WO2017218802A1

WO2017218802A1 - Selective inhibitors of protein arginine methyltransferase 5 (prmt5)

Info

Publication number: WO2017218802A1
Application number: PCT/US2017/037720
Authority: WO
Inventors: Juan Luengo; Krishna Vaddi
Original assignee: Prelude Therapeutics Inc
Current assignee: Prelude Therapeutics Inc
Priority date: 2016-06-15
Filing date: 2017-06-15
Publication date: 2017-12-21
Anticipated expiration: 2018-12-15

Abstract

The disclosure is directed to compounds of Formula (I) and Formula (II). Methods of their use and preparation is other described.

Description

SELECTIVE INHIBITORS OF PROTEIN ARGENTINE METHYLTRANSFERASE 5 (PRMT5)

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 62/350,290, filed June 15, 2016, which is incorporated by reference herein in its entirety.

TECHNICAL FIELD

[0002] The disclosure is directed to PRMT5 inhibitors and methods of their use.

BACKGROUND

[0003] Protein arginine methylation is a common post-translational modification that regulates numerous cellular processes, including gene transcription, mRNA splicing, DNA repair, protein cellular localization, cell fate determination, and signaling. Three types of methyl-arginine species exist: ω NG monomethylarginine (MMA), ω NG, NG asymmetric dimethylarginine (ADMA) and ω NG,N'G symmetric dimethylarginine (SDMA) The formation of methylated arginines is catalyzed by the protein arginine methyl transferases (PRMTs) family of

methyltransferases. Currently, there are nine PRMTs annotated in the human genome The majority of these enzymes are Type I enzymes (PRMT1, -2,-3,-4,-6,-8) that are capable of mono- and asymmetric dimethylation of arginine, with S-adenosylmethionine (SAM) as the methyl donor. PRMT-5, -7 and -9 are considered to be Type II enzymes that catalyze symmetric dimethylation of arginines. Each PRMT species harbors the characteristic motifs of seven beta strand

methyltransferases (Katz et al., 2003), as well as additional "double E" and "THW" sequence motifs particular to the PRMT subfamily.

[0004] PRMT5 is as a general transcriptional repressor that functions with numerous transcription factors and repressor complexes, including BRGl and hBRM, Blimp 1, and Snail. This enzyme, once recruited to a promoter, symmetrically dimethylates H3R8 and H4R3. Importantly, the H4R3 site is a major target for PRMTl methylation (ADMA) and is generally regarded as a transcriptional activating mark. Thus, both H4R3me2s (repressive; me2s indicates SDMA modification) and H4R3me2a (active; me2a indicates ADMA modification) marks are produced in vivo. The specificity of PRMT5 for H3R8 and H4R3 can be altered by its interaction with COPR5 and this could perhaps play an important role in determining PRMT5 corepressor status.

Role of PRMTs in Cancer

[0005] Aberrant expression of PRMTs has been identified in human cancers, and PRMTs are considered to be therapeutic targets. Global analysis of histone modifications in prostate cancer has shown that the dimethylation of histone H4R3 is positively correlated with increasing grade, and these changes are predictive of clinical outcome.

[0006] PRMT5 levels have been shown to be elevated in a panel of lymphoid cancer cell lines as well as mantle cell lymphoma clinical samples. PRMT5 interacts with a number of substrates that are involved in a variety of cellular processes, including RNA processing, signal transduction, and transcriptional regulation. PRMT5 can directly modify histone H3 and H4, resulting in the repression of gene expression. PRMT5 overexpression can stimulate cell growth and induce transformation by directly repressing 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 safeguards proper pre-messenger- RNA splicing as an essential step in lymphomagenesis. Koh et al. Nature 2015, 523 7558; Hsu et al. Nature 2015 525, 384.

[0007] The discovery of cancer dependencies has the potential to inform therapeutic strategies and to identify putative drug targets. Integrating data from comprehensive genomic profiling of cancer cell lines and from functional characterization of cancer cell dependencies, it has been recently discovered that loss of the enzyme methylthioadenosine phosphorylase (MTAP) confers a selective dependence on protein arginine methyltransferase 5 (PRMT5) and its binding partner WDR77. MTAP is frequently lost due to its proximity to the commonly deleted tumor suppressor gene, CDKN2A. Cells harboring MTAP deletions possess increased intracellular concentrations of methylthioadenosine (MTA, the metabolite cleaved by MTAP). Furthermore, MTA specifically inhibits PRMT5 enzymatic activity. Administration of either MTA or a small- molecule PRMT5 inhibitor shows a preferential impairment of cell viability for MTAP-null cancer cell lines compared to isogenic MTAP-expressing counterparts. Together, these findings reveal PRMT5 as a potential vulnerability across multiple cancer lineages augmented by a common "passenger" genomic alteration. Role of PRMT5 in Hemoglobinopathies

[0008] The developmental switch in human globin gene subtype from fetal to adult that begins at birth heralds the onset of the hemoglobinopathies, b-thalassemia and sickle cell disease (SCD). The observation that increased adult globin gene expression (in the setting of hereditary persistence of fetal hemoglobin [HPFH] mutations) significantly ameliorates the clinical severity of thalassemia and SCD has prompted the search for therapeutic strategies to reverse gamma-globin gene silencing. Central to silencing of the gamma-genes is DNA methylation, which marks critical CpG dinucleotides flanking the gene transcriptional start site in adult bone marrow erythroid cells. It has been shown that these marks are established as a consequence of recruitment of the DNA methyltransferase, DNMT3 A to the gamma-promoter by the protein arginine methyltransf erase PRMT5. Zhao et al. Nat Struct Mol Biol. 2009 16, 304. PRMT5-mediated methylation of histone H4R3 recruits DNMT3 A, coupling histone and DNA methylation in gene silencing.

[0009] PRMT5 induces the repressive histone mark, H4R3me2s, which serves as a template for direct binding of DNMT3A, and subsequent DNA methylation. Loss of PRMT5 binding or its enzymatic activity leads to demethylation of the 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 essential for assembly of a multiprotein complex on the gamma-promoter, which 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 developing PRMT5 inhibitors as targeted therapies for thalassemia and SCD.

SUMMARY

0010] The disclosure is directed to compounds of Formula I and Formula II:

wherein

n is 1, 2, 3, 4, 5, or 6;

X is -S-, -SO-, -SC-2-, -0-, -NH-, or -N(Ci-C₆alkyl)-; R¹ is -Ci-Cealkyl, -C(0)-Ci-C₆alkyl, -C(0)-0-Ci-C₆alkyl, -Ci-C₆alk-OH,

-Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₆alk- H₂, -Ci-C₆alk- H-Ci-C₆alkyl, or -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl;

R² is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH,

-Co-Cealk-O-Ci-Cealkyl, -C₀-C₆alk- H₂, -Co-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -C₀-C₆alk- H-C₃-C₆cycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃-C₆Cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN;

R³ is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH,

-C₀-C₆alk-O-Ci-C₆alkyl, -C₀-C₆alk- H₂, -C₀-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-Ci-C₆alkyl, -Co-C₆alk- H-C₃-C₆cycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN;

R⁴ is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH,

-C₀-C₆alk-O-Ci-C₆alkyl, -C₀-C₆alk-NH₂, -C₀-C₆alk-NH-Ci-C₆alkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-Ci-C₆alkyl, -Co-C₆alk-NH-C₃-C₆cycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN;

or R² and R³, together with the atoms to which they are attached, form a

C₃-C₆cycloalkenyl ring;

or R² and R³, together form a triple bond;

or R³ and R⁴, together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring or a heterocycloalkyl ring;

R⁵ is H, Ci-Cealkyl, or Co-C₆alk-C₃-C₆cycloalkyl;

R⁶ is H, Ci-C₆alkyl, or C₀-C₆alk-C₃-C₆cycloalkyl; and

R⁷ and R^r are each independently H, Ci-C₆alkyl, or -Co-C₆alk-OCi-C₆alkyl;

or R⁷ and R⁷ , together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring.

[0011] Stereoisomers of the compounds of Formula I and Formula II, and the

pharmaceutical salts thereof, are also described. Methods of using compounds of Formula I and Formula II are described, as well as pharmaceutical compositions including the compounds of Formula I and Formula II. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 shows the results of dialysis of the compounds of Example 2 and Example 20 with wild type PRMT5.

[0013] Figure 2 shows the results of dialysis of the compounds of Example 2 and Eample 20 with C449S mutant of PRMT5.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0014] The disclosure may be more fully appreciated by reference to the following description, including the following definitions and examples. Certain features of the disclosed compositions and methods which are described herein in the context of separate aspects, may also be provided in combination in a single aspect. Alternatively, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single aspect, may also be provided separately or in any subcombination.

[0015] The term "alkyl," when used alone or as part of a substituent group, refers to a straight- or branched-chain alkyl group having from 1 to 12 carbon atoms ("Ci.i₂"), preferably 1 to 6 carbons atoms ("CiV), in the chain. Examples of alkyl groups include methyl (Me, Cialkyl) ethyl (Et, C₂alkyl), n-propyl (C₃alkyl), isopropyl (C₃alkyl), butyl (C₄alkyl), isobutyl (C₄alkyl), sec-butyl (C₄alkyl), tert-butyl (C₄alkyl), pentyl (C₅alkyl), isopentyl (C₅alkyl), tert-pentyl (C₅alkyl), hexyl (C₆alkyl), isohexyl (C₆alkyl), and the like.

[0016] When a range of carbon atoms is used herein, for example, Ci_-6, all ranges, as well as individual numbers of carbon atoms are encompassed. For example, "Ci_-3" includes Ci_-3, Ci_-2,

[0017] The term "C^alk" refers to a straight chain, branched, or cyclic aliphatic linker having 1, 2, 3, 4, 5, or 6 carbon atoms and includes, for example, -CH₂-, -CH(CH₃)-,— C— _? . CH(CH₃)-CH₂-, and -C(CH₃)₂-. The term "-C₀alk-" refers to a bond.

[0018] The term "cycloalkenyl," refers to cyclic, non-aromatic hydrocarbon groups having from 3 to 10 carbon atoms ("C_3-io"), preferably from 3 to 6 carbon atoms ("C_{3 -}6") and containing at least one double bond. For example, cycloalkenyl moieties include, but are not limited to

cyclopropenyl, cyclobutenyl, and the like.

[0019] The term "cycloalkyl" refers to cyclic, non-aromatic hydrocarbon groups having from 3 to 10 carbon atoms ("C_3-io"), preferably from 3 to 6 carbon atoms ("C_{3 -}6")- Examples of cycloalkyl groups include, for example, cyclopropyl (C₃), cyclobutyl (C₄), cyclopentyl (C₅), cyclohexyl (C₆), 1-methylcyclopropyl (C₄), 2-methylcyclopentyl (C₄), adamantanyl (C₁₀), and the like.

[0020] The term "heterocycloalkyl" refers to any five to ten membered monocyclic or bicyclic, saturated ring structure containing at least one heteroatom selected from the group consisting of O, N and S. The heterocycloalkyl group may be attached at any heteroatom or carbon atom of the ring such that the result is a stable structure. Optionally, the heteroatoms or carbon atoms of the ring may be substituted with, for example, a Ci_-6 alkyl group. Examples of suitable heterocycloalkyl groups include, but are not limited to, azepanyl, aziridinyl, azetidinyl, pyrrolidinyl, dioxolanyl, imidazolidinyl, pyrazolidinyl, piperazinyl, 4-methylpiperazin-l-yl, piped dinyl, dioxanyl, morpholinyl, dithianyl, thiomorpholinyl, oxazepanyl, oxiranyl, oxetanyl, 3 -methyl oxitan-3-yl, quinuclidinyl, tetrahyofuranyl, tetrahydropyranyl, piperazinyl, and the like.

[0021] The term "heteroaryl" refers to a mono-or bicyclic aromoatic ring structure including carbon atoms as well as up to four heteroatoms selected from nitrogen, oxygen, and sulfur. Heteroaryl rings can include a total of 5, 6, 9, or 10 ring atoms ("Cs-io")- Optionally, the

heteroatoms or carbon atoms of the ring may be substituted with, for example, a Ci_-6 alkyl group. Examples of heteroaryl groups include but are not limited to, pyrrolyl, furyl, thiophenyl (thienyl), oxazolyl, imidazolyl, l-methylimidazol-2-yl, purazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, pyrazolyl, l-methylpyrazol-3-yl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl, furazanyl, indolizinyl, indolyl, isoindolinyl, indazolyl, benzofuranyl, benzothiophenyl,

benzimidazolyl, benzthiazolyl, purinyl, quinolizinyl, quinolinyl, isoquinolinyl, isothiazolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, and the like.

[0022] The term "halo" represents chloro, fluoro, bromo, or iodo.

[0023] The term "haloalkyl" refers to an alkyl moiety wherein one or more of the hydrogen atoms has been replaced with one or more halo. One exemplary substitutent is fluoro. Preferred haloalkyl groups of the disclosure include trihalogenated alkyl groups such as trifluoromethyl (CF₃) groups.

[0024] "Pharmaceutically acceptable" means approved or approvable by a regulatory agency of the Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized

pharmacopoeia for use in animals, e.g., in humans. [0025] "Pharmaceutically acceptable salt" refers to a salt of a compound of the disclosure that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic may be inorganic or organic acid addition salts and base addition salts. Specifically, 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, hexanoic 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-(4- hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2 -hydroxy ethanesulfonic acid, benzenesulfonic acid, 4- chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-l-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium,

tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.

[0026] A "pharmaceutically acceptable excipient" refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance, added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of an agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0027] "Subject" includes humans. The terms "human," "patient," and "subject" are used interchangeably herein.

[0028] "Treating" or "treatment" of any disease or disorder refers, in one embodiment, to ameliorating the disease or disorder (i.e., arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment "treating" or "treatment" refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, "treating" or "treatment" refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In yet another embodiment, "treating" or "treatment" refers to delaying the onset of the disease or disorder.

[0029] "Compounds of the present disclosure," and equivalent expressions, are meant to embrace compounds of Formula (I) and/ or (II) as described herein, which expression includes the stereoisomers of compounds of Formula I and/or Formula II, as well as the pharmaceutically acceptable salts, where the context so permits.

[0030] As used herein, the term "isotopic variant" refers to a compound that contains proportions of isotopes at one or more of the atoms that constitute such compound that is greater than natural abundance. For example, an "isotopic variant" of a compound can be radiolabeled, that is, contain one or more non-radioactive isotopes, such as for example, deuterium (²H or D), carbon- 13 (¹³C), nitrogen- 15 (¹⁵N), or the like. It will be understood that, in a compound where such isotopic substitution is made, the following atoms, where present, may vary, so that for example, any hydrogen may be ²H/D, any carbon may be ¹³C, or any nitrogen may be ¹⁵N, and that the presence and placement of such atoms may be determined within the skill of the art.

[0031] It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed "isomers." Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers," for example, diastereomers, enantiomers, and atropisomers. The compounds of this disclosure may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)-or (^-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. Where a chiral center exists in a structure, but no specific stereochemistry is shown for that center, both enantiomers, individually or as a mixture of enantiomers, are encompassed by that structure. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art. [0032] The disclosure is directed to compounds of Formula I or Formula II. In some aspects, the disclosure is directed to compounds of Formula I:

[0033] In other as ects, the disclosure is directed to compounds of Formula II:

[0034] According to the disclosure, n in Formula I and/or Formula II is 0, 1, 2, 3, 4, 5, or 6. In some aspects, n is 0, i.e., is a bond. In other aspects, n is 1, 2, 3, 4, 5, or 6. In other aspects, n is 1, 2, or 3. In other aspects, n is 4, 5, or 6. In some aspects, n is 1. In other aspects, n is 2. In yet other aspects, n is 3. In some aspects, n is 4. In other aspects, n is 5. In still other aspects, n is 6.

[0035] According to the disclosure, X in Formula I and/or Formula II is -S-, -SO-, -SO₂-, - 0-, -NH-, or -N(Ci-C₆alkyl)-. In some aspects, X is -S-, -SO-, or -S0₂-. In other aspects, X is -0-. In yet other aspects, X is H- or -N(Ci-C₆alkyl)-, for example -N(CH₃)-, -N(CH₂CH₃)-, or

-N(iPr)-. In some aspects, X is -S-. In some aspects, X is -SO-. In some aspects, X is -S0₂-. In some aspects, X is -0-. In some aspects, X is - H-. In some aspects, X is -N(Ci-C₆alkyl)-, for example -N(CH₃)-, -N(CH₂CH₃)-, or -N(iPr)-.

[0036] According to the disclosure, R¹ in Formula I and/or Formula II is -Ci-C₆alkyl, - C(0)-Ci-C₆alkyl, -C(0)-0-Ci-C₆alkyl, -Ci-C₆alk-OH, -Ci-C₆alk-0-Ci-C₆alkyl, -d-C₆alk- H₂, - Ci-C₆alk- H-Ci-C₆alkyl, or -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl. In some aspects, R¹ is -Ci- C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, and the like. In some aspects, R¹ is -C(0)-Ci-C₆alkyl, for example, -C(0)-methyl, -C(0)-ethyl, -C(O)- propyl, -C(0)-isopropyl, -C(0)-t-butyl, and the like. In some aspects, R¹ is -C(0)-0-Ci-C₆alkyl, for example, -C(0)-0-methyl, -C(0)-0-ethyl, -C(0)-0-propyl, -C(0)-0-isopropyl, -C(0)-0-t-butyl, and the like. In some aspects, R¹ is -Ci-C₆alk-OH, for example, -CH₂CH₂OH, -CH₂CH₂CH₂OH, and the like. In some aspects, R¹ is -Ci-C₆alk-0-Ci-C₆alkyl, for example, -CH₂CH₂OCH₃, - CH₂CH₂CH₂OCH₃, and the like. In some aspects, R¹ is -Ci-C₆alk- H₂, for example, -CH₂CH₂ H₂, -CH₂CH₂CH₂ H₂, and the like. In some aspects, R¹ is -Ci-Cealk- H-Ci-Cealkyl, for example, - CH₂CH₂ HCH₃, -CH₂CH₂CH₂ HCH₃, and the like. In some aspects, R¹ is -Ci-C₆alk-N(d- C₆alkyl)-Ci-C₆alkyl, for example, -CH₂CH₂N(CH₃)₂, -CH₂CH₂CH₂N(CH₃)₂, and the like.

[0037] According to the disclosure, R² in Formula I is H, halo, -Ci-C₆alkyl, -Ci- C₆haloalkyl, -Co-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk- H₂, - Co-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Co-C₆alk- H-C₃-C₆cycloalkyl, -C₀- C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -C₀-C₆alk-heterocycloalkyl, heteroaryl, or -CN. In some aspects, R² is H.

[0038] In some aspects, R² is halo, for example, F, CI, Br, or I, with F, CI, and Br being preferred and F and CI being more preferred.

[0039] In some aspects, R² is -Ci-C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, and the like.

[0040] In some aspects, R² is -Ci-C₆haloalkyl, for example, -CF₃ or -CHF₂.

[0041] In some aspects, R² is -Co-C₆alk-C₃-C₆cycloalkyl, for example, -C₀alk-C₃- C₆cycloalkyl, -Ci-C₆alk-C₃-C₆cycloalkyl, -Ci-C₅alk-C₃-C₆cycloalkyl, -Ci-C₄alk-C₃-C₆cycloalkyl, - Ci-C₃alk-C₃-C₆cycloalkyl, -Ci-C₂alk-C₃-C₆cycloalkyl, -Cialk-C₃-C₆cycloalkyl, -Coalk-C₃cycloalkyl, -Coalk-C₄cycloalkyl, -Coalk-C₅cycloalkyl, -Coalk-C₆cycloalkyl -Ci-C₆alk-C₃cycloalkyl, -Ci-C₆alk- C₄cycloalkyl, -Ci-dalk-Cscycloalkyl, or -Ci-Cealk-Cecycloalkyl. In some aspects wherein R² is - C₀-C₆alk-C₃-C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R² is -C₀-C₆alk- C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0042] In some aspects, R² is -C₀-C₆alk-OH, for example, -C₀alk-OH, -Ci-C₆alk-OH, -Ci-C₅alk-OH, -d-C₄alk-OH, -C C₃alk-OH, -d-dalk-OH, or -Cialk-OH.

[0043] In some aspects, R² is -Co-C₆alk-0-Ci-C₆alkyl, for example, -Coalk-0-Ci-C₆alkyl, - Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₅alk-0-Ci-C₆alkyl, -Ci-C₄alk-0-Ci-C₆alkyl, -Ci-C₃alk-0-Ci-C₆alkyl, -Ci-C₂alk-0-Ci-C₆alkyl, -Cialk-O-Ci-Cealkyl, -C₀alk-O-Ci-C₅alkyl, -C₀alk-O-Ci-C₄alkyl, -C₀alk- 0-Ci-C₃alkyl_j-Coalk-0-Ci-C₂alkyl, -C₀alk-O-Cialkyl, -d-dalk-O-d-dalkyl, -d-C₆alk-0-Ci- C₄alkyl, -Ci-C₆alk-0-Ci-C₃alkyl_j-Ci-C₆alk-0-Ci-C₂alkyl, or -Ci-Cealk-O-Cialkyl.

[0044] In some aspects, R² is Ci-C₆alk- H2, for example, -C₀alk- H₂, -Ci-C₆alk- H₂, -d- C₅alk- H₂, -Ci-C₄alk- H₂, -d-C₃alk- H₂, -d-C₂alk- H₂, or -dalk- H₂. [0045] In some aspects, R² is -Co-C₆alk- H-Ci-C₆alkyl, for example, -C₀alk- H-Ci- C₆alkyl, -Ci-C₆alk- H-Ci-C₆alkyl, -Ci-C₅alk- H-Ci-C₆alkyl, -Ci-C₄alk- H-Ci-C₆alkyl, -C C₃alk- H-Ci-Cealkyl, -Ci-C₂alk- H-Ci-C₆alkyl, -Cialk-NH-Ci-C₆alkyl, -C₀alk- H-Ci-C₅alkyl, -C₀alk- H-Ci-C₄alkyl, -C₀alk- H-Ci-C₃alkyl, -C₀alk- H-Ci-C₂alkyl, -C₀alk- H-Cialkyl, -Ci-C₆alk- H- Ci-Csalkyl, -Ci-C₆alk- H-Ci-C₄alkyl, -Ci-C₆alk- H-Ci-C₃alkyl, -Ci-C₆alk- H-Ci-C₂alkyl, or -Ci- Cealk- H-Cialkyl.

[0046] In some aspects, R² is -Co-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, for example, -C₀alk- N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₅alk-N(Ci-C₆alkyl)-Ci- Cealkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₂alk-N(Ci- Cealky -Ci-Cealkyl, -Cialk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Coalk-N(Ci-C₆alkyl)-Ci-C₅alkyl, -C₀alk- N(Ci-C₆alkyl)-Ci-C₄alkyl, -C₀alk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -C₀alk-N(Ci-C₆alkyl)-Ci-C₂alkyl, - C₀alk-N(Ci-C₆alkyl)-Cialkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₅alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci- C₄alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₂alkyl, -Ci-C₆alk-N(Ci- C₆alkyl)-Cialkyl, -Coalk-N(Ci-C₅alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -C₀alk-N(Cr C₃alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci-C₂alkyl)-Ci-C₆alkyl, -C₀alk-N(Cialkyl)-Ci-C₆alkyl, -Ci-C₆alk- N(Ci-C₅alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₃alkyl)-Ci- Cealkyl, -Ci-C₆alk-N(Ci-C₂alkyl)-Ci-C₆alkyl, or -Co-C₆alk-N(Cialkyl)-Ci-C₆alkyl.

[0047] In some aspects, R² is -C₀-C₆alk-NH-C₃-C₆cycloalkyl, for example, -C₀alk- H-C₃- C₆cycloalkyl, -Ci-C₆alk- H-C₃-C₆cycloalkyl, -Ci-C₅alk- H-C₃-C₆cycloalkyl, -Ci-C₄alk- H-C₃- Cecycloalkyl, -Ci-C₃alk- H-C₃-C₆cycloalkyl, -Ci-C₂alk- H-C₃-C₆cycloalkyl, -Cialk- H-C₃- C₆cycloalkyl, -C₀alk- H-C₃cycloalkyl, -C₀alk- H-C₄cycloalkyl, -C₀alk- H-C₅cycloalkyl, -C₀alk- H-Cecycloalkyl, -Ci-C₆alk- H-C₃cycloalkyl, -Ci-C₆alk- H-C₄cycloalkyl, -Ci-C₆alk- H- C₅cycloalkyl, or -Ci-C₆alk- H-C₆cycloalkyl. In some aspects wherein R² is -C₀-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R² is -Co-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0048] In some aspects, R² is -Co-C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, for example, - C₀alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃-C₆Cycloalkyl, -Ci-C₅alk-N(Ci- C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -C₁-C₂alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -C₁alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, - Coalk-N(Ci-C₆alkyl)-C₃cycloalkyl, -Coalk-N(Ci-C₆alkyl)-C₄cycloalkyl, -C₀alk-N(Ci-C₆alkyl)- Cscycloalkyl, -C₀alk-N(Ci-C₆alkyl)-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃cycloalkyl, -C C₆alk-N(Ci-C₆alkyl)-C₄cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₅cycloalkyl, -Ci-C₆alk-N(d- C₆alkyl)-C₆cycloalkyl, -C₀alk-N(Ci-C₅alkyl)-C₃-C₆cycloalkyl, -C₀alk-N(Ci-C₄alkyl)-C₃- Cecycloalkyl, -C₀alk-N(Ci-C₃alkyl)-C₃-C₆cycloalkyl, -C₀alk-N(Ci-C₂alkyl)-C₃-C₆cycloalkyl, - C₀alk-N(Cialkyl)-C₃-C₆cycloalkyl, -C₁-C₆alk-N(Ci-C₅alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(d- C₄alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₃alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₂alkyl)-C₃- C₆cycloalkyl, or -Ci-C₆alk-N(Cialkyl)-C₃-C₆cycloalkyl. In some aspects wherein R² is -Co-C₆alk- N(Ci-C6alkyl)-C₃-C6cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R² is -C₀- C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), - OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0049] In some aspects, R² is -Co-Cealk-heterocycloalkyl, for example, -C₀alk- heterocycloalkyl, -Ci-C₆alk-heterocycloalkyl, -Ci-C₅alk-heterocycloalkyl, -Ci-C₄alk- heterocycloalkyl, -Ci-C₃alk-heterocycloalkyl, -Ci-C₂alk-heterocycloalkyl, or -Cialk- heterocycloalkyl. Preferred heterocyloalkyl moieties include, for example piperidinyl, piperazinyl, morpholinyl, aziridinyl, dioxanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, and oxetanyl. In some aspects wherein R² is -C₀-C₆alk-heterocycloalkyl, the heterocycloalkyl is unsubstituted. In other aspects wherein R² is -C₀-C₆alk-heterocycloalkyl, the heterocycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0050] In some aspects, R² is heteroaryl, for example furanyl, imidazolyl, and pyrazolyl. In some aspects wherein R² is heteroaryl, the heteroaryl is unsubstituted. In other aspects wherein R² is heteroaryl, the heteroaryl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0051] In some aspects, R² is -CN.

[0052] According to the disclosure, R³ in Formula I is H, halo, -Ci-C₆alkyl, -Ci- C₆haloalkyl, -Co-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk- H₂, - C₀-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -C₀-C₆alk- H-C₃-C₆cycloalkyl, -C₀- C6alk-N(Ci-C6alkyl)-C3-C6cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN. In some aspects, R³ is H.

[0053] In some aspects, R³ is halo, for example, F, CI, Br, or I, with F, CI, and Br being preferred and F and CI being more preferred.

[0054] In some aspects, R³ is -Ci-C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, and the like.

[0055] In some aspects, R³ is -Ci-C₆haloalkyl, for example, -CF₃ or -CHF₂.

[0056] In some aspects, R³ is -Co-C6alk-C₃-C6cycloalkyl, for example, -C₀alk-C₃- C₆cycloalkyl, -Ci-C6alk-C₃-C6Cycloalkyl, -Ci-C5alk-C₃-C6cycloalkyl, -Ci-C4alk-C₃-C6Cycloalkyl, - Ci-C₃alk-C₃-C₆cycloalkyl, -Ci-C₂alk-C₃-C₆cycloalkyl, -Cialk-C₃-C₆cycloalkyl, -C₀alk-C₃cycloalkyl, -Coalk-C4cycloalkyl, -Coalk-C₅cycloalkyl, -Coalk-C₆cycloalkyl -Ci-C6alk-C₃cycloalkyl, -Ci-C₆alk- C4Cycloalkyl, -Ci-C₆alk-C5Cycloalkyl, or -Ci-Cealk-Cecycloalkyl. In some aspects wherein R3 is - Co-C6alk-C₃-C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R³ is -Co-C₆alk- C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0057] In some aspects, R³ is -C₀-C₆alk-OH, for example, -C₀alk-OH, -Ci-C₆alk-OH, -Ci-C₅alk-OH, -d-C₄alk-OH, -C C₃alk-OH, -d-C₂alk-OH, or -Cialk-OH.

[0058] In some aspects, R³ is -C₀-C₆alk-O-Ci-C₆alkyl, for example, -C₀alk-O-Ci-C₆alkyl, - Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₅alk-0-Ci-C₆alkyl, -Ci-C₄alk-0-Ci-C₆alkyl, -Ci-C₃alk-0-Ci-C₆alkyl, -Ci-C₂alk-0-Ci-C₆alkyl, -Cialk-O-Ci-Cealkyl, -C₀alk-O-Ci-C₅alkyl, -C₀alk-O-Ci-C₄alkyl, -C₀alk- O-Ci-C₃alkyl_j-C₀alk-O-Ci-C₂alkyl, -C₀alk-O-Cialkyl, -Ci-C₆alk-0-Ci-C₅alkyl, -Ci-C₆alk-0-Ci- C₄alkyl, -C₁-C₆alk-0-Ci-C₃alkyl_i-Ci-C₆alk-0-Ci-C₂alkyl, or -Ci-C₆alk-0-Cialkyl.

[0059] In some aspects, R³ is Ci-C₆alk- H₂, for example, -C₀alk- H₂, -Ci-C₆alk- H₂, -Ci- C₅alk- H₂, -Ci-C₄alk- H₂, -Ci-C₃alk- H₂, -Ci-C₂alk- H₂, or -Cialk- H₂.

[0060] In some aspects, R³ is -Co-C₆alk- H-Ci-C₆alkyl, for example, -C₀alk- H-Ci- C₆alkyl,

-Ci-C₆alk- H-Ci-C₆alkyl, -Ci-C₅alk- H-Ci-C₆alkyl, -Ci-C₄alk- H-Ci-C₆alkyl, -Ci-C₃alk- H-Ci- C₆alkyl, -Ci-C₂alk- H-Ci-C₆alkyl, -Cialk- H-Ci-C₆alkyl, -C₀alk- H-Ci-C₅alkyl, -Coalk-NH-Ci- C₄alkyl, -C₀alk- H-Ci-C₃alkyl, -C₀alk- H-Ci-C₂alkyl, -C₀alk- H-Cialkyl, -Ci-C₆alk- H-Ci- Csalkyl, -Ci-C₆alk- H-Ci-C₄alkyl, -Ci-C₆alk- H-Ci-C₃alkyl, -Ci-C₆alk- H-Ci-C₂alkyl, or -Ci- Cealk- H-Cialkyl.

[0061] In some aspects, R³ is -Co-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, for example, -C₀alk- N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₅alk-N(Ci-C₆alkyl)-Ci- Cealkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₂alk-N(Ci- Cealky -Ci-Cealkyl, -Cialk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Coalk-N(Ci-C₆alkyl)-Ci-C₅alkyl, -C₀alk- N(Ci-C₆alkyl)-Ci-C₄alkyl, -C₀alk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -Coalk-N(Ci-C₆alkyl)-Ci-C₂alkyl, - C₀alk-N(Ci-C₆alkyl)-Cialkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₅alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci- C₄alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₂alkyl, -Ci-C₆alk-N(Ci- C₆alkyl)-Cialkyl, -Coalk-N(Ci-C₅alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -C₀alk-N(Cr C₃alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci-C₂alkyl)-Ci-C₆alkyl, -C₀alk-N(Cialkyl)-Ci-C₆alkyl, -Ci-C₆alk- N(Ci-C₅alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₃alkyl)-Ci- Cealkyl, -Ci-C₆alk-N(Ci-C₂alkyl)-Ci-C₆alkyl, or -Co-C₆alk-N(Cialkyl)-Ci-C₆alkyl.

[0062] In some aspects, R³ is -C₀-C₆alk-NH-C₃-C₆cycloalkyl, for example, -C₀alk- H-C₃- Cecycloalkyl, -Ci-C₆alk- H-C₃-C₆cycloalkyl, -Ci-C₅alk- H-C₃-C₆cycloalkyl, -Ci-C₄alk- H-C₃- Cecycloalkyl, -Ci-C₃alk- H-C₃-C₆cycloalkyl, -Ci-C₂alk- H-C₃-C₆cycloalkyl, -Cialk-NH-C₃- C₆cycloalkyl, -C₀alk- H-C₃cycloalkyl, -C₀alk- H-C₄cycloalkyl, -C₀alk- H-C₅cycloalkyl, -C₀alk- H-Cecycloalkyl, -Ci-C₆alk- H-C₃cycloalkyl, -Ci-C₆alk- H-C₄cycloalkyl, -Ci-C₆alk- H- C₅cycloalkyl, or -Ci-C₆alk- H-C₆cycloalkyl. In some aspects wherein R³ is -C₀-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R³ is -Co-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0063] In some aspects, R³ is -Co-C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, for example, - C₀alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃-C₆Cycloalkyl, -Ci-C₅alk-N(Ci- C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -C₁-C₂alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -C₁alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, - C₀alk-N(Ci-C₆alkyl)-C₃cycloalkyl, -C₀alk-N(Ci-C₆alkyl)-C₄cycloalkyl, -C₀alk-N(Ci-C₆alkyl)- Cscycloalkyl, -C₀alk-N(Ci-C₆alkyl)-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃cycloalkyl, -Ci- C₆alk-N(Ci-C₆alkyl)-C₄cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₅cycloalkyl, -Ci-C₆alk-N(Ci- C₆alkyl)-C₆cycloalkyl, -C₀alk-N(Ci-C₅alkyl)-C₃-C₆cycloalkyl, -C₀alk-N(Ci-C₄alkyl)-C₃- Cecycloalkyl, -C₀alk-N(Ci-C₃alkyl)-C3-C6cycloalkyl, -C₀alk-N(Ci-C₂alkyl)-C3-C6cycloalkyl, - Coalk-N(Cialkyl)-C3-C₆cycloalkyl, -Ci-C6alk-N(Ci-C₅alkyl)-C3-C₆cycloalkyl, -Ci-C₆alk-N(d- C₄alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₃alkyl)-C₃-C₆Cycloalkyl, -Ci-Cealk-N(Ci-C₂alkyl)-C₃- C₆cycloalkyl, or -Ci-C₆alk-N(Cialkyl)-C₃-C₆cycloalkyl. In some aspects wherein R³ is -Co-C₆alk- N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R³ is -C₀- C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), - OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0064] In some aspects, R³ is -Co-Cealk-heterocycloalkyl, for example, -C₀alk- heterocycloalkyl, -Ci-C₆alk-heterocycloalkyl, -Ci-C₅alk-heterocycloalkyl, -Ci-C₄alk- heterocycloalkyl, -Ci-C₃alk-heterocycloalkyl, -Ci-C₂alk-heterocycloalkyl, or -Cialk- heterocycloalkyl. Preferred heterocyloalkyl moieties include, for example piperidinyl, piperazinyl, morpholinyl, aziridinyl, dioxanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, and oxetanyl. In some aspects wherein R³ is -C₀-C₆alk-heterocycloalkyl, the heterocycloalkyl is unsubstituted. In other aspects wherein R³ is -Co-Cealk-heterocycloalkyl, the heterocycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0065] In some aspects, R³ is heteroaryl, for example furanyl, imidazolyl, and pyrazolyl. In some aspects wherein R³ is heteroaryl, the heteroaryl is unsubstituted. In other aspects wherein R³ is heteroaryl, the heteroaryl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0066] In some aspects, R³ is -CN.

[0067] According to the disclosure, R⁴ in Formula I is H, halo, -Ci-C₆alkyl, -Ci- C₆haloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk- H₂, - C₀-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -C₀-C₆alk- H-C₃-C₆cycloalkyl, -C₀- C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN. In some aspects, R⁴ is H.

[0068] In some aspects, R⁴ is halo, for example, F, CI, Br, or I, with F, CI, and Br being preferred and F and CI being more preferred. [0069] In some aspects, R⁴ is -Ci-C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, and the like.

[0070] In some aspects, R⁴ is -Ci-C₆haloalkyl, for example, -CF₃ or -CHF₂.

[0071] In some aspects, R⁴ is -Co-C₆alk-C₃-C₆cycloalkyl, for example, -C₀alk-C₃- C₆cycloalkyl, -Ci-C₆alk-C₃-C₆Cycloalkyl, -Ci-C₅alk-C₃-C₆cycloalkyl, -Ci-C₄alk-C₃-C₆Cycloalkyl, - Ci-C₃alk-C₃-C₆cycloalkyl, -Ci-C₂alk-C₃-C₆cycloalkyl, -Cialk-C₃-C₆cycloalkyl, -C₀alk-C₃cycloalkyl, -Coalk-C₄cycloalkyl, -Coalk-C₅cycloalkyl, -Coalk-C₆cycloalkyl -Ci-C₆alk-C₃cycloalkyl, -Ci-C₆alk- C₄Cycloalkyl, -Ci-C₆alk-C₅Cycloalkyl, or -Ci-Cealk-Cecycloalkyl. In some aspects wherein R⁴ is - Co-C₆alk-C₃-C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R⁴ is -Co-C₆alk- C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0072] In some aspects, R⁴ is -C₀-C₆alk-OH, for example, -C₀alk-OH, -Ci-C₆alk-OH, -Ci-C₅alk-OH, -d-dalk-OH, -C C₃alk-OH, -d-C₂alk-OH, or -Cialk-OH.

[0073] In some aspects, R⁴ is -Co-C₆alk-0-Ci-C₆alkyl, for example, -Coalk-0-Ci-C₆alkyl, - Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₅alk-0-Ci-C₆alkyl, -Ci-C₄alk-0-Ci-C₆alkyl, -Ci-C₃alk-0-Ci-C₆alkyl, -Ci-C₂alk-0-Ci-C₆alkyl, -Cialk-O-Ci-Cealkyl, -C₀alk-O-Ci-C₅alkyl, -C₀alk-O-Ci-C₄alkyl, -C₀alk- O-Ci-C₃alkyl_j-C₀alk-O-Ci-C₂alkyl, -C₀alk-O-Cialkyl, -d-C₆alk-0-Ci-C₅alkyl, -d-C₆alk-0-Ci- C₄alkyl, -C₁-C₆alk-0-Ci-C₃alkyl_i-Ci-C₆alk-0-Ci-C₂alkyl ,-Ci-C₆alk-0-Cialkyl.

[0074] In some aspects, R⁴ is Ci-C₆alk- H₂, for example, -C₀alk- H₂, -Ci-C₆alk- H₂, -d- C₅alk- H₂, -Ci-C₄alk- H₂, -Ci-C₃alk- H₂, -Ci-C₂alk- H₂, or -Cialk- H₂.

[0075] In some aspects, R⁴ is -Co-C₆alk- H-Ci-C₆alkyl, for example, -C₀alk- H-Ci- C₆alkyl, -d-C₆alk- H-Ci-C₆alkyl, -Ci-C₅alk- H-Ci-C₆alkyl, -Ci-C₄alk- H-Ci-C₆alkyl, -C C₃alk- H-Ci-Cealkyl, -Ci-C₂alk- H-Ci-C₆alkyl, -Cialk- H-Ci-Cealkyl, -C₀alk- H-Ci-C₅alkyl, -C₀alk- H-Ci-C₄alkyl, -C₀alk- H-Ci-C₃alkyl, -C₀alk- H-Ci-C₂alkyl, or -C₀alk- H-Cialkyl, -Ci-dalk- H-Ci-dalkyl, -Ci-C₆alk- H-Ci-C₄alkyl, -d-C₆alk- H-Ci-C₃alkyl, -Ci-C₆alk- H-Ci-C₂alkyl, or -Ci-dalk- H-Cialkyl.

[0076] In some aspects, R⁴ is -Co-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, for example, -C₀alk- N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₅alk-N(Ci-C₆alkyl)-Ci- C₆alkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Ci-C₂alk-N(Ci- C₆alkyl)-Ci-C₆alkyl, -Cialk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Coalk-N(Ci-C6alkyl)-Ci-C₅alkyl, -C₀alk- N(Ci-C₆alkyl)-Ci-C₄alkyl, -Coalk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -Coalk-N(Ci-C₆alkyl)-Ci-C₂alkyl, - Coalk-N(Ci-C₆alkyl)-Cialkyl, -Ci-C6alk-N(Ci-C6alkyl)-Ci-C₅alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci- C₄alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₃alkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₂alkyl, -Ci-C₆alk-N(Ci- C₆alkyl)-Cialkyl, -Coalk-N(Ci-C₅alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -C₀alk-N(Ci- C₃alkyl)-Ci-C₆alkyl, -Coalk-N(Ci-C₂alkyl)-Ci-C₆alkyl, -C₀alk-N(Cialkyl)-Ci-C₆alkyl, -Ci-C₆alk- N(Ci-C₅alkyl)-Ci-C₆alkyl, -Ci-C₆alk-N(Ci-C₄alkyl)-Ci-C₆alkyl, -C₁-C₆alk-N(Ci-C₃alkyl)-C₁- Cealkyl, -Ci-C₆alk-N(Ci-C₂alkyl)-Ci-C₆alkyl, or -Co-C₆alk-N(Cialkyl)-Ci-C₆alkyl.

[0077] In some aspects, R⁴ is -Co-C₆alk- H-C₃-C₆cycloalkyl, for example, -C₀alk- H-C₃- Cecycloalkyl, -Ci-C₆alk- H-C₃-C₆cycloalkyl, -Ci-C₅alk- H-C₃-C₆cycloalkyl, -Ci-C₄alk- H-C₃- C₆cycloalkyl, -Ci-C₃alk- H-C₃-C₆cycloalkyl, -Ci-C₂alk- H-C₃-C₆cycloalkyl, -Cialk-NH-C₃- C₆cycloalkyl, -C₀alk-NH-C₃cycloalkyl, -C₀alk- H-C₄cycloalkyl, -C₀alk- H-C₅cycloalkyl, -C₀alk- H-Cecycloalkyl, -Ci-C₆alk- H-C₃cycloalkyl, -Ci-C₆alk- H-C₄cycloalkyl, -Ci-C₆alk- H- C₅cycloalkyl, or -Ci-Cealk- H-Cecycloalkyl. In some aspects wherein R⁴ is -Co-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R⁴ is -C₀-C₆alk- H-C₃- C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0078] In some aspects, R⁴ is -Co-C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, for example, - C₀alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₅alk-N(C C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₄alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Ci-C₃alk-N(Ci-C₆alkyl)-C₃- Cecycloalkyl, -Ci-C₂alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, -Cialk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, - C₀alk-N(Ci-C₆alkyl)-C₃cycloalkyl, -C₀alk-N(Ci-C₆alkyl)-C₄cycloalkyl, -C₀alk-N(Ci-C₆alkyl)- Cjcycloalkyl, -C₀alk-N(Ci-C₆alkyl)-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₃cycloalkyl, -C C₆alk-N(Ci-C₆alkyl)-C₄cycloalkyl, -Ci-C₆alk-N(Ci-C₆alkyl)-C₅cycloalkyl, -Ci-C₆alk-N(Ci- C₆alkyl)-C₆cycloalkyl, -C₀alk-N(Ci-C₅alkyl)-C₃-C₆cycloalkyl, -C₀alk-N(Ci-C₄alkyl)-C₃- Cecycloalkyl, -C₀alk-N(Ci-C₃alkyl)-C₃-C₆cycloalkyl, -C₀alk-N(Ci-C₂alkyl)-C₃-C₆cycloalkyl, - C₀alk-N(Cialkyl)-C₃-C₆cycloalkyl, -C₁-C₆alk-N(Ci-C₅alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(d- C₄alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₃alkyl)-C₃-C₆cycloalkyl, -Ci-C₆alk-N(Ci-C₂alkyl)-C₃- C₆cycloalkyl, or -Ci-C₆alk-N(Cialkyl)-C₃-C₆cycloalkyl. In some aspects wherein R⁴ is -Co-C₆alk- N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, the cycloalkyl is unsubstituted. In other aspects wherein R⁴ is -C₀- C₆alk-N(Ci-C₆alkyl)-C₃-C₆cycloalkyl, the cycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), - OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0079] In some aspects, R⁴ is -Co-Cealk-heterocycloalkyl, for example, -C₀alk- heterocycloalkyl, -Ci-Cealk-heterocycloalkyl, -Ci-Csalk-heterocycloalkyl, -Ci-C₄alk- heterocycloalkyl, -Ci-C₃alk-heterocycloalkyl, -Ci-C₂alk-heterocycloalkyl, or -Cialk- heterocycloalkyl. Preferred heterocyloalkyl moieties include, for example, piperidinyl, piperazinyl, morpholinyl, aziridinyl, dioxanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, or oxetanyl. In some aspects wherein R⁴ is -Co-Cealk-heterocycloalkyl, the heterocycloalkyl is unsubstituted. In other aspects wherein R⁴ is -Co-Cealk-heterocycloalkyl, the heterocycloalkyl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., -Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0080] In some aspects, R⁴ is heteroaryl, for example furanyl, imidazolyl, and pyrazolyl. In some aspects wherein R⁴ is heteroaryl, the heteroaryl is unsubstituted. In other aspects wherein R⁴ is heteroaryl, the heteroaryl is substituted with one, two, or three R substitutents independently selected from Ci-C₆alkyl, (e.g., methyl, ethyl, propyl, isopropyl, butyl), -OCi-C₆alkyl (e.g., - Omethyl, -Oethyl, -Opropyl, -Oisopropyl, -Obutyl), and halo (e.g., F or CI).

[0081] In some aspects, R⁴ is -CN.

[0082] In some aspects, R² and R³ are each H and R⁴ is halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk- H₂, -C₀-C₆alk- H- Ci-Cealkyl, -Co-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl, -Co-Cealk- H-Cs-Cec cloalkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-C₃-C₆Cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN.

[0083] In some aspects, R² and R⁴ are each H and R³ is halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk-NH₂, -C₀-C₆alk-NH- Ci-Cealkyl, -Co-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl, -Co-Cealk- H-Cs-Cec cloalkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-C₃-C₆Cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN.

[0084] In some aspects, R³ and R⁴ are each H and R² is halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk-NH₂, -C₀-C₆alk-NH- Ci-Cealkyl, -Co-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl, -Co-Cealk- H-Cs-Cec cloalkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-C₃-C₆Cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN. [0085] In some embodiments of the disclosure wherein the compounds are of Formula I, R² and R³, together with the atoms to which they are attached, form a C₃-C₆cycloalkenyl ring, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl, or cyclohexenyl.

[0086] In some embodiments of the disclosure wherein the compounds are of Formula I, R² and R³ together form a triple bond.

[0087] In some embodiments of the disclosure wherein the compounds are of Formula I, R³ and R⁴, together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring or a heterocycloalkyl ring. In some aspects, R³ and R⁴, together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In other aspects, R³ and R⁴, together with the atom to which they are attached, form a heterocycloalkyl ring, for example, piperidinyl, piperazinyl, morpholinyl, aziridinyl, dioxanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, or oxetanyl.

[0088] In embodiments of the disclosure wherein the compounds are of Formula I and/or Formula II, R⁵ is H, -Ci-C₆alkyl, or C₀-C₆alk-C₃-C₆cycloalkyl. In some aspects, R⁵ is H. In other aspects, R⁵ is -Ci-C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t- butyl, pentyl, and the like. In other aspects, R⁵ is Co-C₆alk-C₃-C₆cycloalkyl, for example, C₀-alk-C₃- C₆cycloalkyl, Ci-C₆alk-C₃-C₆cycloalkyl, Ci-C₅alk-C₃-C₆Cycloalkyl, Ci-C₄alk-C₃-C₆Cycloalkyl, Ci- C₃alk-C₃-C₆cycloalkyl, Ci-C₂alk-C₃-C₆cycloalkyl, Cialk-C₃-C₆cycloalkyl, C₀alk-C₃cycloalkyl, C₀alk-C₄cycloalkyl, C₀alk-C₅cycloalkyl, C₀alk-C₆cycloalkyl, Ci-C₆alk-C₃cycloalkyl, Ci-C₆alk- C₄cycloalkyl, Ci-C₆alk-C₅cycloalkyl, or Ci-Cealk-Cecycloalkyl.

[0089] In embodiments of the disclosure wherein the compounds are of Formula II, R⁶ is H, -Ci-C₆alkyl, or Co-C₆alk-C₃-C₆cycloalkyl. In some aspects, R⁶ is H. In other aspects, R⁶ is -Ci- C₆alkyl, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, and the like. In other aspects, R⁶ is Co-C₆alk-C₃-C₆cycloalkyl, for example, Co-alk-C₃-C₆cycloalkyl, Ci- C₆alk-C₃-C₆cycloalkyl, Ci-C₅alk-C₃-C₆cycloalkyl, Ci-C₄alk-C₃-C₆cycloalkyl, Ci-C₃alk-C₃- C₆cycloalkyl, Ci-C₂alk-C₃-C₆cycloalkyl, Cialk-C₃-C₆Cycloalkyl, Coalk-C₃cycloalkyl, C₀alk- C cycloalkyl, C₀alk-C₅cycloalkyl, C₀alk-C₆cycloalkyl, Ci-C₆alk-C₃cycloalkyl, Ci-C₆alk- C₄cycloalkyl, Ci-C₆alk-C₅cycloalkyl, or Ci-Cealk-Cecycloalkyl.

[0090] In embodiments of the disclosure wherein the compounds are of Formula II, R⁷ and R⁷' are each independently H, Ci-C₆alkyl (e.g., , methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s- butyl, t-butyl, pentyl, and the like), or -C₀-C₆alk-OCi-C₆alkyl (e.g., C₀alk-OCi-C₆alkyl, Ci-C₆alk- OCi-Cealkyl, Ci-C₅alk-OCi-C₆alkyl, Ci-C₄alk-OCi-C₆alkyl, Ci-C₃alk-OCi-C₆alkyl, Ci-C₂alk-OCi- Cealkyl, Cialk-OCi-C₆alkyl, C₀-C₆alk-OCi-C₅alkyl, C₀-C₆alk-OCi-C₄alkyl, C₀-C₆alk-OCi-C₃alkyl, Co-C₆alk-OCi-C₂alkyl, or Co-C₆alk-OCialkyl). In some embodiments, R⁷ is Ci-C₆alkyl or -C₀- C₆alk-OCi-C₆alkyl and R⁷ is H, Ci-C₆alkyl, or -Co-C₆alk-OCi-C₆alkyl. In some aspects, R⁷ and R⁷ are each H. In other aspects, R⁷ and R⁷' are each independently Ci-C₆alkyl. In other aspects, R⁷ and R⁷ are each independently -C₀-C₆alk-OCi-C₆alkyl. In some aspects, R⁷ is Ci-C₆alkyl and R⁷ is H. In other aspects, R⁷ is -Co-C₆alk-OCi-C₆alkyl and R⁷ is H.

[0091] In embodiments of the disclosure wherein the compounds are of Formula II, R⁷ and R⁷ , together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0092] Stereoisomers of compounds of Formula I and/or Formula II are also contemplated.

[0093] Pharmaceutically acceptable salts of the compounds of Formula I and/or Formula II are also within the scope of the disclosure.

Pharmaceutical compositions and methods of administration

[0094] The subject pharmaceutical compositions are typically formulated to provide a therapeutically effective amount of a compound of the present disclosure as the active ingredient, or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof. Where desired, the pharmaceutical compositions contain pharmaceutically acceptable salt and/or coordination complex thereof, and one or more pharmaceutically acceptable excipients, carriers, including inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants.

[0095] The subject pharmaceutical compositions can be administered alone or in combination with one or more other agents, which are also typically administered in the form of pharmaceutical compositions. Where desired, the one or more compounds of the invention and other agent(s) may be mixed into a preparation or both components may be formulated into separate preparations to use them in combination separately or at the same time.

[0096] 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) w/w, w/v or v/v.

[0097] In some embodiments, the concentration of one or more compounds of the 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%, 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) w/w, w/v, or v/v.

[0098] In some embodiments, the concentration of one or more compounds of the invention is in the range from approximately 0.0001%) to approximately 50%, approximately 0.001%) to approximately 40%, approximately 0.01%> to approximately 30%>, approximately 0.02% to approximately 29%, approximately 0.03% to approximately 28%, approximately 0.04% to approximately 27%, approximately 0.05% to approximately 26%, approximately 0.06% to approximately 25%, approximately 0.07% to approximately 24%, approximately 0.08% to approximately 23%, approximately 0.09% to approximately 22%, approximately 0.1% to approximately 21%, approximately 0.2% to approximately 20%, approximately 0.3% to approximately 19%, approximately 0.4% to approximately 18%, approximately 0.5% to approximately 17%, approximately 0.6% to approximately 16%, approximately 0.7% to approximately 15%, approximately 0.8% to approximately 14%, approximately 0.9% to approximately 12%, approximately 1% to approximately 10% w/w, w/v or v/v.

[0099] In some embodiments, the concentration of one or more compounds of the invention is in the range from approximately 0.001%) to approximately 10%, approximately 0.01% to approximately 5%, approximately 0.02% to approximately 4.5%, approximately 0.03% to approximately 4%, approximately 0.04% to approximately 3.5%, approximately 0.05% to approximately 3%, approximately 0.06% to approximately 2.5%, approximately 0.07% to approximately 2%, approximately 0.08% to approximately 1.5%, approximately 0.09% to approximately 1%, approximately 0.1% to approximately 0.9% w/w, w/v or v/v.

[00100] In some embodiments, the amount of one or more compounds of the invention is equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g (or a number in the range defined by and including any two numbers above).

[00101] In some embodiments, the amount of one or more compounds of the invention is more than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, , 0.15 g, 0.2 g, , 0.25 g, 0.3 g, , 0.35 g, 0.4 g, , 0.45 g, 0.5 g, 0.55 g, 0.6 g, , 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5g, 7 g, 7.5g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g (or a number in the range defined by and including any two numbers above).

[00102] In some embodiments, the amount of one or more compounds of the invention is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[00103] The compounds according to the invention are effective over a wide dosage range. For example, in the treatment of adult humans, dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg per day are examples of dosages that may be used. An exemplary dosage is 10 to 30 mg per day. The exact dosage will depend upon the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, and the preference and experience of the attending physician.

[00104] A pharmaceutical composition of the invention typically contains an active ingredient (i.e., a compound of the disclosure) of the present invention or a pharmaceutically acceptable salt and/or coordination complex thereof, and one or more pharmaceutically acceptable excipients, carriers, including but not limited to inert solid diluents and fillers, diluents, sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants.

[00105] Described below are non- limiting exemplary pharmaceutical compositions and methods for preparing the same.

Pharmaceutical compositions for oral administration.

[00106] In some embodiments, the invention provides a pharmaceutical composition for oral administration containing a compound of the invention, and a pharmaceutical excipient suitable for oral administration.

[00107] 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 a second agent; and (iii) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further contains: (iv) an effective amount of a third agent.

[00108] In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral consumption. Pharmaceutical compositions of the invention suitable for oral administration can be presented as discrete dosage forms, such as capsules, cachets, or tablets, or liquids or aerosol sprays each containing a predetermined amount of an active ingredient as a powder or in granules, a solution, or a suspension in an aqueous or nonaqueous liquid, an oil-in- water emulsion, or a water-in-oil liquid emulsion. Such dosage forms can be prepared by any of the methods of pharmacy, but all methods include the step of bringing the active ingredient into association with the carrier, which constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation. For example, a tablet can be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free- flowing form such as powder or granules, optionally mixed with an excipient such as, but not limited to, a binder, a lubricant, an inert diluent, and/or a surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. [00109] This invention further encompasses anhydrous pharmaceutical compositions and dosage forms comprising an active ingredient, since water can facilitate the degradation of some compounds. For example, water may be added (e.g., 5%) in the pharmaceutical arts as a means of simulating long-term storage in order to determine characteristics such as shelf- life or the stability of formulations over time. Anhydrous pharmaceutical compositions and dosage forms of the invention can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms of the invention which contain lactose can be made anhydrous if substantial contact with moisture and/or humidity during manufacturing, packaging, and/or storage is expected. An anhydrous pharmaceutical composition may be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions may be packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastic or the like, unit dose containers, blister packs, and strip packs.

[00110] An active ingredient can be combined in an intimate admixture with a

pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration. In preparing the compositions for an oral dosage form, any of the usual

pharmaceutical media can be employed as carriers, 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 carriers such as starches, sugars, micro-crystalline cellulose, diluents, granulating agents, lubricants, binders, and

disintegrating agents can be used in the case of oral solid preparations, in some embodiments without employing the use of lactose. For example, suitable carriers include powders, capsules, and tablets, with the solid oral preparations. If desired, tablets can be coated by standard aqueous or nonaqueous techniques.

[00111] 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 acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre- gelatinized starch, hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixtures thereof.

[00112] Examples of suitable fillers for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

[00113] Disintegrants may be used in the compositions of the invention to provide tablets that disintegrate when exposed to an aqueous environment. Too much of a disintegrant may produce tablets which may disintegrate in the bottle. Too little may be insufficient for disintegration to occur and may thus alter the rate and extent of release of the active ingredient(s) from the dosage form. Thus, a sufficient amount of disintegrant that is neither too little nor too much to detrimentally alter the release of the active ingredient(s) may be used to form the dosage forms of the compounds disclosed herein. The amount of disintegrant used may vary based upon the type of formulation and mode of administration, and may be readily discernible to those of ordinary skill in the art. About 0.5 to about 15 weight percent of disintegrant, or about 1 to about 5 weight percent of disintegrant, may be used in the pharmaceutical composition. Disintegrants that can be used to form

pharmaceutical compositions and dosage forms of the invention include, but are not limited to, agar- agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium,

crospovidone, polacnlin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums or mixtures thereof.

[00114] Lubricants which can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, or mixtures thereof. Additional lubricants include, for example, a syloid silica gel, a coagulated aerosol of synthetic silica, or mixtures thereof. A lubricant can optionally be added, in an amount of less than about 1 weight percent of the pharmaceutical composition.

[00115] When aqueous suspensions and/or elixirs are desired for oral administration, the active ingredient therein may be combined with various sweetening or flavoring agents, coloring matter or dyes and, if so desired, emulsifying and/or suspending agents, together with such diluents as water, ethanol, propylene glycol, glycerin and various combinations thereof.

[00116] The tablets can be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed. Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin or olive oil.

[00117] Surfactant which can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants may be employed, a mixture of lipophilic surfactants may be employed, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be employed.

[00118] A suitable hydrophilic surfactant may generally have an HLB value of at least 10, while suitable lipophilic surfactants may generally have an HLB value of or less than about 10. An empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of non-ionic amphiphilic compounds is the 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.

[00119] Hydrophilic surfactants are generally considered to be those compounds having an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is not generally applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds having an HLB value equal to or less than about 10. However, HLB value of a surfactant is merely a rough guide generally used to enable formulation of industrial, pharmaceutical and cosmetic emulsions.

[00120] Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidic acid salts; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithins and hydrogenated lecithins; lysolecithins and hydrogenated lysolecithins; phospholipids and derivatives thereof; lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; salts of alkyl sulfates; fatty acid salts; sodium docusate; acyl lactylates; mono- and di- acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

[00121] Within the aforementioned group, ionic surfactants include, by way of example: lecithins, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; salts of alkyl sulfates; fatty acid salts; sodium docusate; acylactylates; mono- and di- acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

[00122] Ionic surfactants may be the ionized forms of lecithin, lysolecithin,

phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine,

lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG- phosphatidylethanolamine, PVP -phosphatidylethanolamine, lactylic esters of fatty acids, stearoyl-2- lactylate, 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, lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, and salts and mixtures thereof.

[00123] Hydrophilic non-ionic surfactants may include, but are not limited to,

alkylglucosides; alkylmaltosides; alkylthioglucosides; lauryl macrogolglycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethylene glycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esters such as polyethylene glycol fatty acids monoesters and polyethylene glycol fatty acids diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylene sterols, derivatives, and analogues thereof; polyoxyethylated vitamins and derivatives thereof; polyoxyethylene- polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of a polyol with at least one member of 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.

[00124] Other hydrophilic-non-ionic surfactants include, without limitation, 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 di oleate, 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, PEG- 35 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, polyglyceiyl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30 soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan 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-lOoleate, Tween 40, Tween 60, sucrose monostearate, sucrose mono laurate, sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG 15-100 octyl phenol series, and poloxamers.

[00125] Suitable lipophilic surfactants include, by way of example only: fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acids esters;

propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and di-glycerides; hydrophobic transesterification products of a polyol with at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; oil- soluble vitamins/vitamin derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or are hydrophobic transesterification products of a polyol with at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides.

[00126] In one embodiment, the composition may include a solubilizer to ensure good solubilization and/or dissolution of the compound of the present invention and to minimize precipitation of the compound of the present invention. This can be especially important for compositions for non-oral use, e.g., 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.

[00127] Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols, such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediols and isomers thereof, glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinylalcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; ethers of polyethylene glycols having an average molecular weight of 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, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam,

dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributylcitrate, acetyl triethylcitrate, acetyl tributyl citrate, tri ethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and isomers thereof, δ-valerolactone and isomers thereof, β-butyrolactone and isomers thereof; and other solubilizers known in the art, such as dimethyl acetamide, dimethyl isosorbide, N-methyl pyrrolidones, monooctanoin, diethylene glycol monoethyl ether, and water.

[00128] Mixtures of solubilizers may also be used. Examples include, but not limited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N- hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol, transcutol, propylene glycol, and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycofurol and propylene glycol.

[00129] The amount of solubilizer that can be included is not particularly limited. The amount of a given solubilizer may be limited to a bioacceptable amount, which may be readily determined by one of skill in the art. In some circumstances, it may be advantageous to include amounts of solubilizers far in excess of bioacceptable amounts, for example to maximize the concentration of the drug, with excess solubilizer removed prior to providing the composition to a subject using conventional techniques, such as distillation or evaporation. Thus, if present, the solubilizer can be in a weight ratio of 10%, 25%o, 50%), 100%o, or up to about 200%> by weight, based on the combined weight of the drug, and other excipients. If desired, very small amounts of solubilizer may also be used, such as 5%>, 2%>, 1%) or even less. Typically, the solubilizer may be present in an amount of about 1%> to about 100%, more typically about 5%> to about 25%> by weight.

[00130] The composition can further include one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, without limitation, detackifiers, anti- foaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscomodulators, tonicifiers, flavorants, colorants, odorants, opacifiers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.

[00131] In addition, an acid or a base may be incorporated into the composition to facilitate processing, to enhance stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine,

ethylenediamine, triethanolamine, triethylamine, triisopropanol amine, trimethylamine,

tris(hydroxymethyl)aminom ethane (TRIS) and the like. Also suitable are bases that 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, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p- toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and the like. Salts of polyprotic acids, such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate can also be used. When the base is a salt, the cation can be any convenient and pharmaceutically acceptable cation, such as ammonium, alkali metals, alkaline earth metals, and the like. Example may include, but not limited to, sodium, potassium, lithium, magnesium, calcium and ammonium.

[00132] Suitable acids are pharmaceutically acceptable organic or inorganic acids.

Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydriodic 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, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenyl sulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid and the like.

Pharmaceutical compositions for injection.

[00133] In some embodiments, the invention provides a pharmaceutical composition for injection containing a compound of the present invention and a pharmaceutical excipient suitable for injection. Components and amounts of agents in the compositions are as described herein.

[00134] The forms in which the novel compositions of the present invention may be incorporated for administration by injection include aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles.

[00135] Aqueous solutions in saline are also conventionally used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and the like (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be employed. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, for the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

[00136] Sterile injectable solutions are prepared by incorporating the compound of the present invention in the required amount in the appropriate solvent with various other ingredients as enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, certain desirable methods of preparation are vacuum-drying and freeze- drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. Pharmaceutical compositions for topical (e.g. transdermal) delivery.

[00137] In some embodiments, the invention provides a pharmaceutical composition for transdermal delivery containing a compound of the present invention and a pharmaceutical excipient suitable for transdermal delivery.

[00138] Compositions of the present invention can be formulated into preparations in solid, semisolid, or liquid forms suitable for local or topical administration, such as gels, water soluble jellies, creams, lotions, suspensions, foams, powders, slurries, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline solutions, dimethylsulfoxide (DMSO)-based solutions. In general, carriers with higher densities are capable of providing an area with a prolonged exposure to the active ingredients. In contrast, a solution formulation may provide more immediate exposure of the active ingredient to the chosen area.

[00139] The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients, which are compounds that allow increased penetration of, or assist in the delivery of, therapeutic molecules across the stratum corneum permeability barrier of the skin. There are many of these penetration- enhancing molecules known to those trained in the art of topical formulation.

[00140] Examples of such carriers and excipients include, but are not limited to, humectants (e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropyl myristate and sodium lauryl sulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes (e.g., menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[00141] Another exemplary formulation for use in the methods of the present invention employs transdermal delivery devices ("patches"). Such transdermal patches may be used to provide continuous or discontinuous infusion of a compound of the present invention in controlled amounts, either with or without another agent.

[00142] The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents. Pharmaceutical compositions for inhalation.

[00143] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described supra. Preferably the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a face mask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner.

Other pharmaceutical compositions.

[00144] Pharmaceutical compositions may also be prepared from compositions described herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural, or intraspinal administration. Preparations for such pharmaceutical compositions are well-known in the art. See, e.g., 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 by reference herein in their entirety.

[00145] Administration of the compounds or pharmaceutical composition of the present invention can be effected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), topical (e.g. transdermal application), rectal administration, via local delivery by catheter or stent or through inhalation. Compounds can also be administered intraadiposally or intrathecally. [00146] The amount of the compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g/day, preferably about 0.05 to about 2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, e.g. by dividing such larger doses into several small doses for administration throughout the day.

[00147] In some embodiments, a compound of the invention is administered in a single dose.

[00148] Typically, such administration will be by injection, e.g., intravenous injection, in order to introduce the agent quickly. However, other routes may be used as appropriate. A single dose of a compound of the invention may also be used for treatment of an acute condition.

[00149] In some embodiments, a compound of the invention is administered in multiple doses. Dosing may be about once, twice, three times, four times, five times, six times, or more than six times per day. Dosing may be about once a month, once every two weeks, once a week, or once every other day. In another embodiment a compound of the invention and another agent are administered together about once per day to about 6 times per day. In another embodiment the administration of a compound of the invention and an agent continues for less than about 7 days. In yet another embodiment the administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.

[00150] Administration of the compounds of the invention may continue as long as necessary. In some embodiments, a compound of the invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a compound of the invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a compound of the invention is administered chronically on an ongoing basis, e.g., for the treatment of chronic effects.

[00151] An effective amount of a compound of the invention may be administered in either single or multiple doses by any of the accepted modes of administration of agents having similar utilities, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.

[00152] The compositions of the invention may also be delivered via an impregnated or coated device such as a stent, for example, or an artery-inserted cylindrical polymer. Such a method of administration may, for example, aid in the prevention or amelioration of restenosis following procedures such as balloon angioplasty. Without being bound by theory, compounds of the invention may slow or inhibit the migration and proliferation of smooth muscle cells in the arterial wall which contribute to restenosis. A compound of the invention may be administered, for example, by local delivery from the struts of a stent, from a stent graft, from grafts, or from the cover or sheath of a stent. In some embodiments, a compound of the invention is admixed with a matrix. Such a matrix may be a polymeric matrix, and may serve to bond the compound to the stent.

Polymeric matrices suitable for such use, include, for example, lactone-based polyesters or copolyesters such as polylactide, polycaprolactonglycolide, polyorthoesters, polyanhydrides, polyaminoacids, polysaccharides, polyphosphazenes, poly (ether-ester) copolymers (e.g. PEO- PLLA); polydimethylsiloxane, poly(ethylene-vinylacetate), acrylate-based polymers or copolymers (e.g. polyhydroxyethyl methylmethacrylate, polyvinyl pyrrolidinone), fluorinated polymers such as polytetrafluoroethylene and cellulose esters. Suitable matrices may be nondegrading or may degrade with time, releasing the compound or compounds. Compounds of the invention may be applied to the surface of the stent by various methods such as dip/spin coating, spray coating, dip-coating, and/or brush-coating. The compounds may be applied in a solvent and the solvent may be allowed to evaporate, thus forming a layer of compound onto the stent. Alternatively, the compound may be located in the body of the stent or graft, for example in microchannels or micropores. When implanted, the compound diffuses out of the body of the stent to contact the arterial wall. Such stents may be prepared by dipping a stent manufactured to contain such micropores or microchannels into a solution of the compound of the invention in a suitable solvent, followed by evaporation of the solvent. Excess drug on the surface of the stent may be removed via an additional brief solvent wash. In yet other embodiments, compounds of the invention may be covalently linked to a stent or graft. A covalent linker may be used which degrades in vivo, leading to the release of the compound of the invention. Any bio-labile linkage may be used for such a purpose, such as ester, amide or anhydride linkages. Compounds of the invention may additionally be administered intravascularly from a balloon used during angioplasty. Extravascular administration of the compounds via the pericard or via advential application of formulations of the invention may also be performed to decrease restenosis.

[00153] A variety of stent devices which may be used as described are disclosed, for example, in the following references, all of which are hereby incorporated by reference: U.S. Pat. No. 5451233; U.S. Pat. No. 5040548; U.S. Pat. No. 5061273; U.S. Pat. No. 5496346; U.S. Pat. No. 5292331; U.S. Pat. No. 5674278; U.S. Pat. No. 3657744; U.S. Pat. No. 4739762; U.S. Pat. No. 5195984; U.S. Pat. No. 5292331 ; U.S. Pat. No. 5674278; U.S. Pat. No. 5879382; U.S. Pat. No. 6344053.

[00154] The compounds of the invention may be administered in dosages. It is known in the art that due to intersubject variability in compound pharmacokinetics, individualization of dosing regimen is necessary for optimal therapy. Dosing for a compound of the invention may be found by routine experimentation in light of the instant disclosure.

[00155] When a compound of the invention is administered in a composition that comprises one or more agents, and the agent has a shorter half- life than the compound of the invention unit dose forms of the agent and the compound of the invention may be adjusted accordingly.

[00156] The subject pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository. The

pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.

[00157] Exemplary parenteral administration forms include solutions or suspensions of active compound in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered, if desired.

Methods of Use

[00158] The method typically comprises administering to a subject a therapeutically effective amount of a compound of the invention. The therapeutically effective amount of the subject combination of compounds may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of proliferation or downregulation of activity of a target protein. The specific dose will vary depending on the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

[00159] As used herein, the term "IC₅₀" refers to the half maximal inhibitory concentration of an inhibitor in inhibiting biological or biochemical function. This quantitative measure indicates how much of a particular inhibitor is needed to inhibit a given biological process (or component of a process, i.e. an enzyme, cell, cell receptor or microorganism) by half. In other words, it is the half maximal (50%) inhibitory concentration (IC) of a substance (50% IC, or IC50). EC50 refers to the plasma concentration required for obtaining 50%> of a maximum effect in vivo.

[00160] In some embodiments, the subject methods utilize a PRMT5 inhibitor with an IC50 value of about or less than a predetermined value, as ascertained in an in vitro assay. In some embodiments, the PRMT5 inhibitor inhibits PRMT5 a with an IC50 value of about 1 nM or less, 2 nM or less, 5 nM or less, 7 nM or less, 10 nM or less, 20 nM or less, 30 nM or less, 40 nM or less, 50 nM or less, 60 nM or less, 70 nM or less, 80 nM or less, 90 nM or less, 100 nM or less, 120 nM or less, 140 nM or less, 150 nM or less, 160 nM or less, 170 nM or less, 180 nM or less, 190 nM or less, 200 nM or less, 225 nM or less, 250 nM or less, 275 nM or less, 300 nM or less, 325 nM or less, 350 nM or less, 375 nM or less, 400 nM or less, 425 nM or less, 450 nM or less, 475 nM or less, 500 nM or less, 550 nM or less, 600 nM or less, 650 nM or less, 700 nM or less, 750 nM or less, 800 nM or less, 850 nM or less, 900 nM or less, 950 nM or less, 1 μΜ or less, 1.1 μΜ or less, 1.2 μΜ or less, 1.3 μΜ or less, 1.4 μΜ or less, 1.5 μΜ or less, 1.6 μΜ or less, 1.7 μΜ or less, 1.8 μΜ or less, 1.9 μΜ or less, 2 μΜ or less, 5 μΜ or less, 10 μΜ or less, 15 μΜ or less, 20 μΜ or less, 25 μΜ or less, 30 μΜ or less, 40 μΜ or less, 50 μΜ, 60 μΜ, 70 μΜ, 80 μΜ, 90 μΜ, 100 μΜ, 200 μΜ, 300 μΜ, 400 μΜ, or 500 μΜ, or less, (or a number in the range defined by and including any two numbers above). [00161] In some embodiments, the PRMT5 inhibitor selectively inhibits PRMT5 a 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 its IC50 value against one, two, or three other PRMTs.

[00162] In some embodiments, the PRMT5 inhibitor selectively inhibits PRMT5 a with an IC50 value that is less than about 1 nM, 2 nM, 5 nM, 7 nM, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 120 nM, 140 nM, 150 nM, 160 nM, 170 nM, 180 nM, 190 nM, 200 nM, 225 nM, 250 nM, 275 nM, 300 nM, 325 nM, 350 nM, 375 nM, 400 nM, 425 nM, 450 nM, 475 nM, 500 nM, 550 nM, 600 nM, 650 nM, 700 nM, 750 nM, 800 nM, 850 nM, 900 nM, 950 nM, 1 μΜ, 1.1 μΜ, 1.2 μΜ, 1.3 μΜ, 1.4 μΜ, 1.5 μΜ, 1.6 μΜ, 1.7 μΜ, 1.8 μΜ, 1.9 μΜ, 2 μΜ, 5 μΜ, 10 μΜ, 15 μΜ, 20 μΜ, 25 μΜ, 30 μΜ, 40 μΜ, 50 μΜ, 60 μΜ, 70 μΜ, 80 μΜ, 90 μΜ, 100 μΜ, 200 μΜ, 300 μΜ, 400 μΜ, or 500 μΜ (or in the range defined by and including any two numbers above), and said IC50 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 a number in the range defined by and including any two numbers above) than its IC50 value against one, two or three other PRMTs.

[00163] The subject methods are useful for treating a disease condition associated with PRMT5. Any disease condition that results directly or indirectly from an abnormal activity or expression level of PRMT5 can be an intended disease condition.

[00164] Different disease conditions associated with PRMT5 have been reported. PRMT5 has been implicated, for example, in a variety of human cancers as well as a number of

hemoglobinopathies.

[00165] Non- limiting examples of such conditions include but are not limited to

Acanthoma, Acinic cell carcinoma, Acoustic neuroma, Acral lentiginous melanoma, Acrospiroma, Acute eosinophilic leukemia, Acute lymphoblastic leukemia, Acute megakaryoblastic leukemia, Acute monocytic leukemia, Acute myeloblasts leukemia with maturation, Acute myeloid dendritic cell leukemia, Acute myeloid leukemia, Acute promyelocytic leukemia, Adamantinoma,

Adenocarcinoma, Adenoid cystic carcinoma, Adenoma, Adenomatoid odontogenic tumor,

Adrenocortical carcinoma, Adult T-cell leukemia, Aggressive K-cell leukemia, AIDS-Related Cancers, AIDS-related lymphoma, Alveolar soft part 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, Bronchial Tumor, 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, Central Nervous System Embryonal Tumor, Cerebellar Astrocytoma, Cerebral Astrocytoma, Cervical Cancer, Cholangiocarcinoma, Chondroma,

Chondrosarcoma, Chordoma, Choriocarcinoma, Choroid plexus papilloma, Chronic Lymphocytic Leukemia, Chronic monocytic leukemia, Chronic myelogenous leukemia, Chronic

Myeloproliferative Disorder, Chronic neutrophilic leukemia, Clear-cell tumor, Colon Cancer, Colorectal 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, Endometrial Uterine Cancer, Endometrioid tumor, Enteropathy-associated T-cell lymphoma, Ependymoblastoma, Ependymoma, Epithelioid sarcoma, Erythroleukemia, Esophageal cancer, Esthesioneuroblastoma, Ewing Family of Tumor, Ewing Family Sarcoma, Ewing's sarcoma, Extracranial Germ Cell Tumor, Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer, Extramammary Paget's disease, Fallopian tube cancer, Fetus in fetu, Fibroma, Fibrosarcoma, Follicular lymphoma, Follicular thyroid cancer, Gallbladder Cancer, Gallbladder cancer, Ganglioglioma, Ganglioneuroma, Gastric Cancer, Gastric lymphoma,

Gastrointestinal cancer, Gastrointestinal Carcinoid Tumor, Gastrointestinal Stromal Tumor, 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, Hairy cell leukemia, Head and Neck Cancer, Head and neck cancer, Heart cancer, Hemangioblastoma, Hemangiopericytoma, Hemangiosarcoma, Hematological malignancy,

Hepatocellular carcinoma, Hepatosplenic T-cell lymphoma, Hereditary breast-ovarian cancer syndrome, Hodgkin Lymphoma, Hodgkin's lymphoma, Hypopharyngeal Cancer, Hypothalamic Glioma, Inflammatory breast cancer, Intraocular Melanoma, Islet cell carcinoma, Islet Cell Tumor, Juvenile myelomonocytic leukemia, Kaposi Sarcoma, Kaposi's sarcoma, Kidney Cancer, Klatskin tumor, Krukenberg tumor, Laryngeal Cancer, Laryngeal cancer, Lentigo maligna melanoma, Leukemia, Leukemia, Lip and Oral Cavity Cancer, Liposarcoma, Lung cancer, Luteoma, Lymphangioma, Lymphangiosarcoma, Lymphoepithelioma, Lymphoid leukemia, Lymphoma, Macroglobulinemia, Malignant Fibrous Histiocytoma, Malignant fibrous histiocytoma, Malignant Fibrous Histiocytoma of Bone, Malignant Glioma, Malignant Mesothelioma, Malignant peripheral nerve sheath tumor, Malignant rhabdoid tumor, Malignant triton tumor, MALT lymphoma, Mantle cell lymphoma, Mast cell leukemia, Mediastinal germ cell tumor, Mediastinal tumor, Medullary thyroid cancer, Medulloblastoma, Medulloblastoma, Medulloepithelioma, Melanoma, Melanoma, Meningioma, Merkel Cell Carcinoma, Mesothelioma, Mesothelioma, Metastatic Squamous Neck Cancer with Occult Primary, Metastatic urothelial carcinoma, Mixed Mullerian tumor, Monocytic leukemia, Mouth Cancer, Mucinous tumor, Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma, Multiple myeloma, Mycosis Fungoides, Mycosis fungoides, Myelodysplasia Disease, Myelodysplasia Syndromes, Myeloid leukemia, Myeloid sarcoma, Myeloproliferative Disease, Myxoma, Nasal Cavity Cancer, Nasopharyngeal Cancer, Nasopharyngeal carcinoma, Neoplasm, Neurinoma, Neuroblastoma, Neuroblastoma, Neurofibroma, Neuroma, Nodular melanoma, Non- Hodgkin Lymphoma, Non-Hodgkin lymphoma, Nonmelanoma Skin Cancer, Non-Small Cell Lung Cancer, Ocular oncology, Oligoastrocytoma, Oligodendroglioma, Oncocytoma, Optic nerve sheath meningioma, Oral Cancer, Oral cancer, Oropharyngeal Cancer, Osteosarcoma, Osteosarcoma, Ovarian Cancer, Ovarian cancer, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor, Ovarian Low Malignant Potential Tumor, Paget's disease of the breast, Pancoast tumor, Pancreatic Cancer, Pancreatic cancer, Papillary thyroid cancer, Papillomatosis, Paraganglioma, Paranasal Sinus Cancer, Parathyroid Cancer, Penile Cancer, Perivascular epithelioid cell tumor, Pharyngeal Cancer,

Pheochromocytoma, Pineal Parenchymal Tumor of Intermediate Differentiation, Pineoblastoma, Pituicytoma, Pituitary adenoma, Pituitary tumor, Plasma Cell Neoplasm, Pleuropulmonary blastoma, Polyembryoma, Precursor T-lymphoblastic lymphoma, Primary central nervous system lymphoma, Primary effusion lymphoma, Primary Hepatocellular Cancer, Primary Liver Cancer, Primary peritoneal cancer, Primitive neuroectodermal tumor, Prostate cancer, Pseudomyxoma peritonei, Rectal Cancer, Renal cell carcinoma, Respiratory Tract Carcinoma Involving the NUT Gene onChromosome 15, Retinoblastoma, Rhabdomyoma, Rhabdomyosarcoma, Richter's transformation, Sacrococcygeal teratoma, Salivary Gland Cancer, Sarcoma, Schwannomatosis, Sebaceous gland carcinoma, Secondary neoplasm, Seminoma, Serous tumor, Sertoli-Leydig cell tumor, Sex cord-stromal tumor, Sezary Syndrome, Signet ring cell carcinoma, Skin Cancer, Small blue round cell tumor, Small cell carcinoma, Small Cell Lung Cancer, Small cell lymphoma, Small intestine cancer, Soft tissue sarcoma, Somatostatinoma, Soot wart, Spinal Cord Tumor, Spinal tumor, Splenic marginal zone lymphoma, Squamous cell carcinoma, Stomach cancer, Superficial spreading melanoma, Supratentorial Primitive Neuroectodermal Tumor, Surface epithelial-stromal tumor, Synovial sarcoma, T-cell acute lymphoblastic leukemia, T-cell large granular lymphocyte leukemia, T-cell leukemia, T-cell lymphoma, T-cell prolymphocytic leukemia, Teratoma, Terminal lymphatic cancer, Testicular cancer, Thecoma, Throat Cancer, Thymic Carcinoma, Thymoma, Thyroid cancer, Transitional Cell Cancer of Renal Pelvis and Ureter, Transitional cell carcinoma, Urachal cancer, Urethral cancer, Urogenital neoplasm, Uterine sarcoma, Uveal melanoma, Vaginal Cancer, Verner Morrison syndrome, Verrucous carcinoma, Visual Pathway Glioma, Vulvar Cancer, Waldenstrom's macroglobulinemia, Warthin's tumor, Wilms' tumor, or any combination thereof.

[00166] In some embodiments, said method is for treating a disease selected from the group consisting of tumor angiogenesis, 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 and ovarian, breast, lung, pancreatic, prostate, colon and epidermoid cancer.

[00167] The examples and preparations provided below further illustrate and exemplify the compounds of the present invention and methods of preparing such compounds. It is to be understood that the scope of the present invention is not limited in any way by the scope of the following examples and preparations. In the following examples molecules with a single chiral center, unless otherwise noted, exist as a racemic mixture. Those molecules with two or more chiral centers, unless otherwise noted, exist as a racemic mixture of diastereomers. Single

enantiomers/diastereomers may be obtained by methods known to those skilled in the art.

[00168] Compounds of the disclosure can be prepared, for example, by reference to the following schemes. Scheme 1

Scheme 2

Compounds 26-33 Example 2. N-(9-((2R,3R,4S,5S)-3,4-dihydroxy-5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H- purin-6-yl)acrylamide (2)

2a 2b Ex. 2 a) N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6 (methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4- yl)-9H-purin-6-yl)acrylamide (2b)

[00169] To a solution of 9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-amine (500.00 mg, 1.48 mmol) and TEA (750.00 mg, 7.42 mmol) in DCM (10 mL) was added acryloyl chloride (220.00 mg, 2.37 mmol) at 0^°C, then the reaction mixture was stirred at 25 °C for 16 hrs. Desired product was detected by LCMS. The reaction mixture was poured into water, extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na₂S0₄. The solvent was concentrated under reduced pressure to give crude product which was purified by silica gel column chromatography (DCM:CH₃OH=60: 1) to give N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-yl)acrylamide (230.00 mg, 39.66% yield). LCMS [M+H]: 392.1 b) N-(9-((2R,3R,4S,5S)-3,4-dihydroxy-5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H-purin-6- yl)acrylamide (2)

[00170] A solution of N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-yl)acrylamide (50.00 mg, 0.18 mmol) in TFA:H₂0 (1 :5, 3 mL) was stirred at 25 ^°C for 1 hour. LCMS showed the reaction was completed. Na₂C0₃ (sat.aq.) was added to adjust the pH of mixture to 7. Then the mixture was extracted with EA (30mL*3). The combined organic layers were washed with brine, dried over anhydrous Na₂S0₄. The solvent was concentrated under reduced pressure to give crude product which was purified by Pre-TLC (DCM:CH₃OH=10: 1) to give N-(9-((2R,3R,4S,5S)-3,4-dihydroxy- 5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)acrylamide (12.00mg, 26.74% yield). LCMS [M+H] :352. 1H MR (400 MHz, OMSO-d₆) δ 1 1.01 (s, 1 H), 8.72 (s, 1 H), 8.71 (s, 1H), 6.73-6.80 (m, 1 H), 6.33-6.37 (m, 1 H), 6.02 (d, J = 5.6 Hz, 1 H), 5.86 (d, J = 10.4 Hz, 1 H), 5.58 (d, J = 5.6 Hz, 1 H), 5.38 (d, J = 4.8 Hz, 1 H), 4.78-4.82 (m, 1 H), 4.09-4.19 (m, 2 H), 2.78-2.93 (m, 2 H), 2.06 (s, 3 H).

Example. 20. N-(9-((2R,3R,4S,5S)-3,4-dihydroxy-5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H- purin-6-yl)methacrylamide (20)

2a 20a Ex. 20 a) N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6-((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4- yl)-9H-purin-6-yl)-N-methacryloylmethacrylamide (20a)

[00171] To a solution of 9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-amine (2a) (220 mg, 0.652 mmol) and TEA (395 mg, 3.91 mmol) in DCM (5 mL) was added methacryloyl chloride (341 mg, 3.2 mmol). The reaction mixture was stirred at 25 °C for 2 hrs. LCMS showed the reaction was completed. The reaction mixture was poured into water, extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na₂SC>4. The solvent was concentrated under reduced pressure to give crude product which was purified by silica gel column chromatography (PE:EA=1 : 1) to give N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-yl)-N- methacryloylmethacrylamide (292 mg, 88% purity, 94.56 % yield ) as thick oil. LCMS [M+H]

:474.2. b) N-(9-((2R,3R,4S,5S)-3,4-dihydroxy-5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H-purin-6- yl)methacrylamide (20) [00172] A solution of N-(9-((3aR,4R,6S,6aS)-2,2-dimethyl-6- ((methylthio)methyl)tetrahydrofuro[3,4-d][l,3]dioxol-4-yl)-9H-purin-6-yl)-N- methacryloylmethacrylamide (270 mg, 0.57 mmol) in TFA:H₂0 (1 : 10, 5 mL) was stirred at 25 °C for 3 hrs. LCMS showed the reaction was completed. Na₂C0₃ (sat. aq.) was added to adjust pH to 9.0 and stirred at 25 °C for 18 hrs. LCMS showed the reaction was completed. The reaction mixture was purified by prep-HRLC (0.1% TFA) eluting with H₂0:CH₃CN from 90: 10 to 5:95 to give N-(9- ((2R,3R,4S,5S)-3,4-dihydroxy-5-((methylthio)methyl)tetrahydrofuran-2-yl)-9H-purin-6- yl)methacrylamide (15 mg) as off-white solid. LCMS [M+H] :366.2. ¹H MR (400 MHz, DMSO- d₆) δ 10.65 (s, 1 H), 8.72 (s, 1 H), 8.68 (s, 1H), 6.02 (m, 2 H), 5.68 (s, 1 H), 5.58 (d, J = 5.6 Hz, 1 H), 5.37 (d, J = 3.2 Hz, 1 H), 4.80 (d, J = 4.8 Hz, 1 H), 4.18 (d, J = 2.8 Hz, 1H), 4.07 (d, J = 3.2 Hz, 1 H), 2.78-2.92 (m, 2 H), 2.06 (s, 3 H), 1.96 (s, 3 H).

[00173] Compounds of the disclosure can be tested for inhibition of PRMT5-mediated H4R3 methylation using the following protocol.

Biochemical Assay Protocol

[00174] Compounds were solubilized and 3-fold diluted in 100% DMSO. These diluted compounds were further diluted in the assay buffer (50 mM Tris-HCl, pH 8.5, 50 mM NaCl, 5 mM MgCl₂, 0.01% Brij35, 1 mM DTT, 1% DMSO) for 10-dose IC₅₀ mode at a concentration 10-fold greater than the desired assay concentration. Standard reactions were performed in a total volume of 50 μΐ in assay buffer, with histone H2A (5 μΜ final) as substrate. To this was added the

PRMT5/MEP50 complex diluted to provide a final assay concentration of 5 nM and the compounds were allowed to preincubate for 15 to 20 minutes at room temperature. The reaction was initiated by adding S-[3 H-methyl]-adenosyl-L-methionine (PerkinElmer) to final concentration of 1 μΜ.

Following a 60 minutes incubation at 30 °C, the reaction was stopped by adding 100 of 20% TCA. Each reaction was spotted onto filter plate (Multi Screen FB Filter Plate, Millipore), and washed 5 times with PBS buffer, Scintillation fluid was added to the filter plate and read in a scintillation counter. IC₅₀ values were determined by fitting the data to the standard 4 parameters with Hill Slope using GraphPad Prism software Table 1. Biochemical and cellular potency (in Granta cell line)

Dialysis protocol with wild type PRMT5 and C449S PRMT5

Wild type PRTMT5 dialysis

[00175] A pre-dialysis sample (total 30 μΐ.) was prepared by incubating 7.5 μΙ_, of 400 nM PRMT5/MEP50 (cat#: HMT-22-148, Reaction Biology Corp.) (100 nM final) with 3 μΐ, of varied concentrations of different inhibitors (500 μΜ Ex. 2, and 250 μΜ Ex. 20) or 3 μΙ_, of 1% DMSO as control - at 37 °C for 30 minutes. 5 μΙ_, of protein/compound complex was then taken out and added to 5 μΐ, of reaction buffer (20 mM Tris, 0.002% Tween 20, 1 mM TCEP pH=8.0) to make the pre- dialysis samples containing 50 nM of PRMT5 or PRMT5/inhibitor complex. The remaining 25 μΙ_, of protein/compound complex was transferred to Slide-A-Lyzer (10K MWCO) mini dialysis devices (Thermo Scientific cat#: PI69570) and dialyzed in 500 mL reaction buffer at room temperature for 1 hour. The post-dialysis sample containing 50 nM of PRMT5 or PRMT5/inhibitor complexes was prepared by transfering 25 μΙ_, of protein/compound complex to 25 μΙ_, of reaction buffer.6 μΙ_, of pre- and post- dialysis samples (final enzyme or enzyme/inhibitor complex concentration is 10 nM) were then added to 27 μΙ_, of substrate mixture (containing 300 nM biotinylated AcH4-23 peptide and 1 μΜ 3H-SAM) to initiate the reaction; the reaction continued at 37 °C for 15 minutes to test the methyltransferase activities.

[00176] As shown in Figure 1, dialysis results indicated Example 2 and Example 20 may covalently modify PRMT5. Both Example 2 and Example 20 inhibited 80% of methylation activity of PRMT5 before dialysis, and after dialysis the enzymatic activity did not recover which suggested a covalent modification happened to target enzyme.

C449S mutant PRMT5 dialysis

[00177] A pre-dialysis sample (total 30 μΐ.) was prepared by incubating 7.5 μΙ_, of 400 nM PRMT5 C449S mutant (cat#: HMT-22-434, Reaction Biology Corp.) (100 nM final) with 3 μΐ, of varied concentrations of different inhibitors (500 μΜ Ex. 2, and 250 μΜ Ex. 20) or 3 μL· of 1% DMSO as control - at 37 °C for 30 minutes. 10 μΙ_, of protein/ compound complex was then taken out and added to 10 μΐ, of reaction buffer (20 mM Tris, 0.002% Tween 20, 1 mM TCEP pH=8.0) to make the pre-dialysis samples contain 50 nM of PRMT5 or PRMT5/inhibitor complex. The remaining 20 μΙ_, of protein/compound complex was transferred to Slide-A-Lyzer (10K MWCO) mini dialysis devices (Thermo Scientific cat#: PI69570) and dialyzed in 500 mL reaction buffer at room temperature for 1 hour. All protein/compound complex mixtures were transferred to microcentrifuge tube and an appropriate amount of reaction buffer was added to make it 40 μΙ_, as the post-dialysis sample containing 50 nM of PRMT5 or PRMT5/inhibitor complexes. 6 μΙ_, of pre- and post- dialysis samples (final enzyme or enzyme/inhibitor complex concentration is 10 nM) was then added to 24 μΙ_, of substrate mixture (containing 300 nM biotinylated AcH4-23 peptide and 1 μΜ 3H-SAM) to initiate the reaction; the reaction continued at 37 °C for 15 minutes to test the methyltransferase activities.

[00178] As shown in Figure 2, dialysis results of Example 2 and Example 20 with C449S mutant of PRMT5 indicated these two molecules may modify C449 in wild type PRMT5. Before dialysis, both Example 2 and Example 20 only inhibited 50% activity of the mutant, and after dialysis, the methylation activity of enzymes recovered to 90%.

[00179] Compounds of the disclosure include, for example, the compounds shown in

Table 2:

Table 2.

Ex. No. Compound Name

85 N

III o N^N ^H¾ PH 3 -( 1 -cyano-2-methoxy ethyl)- 1 -(9- ((2R,3R,4S,5S)-3,4-dihydroxy-5- ((methylthio)methyl)tetrahydrofuran-2-yl)-9H- purin-6-yl)- 1 -methylurea

86

l-(l-cyanocyclopropyl)-3-(9-((2R,3R,4S,5S)- 3,4-dihydroxy-5-

((methylthio)methyl)tetrahydrofuran-2-yl)-9H- purin-6-yl)urea

HO

87

N-(9-((2R,3R,4S,5S)-3,4-dihydroxy-5-(((2-

(methylamino)ethyl)thio)methyl)tetrahydrofuran-

H 2-yl)-9H-purin-6-yl)-N-methylacrylamide

88

(E)-N-cyclopropyl-N-(9-((2R,3R,4S,5S)-3,4- dihydroxy-5-

((methylthio)methyl)tetrahydrofuran-2-yl)-9H- purin-6-yl)-4-(dimethylamino)but-2-enamide

OH

89 1 W ^H¾ PH N-(9-((2R,3R,4S,5S)-5-(((2- (dimethylamino)ethyl)thio)methyl)-3,4-

1 dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)- N-methy lpropi ol ami de

90 0 N^N ^H¾. ,PH 3-(cyanomethyl)-l-(9-((2R,3R,4S,5R)-5- (ethoxymethyl)-3,4-dihydroxytetrahydrofuran-2- yl)-9H-purin-6-yl)- 1 -methylurea

91 0 N^N ^H¾ ,PH 3-(9-((2R,3R,4S,5R)-5-((2- aminoethoxy)methyl)-3,4- dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)- 1 -(cyanom ethyl)- 1 -isopropylurea

92 0

3 -( 1 -cy anoethyl)- 1 -(cy clopropylmethyl)- 1 -(9- ((2R,3R,4S,5R)-3,4-dihydroxy-5-((2- hydroxyethoxy)methyl)tetrahydrofuran-2-yl)- 9H-purin-6-yl)urea

HO ^0H

The disclosure is directed to the following aspects:

As ect 1. A compound of Formula I or Formula II:

wherein

n is 0, 1, 2, 3, 4, 5, or 6;

X is -S-, -SO-, S0₂-, -0-, - H-, or -N(Ci-C₆alkyl)-;

R¹ is -Ci-Cealkyl, -C(0)-Ci-C₆alkyl, or -C(0)-0-Ci-C₆alkyl;

R² is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cecycloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -Co-C₆alk- H₂, -Co-C₆alk- H-Ci-C₆alkyl,

-Co-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl, -Co-Cealk- H-Cs-Cecycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃-C₆Cycloalkyl, -Co-Cealk-heterocycloalkyl,

heteroaryl, or -CN;

R³ is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cecycloalkyl, -C₀-C₆alk-OH,

-Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk- H₂, -Co-C₆alk- H-Ci-C₆alkyl, -C₀-C₆alk-N(C C₆alkyl)-Ci-C₆alkyl, -Co-C₆alk-NH-C₃-C₆cycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -C₀-C₆alk-heterocycloalkyl, heteroaryl, or -CN;

R⁴ is H, halo, -Ci-C₆alkyl, -Ci-C₆haloalkyl, -Co-Cealk-Cs-Cecycloalkyl, -C₀-C₆alk-OH,

-Co-Cealk-O-Ci-Cealkyl, -C₀-C₆alk-NH₂, -C₀-C₆alk-NH-Ci-C₆alkyl, -C₀-C₆alk-N(C C₆alkyl)-Ci-C₆alkyl, -Co-C₆alk-NH-C₃-C₆cycloalkyl, -Co-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -C₀-C₆alk-heterocycloalkyl, heteroaryl, or -CN;

or R² and R³, together with the atoms to which they are attached, form a

C₃-C₆cycloalkenyl ring;

or R² and R³, together form a triple bond;

or R³ and R⁴, together with the atom to which they are attached, form a C₃-C₆cycloalkyl rin or a heterocycloalkyl ring;

R⁵ is H, Ci-Cealkyl, or Co-Cealk-Cs-Cec cloalkyl;

R⁶ is H, Ci-Cealkyl, or Co-Cealk-Cs-Cec cloalkyl;

R⁷ and R⁷' are each independently H, Ci-C₆alkyl, or -C₀-C₆alk-OCi-C₆alkyl;

or R⁷ and R⁷', together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring;

or a stereoisomer thereof;

or a pharmaceutically acceptable salt thereof.

Aspect 2. The compound of Aspect 1, wherein n is i .

Aspect 3. The compound of any one of the preceding Aspects, wherein X is -S-, -0-, or -NH-.

Aspect 4. The compound of any one of the preceding Aspects, wherein R¹ is -Ci-C₆alkyl.

Aspect 5. The compound of any one of Aspects 1 to 3, wherein R¹ is -C(0)-Ci-C₆alkyl.

Aspect 6. The compound of any one of Aspects 1 to 3, wherein R¹ is -C(0)-0-Ci-C₆alkyl. Aspect 7. The compound of any one of the preceding Aspects, which is a compound of Formula II.

Aspect 8. The compound of any one of Aspects 1 to 6, which is a compound of Formula I.

Aspect 9. The compound of Aspect 8, wherein R² is H.

Aspect 10. The compound of any one of Aspects 8 or 9, wherein R⁴ is H.

Aspect 1 1. The compound of any one of Aspects 8 to 10, wherein R³ is H.

Aspect 12. The compound of any one of Aspects 8 to 10, wherein R³ is halo, -Ci-C₆alkyl,

-Ci-Cehaloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -Co-C₆alk- H₂, -Co-Cealk- H-Ci-Cealkyl, -Co-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl,

-Co-C₆alk- H-C₃-C₆cycloalkyl, -C₀-C₆alk-heterocycloalkyl, heteroaryl, or -CN.

Aspect 13. The compound of any one of Aspects 8, 9, 1 1, or 12, wherein R⁴ is halo, -Ci-C₆alkyl, -Ci-Cehaloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -Co-C₆alk- H₂, -Co-Cealk-NH-Ci-Cealkyl, -Co-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl,

Aspect 14. The compound of any one of Aspects 8 or 10 to 13, wherein R² is halo, -Ci-C₆alkyl, -Ci-Cehaloalkyl, -Co-Cealk-Cs-Cec cloalkyl, -C₀-C₆alk-OH, -Co-C₆alk-0-Ci-C₆alkyl, -C₀-C₆alk-NH₂, -C₀-C₆alk-NH-Ci-C₆alkyl, -Co-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl,

-Co-Cealk- H-Cs-Cec cloalkyl, -Co-Cealk-NCCi-Cealky -Cs-Cec cloalkyl,

-C₀-C₆alk-heterocycloalkyl, -C₀-C₆alk-heterocycloalkyl, heteroaryl, or -CN.

Aspect 15. The compound of any one of Aspects 8, 10, or 13, wherein R² and R³, together with the atoms to which they are attached, form a C₃-C₆cycloalkenyl ring.

Aspect 16. The compound of any one of Aspects 8, 10, or 13, wherein R² and R³ together form a triple bond.

Aspect 17. The compound of any one of Aspects 8, 9, or 14, wherein R³ and R⁴, together with the atom to which they are attached, form a C₃-C₆cycloalkyl ring or a heterocycloalkyl ring. Aspect 18. A pharmaceutical composition comprising a compound according to any one of the preceding Aspects and a pharmaceutically acceptable excipient.

Aspect 19. A method of inhibiting a protein arginine methyltransferase 5 (PRMT5) enzyme, comprising: contacting the PRMT5 enzyme with an effective amount of a compound of any one of any one of Aspects 1 to 17.

Aspect 20. A method of disease or disorder associated with aberrant PRMT5 activity in a subject comprising administering to the subject, a compound of any one of Aspects 1 to 17.

Aspect 21. The method of Aspects 20, 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), 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 cancer, or hemoglobinopathies such as b-thalassemia and sickle cell disease (SCD).

Claims

What is Claimed:

1. A com ound of Formula I or Formula II:

wherein

n is 1, 2, 3, 4, 5, or 6;

X is -S-, -SO-, S0₂-, -0-, - H-, or -N(Ci-C₆alkyl)-;

R¹ is -Ci-Cealkyl, -C(0)-Ci-C₆alkyl, -C(0)-0-Ci-C₆alkyl, -Ci-C₆alk-OH,

-Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₆alk- H₂, -Ci-C₆alk- H-Ci-C₆alkyl, or

-Ci-C₆alk-N(Ci-C₆alkyl)-Ci-C₆alkyl;

-Co-Cealk-O-Ci-Cealkyl, -Co-C₆alk- H₂, -Co-Cealk- H-Ci-Cealkyl,

-Co-Cealk-NCCi-Cealky -Ci-Cealkyl, -C₀-C₆alk-NH-C₃-C₆Cycloalkyl,

-C₀-C₆alk-N(C₁-C₆alkyl)-C₃-C₆cycloalkyl, -C₀-C₆alk-heterocycloalkyl,

heteroaryl, or -CN;

-Co-Cealk-O-Ci-Cealkyl, -C₀-C₆alk- H₂, -Co-Cealk-NH-Ci-Cealkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-Ci-C₆alkyl, -C₀-C₆alk- H-C₃-C₆cycloalkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN;

-Co-Cealk-O-Ci-Cealkyl, -C₀-C₆alk-NH₂, -Co-C₆alk-NH-Ci-C₆alkyl, -C₀-C₆alk-N(Ci- C₆alkyl)-Ci-C₆alkyl, -C₀-C₆alk-NH-C₃-C₆cycloalkyl, -C₀-C₆alk-N(Ci-C₆alkyl)-C₃- C₆cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN;

or R² and R³, together with the atoms to which they are attached, form a

C₃-Cecycloalkenyl ring;

or R² and R³, together form a triple bond;

R⁵ is H, Ci-Cealkyl, or Co-C₆alk-C₃-C₆cycloalkyl; R⁶ is H, Ci-Cealkyl, or Co-C₆alk-C3-C₆cycloalkyl;

or a stereoisomer thereof;

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein n is 1.

3. The compound of any one of the preceding claims, wherein X is -S-, -0-, or - H-.

4. The compound of any one of the preceding claims, wherein R¹ is -Ci-C₆alkyl.

5. The compound of any one of claims 1 to 3, wherein R¹ is -C(0)-Ci-C₆alkyl or -C(0)-0-C₁- C₆alkyl.

6. The compound of any one of claims 1 to 3, wherein R¹ is -Ci-C₆alk-OH,

-Ci-C₆alk-0-Ci-C₆alkyl, -Ci-C₆alk- H₂, -Ci-C₆alk- H-Ci-C₆alkyl, or

-Ci-Cealk-NCCi-Cealky -Ci-Cealkyl.

7. The compound of any one of the preceding claims, which is a compound of Formula II.

8. The compound of any one of claims 1 to 6, which is a compound of Formula I.

9. The compound of claim 8, wherein R² is H.

10. The compound of any one of claims 8 or 9, wherein R⁴ is H.

11. The compound of any one of claims 8 to 10, wherein R³ is H.

12. The compound of any one of claims 8 to 10, wherein R³ is halo, -Ci-C₆alkyl,

13. The compound of any one of claims 8, 9, 1 1, or 12, wherein R⁴ is halo, -Ci-C₆alkyl, -Ci-Cehaloalkyl, -C₀-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH, -C₀-C₆alk-O-Ci-C₆alkyl, -C₀-C₆alk- H₂, -Co-C₆alk- H-Ci-C₆alkyl, -C₀-C6alk-N(Ci-C6alkyl)-Ci-C₆alkyl,

-Co-C₆alk- H-C₃-C₆cycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN.

14. The compound of any one of claims 8 or 10 to 13, wherein R² is halo, -Ci-C₆alkyl,

-Ci-Cehaloalkyl, -C₀-C₆alk-C₃-C₆cycloalkyl, -C₀-C₆alk-OH, -C₀-C₆alk-O-Ci-C₆alkyl, -C₀-C₆alk- H₂, -Co-C₆alk- H-Ci-C₆alkyl, -Co-Cealk-NCCi-Cealky -Ci-Cealkyl,

-Co-Cealk- H-Cs-Cecycloalkyl, -Co-Cealk-NCCi-Cealky -Cs-Cecycloalkyl,

-Co-Cealk-heterocycloalkyl, -Co-Cealk-heterocycloalkyl, heteroaryl, or -CN.

15. The compound of any one of claims 8, 10, or 13, wherein R² and R³, together with the atoms to which they are attached, form a C₃-C₆cycloalkenyl ring.

16. The compound of any one of claims 8, 10, or 13, wherein R² and R³ together form a triple bond.

17. The compound of any one of claims 8, 9, or 14, wherein R³ and R⁴, together with the atom to which they are attached, form a C3-C₆cycloalkyl ring or a heterocycloalkyl ring.

18. A pharmaceutical composition comprising a compound according to any one of the

preceding claims and a pharmaceutically acceptable excipient.

19. A method of inhibiting a protein arginine methyltransferase 5 (PRMT5) enzyme,

comprising: contacting the PRMT5 enzyme with an effective amount of a compound of any one of any one of claims 1 to 17.

20. A method of disease or disorder associated with aberrant PRMT5 activity in a subject

comprising administering to the subject, a compound of any one of claims 1 to 17.

21. The method of claim 20, 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), 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 cancer, or hemoglobinopathies such as b-thalassemia and sickle cell disease (SCD).

22. A compound having the structural formula:

or a stereoisomer thereof; or a pharmaceutically acceptable salt thereof.

23. A compound having the structural formula:

or a stereoisomer thereof; or a pharmaceutically acceptable salt thereof.