyl ether (MTBE), toluene, ethyl acetate, acetone, methyl ethyl ketone (MEK), isopropanol (IPA), tetrahydrofuran (THF), acetonitrile, and isopropyl acetate (IPAc) were used as antisolvents. Experiments that do not produce particulate solids when antisolvent is added have not been further studied. The results are presented in the table below. eleven.

Table 11

Antisolvent (ml) Solvent (ml) Resulting polymorphic form MTBE (1,2) MeOH (0.6) IV IPAc (1,2) MeOH (0.6) IV IPA (2.5) MeOH (0.6) IV EtOAc (1.5) MeOH (0.6) IV THF (2.8) MeOH (0.6) IV IEC (2.8) MeOH (0.6) I Acetone (2.8) MeOH (0.6) I Toluene (3.0) MeOH (0.6) n/a Acetonitrile (3.0) MeOH (0.6) n/a MTBE (4.0) EtOH (2.5) IV Heptane (4.0) EtOH (2.5) IV Hexanes (4.0) EtOH (2.5) IV Toluene (4.5) EtOH (2.5) N/A Cyclohexane (4.0) EtOH (2.5) IV Ethyl acetate (4.0) EtOH (2.5) N/A Heptane (3.5) IPA (3.0) IV Hexanes (3.5) IPA (3.0) IV MTBE (5.0) IPA (3.0) N/A THF (4.0) Water (2.5) N/A IP A (4.0) Water (2.5) N/A 1-propanol (4.0) Water (2.0) N/A Acetone (4.0) Water (2.0) N/A Acetonitrile (4.0) Water (2.5) N/A IEC (4.0) Water (2.0) N/A 1,4-dioxane (4.0) Water (2.0) N/A

Reverse addition experiments.

Saturated or nearly saturated solutions of Formula IA were prepared from Formula IA-Form I in solvent. This solution was then added to a larger volume of mixed antisolvent. Hexane, heptane, MTBE, toluene, ethyl acetate, acetone, MEK, IPA, THF, acetonitrile, and IPAc were used as antisolvents. Experiments that do not produce particulate solids after addition to the antisolvent have not been studied further. The results are presented in the table below. 12.

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Table 12

Antisolvent (ml) Solvent (ml) Resulting polymorphic form MTBE (2.8) MeOH (0.5) IV IP Ac (2.5) MeOH (0.5) IV IP A (3.0) MeOH (0.8) IV EtOAc (2.5) MeOH (0.6) IV THF (2.5) MeOH (0.6) IV Toluene (3.0) MeOH (0.8) N/A IEC (2.8) MeOH (0.6) N/A Acetone (3.0) MeOH (0.6) I MTBE (4.3) EtOH (2.5) IV Heptane (4.0) EtOH (2.5) IV Hexanes (4.0) EtOH (2.5) IV IP Ac (4.0) EtOH (2.5) IV Toluene (4.5) EtOH (2.5) N/A Cyclohexane (4.0) EtOH (2.5) IV Ethyl acetate EtOH (2.5) N/A Heptanes (6.5) IP A (3.0) IV Hexanes (6.5) IP A (3.0) IV MTBE (8.0) IP A (3.0) IV THF (4.0) Water (2.5) N/A IP A (4.0) Water (2.5) N/A 1-propanol (4.0) Water (2.0) N/A Acetone (4.0) Water (2.0) N/A Acetonitrile (4.0) Water (2.5) N/A IEC (4.0) Water (2.0) N/A 1,4-dioxane (4.0) Water (2.0) N/A

Cooling of saturated solutions.

Saturated or nearly saturated solutions of Formula IA-Form I in methanol or ethanol were prepared at room temperature and quenched at about -40°C. A saturated solution in water was prepared at 35°C and quenched at about 5°C. Both experiments were designed to induce precipitation of higher energy forms. The results of these experiments are given below in table. 13.

Table 13

Solvent (ml) Resulting polymorphic form MeOH N/A EtOH II water III

Competitive suspension experiment

To evaluate the transformation of the solid forms of Formula IA, a competition suspension experiment was performed as follows, Formula IA was added to the solvent mixture until a saturated solution was formed. An additional 8 mg of Formula IA-Form I was then added in addition to 8 mg each of Formula IA-Form IIa, Formula IA-Form III, and Formula IA-Form IV. The suspension was stirred and analyzed by XRPD at various time points, including overnight and 24 hours. The results are presented in table. 14.

- 37 044352

Table 14

Solvent Resulting polymorphic form Solid form (after overnight) Solid form (after 24 hours) MeOH/IPA (40/60) I I MeOH/IPA (20/80) I I MeOH/IPA/heptane (30/50/20) I I Ethyl acetate n/a n/a Isopropyl acetate n/a n/a isopropanol IV IV water n/a n/a

Typical synthesis procedures.

Formula IA-Form II.

Approximately 50 mg of drug was added to approximately 3 mL of saturated drug prepared in ethanol, followed by stirring at 25 ± 1°C for 2 days, filtered, air-dried in a fume hood for 24 hours, and analyzed by XRPD at form of formula IA-form II. See fig. 4, fig. 5 3.0 ml of a saturated solution in ethanol was cooled simultaneously with quenching to -20°C and kept at temperature for 30 minutes to obtain a suspension, which was filtered and air-dried. The solid was analyzed by XRPD and partitioned as Formula IA-Form II.

Formula IA-Pa form.

Formula IA-form Pa was prepared by drying formula IA-form II under vacuum at 4850°C for 24 hours.

Formula IA-Form III.

Approximately 50 mg of drug was added to approximately 3 mL of saturated drug prepared in water, followed by stirring at 25 ± 1°C for 2 days, filtered, air-dried in a fume hood for 24 hours, and analyzed by XRPD at form of formula IA-form III. See fig. 8, 10.

3.0 ml of a saturated solution in water was cooled simultaneously with quenching to 2-3°C and kept at the specified temperature for 1.0 hour to obtain a suspension, which was filtered and dried in air. The solid was analyzed by XRPD and partitioned as Formula IA-Form III. See fig. 9.

Formula IA-Form IV.

To 2.5 ml of ethyl acetate, 0.6 ml of drug solution prepared in MeOH (50 mg/ml) was added, followed by stirring for about 5 min, from which the solid was filtered and analyzed by XRPD as Formula IA-Form IV. See fig. 12 and 13.

To 0.6 ml of drug solution prepared in MeOH (50 mg/ml), 1.5 ml of ethyl acetate was added, followed by stirring for approximately 5 minutes, from which the solid was filtered and analyzed by XRPD as Formula IA-Form IV.

Example 8. Synthesis of phosphate salt (2S,3S,4R,5R)-2-((R)-6-chloroisoxpoman-l-yl)-5-(4-methyl-7Hpyrrolo[2,3-b]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IB).

To 50 mg of the free base of formula I (0.125 mmol) were added 1.5 ml of isopropanol (IPA) and 1.0 ml of dichloromethane (DCM) and the mixture was stirred to obtain a clear solution. Phosphoric acid (1 M solution in IPA; 0.15 mL, 0.15 mmol, 1.20 eq) was added and the resulting mixture was stirred at room temperature for 40 min. DCM was removed at 40°C to obtain a suspension. The suspension was stirred at 65°C for 60 minutes and then cooled to room temperature and stirred for 2 hours. The mixture was filtered and washed with methyl tert-butyl ether (MTBE). The filter cake was dried at 45-48°C under vacuum overnight to obtain the phosphate salt (formula III).

The XRPD of this phosphate salt is shown in FIG. 14V.

DSC of this phosphate salt is shown in FIG. 15V.

The TGA of this phosphate salt is shown in FIG. 16V.

Example 9. Synthesis of phosphate salt (2S,3S,4R,5R)-2-((R)-6-chloroisoxpoman-l-yl)-5-(4-methyl-7Hpyrrolo[2,3-0]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IB).

To 100.6 mg of the free base of formula I (0.25 mmol, 1.0 eq.) was added 4.0 ml of EtOH. The resulting mixture was stirred for 5 minutes. 263 μl of 1.0 M H ₃ PO4 was added to ΙΡΑ (0.263 mmol, 1.06 eq.). The resulting mixture was stirred continuously overnight and then filtered to obtain a solid. The filter cake was dried at 40°C under vacuum overnight to obtain 86.7 mg (69.3%) of salt. The purity of the salt was 98.5% by HPLC. Comparison of the HPLC peak area of the salt and the free base indicated that the ratio of free base to phosphoric acid was approximately 2:1.

The XRPD of this phosphate salt is shown in FIG. 14A.

DSC of this phosphate salt is shown in FIG. 15 A.

The TGA of this phosphate salt is shown in FIG. 16A.

Example 10. Synthesis of tartrate salt (2S,3S,4R,5R)-2-((R)-6-chloroisochroman-1-yl)-5-(4-methyl-7Hpyrrolo[2,3-d]pyrimidin-7 -yl)tetrahydrofuran-3,4-diol (formula IC).

To 40.5 mg of Z-tartaric acid (0.263 mmol, 1.05 eq.) was added 100.2 mg (0.25 mmol, 1.0 eq.) of formula I. 4.0 ml of 2-butanone and the resulting mixture was continuously stirred overnight. The mixture was filtered to obtain a solid, which was washed with 2.5 ml of MTBE. The filter cake was dried at 40° C. under vacuum overnight to obtain 68.8 mg (50.0%) of the tartrate salt.

The stoichiometric salt ratio between formula I and tartaric acid was determined using its Ή NMR spectrum (Figure 20) as 2:1 (400 MHz in DMSO-d ₆ ).

The XRPD of this tartrate salt is shown in FIG. 17.

DSC of this tartrate salt is shown in FIG. 18.

The TGA of this tartrate salt is shown in FIG. 19.

The 1H NMR spectrum of the tartrate salt is shown in FIG. 20.

Instrumental methods.

X-ray powder diffraction (XRPD).

XRPD samples can also be obtained using the Rigaku MiniFlex X-ray powder diffractometer (XRPD). X-rays come from copper (Cu) at 1.54056 A with a Kβ filter. X-ray power: 30 kV, 15 mA.

Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC).

TGA profiles can be collected using the Q500 TGA from TA Instruments using a scan rate of 20°C/min.

DSC can also be obtained using TA Instrument differential scanning calorimetry, model Q20 with autosampler, using a scan rate of 10°C/min and a nitrogen gas flow of 50 ml/min.

Biochemical analysis protocol.

Compounds were solubilized and diluted 3 times in 100% DMSO. These diluted compounds were further diluted in assay buffer (20 mM Tris-HCl, pH 8.0, 50 mM NaCl, 0.002% Tween20, 1 mM TCEP, 1% DMSO) for 10 doses of IC ₅₀ regimen at 10 times the required concentration concentration of analysis.

Standard reactions were carried out in a total volume of 30 μl in assay buffer with 300 nM histone H4 based on AcH 4-23 (Anaspec: AS-65002) as substrate. To this was added the PRMT5/MEP50 complex, diluted to give a final assay concentration of 2.5 nM, and the compounds were allowed to pre-incubate for 20 min at 37°C. The reaction mixture was initiated by adding S-[3H-methyl]adenosyl-L-methionine (PerkinElmer: NET155001MC) to a final concentration of 1 μM. After a 30-minute incubation at 37°C, the reaction was stopped by adding 25 μl of 8 M guanidine HCl. YSI SPA streptavidin beads (Perkinelmer: RPNQ0012) were prepared at 0.3 mg/ml in assay buffer. To each reaction, 150 μl of SPA bead suspension was added and incubated with shaking at room temperature for 30 min. The plate was centrifuged at 100xg for 30 s before reading in a scintillation counter. IC ₅₀ values were determined by fitting the data to standard 4 Hill Slope parameters using GraphPad Prism software. See table. 15 below (PRMT5 IC ₅₀ ).

Cellular analysis protocol.

Cell processing and Western blotting to detect symmetric di-methylarginine (sDMA) labels.

Compound Titration and Cell Culture: Compounds were dissolved in DMSO to make a 10 mM stock solution and further 3-fold dilutions were made to make working stock solutions at 1 mM on top. Granta-519 cells were maintained in PRMI 1640 (Corning Cellgro, Cat. No.: 10-040-CV) supplemented with 10% v/v FBS (GE Healthcare, Cat. No.: SH30910.03) and U-87 MG cells maintained in DMEM (Corning Cellgro, Cat. No. 10-013-CV) with 10% FBS and 2 mM glutamine (Corning Cellgro, Cat. No. 25005CV).

Western blot analysis was used to determine enzyme inhibition IC ₅₀ values in Granta-519 and U-87 MG cells. On the day before the experiment, Granta-519 cells were passed to a density of 0.5x106 cells/ml. U-87 MG cells were trypsinized and 4x105 cells were seeded into 6-well plates and allowed to grow overnight. The next day, Granta-519 cells were centrifuged at 1500 rpm for 4 min, resuspended in fresh medium at 0.5x106 cells/ml and 3 ml

- 39 044352 cultures (1.5x106 cells) were seeded in a 6-well plate. Eight-point 3-fold serial dilutions of working stock solutions of the compound were added to the cells (3 μL dilution

1:1000, DMSO concentration was 0.1%; final upper concentration of 1 µM) and incubated for 3 days. Cells incubated with DMSO were used as vehicle controls.

Cells were harvested after 3 days, resuspended in 15 μl of PBS, lysed in 4% SDS and homogenized by passing through a homogenizer column (Omega Biotek, catalog no.: HCR003). Total protein concentrations were determined using the BCA assay (ThermoFisher Scientific, Cat#: 23225). Lysates were mixed with 5x Laemmi buffer solution and boiled for 5 min. Forty μg of total protein was separated on SDS-PAGE gels (Bio-Rad, Cat. No.: 4568083, 4568043), transferred to a PVDF membrane, blocked with 5% dry milk (Bio-Rad, Cat. No.: 1706404) in TBS with 0. 1 v/v% Tween 20 (TBST) for 1 h at room temperature (RT) and incubated with primary antibodies (sDMA: Cell signaling, catalog no.: 13222, 1:3000; P-Actin: sigma, no. catalog: 1:5000) in 5% milk powder in TBST at 4°C overnight. The next day, membranes were washed with TBST, 5x5 min and incubated with HRP-conjugated secondary antibody (GE Healthcare; Cat#: NA934-1ML, NA931-1ML; 1:5000) for 2 h at room temperature followed by a 5x5 min wash TBST and incubation with ECL substrates (Bio-Rad, catalog no.: 1705061, 1705062). The chemiluminescent signal was captured using a Fluochem HD2 Cell Imaging System (Proteinsimple). SmD3me2s bands were quantified using ImageJ. Signals were normalized using P-actin and DMSO controls. _IC50 values were calculated using Graphpad Prism ([inhibitor] versus normalized response - modulated slope). See table. 15, below (sDMA IC ₅₀ ).

Cell proliferation assay to determine IC ₅₀ in Granta-519 and U-87 MG cells.

On the day before the experiment, Granta-519 cells were passed to a density of 0.5 x10 ⁶ cells/ml. U87 MG cells were trypsinized and 2000 cells were seeded into 96-well plates and allowed to grow overnight. On the day of the experiment (day 0), Granta-519 cells were centrifuged at 1500 rpm for 4 min, resuspended in fresh medium to 0.5 x 10 ⁶ cells/ml, and 190 μl of cells were added to 96-well plates. For U-87 MG cells, the old medium was removed and replaced with 190 μl of fresh medium. Working stock solutions were first diluted at 1:50 with fresh media in a 96-well plate, and 10 μL of diluted drugs were added to 96-well plates containing cells and incubated for 3 days. DMSO was used as vehicle control.

After one day, 3.50 μl of Granta-519 cells were transferred to a new 96-well plate and 140 μl of fresh medium was added. For U-87 MG cells, the old medium was removed and replaced with 190 μl of fresh medium. Working stock solutions were diluted at 1:50 with fresh medium and 10 μl of diluted drugs were added to the cells and incubated for 3 additional days. The same process was repeated on day 6. Cells were left to grow for an additional 4 days.

On day 10, 100 μl of Granta-519 cells was transferred to a new 96-well plate and 10 μl of Cell Counting Kit-8 solution (CCK-8, Jojindo, CK04-13) was added. For U-87 MG cells, the old medium was removed and replaced with 100 μl of fresh medium and 10 μl of CCK-8 solution was added. The plates were incubated in a CO2 incubator for 2 h (Granta-519 cells) or 30 min (U-87 MG cells) and OD ₄₅₀ values were measured using a plate reader (iMark microplate reader, Bio-Rad). The percentage of viable cells relative to DMSO vehicle control was calculated and plotted in Graphpad Prism ([inhibitor] versus normalized response - modulated slope) to determine proliferation IC50 values at day 10. See table. 15 below (proliferative IC50).

Table 15

Biochemical and cellular activity in the U-87 MG cell line

[Granta-519 cell line]

Note No. PRMT5 IC50 µM PRMT5 IC50 N sDMA IC50 µM sDMA IC ₅₀ N Prolife. IC50 μΜ Prolife. IC ₅₀ N Formula I (free base) 0.0048 2 0.0176 2 0.054 3

In some embodiments, the invention provides the following aspects: Aspect 1. A pharmaceutically acceptable salt of a compound of Formula I:

but ⁿ ° (I).

Aspect 2. The pharmaceutically acceptable salt according to aspect 1, wherein the salt is a hydrochloride salt, Formula IA.

Aspect 3. Crystalline form of the hydrochloride salt of aspect 2.

Aspect 4. The crystalline form of aspect 3, wherein said crystalline form is Formula IA-Form I.

Aspect 5. The crystalline form of aspect 3 or 4, characterized by substantially the same powder X-ray diffraction pattern as shown in FIG. 1.

Aspect 6. A crystalline form of Aspect 3, 4 or 5, characterized by a powder X-ray diffraction pattern containing a peak at 23.8 degrees ±0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka).

Aspect 7. A crystalline form as defined in any of Aspects 3-6, characterized by a powder X-ray diffraction pattern containing peaks at 21.2 and 23.8 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms ( Cu Ka).

Aspect 8. A crystalline form as defined in any of Aspects 3-7, characterized by an X-ray powder diffraction pattern containing peaks at 21.2, 23.8, 27.0 and 32.5 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstrom (Cu Ka).

Aspect 9. A crystalline form as defined in any one of aspects 3-8, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 2 when heated at a rate of 10°C/min.

Aspect 10. The crystalline form of any one of aspects 3-9, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 244°C when heated at a rate of 10°C/min.

Aspect 11. A crystalline form as defined in any one of aspects 3-10 having a thermogravimetric analysis profile substantially as shown in FIG. 3 when heated at a rate of 20°C/min.

Aspect 12. The crystalline form of aspect 3, wherein said crystalline form is Formula IA-Form II.

Aspect 13. The crystalline form of aspect 3 or 12, characterized by substantially the same powder X-ray diffraction pattern as shown in FIG. 4.

Aspect 14. A crystalline form of Aspect 3, 12, or 13, characterized by a powder X-ray diffraction pattern containing a peak at 25.5 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 15. A crystalline form as defined in any of Aspects 3 or 12-14, characterized by an X-ray powder diffraction pattern containing peaks at 14.8, 17.5 and 25.5 degrees ± 0.2 degrees 2theta on a 2-theta scale with lambda=1 .54 angstrom (Cu Ka).

Aspect 16. The crystalline form of any of Aspects 3 or 12-15, characterized by an X-ray powder diffraction pattern containing peaks at 14.8, 17.5, 18.4, 24.0, 25.5, 28.0, and 28.7 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 17. A crystalline form according to any one of aspects 3 or 12-16, characterized by substantially the same thermogravimetric analysis profile as shown in FIG. 5 when heated at a rate of 20°C/min.

Aspect 18. The crystalline form of aspect 3, wherein said crystalline form is Formula IA-Form IIa.

Aspect 19. The crystalline form of aspect 3 or 18, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 6.

Aspect 20. A crystalline form of Aspect 3, 18, or 19, characterized by a powder X-ray diffraction pattern containing a peak at 26.1 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 21. A crystalline form as defined in any of Aspects 3 or 18-20, characterized by an X-ray powder diffraction pattern containing peaks at 14.0, 14.9 and 26.1 degrees ± 0.2 degrees 2 theta on a 2-theta scale with lambda=1 .54 angstrom (Cu Ka).

Aspect 22. The crystalline form of any of Aspects 3 or 18-21, characterized by an X-ray powder diffraction pattern containing peaks at 12.5, 14.0, 14.9, 18.4 and 26.1 degrees ± 0.2 degrees 2- theta on a 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 23. A crystalline form as defined in any one of aspects 3 or 18-22, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 7 when heated at a rate of 10°C/min.

Aspect 24. The crystalline form of any one of aspects 3 or 18-23, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 199°C when heated at a rate of 10°C/min.

Aspect 25. The crystalline form of aspect 3, wherein said crystalline form is Formula IA-Form III.

Aspect 26. The crystalline form of aspect 3 or 25, characterized by substantially the same powder X-ray diffraction pattern as shown in FIG. 8.

Aspect 27. A crystalline form of Aspect 3, 25, or 26, characterized by a powder X-ray diffraction pattern containing a peak at 8.1 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 28. A crystalline form as defined in any of Aspects 3 or 25-27, characterized by a powder X-ray diffraction pattern containing peaks at 8.1 and 23.3 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstrom (Cu Ka).

Aspect 29. The crystalline form of any of Aspects 3 or 25-28, characterized by an X-ray powder diffraction pattern containing peaks at 8.1, 12.5, 13.7, 14.5, 16.2, 18.8, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka).

Aspect 30. The crystalline form of any one of aspects 3 or 25-29, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 9 when heated at a rate of 10°C/min.

Aspect 31. The crystalline form of any one of aspects 3 or 25-30, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 121°C when heated at a rate of 10°C/min.

Aspect 32. A crystalline form according to any one of aspects 3, 25-31, characterized by substantially the same thermogravimetric analysis profile as shown in FIG. 10 when heating at a rate of 20°C/min.

Aspect 33. The crystalline form of aspect 3, wherein said crystalline form is Formula IA-Form IV.

Aspect 34. The crystalline form of aspect 3 or 33, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. eleven.

Aspect 35. A crystalline form as defined in any of Aspects 3, 33, or 34, characterized by a powder X-ray diffraction pattern containing a peak at 4.0 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka ).

Aspect 36. The crystalline form of any of Aspects 3 or 33-35, characterized by an X-ray powder diffraction pattern containing peaks at 4.0 and 22.7 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstrom (Cu Ka).

Aspect 37. A crystalline form as defined in any of Aspects 3 or 33-36, characterized by an X-ray powder diffraction pattern containing peaks at 4.0, 22.7 and 27.8 degrees ± 0.2 degrees 2theta on a 2-theta scale with lambda=1 .54 angstrom (Cu Ka).

Aspect 38. The crystalline form of any one of aspects 3 or 33-37, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 12 when heated at a rate of 10°C/min.

Aspect 39. The crystalline form of any one of aspects 3 or 33-38, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 169°C when heated at a rate of 10°C/min.

Aspect 40. A crystalline form as defined in any one of aspects 3, 33-39, having a thermogravimetric analysis profile substantially as shown in FIG. 13 when heated at a rate of 20°C/min.

Aspect 41. The pharmaceutically acceptable salt according to aspect 1, wherein the salt is a phosphate salt, formula III.

Aspect 42. Crystalline form of the phosphate salt of aspect 41.

Aspect 43. The crystalline form of aspect 42, wherein said crystalline form is Formula III-Form I.

Aspect 44. The crystalline form of aspect 42 or 43, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 14A.

Aspect 45. A crystalline form as defined in any one of aspects 42-44, characterized by a powder X-ray diffraction pattern containing a peak at 24.9 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka).

Aspect 46. A crystalline form as defined in any one of aspects 42-45, characterized by an X-ray powder diffraction pattern containing peaks at 18.2, 19.6, 24.9 degrees ± 0.2 degrees 2-theta

- 42 044352 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 47. A crystalline form as defined in any one of aspects 42-46, characterized by an X-ray powder diffraction pattern containing peaks at 18.2, 19.6, 24.9, 25.7 and 27.0 degrees ± 0.2 degrees

2-theta on a 2-theta scale with lambda=1.54 angstroms (Cu Ka).

Aspect 48. The crystalline form of any one of aspects 42-47, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 15A when heating at a rate of 10°C/min.

Aspect 49. The crystalline form of any one of aspects 42-48, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 201° C. when heated at a rate of 10° C./min.

Aspect 50. A crystalline form as defined in any one of aspects 42-49 having a thermogravimetric analysis profile substantially as shown in FIG. 16A when heating at a speed of 20°C/min.

Aspect 51. The crystalline form of aspect 42, wherein said crystalline form is Formula III-Form II.

Aspect 52. The crystalline form of aspect 42 or 51, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 14V.

Aspect 53. The crystalline form of any of Aspects 42 or 51, 52, characterized by a powder X-ray diffraction pattern containing a peak at 24.6 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka ).

Aspect 54. The crystalline form of any of Aspects 42 or 51-53, characterized by a powder X-ray diffraction pattern containing peaks at 19.3, 24.6 and 27.4 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda =1.54 angstrom (Cu Ka).

Aspect 55. The crystalline form of any one of Aspects 42 or 51-54, characterized by an X-ray powder diffraction pattern containing peaks at 19.3, 22.3, 23.6, 24.6 and 27.4 degrees ± 0.2 degrees 2- theta on a 2-theta scale with lambda = 1.54 angstroms (Cu Ka).

Aspect 56. The crystalline form of any one of aspects 42 or 51-55, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 15V when heating at a rate of 10°C/min.

Aspect 57. The crystalline form of any one of aspects 42 or 51-56, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 229° C. when heated at a rate of 10° C./min.

Aspect 58. The crystalline form of any one of aspects 42 or 51-57 having a thermogravimetric analysis profile substantially as shown in FIG. 16V when heating at a rate of 20°C/min.

Aspect 59. The pharmaceutically acceptable salt of Aspect 1, wherein the salt is a tartrate salt, Formula IC.

Aspect 60. The crystalline form of aspect 59, wherein said tartrate salt is crystalline.

Aspect 61. The crystalline form of aspect 59 or 60, characterized by substantially the same powder X-ray diffraction pattern as shown in FIG. 17.

Aspect 62. A crystalline form as defined in any one of aspects 59-61, characterized by a powder X-ray diffraction pattern containing a peak at 18.4 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka).

Aspect 63. A crystalline form as defined in any of Aspects 59-62, characterized by an X-ray powder diffraction pattern containing peaks at 18.4, 19.9 and 21.5 degrees ± 0.2 degrees 2-theta on a 2-theta scale with lambda=1 .54 angstrom (Cu Ka).

Aspect 64. A crystalline form as defined in any one of Aspects 59-63, characterized by an X-ray powder diffraction pattern containing peaks at 18.4, 19.4, 19.9, 21.5 and 26.3 degrees ± 0.2 degrees 2-theta scale 2-theta with lambda = 1.54 angstroms (Cu Ka).

Aspect 65. A crystalline form as defined in any one of aspects 59-64, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 18 when heated at a rate of 10°C/min.

Aspect 66. The crystalline form of any one of aspects 59-65, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 190° C. when heated at a rate of 10° C./min.

Aspect 67. A crystalline form as defined in any one of aspects 59-66 having a thermogravimetric analysis profile substantially as shown in FIG. 19 when heated at a rate of 20°C/min.

Aspect 68. A pharmaceutical composition comprising a pharmaceutically acceptable salt and/or crystalline form according to any one of aspects 1-67 and a pharmaceutically acceptable excipient.

-

Claims (62)

Aspect 69: Method of treating a disease or disorder associated with aberrant activity PRMT5 in a subject, comprising administering to the subject a pharmaceutically acceptable salt and/or crystalline form of any one of aspects 1-67. Aspect 70. The method of aspect 69, wherein the disease or disorder associated with aberrant PRMT5 activity is breast cancer, lung cancer, pancreatic cancer, prostate cancer, colon cancer, ovarian cancer, uterine cancer, cervical cancer , leukemia such as acute myeloid leukemia (AML), chronic lymphocytic leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia, myeloproliferative disorders, acute myelocytic leukemia (AML), chronic myelogenous leukemia z (HML), mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma (MM), myelodysplastic syndrome (MDS), squamous cell carcinoma, hemoglobinopathies such as bthalassemia and sickle cell anemia (SCA), cancers with deletion of the CDKN2A gene; types of cancer with deletion of the 9P gene; types of cancer with deletion of the MTAP gene; cancers caused by splice gene mutations, glioblastoma, non-small cell lung cancer (NSCLC), head and neck cancer, bladder cancer, or hepatocellular carcinoma. CLAIM 1. Pharmaceutically acceptable salt of the compound of formula I: CH ₃ N |HA Cl BUT - (I). 2. The pharmaceutically acceptable salt according to claim 1, wherein the salt is a hydrochloride salt, Formula IA. 3. Crystalline form of the hydrochloride salt according to claim 2. 4. The crystalline form according to claim 3, wherein said crystalline form is Formula IA-Form I. 5. The crystalline form of claim 3 or 4, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 1. 6. The crystalline form of claim 3, 4 or 5, characterized by a powder X-ray diffraction pattern containing a peak at 23.8 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 7. The crystalline form according to any one of claims 3 to 6, characterized by a powder X-ray diffraction pattern containing peaks at 21.2 and 23.8 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms ( Cu Ka). 8. The crystalline form according to any one of claims 3 to 7, characterized by a powder X-ray diffraction pattern containing peaks at 21.2, 23.8, 27.0 and 32.5 degrees ± 0.2 degrees 2-theta on the 2theta scale with lambda =1.54 angstrom (Cu Ka). 9. A crystalline form according to any one of claims 3 to 8, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 2 when heated at a rate of 10°C/min. 10. The crystalline form according to any one of claims 3 to 9, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 244°C when heated at a rate of 10°C/min. 11. A crystalline form according to any one of claims 3 to 10, characterized by substantially the same thermogravimetric analysis profile as shown in FIG. 3 when heated at a rate of 20°C/min. 12. The crystalline form according to claim 3, wherein said crystalline form is Formula IA-Form II. 13. The crystalline form of claim 3 or 12, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 4. 14. The crystalline form of claim 3, 12 or 13, characterized by a powder x-ray diffraction pattern containing a peak at 25.5 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). - 44 044352 15. The crystalline form according to any one of claims 3 or 12-14, characterized by a powder x-ray diffraction pattern containing peaks at 14.8, 17.5 and 25.5 degrees ± 0.2 degrees 2-theta on the scale 2-theta with lambda=1.54 angstroms (Cu Ka). 16. Crystalline form according to any one of claims 3 or 12-15, characterized by a powder x-ray diffraction pattern containing peaks at 14.8, 17.5, 18.4, 24.0, 25.5, 28.0 and 28.7 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 17. The crystalline form according to any one of claims 3 or 12-16, characterized by essentially the same thermogravimetric analysis profile as shown in FIG. 5 when heated at a rate of 20°C/min. 18. The crystalline form according to claim 3, wherein said crystalline form is Formula IA-Form IIa. 19. The crystalline form of claim 3 or 18, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 6. 20. The crystalline form of claim 3, 18 or 19, characterized by a powder x-ray diffraction pattern containing a peak at 26.1 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 21. The crystalline form according to any one of claims 3 or 18-20, characterized by an X-ray powder diffraction pattern containing peaks at 14.0, 14.9 and 26.1 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda =1.54 angstrom (Cu Ka). 22. The crystalline form according to any one of claims 3 or 18-21, characterized by a powder X-ray diffraction pattern containing peaks at 12.5, 14.0, 14.9, 18.4 and 26.1 degrees ± 0.2 degrees 2- theta on a 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 23. The crystalline form according to any one of claims 3 or 18-22, characterized by essentially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 7 when heated at a rate of 10°C/min. 24. The crystalline form according to any one of claims 3 or 18-23, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 199°C when heated at a rate of 10°C/min. 25. The crystalline form according to claim 3, wherein said crystalline form is Formula IA-Form III. 26. The crystalline form of claim 3 or 25, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 8. 27. The crystalline form of claim 3, 25 or 26, characterized by a powder x-ray diffraction pattern containing a peak at 8.1 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 28. The crystalline form according to any one of claims 3 or 25-27, characterized by a powder x-ray diffraction pattern containing peaks at 8.1 and 23.3 degrees ± 0.2 degrees 2-theta on a 2-theta scale with lambda = 1.54 angstrom (Cu Ka). 29. Crystalline form according to any one of claims 3 or 25-28, characterized by a powder X-ray diffraction pattern containing peaks at 8.1, 12.5, 13.7, 14.5, 16.2, 18.8, 23.3 and 24.5 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda=1.54 angstroms (Cu Ka). 30. The crystalline form according to any one of claims 3 or 25-29, characterized by essentially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 9 when heated at a rate of 10°C/min. 31. The crystalline form according to any one of claims 3 or 25-30, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 121°C when heated at a rate of 10°C/min. 32. The crystalline form according to any one of claims 3, 25-31, characterized by essentially the same thermogravimetric analysis profile as shown in FIG. 10 when heating at a rate of 20°C/min. 33. The crystalline form according to claim 3, wherein said crystalline form is Formula IA-Form IV. 34. The crystalline form of claim 3 or 33, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. eleven. 35. The crystalline form according to any one of claims 3, 33 or 34, characterized by a powder x-ray diffraction pattern containing a peak at 4.0 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka ). 36. The crystalline form according to any one of claims 3 or 33-35, characterized by a powder x-ray diffraction pattern containing peaks at 4.0 and 22.7 degrees ± 0.2 degrees 2-theta on a 2-theta scale with lambda = 1.54 angstrom (Cu Ka). 37. Crystalline form according to any one of claims 3 or 33-36, characterized by powder diffraction - 45 044352 tion radiograph containing peaks at 4.0, 22.7 and 27.8 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 38. The crystalline form according to any one of claims 3 or 33-37, characterized by essentially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 12 when heated at a rate of 10°C/min. 39. The crystalline form according to any one of claims 3 or 33-38, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 169°C when heated at a rate of 10°C/min. 40. The crystalline form according to any one of claims 3, 33-39, characterized by essentially the same thermogravimetric analysis profile as shown in FIG. 13 when heated at a rate of 20°C/min. 41. The pharmaceutically acceptable salt according to claim 1, wherein the salt is a phosphate salt. 42. Crystalline form of the phosphate salt according to claim 41. 43. The crystalline form according to claim 42, wherein said crystalline form is Formula III-Form I. 44. The crystalline form of claim 42 or 43, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 14A. 45. The crystalline form according to any one of claims 42 to 44, characterized by a powder x-ray diffraction pattern containing a peak at 24.9 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka). 46. The crystalline form according to any one of claims 42-45, characterized by a powder X-ray diffraction pattern containing peaks at 18.2, 19.6, 24.9 degrees ± 0.2 degrees 2-theta on a 2-theta scale with lambda=1 .54 angstrom (Cu Ka). 47. The crystalline form according to any one of claims 42 to 46, characterized by a powder x-ray diffraction pattern containing peaks at 18.2, 19.6, 24.9, 25.7 and 27.0 degrees ± 0.2 degrees 2-theta scale 2-theta with lambda = 1.54 angstroms (Cu Ka). 48. The crystalline form of any one of claims 42 to 47, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 15A when heating at a rate of 10°C/min. 49. The crystalline form according to any one of claims 42 to 48, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 201°C when heated at a rate of 10°C/min. 50. A crystalline form according to any one of claims 42 to 49, characterized by substantially the same thermogravimetric analysis profile as shown in FIG. 16A when heating at a speed of 20°C/min. 51. The crystalline form according to claim 42, wherein said crystalline form is Formula III-Form II. 52. The crystalline form of claim 42 or 51, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 14V. 53. The crystalline form according to any one of claims 42 or 51, 52, characterized by a powder x-ray diffraction pattern containing a peak at 24.6 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda = 1.54 angstroms (Cu Ka ). 54. The crystalline form according to any one of claims 42 or 51-53, characterized by an X-ray powder diffraction pattern containing peaks at 19.3, 24.6 and 27.4 degrees ± 0.2 degrees 2-theta on the 2-theta scale with lambda =1.54 angstrom (Cu Ka). 55. The crystalline form according to any one of claims 42 or 51-54, characterized by a powder x-ray diffraction pattern containing peaks at 19.3, 22.3, 23.6, 24.6 and 27.4 degrees ± 0.2 degrees 2theta scale 2-theta with lambda = 1.54 angstroms (Cu Ka). 56. The crystalline form according to any one of claims 42 or 51-55, characterized by substantially the same differential scanning calorimetry (DSC) thermogram as shown in FIG. 15V when heating at a rate of 10°C/min. 57. The crystalline form according to any one of claims 42 or 51-56, characterized by a differential scanning calorimetry (DSC) thermogram containing an endothermic peak at about 229°C when heated at a rate of 10°C/min. 58. A crystalline form according to any one of claims 42, 51-57, characterized by substantially the same thermogravimetric analysis profile as shown in FIG. 16V when heating at a rate of 20°C/min. 59. The pharmaceutically acceptable salt according to claim 1, wherein the salt is a tartrate salt. 60. Crystalline form of tartrate salt according to claim 59. 61. The crystalline form of claim 60, characterized by a powder X-ray diffraction pattern substantially as shown in FIG. 17. 62. The crystalline form according to any one of claims 60, 61, characterized by a powder x-ray diffraction pattern containing a peak at 18.4 degrees ± 0.2 degrees 2-theta on a 2-theta scale with lambda -