NZ716427B2 - Kdm1a inhibitors for the treatment of disease - Google Patents

Abstract

Disclosed herein are new cyclopropylamine derivatives and compositions as pharmaceuticals for the treatment of diseases. Methods of inhibition of KDM1A, methods of increasing gamma globin gene expression, and methods to induce differentiation of cancer cells in a human or animal subject are also provided for the treatment of diseases such as acute myelogenous leukemia

Description

KDMlA INHIBITORS FOR THE TREATMENT OF DISEASE This application claims the benefit of priority of United States provisional applications no. 61/862,759, filed August 6, 2013, and 61/954,276, filed March 17, 2014, the disclosures of which are hereby incorporated by reference as if written herein in their entireties.

Field of the Disclosure The present disclosure relates to new compounds and compositions and their application as ceuticals for the ent of diseases.

Detailed Description Inhibiting the enzyme KDMlA (also known as lysine-specific demethylase 1, LSD 1, Flavin-containing Amine Oxidase Domain-Containing n, AOF2, BRAF35- HDAC Complex Protein BHC110, FAD-Binding Protein BRAF35-HDAC x), may alter gene expression in cells sufficient to restore their proper physiologic function or that of the tissue, organ or the patient as a whole. This may be achieved either by enhancing transcription of a gene or genes that are pathologically silenced, e. g., as is the case in some cancer cells and heritable diseases, or decreasing transcription of a gene or genes participating in the pathological state. As such, inhibiting KDMlA would be useful for the treatment of diseases such as cancer and heritable diseases such as Wilson disease, cardiomyopathies, and hemoglobinopathies.

Gene expression is regulated through the recruitment of the RNA polymerase II transcription apparatus to the DNA template. The probability of this large multi-protein complex arriving near or at the start of DNA ription and progressing h the entire coding region of a gene is determined in part by specific DNA sequences called promoters and enhancers, modifications of DNA sequence in the vicinity of the start of ription, proteins bound to DNA and the topology of the DNA template itself. Factors enhancing the probability of RNA synthesis of protein-coding genes are known as ription factors some of which participate in the transcription of all n-coding genes and some of which are specific for the transcription of individual genes.

One major mechanism of transcription control ts of limiting the physical accessibility of the transcriptional regulatory regions to proteins that can activate or complete transcription; proteins bound to promoter or er DNA sequences can occlude activating s from binding to these DNA sequences resulting in fewer transcription initiations or extension of the activated progressing RNA polymerase complex. Likewise, topological constraints that do not allow the template DNA to unwind sufficiently to permit the steady progression of RNA polymerase on the template also serve to limit transcription rates.

The most important general factors influencing RNA synthesis using a DNA template in vivo are modifications of histones ns that l among other s the topology of the DNA template for transcription and its accessibility by the RNA polymerase complex. A small family of histone proteins — H2A, H2B, H3 and H4 — combines to create a scaffold called the histone octamer upon which DNA is spatially and topologically organized into a regular repetitive structure called the nucleosome along the length of DNA. The conglomerate of histones, other proteins, various RNAs and DNA is called chromatin. Both DNA and es are ally modified in such a way as to attract and bind or repel other proteins with the effect of enhancing or repressing transcription.

The modification ofDNA and associated RNAs and proteins that influence the regulation of transcription and replication that does not involve substitution of the canonical DNA bases is termed epigenetic. These epigenetic influences e reversible chemical modifications of the four DNA bases themselves or post-translational chemical changes to the chromatin proteins and RNDs that associate with DNA. These epigenetic processes can play a pivotal role in activating or silencing the expression of a gene; in addition, the epigenetic modifications can be maintained for the life of an organism or can be dynamically modified in response to specific biochemical signals that originate either internally within the cell or extracellularly. These chromatin tions can happen quickly or be very stable, e. g., during the hormonal induction of gene expression, chromatin structure at a c locus can change radically within seconds to permit maximal transcription or chromatin structure can be modified to fully suppress gene expression, a state of chromatin which can be stably maintained over multiple cell divisions and even transgenerationally.

The methylation of cytosine at the 5’ position is a common DNA base modification that is in turn recognized by a class of proteins most often associated with transcriptional sion. Similarly, histone proteins are chemically modified but with a wider variety of chemical adducts each of which either alone or in combination es or represses transcription of nearby genes. These histone ations e, among others methylation, acetylation, ation, phosphorylation, ubiquitylation, and myristoylation are recognized by other chromatin-associated proteins that in turn influence transcription rates and DNA replication. The dynamic state of gene expression and the associated chromatin states imply that e ations are not permanent but instead are added and removed according to the needs of the cell for specific gene products at specific times during ontogeny, adult life and the changing influences of the environment. Indeed, the specific chemical ations of histones are each made by s of s acting at specific sites. These histone-modifying enzymes are in turn subject to tight regulation. These enzymes can potentially be targeted by compounds that inhibit their ty with the uence of altering gene expression in a therapeutic manner. s in the state of histone methylation are now known to play critical roles in normal regulation of the cell cycle and growth, the response to DNA damage and stress, and pre-natal pment including differentiation. ogical states such as cancer are associated with altered patterns of histone modifications and dysregulated histone-modifying proteins including chromatin-modifying enzymes. The need to closely regulate histone modifications is evidenced by the association of histone methylation status with human morbidity including .

Histone methylation can occur on any of the three basic amino acid residues — lysine (K), arginine (R), and histidine (H). Methylation of histone H3 on lysines at positions 4 (H3K4), 9 (H3K9), 27 (H3K27), 36 (H3K36) and 79 (H3K79) are among the best studied of histone ations that influence gene expression. Lysine tri-methylation (Kme3) on histone 3 (H3) at position 4 (H3K4me3) is a histone mark generally associated with activation of gene expression while H3K9mel or H3K27me3 are associated with the repression of gene transcription. H3K4melis associated with DNA enhancers of gene transcription while H3K4me3 is associated with gene promoter activity. Likewise, loss of the methyl group at H3K4 is associated with repression of gene expression. Thus, the addition and removal of methyl groups at H3K4 constitutes a gene transcription switch. It is also evident that lysine can be modified with a mono-, di- or tri-methyl groups, each modification having a different biological effect through the attraction of different proteins recognizing those specific methylation ations at that site.

A critical aspect of the regulation of the state of histone methylation is the recruitment of methyltransferases and demethylases to specific genetic loci. DNA sequence— speciflc binding proteins including transcription factors are one class of proteins sible for this recruitment h the assemblage of n xes that bind these methyl- transferring enzymes. A well-studied example is the Drosophila melanogaster trithrorax group (TrxG) response elements (TREs) which t the H3K4 methyltransferase, TRX, to specific genes via ription factors that recognize the TRE DNA sequence.

The e ation marks are recognized by methyl-binding s in a diverse group of proteins; these domains include PHD s, WD40 and ankyrin repeats, CW and PWWP domains, and the Royal superfamily of proteins. These proteins, in turn, determine which additional activities are recruited into chromatin sites and ultimately the state of transcription at a given locus. Indeed, depending on which methyl-recognition n binds the marked histone, the same methyl-lysine modification can have opposing effects on transcription. H3K4me2 and H3K4me3 are associated with transcriptional activa- tion, but when bound by the PHD-domain-containing co-repressor protein Inhibitor of Growth family member 2 , an associated histone deacetylase complex is stabilized repressing gene expression. Thus, these effector proteins recognizing the methyl-lysine histone modifications significantly influence the level of transcriptional activity.

The ability to alter gene expression selectively by modifying the state of chromatin allows a novel therapeutic strategy to induce or de—repress the expression of genes that can provide a benefit, especially for genes whose sion has been suppressed by pathological mechanism as in the case of some cancers or suppressed by physiologic mechanism but who de-repression can phenotypically suppress mutations in paraologous genes with complementary function.

Many genes within a genome are members of gene families as a consequence of gene duplication. These genes are termed gs of one another. Following gene duplication, patterns of expression of two genes will evolve in a distinct manner in part to control the effects of gene . Following gene duplication, random genetic drift arising from naturally occurring mutations and the uent selection of nucleotide sequence is ly observed first in non—coding regions of duplicated genes, often in transcriptional regulatory regions. DNA changes in regulatory sequences can influence any or all aspects of gene expression: the ude of expression, its developmental timing, induction by i outside the cell including hormonal or metabolic signals, and the cell type in which expression is restricted. In instances in which the duplication is recent in evolutionary time or where natural selection has maintained a high degree of protein—coding sequence rity, the gene product of one paralog, gene A, can complement the pathological loss or silencing of the other paralog, gene B, if sion of gene A is not limiting in the same cell.

Altering patterns of gene expression may offer profound therapeutic benefits for genetic conditions in which enhanced expression of a paralogous gene es" a phenotype caused by a on in a g. This might be called autologous gene complementation.

In the case of Wilson disease caused by mutations in ATP7B, enhanced expression by pharmacologic induction ofATP7A, a closely related copper transporter protein, might rescue mutations in ATP7B, another copper transporter. The basic function of each copper transporter protein has been preserved but following the ation of the common ancestral gene, the expression of these two genes has been separated spatially, one confined to inal enterocytes, the other to hepatocytes. This is one of many examples of paralogous gene in which one gene can ment the loss of the second if appropriately expressed in the same cell or tissue.

A notable example of a paralogous gene family is the well-studied alpha and beta family of globin genes coding for the alpha and beta subunits of hemoglobin. Five beta-like genes each g by gene duplication are arrayed next to each other on chromosome 16 with each gene being transcribed in atemporally-specif1c manner throughout the 9 months of human embryonic and fetal development. The five beta-like globin proteins share a high degree of protein sequence similarity, so much so that genetic mutations inactivating the adult beta globin gene can be ally silent if sion of any one of the other 4 subunit members of the beta—like globin family is adequate. Activation of expression and subsequent riptional silencing of each specific embryonic and fetal beta-like globin gene is regulated in part by epigenetic mechanisms. The rescue of ons in the beta globin gene, mutations which are responsible for diseases such as thalassemia major or sickle cell anemia, by transcriptional induction of one or more of the other beta-like genes through the pharmacologic manipulation of etic silencing would be clinically beneficial.

Autologous tion with a pharmacologic agent of a functionally complementary paralog of a mutated or pathologically silenced gene may be a more successful eutic strategy than replacing or repairing the mutated gene with a wild-type (normal) copy.

Interest in influencing the activity of histone modifications for therapeutic effect derive from observations that the expression of specific genes under epigenetic control could be altered by altering epigenetic marks such as methylation. In the case of cancer, loss of specific histone methylation marks concomitant with overexpression of histone ylases is associated with the recurrence of those cancers with attendant poorer outcomes. These studies suggest that specific tumor suppressor genes are silenced by loss of methylation modifications that in turn enhance the survival and growth ial of neoplastic cells. This had led to the ition that inhibition of histone demethylase activity might have therapeutic value.

KDMlA (also known as Lysine-Specific Demethylase l (LSDl) or AOF2 or BHCl 10) was the first enzyme with specific lysine demethylase activity to be described demonstrating unequivocally that histone modifications are reversible rather than ent.

Among its demethylase substrates, KDMlA is a histone H3 lysine demethylase that catalyzes the ive demethylation of H3K4mel or me2 and H3K9mel or me2 but not the substrate H3K4me3. The enzyme also demethylates non-histone proteins such as p53 and Gfll.

KDMlA contains an amine oxidase domain that demethylates H3Kme substrate in a flavin e eotide (FAD)-dependent manner similar to other monoamine (MAO) and polyamine oxidase inhibitors. Indeed, non-specific inhibitors of MAO enzymes can inhibit the ylase activity of KDMlA KDMlA is over-expressed in many human cancers including Wilm's tumor, small-cell lung, bladder, prostate, , head & neck, colon, and ovarian cancer and associated with more frequent relapses. KDMlA is required for transcriptional regulation mediated by the androgen receptor in prostate cancer, the estrogen receptor in breast carcinomas, and the TLX receptor in neuroblastoma. Knockdown of KDMlA sion decreases proliferation of cancer cells. KDMlA is also overexpressed in cancer cells that are nuclear hormone receptor-independent including ER-negative breast. Potent, selective small molecule tors of KDMlA should be useful for treatment of these and other cancers in which KDMlA ty is overabundant.

The structure and state of chromatin can also influence the y of a pathogenic virus to insert into host DNA, undergo transcription and replicate. Infection by the alpha herpes viruses herpes simplex virus (HSV) and varicella—zoster virus (VSV) effect the remodeling of chromatin after infection of host cells to counter the rapid deposition of nucleosomes containing histones with transcriptional repressive marks by ing virus- encoded transcription factors to recruit the host HCF-l co-activator complex that contains KDMlA and the histone H3K4 methyltransferases Setl or MLL family members. It has been demonstrated that inhibition of KDMlA in cells infected with HSVl inhibits HSV IE gene expression, suppresses lytic infection and reduces viral loads. Similarly, inhibiting KDMlA causes a decrease in the sion of the immediate early genes in cells infected with human cytomegalovirus and adenovirus suggesting a broader role for KDMlA in viral pathogenesis.

The ce KDMlA activity has on the ription of specific genes is dependent on recruitment of KDMlA to a specific gene promoter region via DNA binding proteins. In the case of androgen-dependent gene expression, KDMlA associates with the androgen d receptor which specifically s DNA binding sites in the promoters of androgen—responsive genes. Thus, proteins that bind KDMlA determine where along the chromosome the demethylase ty is targeted. Many ns have been ed to interact with KDMlA including the CoREST, CtBP, NuRD, BRAF35 complexes, DNMTl, MTAl/2, Mi2beta, RbAp46/48, HDACl, 2, and 3, TIFlbeta, Blimp-l, ZNF217 and ZNF198, a subset of which form larger and in some cases complexes that mutually e one another. The KDMlA/CoREST complex which may also include DNMTl and NuRD among other factors is particularly important for the repression of expression of specific genes.

KDMlA is recruited to the promoter region of genes through site—specific transcription factors. Such factors include among others the androgen receptor, the estrogen receptor alpha, Snaill, Slug, HIV Tat, ZEBl, RBP-J, PITl, REST, NR2Cl, NR2C2 and isoforms of Gfllb. These transcription factors can recruit KDMlA to participate in activation of gene expression or silencing of gene expression ing on the cell type and the specific transcription factors.

Many of the enzyme activities that regulate the state of tin are influenced allosterically or require as co—factors metabolic intermediates, mediators or end—products of cell metabolism. These intermolecular relationships between gene expression and metabolism provide cells with signaling pathways connecting the external and internal cellular environment including nutrients with mechanisms modulating gene expression. This cellular sensing can alter both short and long term adjustments to gene sion patterns constituting an epigenetic memory of historical metabolic states and nmental conditions. For example, beta-hydroxybutyrate, a product of long chain fatty acid metabolism and a major source of energy for mammals during starvation or prolonged exertion, inhibits class I histone deacetylases (HDAC) but not class 2b HDAC. Thus the s of starvation and nutrient loss can be epigenetically coded and preserved. Acetyl— coenzyme A, nicotinamide adenine eotide (NAD) and ketoglutarate also influence histone methylation and acetylation states.

Flavin adenine dinucleotide (FAD) is a required co-factor for KDMlA. FAD, in conjunction with NAD and NADP act as cellular redox sensors. KDMlA temporarily converts FAD to FADH after which an electron acceptor, likely 02 and others, completes the catalytic cycle by regenerating FAD and H202. Thus, the cellular redox state influences KDMlA activity both by its ability to oxidize FADH and other electron acceptors. In a general sense, chromatin states, hence gene expression, can be altered by the variable concentrations of metabolic intermediates and in the specific case of KDMlA that activity is entirely dependent on FAD whose tration fluctuates as a function of the energetic economy of the cell. In addition, it has been shown that inhibition of KDMlA can lower serum glucose, reduced hepatic glycogen, and is a powerful insulin secretogogue.

Pharmaceutical manipulation of KDMlA ty may thus prove useful for the ent of diseases that represent pathological aberrations of the energy status of the cell including metabolic syndrome, dyslipidemias, diabetes, obesity, anorexia, failure to thrive, ia, lipodystrophies, and steatohepatitis.

The steroid hormones iol and testosterone and related compound play a key role in both normal development and in pathological states such as breast and prostate cancer in which tumor cell growth is dependent on hormonal signaling. The ical effects of steroid hormones are mediated by structurally and functionally distinct ligand-binding ors that function as a transcription factor ted to a specific DNA binding site. The ligand-bound steroid receptors act as the principal transcriptional regulator of hormone effects. Transcriptional activation of gene expression for all steroid-dependent hormones is dependent on chromatin structure and the presence of co—factors. The estrogen receptor employs, for example, the co-factors SRCl, SRC2, AIBl, PELPl, CBP, p300, PCAF, CARMl, PRMTl and co-repressors such as NCoR, SMRT and MTAl. The transcriptional response to hormone stimulation is dependent on the ction of these co-factors and repressors as well as the state of chromatin, ally modification of histones by histone- ing enzymes associated with the co-regulators. Both enic and androgenic hormone stimulation induces several histone modifications at the promoters of target genes that alter the acetylation, phosphorylation and methylation state of local histones. To affect the maximal rate of transcription for a e-responsive gene, KDMlA activity is required. Thus, KDMAl should prove useful as a therapeutic target of ceuticals in blunting or ablating the hormone-dependence of tumor cells. This same therapeutic logic applies to other ligand-dependent transcription factors whose riptional activation is partly or wholly ent on KDMlA activity to alter chromatin states sufficiently to facilitate transcription — examples of these would include the vitamin D, retinoid and lipid- activated receptors.

Numerous therapeutic agents have been identified that have the effect of altering gene expression acting either directly on proteins, generally enzymes, that alter chromatin states or indirectly. Though the precise mechanisms of their action have not all been fully elucidated, those mechanism can be inferred from our understanding of the protein complexes that participate in the activation of specific gene expression. These agents include cytadine and 5’-aza -2’ deoxycytidine (decitabine) which inhibit DNMTl or other DNA methyltransferases known to be present and active at promoter sites of silenced genes such as gamma globin er; vorinostat and panobinostat or other inhibitors of histone deacetylase (HDAC) enzymes; yurea (HU), valproate and sodium butyrate and its analogues each of which may interfere with the activity of orphan nuclear receptors. All of these agents enjoy some clinical use principally in the management of neoplasic e.

Though some clinical utility of these agents for other disease states has been demonstrated, these agents have not been widely adopted because of their modest therapeutic effects and their toxicity.

The use of agents that inhibit any enzymatic activity resident in the protein complex bound to gene er has the potential to disrupt the sion of gamma globin gene expression and result in increased levels of fetal hemoglobin also known as obin F (HbF). Such targets e any of the interfaces of the specific protein-protein contacts, for example, the NuRD complex and KDMlA; the DNA binding recognition domains of, for example, NR2Cl and NR2C2; the ligand binding domains of, for example, NR2Cl and NR2C2; the enzyme activities such as lysine demethylase, for example, KDMlA; histone deacetylases , for example HDACl, 2, or 3; DNA methyltransferases, for example, DNMTl.

There remains a need for compositions and methods for altering gene expression in cells and tissues sufficient to restore the cell or tissue to normal physiologic function including, e. g., appropriate apoptosis in the case of cancer, or to alter the ogical phenotype of the cell, tissue, organ or organism by inducing the expression of one or more genes sufficiently to suppress the pathological state.

Accordingly, the inventors herein disclose new compounds, compositions and methods for treating diseases ated with KDMlA activity.

Certain embodiments of the invention provide compounds of the formula (1): 0= oW nVY z R5 R1 N R4 \/ ‘R2 or a salt thereof, wherein: Y is chosen from a bond, NR", 0, C(O)NH, NHC(O), S, 802, and CH2; Z is chosen from a bond, NR", 0, C(O)NH, NHC(O), S, 802, and CH2; m is an integer from 0 to 5; n is an r from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, ulfonylalkyl, alkyl, aryl, arylalkyl, cycloalkyl, lkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl and R1 and R2, together with the en to which they attach, form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which may be optionally substituted with between 0 and 3 R6 groups; R3 is chosen from alkylamino, lkylamino, arylamino, heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which may be ally substituted with between 0 and 3 R6 groups; R4, R43, and R4b are independently chosen from hydrogen, alkyl, alkenyl, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which may be optionally substituted with between 0 and 3 R6 groups; each R6 is ndently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, arylalkyl, cyano, alkoxy, amino, alkylamino, dialkylamino, COR7, SOzR7, NHSOzR7, NHSOzNHR7, NHCOR7, NHCONHR7, CONHR7, and CONR7R8; and R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which may be optionally substituted with lower alkyl. [030a] In a particular aspect, the present invention provides a compound of Formula I: O Y Z R5 ( )m ( )n R1 N R4 or a salt thereof, wherein: Y is chosen from a bond, NR4a, O, C(O)NH, NHC(O), S, SO2, and CH2; Z is chosen from a bond, NR4b, O, C(O)NH, NHC(O), S, SO2, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, , biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl or R1 and R2, together with the nitrogen to which they attach, form a nitrogen-containing heterocycloalkyl ring, which is optionally substituted with n 0 and 3 R 6 groups; R3 is chosen from alkylamino, cycloalkylamino, ino, heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, aryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which is optionally substituted with between 0 and 3 R6 groups; R4, R4a, and R4b are independently chosen from en, alkyl, alkenyl, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which is optionally substituted with between 0 and 3 R6 ; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, l, cycloalkyl, haloalkyl, haloalkoxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, heteroarylalkyl, cyano, alkoxy, amino, alkylamino, lamino, COR7, SO2R7, NHSO2R7, NHSO2NHR7, NHCOR7, NHCONHR7, CONHR7, and CONR7R8; and 11 [FOLLOWED BY PAGE 11a] R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which is optionally substituted with lower alkyl. [030b] In another particular , the present invention provides a compound chosen from: , , , , , , , , , , 11 a [FOLLOWED BY PAGE 11b] , , , , , , , , , , 11 b WED BY PAGE 11c] , , , , , , , , , , 11 c WED BY PAGE 11d] , , , and , or a salt thereof.

In some embodiments, the compound has Formula IIa or IIb: R3 R3 O R4 R5 NH NH O Y Z O Y Z ( )m ( )n ( )m ( )n R1 N R4 R5 R1 N R2 R2 (IIa) (IIb) or a salt thereof, n: Y is chosen from a bond, NR4a, O, C(O)NH, , S, SO2, and CH2; Z is chosen from a bond, NR4b, O, C(O)NH, NHC(O), S, SO2, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl and R1 and R2, together with the nitrogen to which they attach, form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which may be optionally tuted with between 0 and 3 R6 groups; R3 is chosen from mino, cycloalkylamino, arylamino, heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, 11 d [FOLLOWED BY PAGE 11e] heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which may be optionally substituted with between 0 and 3 R6 groups; R4, R4a , and R4b are independently chosen from hydrogen, alkyl, l, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which may be optionally substituted with between 0 and 3 R6 groups; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, koxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, heteroarylalkyl, cyano, alkoxy, amino, alkylamino, dialkylamino, COR7, SO2R7, NHSO2R7, NHSO2NHR 7, NHCOR 7, NHCONHR7, CONHR7, and 8; and R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a en-containing heterocycloalkyl or heteroaryl ring, which may be optionally substituted with lower alkyl.

In some embodiments, the compound has Formula IIIa or IIIb: [FOLLOWED BY PAGE 12] 11 e (111a) (Illb) or a salt thereof, wherein: Y is chosen from a bond, NR", 0, C(O)NH, , S, 802, and CH2; Z is chosen from a bond, NR", 0, C(O)NH, NHC(O), S, 802, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and cycloalkylalkyl and R1 and R2, together with the nitrogen to which they attach, form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which may be optionally substituted with n 0 and 3 R6 groups; R3 is chosen from alkylamino, cycloalkylamino, arylamino, heteroarylamino, cycloalkylamino, lkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which may be optionally substituted with between 0 and 3 R6 groups; R4, R43, and R4b are independently chosen from en, alkyl, alkenyl, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which may be optionally substituted with between 0 and 3 R6 groups; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, heteroarylalkyl, cyano, alkoxy, amino, alkylamino, lamino, COR7, SOzR7, NHSOzR7, NHSOzNHR7, NHCOR7, NHCONHR7, CONHR7, and CONR7R8; and R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a en-containing heterocycloalkyl or heteroaryl ring, which may be optionally substituted with lower alkyl.

In certain embodiments, Z is NR".

In certain ments, R4b is chosen from methyl and hydrogen.

In certain ments, the alkyl, whether by itself or as a named part of another clic substituent, is C1-C8 alkyl.

In certain embodiments, R3 is chosen from aryl, arylalkyl, heteroaryl, and heteroarylalkyl, any of which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, R3 is chosen from aryl and heteroaryl, any of which may be optionally substituted with n 0 and 3 R6 groups. [03 8] In certain embodiments, m is an integer from 0 to 1; Y is chosen from NR", 0, S, S02, and CH2; n is an integer from 1 to 3; and R4a is chosen from hydrogen and alkyl.

In n ments, m is 0; Y is CH2; and n is an integer from 1 to 3.

In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

In certain embodiments, R3 is 5-6 membered monocyclic or 8—12 membered bicyclic heteroaryl, in which n one and five ring members may be atoms chosen from N, O, and S, and which may be optionally substituted with n 0 and 3 R6 groups.

In certain embodiments, R3 is 5-6 membered monocyclic heteroaryl, in which between one and four ring s may be heteroatoms chosen from N, O, and S, and which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, each R6 is chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

In certain embodiments, R3 is chosen from O O\ \ A N NI N Q I N\Q HN\N \N\N HN\N / :3 / :3 "$.63 V53 / :1 RA; vs vs. 9%: /\N //\NH ,N\N N "M763 "v? /Nv\x3 "$53 /"N43 In certain embodiments, R4 is hydrogen.

In certain embodiments, R4 is methyl.

In certain embodiments, the nitrogen-containing heterocycloalkyl or heteroaryl ring formed by R1 and R2 together with the nitrogen to which they are attached contains 3 to eight atoms.

In certain embodiments, R1 and R2 are taken together to form a nitrogen-containing heterocycloalkyl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is chosen from: \\ // \ /S\ 023 s o K/Ng‘: K/Nrei and K/Nf‘e‘ 1 ' ' ncerta1n embd'0 1ments,then1trogen-conta1n1n"gheterocylc oalkylformedble and R2 er with the nitrogen to which they are attached is chosen from: \\ // 04,51, \NOf, /S‘Na??? 023Q95, and K/qua0 In certain embodiments, the en-containing heterocycloalkyl formed by R1 and R2 together With the nitrogen to which they are attached is g In certain embodiments, the en-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is {e In certain embodiments, the n1trogen-conta1n1ng heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is \\ ’/ /S\N/W K/N cI: . In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are ed is K/N52 In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is K/Ng In certain embodiments, n is 2 or 3.

In n embodiments, R1 and R2 are taken together with the nitrogen to which they are attached form a nitrogen-containing heteroaryl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, the nitrogen-containing heteroaryl is chosen from pyrrole, imidazole, and pyrazole.

In certain embodiments, R5 is aryl, which may be optionally substituted with between 0 and 3 R6 .

In n embodiments, R5 is phenyl, which may be ally substituted with between 0 and 3 R6 groups.

In certain embodiments, n is 2 or 3.

In certain embodiments, R5 is aryl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, R5 is a 5—6 ed monocyclic or 8—12 membered bicyclic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which may be optionally substituted with between 0 and 3 R6 .

In certain embodiments, R5 is a 5—6 membered monocyclic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which may be optionally substituted with l or 2 R6 groups.

In certain embodiments, R5 is chosen from: XUIU; U; U; U; U; U; U;O N\o HN\N \N\ HN\ "Ck "O;43%NH ’N'x/NEiN\ N§N In certain embodiments, n is 2 or 3.

In certain embodiments, R3 is aryl, ally substituted with between 0 and 3 R6 groups.

In certain embodiments, R3 is chosen from phenyl and biphenyl, either of which may be optionally substituted with n 0 and 3 R6 groups.

In certain embodiments, m is an integer from 0 to 1; Y is chosen from NR", 0, S, S02, and CH2; n is an integer from 1 to 3; and R4a is chosen from hydrogen and alkyl.

In certain embodiments, mis 0; Y is CH2; and n is an integer from 1 to 3.

In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

In certain embodiments, R6 is chosen from lower alkyl, halogen, lower , OCF3 and CF3.

In n embodiments, R4 is hydrogen.

In certain embodiments, R4 is methyl.

In n embodiments, n is 2 or 3.

In certain embodiments, the en-containing heterocycloalkyl or heteroaryl ring formed by R1 and R2 together with the nitrogen to which they are attached contains 3 to eight atoms.

In n embodiments, R1 and R2 are taken together to form a nitrogen-containing heterocycloalkyl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is chosen from: @3st HN/WLNHAJHCEOG, 0%, Gigi,,NH HN \N S \\ ’/ /S‘Nmot. ON,s O and ON," In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is chosen from: Gigi, \Na}; /S‘Na; 023 O K/Ng and a5" I ' bd' ' ncerta1n em 0 1ments,then1trogen-conta1n1n"gheterocylc oalkylformedbyR1 and R2 together with the nitrogen to which they are attached is Oii. In n embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 er with the nitrogen to which they are attached is A; In certa1n embod1ments, the n1trogen-conta1n1ng. . . . . heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is \\ // N ~,5? . In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are attached is K/Ni.

In certain embodiments, the nitrogen-containing heterocycloalkyl formed by R1 and R2 together with the nitrogen to which they are ed is N531.

In certain embodiments, n is 2 or 3.

In certain embodiments, R1 and R2 taken together form a nitrogen-containing heteroaryl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, the nitrogen-containing heteroaryl is chosen from pyrrole, imidazole, and pyrazole.

In certain embodiments, R5 is aryl, which may be optionally substituted with between 0 and 3 R6 groups, each of which is independently chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

In certain embodiments, R5 is phenyl, which may be ally substituted with n 0 and 3 R6 groups, each of which is independently chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

In certain embodiments, n is 2 or 3.

In certain embodiments, R5 is heteroaryl, which may be optionally substituted with between 0 and 3 R6 groups.

In certain embodiments, R5 is a 5—6 membered monocyclic or 8—12 ed bicyclic aryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which may be optionally substituted with between 0 and 3 R6 groups, each of which is independently chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

In certain embodiments, R5 is a 5—6 membered monocyclic heteroaryl, in which between one and five ring s may be atoms chosen from N, O, and S, and which may be optionally substituted with l or 2 R6 groups, each of which is independently, if present, a lower alkyl .

In certain embodiments, R5 is chosen from: CT {5 pN NIAN / o N\Q HN\N \N\N HN\N / 5i, / g:Nyyi, Mg: Q; (A; 8);: K}; K}; "Na;\N Nx/EA 45"; AK);//\NH N§N N§N In certain embodiments, wherein n is 2 or 3.

Also provided are embodiments wherein any embodiment above in aphs — [087] above may be combined with any one or more of these embodiments, provided the combination is not mutually ive. As used herein, two embodiments are "mutually exclusive" when one is defined to be something which cannot overlap with the other. For example, an embodiment wherein Y is CH2 is mutually exclusive with an embodiment wherein Y is NR4b. However, an embodiment wherein R1 and R2 are taken together to form a nitrogen-containing cycloalkyl is not mutually ive with an embodiment wherein R5 is phenyl optionally substituted with fluorine.

In accordance with another aspect of the invention, a compound as sed herein is provided for use as a medicament.

In accordance with another aspect of the invention, a nd as disclosed herein is provided for use in the manufacture of a medicament for the prevention or treatment of a disease or condition chosen from sickle cell disease, thalassemia major, and other beta— hemoglobinopathies.

In accordance with another aspect of the invention, a pharmaceutical composition is provided which ses a nd as disclosed , together with a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutical composition is ated for oral administration.

In some embodiments, the pharmaceutical composition additionally comprises another therapeutic agent.

In accordance with another aspect of the invention, a method of inhibiting KDMlA is provided, comprising contacting KDMlA with a compound as disclosed herein.

In accordance with another aspect of the invention, a method of treating a globin- mediated disease is provided; comprising the administration of a therapeutically effective amount of a nd as disclosed herein.

In some embodiments, the disease is chosen from ysplastic Syndrome (MDS), Acute Myelogenous Leukemia (AML), and Chronic Myelogenous Leukemia (CML).

In accordance with another aspect of the invention, a method for achieving an effect in a patient is provided; sing the administration of a therapeutically effective amount of a compound as disclosed herein; wherein the effect is chosen from an elevation of red blood cell count, an elevation of the red blood cell count of red cells containing fetal hemoglobin, an elevation in the total concentration of fetal hemoglobin in red cells, an elevation in the total tration of fetal hemoglobin in reticulocytes, an increase in the transcription of the gamma globin gene in bone marrow—derived red cell precursors, e.g., pro— erythroblasts, a reduction in the number of sickle cell crises a t experiences over a unit period of time, a halt to or prevention of tissue damage e. g. in the heart, spleen, brain or kidney caused by sickling cells, a reduction in the proportion of red cells that undergo sickling under physiological conditions of relative hypoxia as measured using patient blood in an in vitro assay, an increase in the amount of histone 3 lysine methylation at lysine position 4 (H3K4mel and H3K4me2), and/or a decrease in the amount of histone 3 methylation at lysine position 9 (H3K9mel or H3K4me2) near or at the gamma globin promoter as assayed by ChIP using cells derived from a treated patient.

In accordance with another aspect of the invention, a method of ting at least one KDMlA function is provided; comprising the step of contacting KDMlA with a compound as disclosed herein; wherein the inhibition is measured by phenotype of red cells or their precursors either cultured or in vivo in humans or mouse or transgenic mice ning the human beta globin locus or portions thereof, the ability of cancer cells to proliferate, the expression of specific genes known to be regulated by KDMlA activity such as gamma globin, a change in the histone methylation states, a change in the methylation state of proteins known to be demethylated by KDMlA such as G9a or 1, expression of KDMlA-regulated genes, or g of KDMlA with a natural binding partner such as , DNMTl or HDACs.

Inhibition of LSDl ty alone may be sufficient therapy for the treatment of some diseases; for other such as cancer, combination therapies are often additive or synergistic in their therapeutic effects and may even be necessary to e the full clinical benefit d. There is specific scientific ce to rationalize the combination of an inhibitor of LSDl with all-trans retinoic acid , c trioxide, inhibitors of DNA methytransferases such as 5’-azacytidine or 5’-aza 2’-deoxycytidine, inhibitors of NFKB signaling such as sulindac or conventional anti-neoplastic agents such as anthracyclines or nucleoside analogues such as cytosine arabinoside. Likewise, agents that induce ia stem cells into the cell cycle (G-CSF, GM-CSF, stem cell factor, thrombopoietin (TPO)) or agents that negate the contributory role cytokines (TPO, CCL3 (MIP-1)) play in remodeling the niche of cancer stem cells may be useful as part of a combination including an LSDl inhibitor.

Abbreviations and Definitions To facilitate understanding of the disclosure, a number of terms and abbreviations as used herein are defined below as follows: When introducing elements of the present disclosure or the preferred embodiment(s) thereof, the articles "a", "an", "the" and "said" are intended to mean that there are one or more of the elements. The terms "comprising", "including" and "having" are intended to be inclusive and mean that there may be additional elements other than the listed elements.

The term "and/or" when used in a list of two or more items, means that any one of the listed items can be employed by itself or in combination with any one or more of the listed items. For example, the expression "A and/or B" is intended to mean either or both of A and B, i.e. A alone, B alone or A and B in combination. The expression "A, B and/or C" is intended to mean A alone, B alone, C alone, A and B in combination, A and C in combination, B and C in ation or A, B, and C in combination.

The term "about," as used herein when referring to a measurable value such as an amount of a compound, dose, time, temperature, and the like, is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% from the specified .

A "therapeutically effective " of a drug is an amount of drug or its ceutically acceptable salt that eliminates, alleviates, or provides relief of the symptoms of the disease for which it is administered.

A "subject in need thereof’ is a human or non-human animal that ts one or more ms or indicia of a disease.

When ranges of values are disclosed, and the notation "from m to n2" or en n1 and n2" is used, where m and n2 are the numbers, then unless otherwise specified, this notation is intended to include the numbers themselves and the range n them. This range may be integral or continuous n and including the end values. By way of example, the range "from 2 to 6 carbons" is intended to include two, three, four, five, and six carbons, since carbons come in integer units. e, by way of example, the range "from 1 to 3 uM (micromolar)," which is intended to include 1 "M, 3 "M, and everything in between to any number of significant figures (e.g., 1.255 uM, 2.1 uM, 2.9999 uM, etc.).

When n is set at 0 in the t of "0 carbon atoms", it is intended to indicate a bond or null.

The term "alkylsulfonyl" as used herein, means an alkyl group, as d herein, ed to the parent molecular moiety h a sulfonyl group, as defined herein.

Representative examples of alkylsulfonyl include, but are not limited to, methylsulfonyl and ethylsulfonyl.

The term "alkylsulfonylalkyl" as used herein, means an ulfonyl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of alkylsulfonylalkyl include, but are not limited to, methylsulfonylmethyl and ethylsulfonylmethyl.

The term "acyl," as used herein, alone or in combination, refers to a carbonyl attached to an alkenyl, alkyl, aryl, cycloalkyl, heteroaryl, heterocycle, or any other moiety where the atom attached to the carbonyl is carbon. An "acetyl" group refers to a —C(O)CH3 group. An "alkylcarbonyl" or "alkanoyl" group refers to an alkyl group attached to the parent molecular moiety through a carbonyl group. Examples of such groups include methylcarbonyl and ethylcarbonyl. Examples of acyl groups include formyl, alkanoyl and aroyl.

The term "alkenyl," as used herein, alone or in combination, refers to a straight- chain or branched-chain arbon group having one or more double bonds and containing from 2 to 20 carbon atoms. In certain embodiments, said alkenyl will se from 2 to 6 carbon atoms. The term "alkenylene" refers to a carbon—carbon double bond system attached at two or more positions such as lene [(—CH=CH—), —)]. Examples of suitable alkenyl groups include ethenyl, propenyl, 2-methylpropenyl, tadienyl and the like.

Unless otherwise specified, the term "alkenyl" may include "alkenylene" groups.

The term "alkoxy," as used herein, alone or in combination, refers to an alkyl ether group, wherein the term alkyl is as defined below. Examples of suitable alkyl ether groups include methoxy, , n—propoxy, isopropoxy, n—butoxy, iso—butoxy, sec—butoxy, tert-butoxy, and the like.

The term "alkyl," as used herein, alone or in combination, refers to a straight- chain or ed-chain alkyl group containing from 1 to 20 carbon atoms. In certain embodiments, said alkyl will comprise from 1 to 10 carbon atoms. In further embodiments, said alkyl will comprise from 1 to 6 carbon atoms. Alkyl groups may be optionally substituted as defined herein. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n—butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl, noyl and the like. The term "alkylene," as used herein, alone or in combination, refers to a saturated aliphatic group derived from a straight or ed chain saturated hydrocarbon attached at two or more positions, such as methylene (—CH2—). Unless otherwise specified, the term "alkyl" may include ene" groups.

The term "alkylamino," as used herein, alone or in combination, refers to an alkyl group attached to the parent molecular moiety through an amino group. Suitable alkylamino groups may be mono- or dialkylated, forming groups such as, for example, N—methylamino, N—ethylamino, N,N—dimethylamino, N,N—ethylmethylamino and the like.

The term "alkylidene," as used herein, alone or in combination, refers to an alkenyl group in which one carbon atom of the carbon—carbon double bond belongs to the moiety to which the alkenyl group is attached.

The term "alkylthio," as used herein, alone or in combination, refers to an alkyl thioether (R—S—) group wherein the term alkyl is as d above and wherein the sulfur may be singly or doubly oxidized. Examples of suitable alkyl thioether groups e methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, iso-butylthio, sec—butylthio, tert-butylthio, methanesulfonyl, ethanesulfinyl, and the like.

The term yl," as used herein, alone or in combination, refers to a straight- chain or branched-chain hydrocarbon group haVing one or more triple bonds and containing from 2 to 20 carbon atoms. In certain embodiments, said alkynyl comprises from 2 to 6 carbon atoms. In further embodiments, said alkynyl comprises from 2 to 4 carbon atoms.

The term "alkynylene" refers to a carbon-carbon triple bond attached at two positions such as ethynylene (—CEC—). Examples of alkynyl groups include l, propynyl, hydroxypropynyl, butyn-l-yl, butynyl, pentyn-l-yl, 3-methylbutyn-l-yl, hexynyl, and the like. Unless otherwise ied, the term yl" may include "alkynylene" groups.

The terms "amido" and "carbamoyl,"as used herein, alone or in ation, refer to an amino group as bed below attached to the parent molecular moiety through a carbonyl group, or Vice versa. The term "C—amido" as used herein, alone or in combination, refers to a —C(=O)—NR2 group with R as defined . The term do" as used herein, alone or in combination, refers to a RC(=O)NH- group, with R as d herein. The term WO 21128 "acylamino" as used herein, alone or in combination, embraces an acyl group attached to the parent moiety through an amino group. An example of an "acylamino" group is acetylamino (CH3C(O)NH—).

The term ," as used herein, alone or in combination, refers to —NRR’, wherein R and R’ are independently chosen from hydrogen, alkyl, hydroxyalkyl, acyl, heteroalkyl, aryl, cycloalkyl, heteroaryl, and heterocycloalkyl, any of which may themselves be optionally substituted. Additionally, R and R’ may combine to form heterocycloalkyl, either of which may be optionally substituted.

The term "amino acid", as used herein, alone or in combination, refers to a — NHCHRC(O)O— group, which may be ed to the parent molecular moiety to give either an N—terminus or C—terminus amino acid, wherein R is independently chosen from hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, aminoalkyl, amido, lkyl, carboxyl, carboxylalkyl, guanidinealkyl, hydroxyl, thiol, and thioalkyl, any of which themselves may be optionally tuted. The term C-terminus, as used herein, alone or in combination, refers to the parent molecular moiety being bound to the amino acid at the amino group, to give an amide as described herein, with the carboxyl group d, resulting in a terminal yl group, or the corresponding carboxylate anion. The term N—terminus, as used herein, alone or in combination, refers to the parent molecular moiety being bound to the amino acid at the carboxyl group, to give an ester as described herein, with the amino group d resulting in a terminal secondary amine, or the corresponding ammonium cation. In other words, C—terminus refers to —NHCHRC(O)OH or to —NHCHRC(O)O' and N—terminus refers to H2NCHRC(O)O— or to H3N+CHRC(O)O—.

The term "aryl", as used herein, alone or in combination, means a carbocyclic aromatic system containing one, two or three rings wherein such polycyclic ring systems are fused together. The term "aryl" embraces aromatic groups such as phenyl, naphthyl, anthracenyl, and phenanthryl.

The term "arylalkenyl" or enyl," as used herein, alone or in combination, refers to an aryl group attached to the parent molecular moiety through an alkenyl group.

The term "arylalkoxy" or "aralkoxy," as used herein, alone or in combination, refers to an aryl group attached to the parent lar moiety through an alkoxy group.

The term "arylalkyl" or "aralkyl," as used herein, alone or in combination, refers to an aryl group attached to the parent molecular moiety through an alkyl group. 2014/049906 The term "arylalkynyl" or "aralkynyl," as used herein, alone or in combination, refers to an aryl group attached to the parent molecular moiety through an alkynyl group.

The term "arylalkanoyl" or "aralkanoyl" or "aroyl," as used herein, alone or in combination, refers to an acyl group derived from an aryl-substituted carboxylic acid such as benzoyl, naphthoyl, phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4- phenylbutyryl, (2-naphthyl)acetyl, 4-chlorohydrocinnamoyl, and the like.

The term aryloxy as used herein, alone or in combination, refers to an aryl group ed to the parent molecular moiety through an oxy.

N/"4 The term azetidine, as used herein, alone or in combination, refers to an D group.

The term pyrrolidine, as used herein, alone or in combination, refers to a "‘72 group.

The term olidine, as used herein, alone or in ation, refers to a "‘71 C"N) H group.

The term pyrazolidine, as used herein, alone or in combination, refers to a "‘74.

I\l\/ group.

The term thiomorpholine, as used herein, alone or in combination, refers to a The term pyrrole, as used herein, alone or in combination, refers to a group.

/ C‘" The term pyrazole, as used herein, alone or in combination, refers to a CN group.

The terms "benzo" and " as used herein, alone or in combination, refer to the divalent group C6H4= derived from benzene. Examples include benzothiophene and benzimidazole.

The term "biphenyl" as used herein refers to two phenyl groups connected at one carbon site on each ring.

The term "carbamate," as used herein, alone or in combination, refers to an ester of ic acid (—NHCOO—) which may be attached to the parent molecular moiety from either the nitrogen or acid end, and which may be optionally substituted as defined herein.

The term "O-carbamyl" as used herein, alone or in combination, refers to a —OC(O)NRR’ group, with R and R’ as defined herein.

The term "N—carbamyl" as used herein, alone or in combination, refers to a ROC(O)NR’— group, with R and R’ as defined herein.

The term "carbonyl," as used herein, when alone es formyl [—C(O)H] and in combination is a —C(O)— group.

The term xyl" or "carboxy," as used herein, refers to —C(O)OH or the corresponding "carboxylate" anion, such as is in a carboxylic acid salt. An "O-carboxy" group refers to a RC(O)O— group, where R is as defined herein. A "C—carboxy" group refers to a —C(O)OR groups where R is as d .

The term "cyano," as used herein, alone or in combination, refers to —CN.

The term "cycloalkyl," or, alternatively, "carbocycle," as used herein, alone or in combination, refers to a saturated or partially saturated monocyclic, bicyclic or tricyclic alkyl group wherein each cyclic moiety contains from 3 to 12 carbon atom ring members and which may optionally be a benzo fused ring system which is optionally substituted as defined herein. In certain embodiments, said cycloalkyl will comprise from 5 to 7 carbon atoms.

Examples of such lkyl groups include cyclopropyl, cyclobutyl, entyl, exyl, cycloheptyl, tetrahydronapthyl, indanyl, octahydronaphthyl, 2,3-dihydro-1H- indenyl, adamantyl and the like. "Bicyclic" and clic" as used herein are intended to include both fused ring systems, such as decahydronaphthalene, octahydronaphthalene as well as the multicyclic (multicentered) saturated or partially unsaturated type. The latter type of isomer is exemplified in l by, bicyclo[1,1,1]pentane, camphor, adamantane, and bicyclo[3 ,2,1]octane.

The term "ester," as used herein, alone or in combination, refers to a carboxy group bridging two es linked at carbon atoms.

The term "ether," as used , alone or in combination, refers to an oxy group bridging two moieties linked at carbon atoms.

The term "guanidine", as used herein, alone or in combination, refers to — NHC(=NH)NH2, or the corresponding guanidinium cation.

The term "halo," or "halogen," as used herein, alone or in combination, refers to fluorine, chlorine, bromine, or .

The term lkoxy," as used herein, alone or in combination, refers to a haloalkyl group attached to the parent molecular moiety through an oxygen atom.

The term "haloalkyl," as used herein, alone or in combination, refers to an alkyl group haVing the meaning as defined above wherein one or more hydrogen atoms are replaced with a halogen. Specifically embraced are monohaloalkyl, dihaloalkyl and polyhaloalkyl groups. A monohaloalkyl group, for one example, may have an iodo, bromo, chloro or fluoro atom within the group. Dihalo and polyhaloalkyl groups may have two or more of the same halo atoms or a combination of ent halo groups. Examples of kyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl and ropropyl.

"Haloalkylene" refers to a haloalkyl group attached at two or more positions. Examples e fluoromethylene (—CFH—), difluoromethylene (—CFz —), chloromethylene (—CHCl—) and the like.

The term "heteroalkyl," as used herein, alone or in combination, refers to a stable ht or branched chain, or cyclic hydrocarbon group, or combinations thereof, fully saturated or containing from 1 to 3 degrees of unsaturation, consisting of the stated number of carbon atoms and from one to three heteroatoms chosen from O, N, and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The atom(s) O, N and S may be placed at any interior position of the heteroalkyl group. Up to two heteroatoms may be consecutive, such as, for e, —CH2—NH—OCH3.

WO 21128 2014/049906 The term "heteroaryl," as used herein, alone or in combination, refers to a 3 to 7 membered unsaturated heteromonocyclic ring, or a fused clic, ic, or tricyclic ring system in which at least one of the fused rings is ic, which contains at least one atom chosen from O, S, and N. In certain embodiments, said heteroaryl will comprise from 5 to 7 carbon atoms. The term also embraces fused polycyclic groups wherein heterocyclic rings are fused with aryl rings, wherein heteroaryl rings are fused with other aryl rings, wherein heteroaryl rings are fused with heterocycloalkyl rings, or wherein heteroaryl rings are fused with cycloalkyl rings. Examples of heteroaryl groups include pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, nyl, zinyl, triazolyl, l, furanyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, isothiazolyl, l, isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, quinoxalinyl, quinazolinyl, indazolyl, benzotriazolyl, ioxolyl, benzopyranyl, benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzofuranyl, benzothienyl, chromonyl, coumarinyl, benzopyranyl, ydroquinolinyl, tetrazolopyridazinyl, tetrahydroisoquinolinyl, thienopyridinyl, furopyridinyl, pyrrolopyridinyl, azepinyl, diazepinyl, benzazepinyl, and the like. Exemplary tricyclic heterocyclic groups include carbazolyl, olyl, phenanthrolinyl, dibenzofuranyl, acridinyl, phenanthridinyl, xanthenyl and the like.

The term "heteroarylalkyl" as used herein alone or as part of another group refers to alkyl groups as defined above having a heteroaryl substituent.

The terms "heterocycloalkyl" and, interchangeably, "heterocycle," as used herein, alone or in combination, each refer to a saturated, partially unsaturated, or fully unsaturated monocyclic, bicyclic, or tricyclic heterocyclic group containing at least one heteroatom as a ring member, wherein each said heteroatom may be independently chosen from nitrogen, oxygen, and sulfur. In certain embodiments, said ycloalkyl will comprise from 1 to 4 heteroatoms as ring members. In r embodiments, said hetercycloalkyl will comprise from 1 to 2 heteroatoms as ring members. In certain embodiments, said hetercycloalkyl will comprise from 3 to 8 ring members in each ring. In further embodiments, said ycloalkyl will comprise from 3 to 7 ring members in each ring. In yet further embodiments, said hetercycloalkyl will comprise from 5 to 6 ring members in each ring.

"Heterocycloalkyl" and "heterocycle" are intended to include sulfones, sulfoxides, N—oxides of tertiary nitrogen ring members, and carbocyclic fused and benzo fused ring systems; additionally, both terms also include systems where a heterocycle ring is fused to an aryl group, as defined herein, or an additional heterocycle group. Examples of heterocycle groups 2014/049906 include aziridinyl, azetidinyl, 1,3-benzodioxolyl, dihydroisoindolyl, dihydroisoquinolinyl, dihydrocinnolinyl, dihydrobenzodioxinyl, dihydro [ 1 ,3 ] oxazolo [4,5 -b]pyridinyl, benzothiazolyl, dihydroindolyl, dihy-dropyridinyl, 1,3-dioxanyl, 1,4-dioxanyl, 1,3- dioxolanyl, imidazolidinyl, isoindolinyl, morpholinyl, oxazolidinyl, isoxazolidinyl, piperidinyl, piperazinyl, methylpiperazinyl, N—methylpiperazinyl, pyrrolidinyl, lidinyl, tetrahydrofuranyl, tetrahydropyridinyl, rpholinyl, thiazolidinyl, diazepanyl, and the like. The heterocycle groups may be optionally substituted unless specifically prohibited.

The term "hydrazinyl" as used herein, alone or in combination, refers to two amino groups joined by a single bond, i.e., —N—N—.

The term "hydroxy," as used herein, alone or in combination, refers to —OH.

The term "hydroxyalkyl," as used herein, alone or in combination, refers to a hydroxy group attached to the parent molecular moiety through an alkyl group.

The term "hydroxamic acid", as used , alone or in combination, refers to — C(=O)NHOH, wherein the parent molecular moiety is attached to the amic acid group by means of the carbon atom.

The term "imino," as used herein, alone or in ation, refers to =N—.

The term "iminohydroxy," as used herein, alone or in combination, refers to =N(OH) and =N—O—.

The phrase "in the main chain" refers to the longest contiguous or adjacent chain of carbon atoms starting at the point of ment of a group to the compounds of any one of the formulas disclosed herein.

The term "isocyanato" refers to a —NCO group.

The term "isothiocyanato" refers to a —NCS group.

The phrase "linear chain of atoms" refers to the longest straight chain of atoms independently selected from carbon, en, oxygen and .

The term "lower," as used herein, alone or in a combination, where not otherwise specifically defined, means containing from 1 to and including 6 carbon atoms.

The term "lower aryl," as used herein, alone or in ation, means phenyl or naphthyl, which may be optionally substituted as provided.

The term "lower heteroaryl," as used herein, alone or in combination, means either 1) monocyclic heteroaryl sing five or six ring members, of which between one and four said members may be heteroatoms chosen from O, S, and N, or 2) bicyclic heteroaryl, n each of the fused rings comprises five or six ring members, comprising between them one to four heteroatoms chosen from O, S, and N.

The term "lower cycloalkyl," as used herein, alone or in combination, means a monocyclic cycloalkyl having between three and six ring members. Lower cycloalkyls may be unsaturated. Examples of lower cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term "lower cycloalkyl," as used herein, alone or in combination, means a monocyclic heterocycloalkyl having between three and six ring members, of which between one and four may be heteroatoms chosen from O, S, and N. Examples of lower heterocycloalkyls include pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, and linyl. Lower heterocycloalkyls may be unsaturated.

The term "lower " as used herein, alone or in combination, refers to — NRR’, wherein R and R’ are independently chosen from hydrogen, lower alkyl, and lower heteroalkyl, any of which may be optionally substituted. Additionally, the R and R’ of a lower amino group may combine to form a f1ve- or six-membered heterocycloalkyl, either of which may be optionally substituted.

The term "mercaptyl" as used herein, alone or in combination, refers to an RS— group, where R is as defined herein.

The term "nitro," as used herein, alone or in combination, refers to —N02. [0171 ] The terms "oxy" or "oxa," as used herein, alone or in combination, refer to —O—.

The term "oxo," as used herein, alone or in combination, refers to :0.

The term loalkoxy" refers to an alkoxy group where all of the hydrogen atoms are replaced by halogen atoms.

The term loalkyl" as used herein, alone or in ation, refers to an alkyl group where all of the hydrogen atoms are replaced by halogen atoms.

The term "phosphonate," as used herein, alone or in combination, refers to a — P(=O)(OR)2 group, n R is chosen from alkyl and aryl. The term "phosphonic acid", as used , alone or in combination, refers to a —P(=O)(OH)2 group.

The term "phosphoramide", as used herein, alone or in combination, refers to a — P(=O)(NR)3 group, with R as defined herein.

The terms "sulfonate," "sulfonic acid," and "sulfonic," as used herein, alone or in combination, refer to the —SO3H group and its anion as the sulfonic acid is used in salt formation.

The term "sulfanyl," as used herein, alone or in combination, refers to —S—.

The term "sulfinyl," as used herein, alone or in combination, refers to —S(O)—.

The term "sulfonyl," as used herein, alone or in combination, refers to —S(O)2—.

The term "N—sulfonamido" refers to a RS(=O)2NR’— group with R and R’ as defined herein.

The term fonamido" refers to a —S(=O)2NRR’, group, with R and R’ as defined herein.

The terms "thia" and "thio," as used herein, alone or in combination, refer to a — S— group or an ether wherein the oxygen is replaced with sulfur. The oxidized derivatives of the thio group, namely sulfinyl and yl, are included in the definition of thia and thio.

The term "thiol," as used herein, alone or in combination, refers to an —SH group.

The term "thiocarbonyl," as used herein, when alone es thioformyl —C(S)H and in combination is a —C(S)— group.

The term "N—thiocarbamyl" refers to an ROC(S)NR’— group, with R and R’as d herein.

The term ocarbamyl" refers to a —OC(S)NRR’, group with R and R’as defined herein.

The term "thiocyanato" refers to a —CNS group.

The term "trihalomethoxy" refers to a X3CO— group where X is a halogen.

Any definition herein may be used in combination with any other definition to be a composite structural group. By convention, the trailing element of any such definition is that which attaches to the parent moiety. For example, the composite group alkylamido would represent an alkyl group attached to the parent molecule through an amido group, and the term alkoxyalkyl would represent an alkoxy group attached to the parent molecule through an alkyl group.

When a group is defined to be "null," what is meant is that said group is absent.

Similarly, when a designation such as "n" which may be chosen from a group or range of integers is designated to be 0, then the group which it designates is either , if in a terminal on, or condenses to form a bond, if it falls between two other groups.

The term "optionally tuted" means the anteceding group may be substituted or unsubstituted. When substituted, the substituents of an "optionally substituted" group may include, without limitation, one or more substituents independently selected from the ing groups or a particular designated set of groups, alone or in combination: lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl, lower heterocycloalkyl, lower kyl, lower kenyl, lower kynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl, phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, lower acyloxy, carbonyl, carboxyl, lower alkylcarbonyl, lower carboxyester, lower carboxamido, cyano, hydrogen, halogen, hydroxy, amino, lower alkylamino, ino, amido, nitro, thiol, lower alkylthio, lower haloalkylthio, lower perhaloalkylthio, arylthio, sulfonate, sulfonic acid, trisubstituted silyl, N3, SH, SCH3, C(O)CH3, COzCH3, COzH, pyridinyl, thiophene, furanyl, lower carbamate, and lower urea. Two substituents may be joined together to form a fused f1ve—, siX—, or seven—membered carbocyclic or heterocyclic ring consisting of zero to three heteroatoms, for example forming methylenedioxy or ethylenedioxy. An optionally substituted group may be unsubstituted (e.g., 3), fully substituted (e.g., —CF2CF3), monosubstituted (e. g., —CH2CH2F) or substituted at a level re in-between fully substituted and monosubstituted (e. g., —CH2CF3). Where tuents are recited without qualification as to substitution, both substituted and tituted forms are encompassed. Where a tuent is qualified as "substituted," the substituted form is specifically intended. Additionally, different sets of optional substituents to a particular moiety may be defined as needed; in these cases, the optional substitution will be as defined, often immediately following the phrase, "optionally substituted with." The term R or the term R’, appearing by itself and without a number ation, unless otherwise defined, refers to a moiety chosen from hydrogen, alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl and cycloalkyl, any of which may be optionally tuted. Such R and R’ groups should be understood to be optionally substituted as defined herein. Whether an R group has a number designation or not, every R group, including R, R’ and RH where n=(l, 2, 3, ...n), every substituent, and every term should be tood to be independent of every other in terms of selection from a group. Should any variable, substituent, or term (e.g. aryl, heterocycle, R, etc.) occur more than one time in a formula or generic structure, its definition at each occurrence is independent of the definition at every other occurrence. Those of skill in the art will further recognize that certain groups may be attached to a parent molecule or may occupy a position in a chain of elements from either end as written. Thus, by way of example only, an unsymmetrical group such as — C(O)N(R)— may be attached to the parent moiety at either the carbon or the nitrogen. tric s exist in the nds disclosed herein. These centers are designated by the s "R" or "S," depending on the configuration of substituents around the chiral carbon atom. It should be understood that the invention encompasses all stereochemical isomeric forms, ing diastereomeric, enantiomeric, and epimeric forms, as well as d—isomers and 1—isomers, and mixtures thereof. Individual stereoisomers of compounds can be prepared synthetically from commercially available ng als which contain chiral centers or by preparation of mixtures of enantiomeric products followed by separation such as conversion to a mixture of diastereomers followed by separation or recrystallization, chromatographic techniques, direct separation of enantiomers on chiral chromatographic columns, or any other appropriate method known in the art. Starting compounds of particular stereochemistry are either commercially available or can be made and resolved by techniques known in the art. Additionally, the compounds disclosed herein may exist as geometric isomers. The present invention includes all cis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers as well as the appropriate mixtures thereof.

Additionally, compounds may exist as tautomers; all tautomeric isomers are provided by this invention. Additionally, the compounds disclosed herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable ts such as water, ethanol, and the like.

In general, the solvated forms are considered lent to the unsolvated forms.

The term "bond" refers to a covalent e between two atoms, or two moieties when the atoms joined by the bond are considered to be part of larger substructure. A bond may be single, double, or triple unless otherwise specified. A dashed line between two atoms in a drawing of a molecule indicates that an additional bond may be present or absent at that position.

The term "disease" as used herein is intended to be lly synonymous, and is used interchangeably with, the terms "disorder" and "condition" (as in medical condition), in that all reflect an abnormal condition of the human or animal body or of one of its parts that impairs normal functioning, is typically sted by distinguishing signs and symptoms, and causes the human or animal to have a reduced on or quality of life.

The term nation therapy" means the administration of two or more therapeutic agents to treat a therapeutic condition or disorder described in the present disclosure. Such administration encompasses inistration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule having a fixed ratio of active ingredients or in multiple, separate capsules for each active ingredient. In addition, such administration also encompasses use of each type of therapeutic agent in a sequential manner.

In either case, the ent n will provide beneficial effects of the drug combination in treating the conditions or disorders described herein.

The phrase "therapeutically effective" is intended to qualify the amount of active ingredients used in the treatment of a disease or er. This amount will achieve the goal of reducing or eliminating the said disease or disorder.

The term "therapeutically acceptable" refers to those nds (or salts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitable for use in contact with the tissues of patients without undue toxicity, irritation, and allergic response, are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use.

As used herein, reference to "treatment" of a patient is intended to include prophylaxis. The term "patient" means all s including humans. Examples of patients include humans, cows, dogs, cats, goats, sheep, pigs, and rabbits. Preferably, the patient is a human.

The term "prodrug" refers to a compound that is made more active in vivo.

Certain compounds disclosed herein may also exist as prodrugs, as described in Hydrolysis in Drug and Prodrug Metabolism: try, Biochemistry, and logy (Testa, d and Mayer, Joachim M. Wiley-VHCA, Zurich, Switzerland 2003). Prodrugs of the compounds bed herein are structurally ed forms of the compound that readily undergo chemical changes under physiological conditions to e the compound.

Additionally, prodrugs can be converted to the compound by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly ted to a compound when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent. Prodrugs are often useful because, in some situations, they may be easier to administer than the compound, or parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. A wide variety of prodrug tives are known in the art, such as those that rely on hydrolytic cleavage or oxidative activation of the prodrug. An example, without limitation, of a prodrug would be a compound which is administered as an ester (the "prodrug"), but then is lically hydrolyzed to the carboxylic acid, the active entity. Additional examples include peptidyl derivatives of a compound.

The compounds disclosed herein can exist as therapeutically acceptable salts. The present invention es compounds listed above in the form of salts, including acid addition salts. Suitable salts include those formed with both organic and inorganic acids.

Such acid addition salts will normally be pharmaceutically acceptable. However, salts of non-pharmaceutically acceptable salts may be of utility in the preparation and purification of the compound in question. Basic addition salts may also be formed and be ceutically acceptable. For a more complete discussion of the preparation and selection of salts, refer to Pharmaceutical Salts: Properties, ion, and Use (Stahl, P. Heinrich. Wiley-VCHA, Zurich, Switzerland, 2002).

The term "therapeutically acceptable salt," as used herein, represents salts or zwitterionic forms of the compounds disclosed herein which are water or oil—soluble or dispersible and therapeutically able as defined herein. The salts can be prepared during the final isolation and purification of the compounds or separately by reacting the appropriate compound in the form of the free base with a suitable acid. Representative acid addition salts include e, adipate, alginate, L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, te, camphorate, camphorsulfonate, citrate, onate, formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate), lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nate, 2-naphthalenesulfonate, oxalate, e, pectinate, persulfate, 3-phenylproprionate, phosphonate, picrate, pivalate, nate, pyroglutamate, succinate, ate, tartrate, rate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, para-toluenesulfonate (p-tosylate), and undecanoate. Also, basic groups in the nds disclosed herein can be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, es, and s; and benzyl and phenethyl bromides. Examples of acids which can be employed to form therapeutically acceptable addition salts include inorganic acids such as hloric, hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic, maleic, succinic, and citric. Salts can also be formed by coordination of the compounds with an alkali metal or alkaline earth ion. Hence, the present invention contemplates sodium, potassium, magnesium, and calcium salts of the compounds disclosed herein, and the like.

Basic addition salts can be prepared during the final isolation and purification of the compounds by reaction of a carboxy group with a suitable base such as the hydroxide, carbonate, or bicarbonate of a metal cation or with ammonia or an organic primary, secondary, or tertiary amine. The cations of therapeutically acceptable salts include lithium, sodium, potassium, calcium, magnesium, and aluminum, as well as nontoxic quaternary amine cations such as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, methylaniline, N—methylpiperidine, N—methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, benzylphenethylamine, l-ephenamine, and N,N'—dibenzylethylenediamine. Other representative organic amines useful for the formation of base on salts include ethylenediamine, lamine, diethanolamine, piperidine, and piperazine.

A salt of a compound can be made by reaction of the appropriate nd, in the form of the free base, with the appropriate acid.

The compounds disclosed herein can exist as polymorphs and other distinct solid forms such as solvates, hydrates, and the like. A compound may be a polymorph, solvate, or hydrate of a salt or of the free base or acid.

While it may be possible for the compounds disclosed herein to be administered as the raw chemical, it is also possible to present them as pharmaceutical formulations (equivalently, "pharmaceutical compositions"). Accordingly, provided herein are pharmaceutical formulations which comprise one or more of certain compounds sed herein, or one or more pharmaceutically acceptable salts, esters, prodrugs, amides, or solvates thereof, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient f. Proper formulation is dependent upon the route of administration chosen. Any of the well-known techniques, rs, and ents may be used as suitable and as understood in the art; e.g., in Remington’s Pharmaceutical Sciences.

The pharmaceutical compositions disclosed herein may be manufactured in any manner known in the art, e. g., by means of conventional mixing, dissolving, granulating, - making, levigating, emulsifying, ulating, ping or ssion ses.

The ations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intraarticular, intraadiposal, intraarterial, intracranial, intralesional, intranasal, cular, intrapericardial, eritoneal, intrapleural, intraprostatical, intrarectal, intrathecal, intratracheal, intratumoral, intraumbilical, intravaginal, esicular, intravitreal, and intramedullary), intraperitoneal, rectal, topical (including, without limitation, dermal, , sublingual, vaginal, rectal, nasal, otic, and ocular), local, mucosal, sublingual, subcutaneous, transmucosal, transdermal, transbuccal, transdermal, and vaginal; liposomal, in , in lipid compositions, via a catheter, via a lavage, via continuous infusion, via on, via inhalation, via injection, via local delivery, via localized perfusion, bathing target cells directly, or any combination thereof. Administration although the most suitable route may depend upon for example the condition and disorder of the recipient. The formulations may conveniently be ted in unit dosage form and may be prepared by any of the s well known in the art of cy. Typically, these methods include the step of ng into ation a compound disclosed herein or a pharmaceutically acceptable salt, ester, amide, prodrug or solvate thereof ("active ingredient") with the carrier which constitutes one or more accessory ients. In l, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the d formulation.

Formulations of the compounds disclosed herein suitable for oral administration may be presented as discrete units such as hard or soft capsules, wafers, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a syrup, elixir, on, or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in—water liquid on, a water—in—oil liquid emulsion, or a compound dispersed in a liposome. The active ingredient may also be ted as a bolus, electuary or paste.

Pharmaceutical preparations that can be used orally include tablets, push—fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets may be made by compression or g, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders, inert diluents, or ating, surface active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated to provide delayed, slowed, or controlled release or absorption of the active ingredient therein. Compositions may further comprise an agent that enhances lity or dispersability. All ations for oral stration should be in dosages suitable for such administration. The it capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid in, or liquid polyethylene glycols. In addition, stabilizers may be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene , and/or titanium e, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or ts may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses. ing on the route of administration, the compounds, or granules or les thereof, may be coated in a material to protect the compounds from the action of acids and other natural conditions that may inactivate the compounds.

The compounds may be formulated for eral administration by injection, e.g., by bolus injection or continuous infusion, either to the body or to the site of a disease or wound. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi—dose containers, with an added vative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. The formulations may be presented in unit-dose or multi—dose containers, for example sealed es and vials, and may be stored in powder form or in a freeze-dried (lyophilized) condition requiring only the on of the sterile liquid carrier, for example, saline or sterile pyrogen-free water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

Formulations for parenteral administration include aqueous and non—aqueous (oily) sterile injection solutions of the active compounds which may contain idants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non—aqueous e suspensions which may include suspending agents and thickening . Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the sion, such as sodium carboxymethyl cellulose, sorbitol, or n. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. To administer the therapeutic compound by other than parenteral administration, it may be necessary to coat the compound with, or co—administer the compound with a material to t its inactivation (for e, via liposomal ation).

In addition to the formulations described previously, the compounds may also be formulated as a depot ation. Such long acting formulations may be administered by implantation (for example aneously or intramuscularly) or by intramuscular ion.

Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles, or gels formulated in conventional manner. Such compositions may comprise the active ingredient in a flavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such as suppositories or ion enemas, e.g., containing conventional suppository bases such as cocoa butter, polyethylene glycol, or other glycerides.

Certain compounds disclosed herein may be administered topically, that is by non— systemic administration. This includes the application of a compound disclosed herein ally to the epidermis or the buccal cavity and the lation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream.

In contrast, systemic administration refers to oral, enous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid or semi-liquid preparations le for penetration through the skin to the site of inflammation such as gels, liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. The active ingredient for topical administration may comprise, for e, from 0.001% to 10% W/W (by ) of the formulation. In certain embodiments, the active ingredient may comprise as much as 10% w/w. In other embodiments, it may comprise less than 5% w/w. In certain embodiments, the active ingredient may comprise from 2% w/w to % W/W. In other embodiments, it may comprise from 0.1% to 1% w/w of the formulation.

Topical ophthalmic, otic, and nasal formulations disclosed herein may comprise excipients in addition to the active ingredient. Excipients commonly used in such formulations e, but are not limited to, tonicity agents, preservatives, chelating agents, ing agents, and surfactants. Other excipients comprise solubilizing agents, stabilizing agents, comfort-enhancing agents, polymers, emollients, pH-adjusting agents and/or lubricants. Any of a variety of excipients may be used in formulations disclosed herein including water, mixtures of water and water-miscible solvents, such as C1—C7—alkanols, vegetable oils or mineral oils comprising from 0.5 to 5% non—toxic water—soluble polymers, natural products, such as alginates, pectins, tragacanth, karaya gum, guar gum, xanthan gum, carrageenan, agar and acacia, starch derivatives, such as starch acetate and hydroxypropyl starch, and also other tic products such as polyvinyl alcohol, nylpyrrolidone, nyl methyl ether, polyethylene oxide, preferably cross-linked polyacrylic acid and mixtures of those products. The concentration of the ent is, typically, from 1 to 100,000 times the tration of the active ingredient. In preferred embodiments, the excipients to be included in the formulations are typically selected because of their inertness towards the active ient ent of the formulations.

Relative to ophthalmic, otic, and nasal formulations, suitable tonicity-adjusting agents e, but are not limited to, mannitol, sodium chloride, glycerin, sorbitol and the like. Suitable buffering agents include, but are not limited to, phosphates, borates, acetates and the like. Suitable tants include, but are not limited to, ionic and nonionic surfactants h nonionic tants are preferred), RLM 100, POE 20 cetylstearyl ethers such as ® CS20 and poloxamers such as Pluronic® F68.

The formulations set forth herein may comprise one or more vatives.

Examples of such preservatives include p—hydroxybenzoic acid ester, sodium perborate, sodium chlorite, alcohols such as chlorobutanol, benzyl alcohol or phenyl ethanol, guanidine derivatives such as polyhexamethylene biguanide, sodium perborate, polyquatemium—l, amino alcohols such as AMP-95, or sorbic acid. In certain embodiments, the formulation may be self—preserved so that no preservation agent is required.

In certain topical ments, formulations are prepared using a buffering system that maintains the formulation at a pH of about 4.5 to a pH of about 8. In further embodiments, the pH is from 7 to 8.

Gels for topical or transdermal administration may comprise, generally, a mixture of volatile solvents, nonvolatile solvents, and water. In certain ments, the volatile solvent component of the buffered solvent system may include lower (Cl-C6) alkyl alcohols, lower alkyl glycols and lower glycol polymers. In further embodiments, the volatile solvent is ethanol. The le solvent component is thought to act as a penetration enhancer, while also producing a cooling effect on the skin as it evaporates. The nonvolatile solvent portion of the buffered solvent system is selected from lower ne glycols and lower glycol polymers. In certain embodiments, propylene glycol is used. The atile solvent slows the evaporation of the volatile solvent and reduces the vapor pressure of the buffered solvent system. The amount of this nonvolatile solvent component, as with the volatile solvent, is determined by the pharmaceutical compound or drug being used. When too little of the nonvolatile solvent is in the system, the pharmaceutical compound may crystallize due to evaporation of volatile solvent, while an excess may result in a lack of bioavailability due to poor release of drug from solvent mixture. The buffer component of the buffered solvent system may be ed from any buffer commonly used in the art; in certain embodiments, water is used. A common ratio of ients is about 20% of the nonvolatile solvent, about 40% of the le solvent, and about 40% water. Several optional ingredients can be added to the topical ition. These include, but are not d to, ors and gelling agents.

Appropriate g agents can include, but are not limited to, semisynthetic cellulose tives (such as hydroxypropylmethylcellulose) and synthetic polymers, galactomannan polymers (such as guar and derivatives thereof), and cosmetic agents.

Lotions include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution ally ning a bactericide and may be prepared by methods r to those for the preparation of drops. Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.

Creams, ointments or pastes are semi-solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non—aqueous fluid, with the aid of suitable machinery, with a greasy or easy base. The base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as stearic or oleic acid together with an alcohol such as propylene glycol or a macrogel. The formulation may incorporate any suitable surface active agent such as an anionic, ic or nic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.

Drops may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a le aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and, in certain embodiments, including a e active agent. The resulting solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98—1000C for half an hour. Alternatively, the on may be sterilized by tion and transferred to the container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%). Suitable solvents for the preparation of an oily solution include ol, d alcohol and ene glycol.

Formulations for topical administration in the mouth, for example buccally or gually, include lozenges sing the active ingredient in a flavored basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as n and glycerin or sucrose and acacia.

For administration by inhalation, compounds may be iently delivered from an insufflator, nebulizer pressurized packs or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized l, the dosage unit may be determined by providing a valve to deliver a metered amount. Alternatively, for administration by inhalation or insufflation, the compounds according to the ion may take the form of a dry powder composition, for example, a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, in for example, capsules, dges, gelatin or blister packs from which the powder may be administered with the aid of an inhalator or insufflator.

The therapeutic compound may also be stered pinally or intracerebrally. Dispersions for these types of strations can be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of e and use, these preparations may contain a preservative to prevent the growth of microorganisms.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous ation of sterile injectable solutions or dispersion. In all cases, the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (such as, glycerol, ene glycol, and liquid polyethylene glycol, and the like), le mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the nance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of rganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition.

Sterile inj ectable solutions can be prepared by incorporating the therapeutic compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by ed sterilization. lly, dispersions are prepared by incorporating the therapeutic compound into a sterile carrier that contains a basic dispersion medium and ed other ingredients to be pharmacologically sound. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient (i.e., the therapeutic compound) plus any additional desired ingredient from a previously sterile—filtered solution thereof It is especially advantageous to formulate eral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be d; each unit ning a predetermined quantity of therapeutic compound ated to produce the desired therapeutic effect in association with the required ceutical carrier. The specification for the dosage unit forms of the invention are dictated by and ly dependent on (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such a therapeutic compound for the treatment of a selected condition in a patient.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations described above may include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.

Compounds may be administered at a dose of from 0.1 to 500 mg/kg per day. The dose range for adult humans is generally from 5 mg to 2 g/day. Tablets or other forms of presentation provided in discrete units may conveniently contain an amount of one or more nds which is effective at such dosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 10 mg to 200 mg. red unit dosage ations are those containing an effective dose, as herein below recited, or an appropriate on thereof, of the active ingredient. In certain ments, a formulation disclosed herein is administered once a day. However, the formulations may also be formulated for administration at any frequency of administration, ing once a week, once every 5 days, once every 3 days, once every 2 days, twice a day, three times a day, four times a day, five times a day, six times a day, eight times a day, every hour, or any greater frequency. Such dosing frequency is also maintained for a varying duration of time depending on the therapeutic regimen. The duration of a particular therapeutic regimen may vary from one-time dosing to a regimen that extends for months or years. The formulations are administered at g dosages, but typical dosages are one to two drops at each administration, or a comparable amount of a gel or other formulation. One of ordinary skill in the art would be familiar with determining a therapeutic n for a specific indication. [023 6] The amount of active ingredient that may be combined with the carrier materials to e a single dosage form will vary depending upon the host treated and the particular mode of administration. Similarly, the precise amount of compound administered to a patient will be the responsibility of the attendant physician. The specific dose level for any particular patient will depend upon a variety of factors ing the activity of the ic compound ed, the age, body weight, general health, sex, diets, time of administration, route of administration, rate of excretion, drug combination, the precise disorder being treated, and the severity of the indication or condition being treated. In addition, the route of stration may vary depending on the condition and its ty. [023 7] In certain instances, it may be appropriate to ster at least one of the compounds described herein (or a pharmaceutically acceptable salt, ester, or prodrug thereof) in ation with another therapeutic agent. By way of example only, if one of the side effects experienced by a patient upon receiving one of the compounds herein is inflammation, then it may be appropriate to administer an anti-inflammatory agent in combination with the initial therapeutic agent. Alternatively, by way of example only, the therapeutic effectiveness of one of the compounds bed herein may be ed by administration of an adjuvant (i.e., by itself the adjuvant may only have minimal therapeutic benefit, but in combination with another therapeutic agent, the l therapeutic benefit to the patient is enhanced).

There is even the possibility that two compounds, one of the compounds described herein and a second compound may together achieve the desired therapeutic effect that neither alone could achieve. Alternatively, by way of example only, the benefit experienced by a patient may be increased by administering one of the compounds described herein with another therapeutic agent (which also includes a therapeutic regimen) that also has eutic benefit. By way of example only, in a ent for acute myelogenous leukemia or sickle cell anemia involving administration of one of the compounds described herein, increased therapeutic benefit may result by also providing the patient with another therapeutic agent for sickle cell anemia or for acute myelogenous leukemia. In any case, less of the disease, disorder or condition being treated, the overall benefit experienced by the patient may simply be additive of the two therapeutic agents or the two agents may have synergistic therapeutic effects in a patient.

Effective combination therapy may be achieved with a single composition or pharmacological formulation that includes both agents, or with two distinct compositions or formulations, at the same time, wherein one composition includes a compound of the present disclosure, and the other es the second s). Alternatively, the therapy may precede or follow the other agent treatment by als ranging from minutes to months.

Administration of the compounds of the present sure to a patient will follow general protocols for the administration of pharmaceuticals, taking into account the toxicity, if any, of the drug. It is expected that the treatment cycles would be repeated as necessary. [023 9] Specific, non-limiting examples of possible combination therapies include use of certain compounds of the invention with the following agents and classes of agents: agents that inhibit DNA methyltransferases such as decitabine or 5’-aza-cytadine; agents that inhibit the activity of histone deacetylases, histone oylases, histone de—ubiquitinases, or histone phosphatases such as hydroxyurea; antisense RNAs that might t the expression of other components of the protein complex bound at the DR site in the gamma globin er; agents that inhibit the action of Klfl or the expression ofKLF1 ; agents that inhibit the action of Bell la or the expression of BCLI IA; and agents that inhibit cell cycle progression such as yurea, ara—C or daunorubicin; agents that induce entiation in leukemic cells such as all-trans retinoic acid (ATRA).

Thus, in another aspect, the present invention provides methods for treating diseases or disorders in a human or animal subject in need of such treatment comprising administering to said subject an amount of a compound disclosed herein effective to reduce or t said disorder in the subject in combination with at least one additional agent for the treatment of said disorder that is known in the art.

Used either as a monotherapy or in combination with other agents, the compounds disclosed herein are useful in the prevention and/or treatment of beta—hemoglobinopathies such as thalassemia major, sickle cell disease, obin E/thalassemia, and thalassemia intermedia.

The compounds disclosed herein can be used in the treatment of diseases in which an increase in transcription h the manipulation of epigenetic regulatory factors such as inhibition of KDMlA would be beneficial to the t. This applies to diseases in which but not d to loss of on mutations, mutations resulting in haploinsuff1ciency, deletions and duplications of genetic material or epigenetic regulatory mechanisms have altered the normal expression pattern of a gene or genes that has the effect of altering the dose of a gene product(s). Such diseases may include diseases both acquired and hereditary in which the expression of, for example, cytokines affecting immune function, are d, X- linked mental retardation and other forms of compromised cognitive or motor on such as Alzheimer and Parkinson disease whether they are the ed or tary forms, lipid disorders such as elevated cholesterol, low density lipoprotein, very low density lipoprotein or triglycerides, both type one and type two diabetes, and Mendelian genetic es.

Other disorders or conditions that can be advantageously treated by the compounds disclosed herein include inflammation and inflammatory conditions.

Inflammatory conditions include, without limitation: arthritis, including sub-types and related conditions such as rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus, juvenile arthritis, acute rheumatic arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, and pyogenic arthritis; osteoporosis, tendonitis, bursitis, and other related bone and joint disorders; gastrointestinal ions such as reflux esophagitis, diarrhea, inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome, ulcerative colitis, acute and c inflammation of the pancreas; ary inflammation, such as that associated with viral infections and cystic fibrosis; skin-related conditions such as psoriasis, eczema, burns, sunburn, dermatitis (such as contact dermatitis, atopic itis, and allergic dermatitis), and hives; pancreatitis, hepatitis, pruritus and vitiligo. In addition, compounds of invention are also useful in organ transplant patients either alone or in combination with tional immunomodulators.

Autoimmune ers may be ameliorated by the treatment with compounds disclosed herein. Autoimmune disorders include Crohn’s disease, ulcerative colitis, dermatitis, dermatomyositis, diabetes mellitus type 1, Goodpasture's syndrome, Graves' e, Guillain-Barré syndrome (GBS), autoimmune encephalomyelitis, Hashimoto's disease, idiopathic thrombocytopenic purpura, lupus erythematosus, mixed connective tissue disease, multiple sclerosis (MS), myasthenia gravis, narcolepsy, pemphigus vulgaris, pernicious , psoriasis, psoriatic tis, polymyositis, primary biliary cirrhosis, rheumatoid arthritis, Sjogren's syndrome, scleroderma, temporal arteritis (also known as "giant cell arteritis"), vasculitis, and Wegener's granulomatosis.

The compounds disclosed herein are also useful for the treatment of organ and tissue injury associated with severe burns, sepsis, trauma, wounds, and hage— or resuscitation—induced nsion, and also in such diseases as vascular diseases, migraine headaches, teritis nodosa, thyroiditis, aplastic , n's disease, sclerodoma, rheumatic fever, type I diabetes, neuromuscular on disease ing enia gravis, white matter disease including multiple sclerosis, sarcoidosis, nephritis, nephrotic syndrome, Behcet's syndrome, ositis, gingivitis, periodontis, ng occurring after injury, chemias including myocardial ischemia, cardiovascular ischemia, and ia secondary to cardiac arrest, and the like.

The compounds disclosed herein are also useful for the treatment of certain diseases and disorders of the nervous system. Central nervous system ers in KDMlA inhibition is useful include cortical dementias including Alzheimer's disease, central nervous system damage resulting from stroke, ischemias including cerebral ia (both focal ischemia, thrombotic stroke and global ischemia (for example, secondary to cardiac arrest), and trauma. Neurodegenerative disorders in which KDMlA tion is useful include nerve degeneration or nerve necrosis in disorders such as hypoxia, hypoglycemia, epilepsy, and in cases of central nervous system (CNS) trauma (such as spinal cord and head injury), hyperbaric oxygen—induced convulsions and toxicity, dementia e.g., pre—senile dementia, and AIDS-related dementia, ia, Sydenham's chorea, Huntington's disease, Parkinson’s Disease, amyotrophic lateral sclerosis (ALS), Korsakoffs disease, cognitive disorders relating to a cerebral vessel disorder, hypersensitivity, sleeping disorders, schizophrenia, depression, depression or other symptoms associated with Premenstrual Syndrome (PMS), and anxiety.

Still other disorders or conditions advantageously treated by the compounds disclosed herein include the prevention or treatment of hyperproliferative diseases, especially cancers, either alone or in ation with standards of care especially those agents that target tumor growth by re-instating tumor suppressor genes in the malignant cells.

Hematological and non-hematological malignancies which may be treated or prevented include but are not limited to multiple myeloma, acute and chronic leukemias and hematopoietic erative and stic disorders including Myelodysplastic Syndrome (MDS), Acute Myelogenous Leukemia (AML), Acute Lymphocytic Leukemia (ALL), Chronic cytic Leukemia (CLL), and Chronic Myelogenous Leukemia (CML), lymphomas, including Hodgkin’s lymphoma and non-Hodgkin’s lymphoma (low, intermediate, and high grade), as well as solid tumors and malignancies of the brain, head and neck, breast, lung (including non-small-cell lung cancer), reproductive tract, upper digestive tract, pancreas, liver, renal system, r, prostate and colorectal. The t compounds and methods can also be used to treat fibrosis, such as that which occurs with radiation y. The present nds and methods can be used to treat subjects having or prevent the progression of atous polyps, including those with familial adenomatous polyposis (FAP) or sarcoidosis. Non—cancerous proliferative disorders additionally include psoriasis, eczema, and dermatitis.

The present nds may also be used in co—therapies, partially or completely, in place of other conventional anti-inflammatory ies, such as er with ds, NSAIDs, COX—2 selective inhibitors, 5-lipoxygenase inhibitors, LTB4 antagonists and LTA4 hydrolase inhibitors. The compounds disclosed herein may also be used to prevent tissue damage when therapeutically combined with antibacterial or antiviral agents.

The compounds disclosed herein are also useful for the treatment of treat metabolic disorders. KDMlA, using flavin adenosine dinucleotide (FAD) as a cofactor, epigenetically regulates energy-expenditure genes in adipocytes depending on the cellular FAD availability. Additionally, loss of KDMlA function induces a number of tors of energy expenditure and mitochondrial metabolism resulting in the activation of mitochondrial respiration. Furthermore, in the adipose s from mice fed a high-fat diet, expression of KDMlA-target genes is reduced.

Metabolic syndrome (also known as metabolic syndrome X) is characterized by having at least three of the following symptoms: insulin resistance; nal fat - in men this is defined as a 40 inch waist or larger, in women 35 inches or larger; high blood sugar levels - at least 110 milligrams per deciliter ) after fasting; high triglycerides - at least 150 mg/dL in the blood stream; low HDL- less than 40 mg/dL; pro-thrombotic state (e. g., high fibrinogen or plasminogen activator inhibitor in the blood); or blood pressure of 130/85 mmHg or higher. A connection has been found between metabolic me and other conditions such as obesity, high blood pressure and high levels of LDL cholesterol, all of which are risk factors for cardiovascular diseases. For example, an increased link between metabolic syndrome and sclerosis has been shown. People with metabolic syndrome are also more prone to developing type 2 diabetes, as well as PCOS (polycystic ovarian syndrome) in women and prostate cancer in men.

As described above, insulin resistance can be manifested in l ways, including type 2 es. Type 2 diabetes is the condition most obviously linked to insulin resistance. Compensatory hyperinsulinemia helps maintain normal glucose levels often for decades before overt es develops. Eventually the beta cells of the pancreas are unable to overcome insulin resistance h ecretion. Glucose levels rise and a diagnosis of es can be made. Patients with type 2 diabetes remain hyperinsulinemic until they are in an advanced stage of disease. As bed above, insulin resistance can also correlate with hypertension. One half of patients with essential hypertension are insulin resistant and hyperinsulinemic, and there is evidence that blood pressure is linked to the degree of insulin resistance. Hyperlipidemia, too, is ated with insulin resistance. The lipid profile of patients with type 2 diabetes includes increased serum very-low—density lipoprotein (VLDL) cholesterol and triglyceride levels and, sometimes, a decreased nsity lipoprotein (LDL) cholesterol level. Insulin resistance has been found in persons with low levels of high— density lipoprotein HDL). n levels have also been linked to VLDL synthesis and plasma triglyceride .

Specific metabolic diseases and symptoms to be treated by the compounds, compositions, and methods disclosed herein are those mediated at least in part by KDMlA.

Accordingly, disclosed herein are methods: for treating insulin resistance in a subject; for reducing glycogen accumulation in a subject; for raising HDL or HDLc, lowering LDL or LDLc, shifting LDL particle size from small dense to normal LDL, lowering VLDL, lowering triglycerides, or inhibiting cholesterol absorption in a subject; for reducing insulin resistance, enhancing glucose utilization or lowering blood re in a subject; for reducing visceral fat in a subject; for reducing serum transaminases in a subject; for inducing mitochondrial respiration in a subject; or for treating e; all comprising the stration of a therapeutic amount of a compound as described herein, to a patient in need thereof. In further embodiments, the disease to be treated may be a metabolic e.

In further ment, the metabolic disease may be selected from the group consisting of: obesity, diabetes mellitus, especially Type 2 diabetes, hyperinsulinemia, glucose intolerance, metabolic syndrome X, dyslipidemia, hypertriglyceridemia, hypercholesterolemia, and hepatic steatosis. In other embodiments, the disease to be treated may be ed from the group ting of: cardiovascular diseases including vascular disease, atherosclerosis, coronary heart disease, cerebrovascular disease, heart e and peripheral vessel disease.

In preferred embodiments, the methods above do not result in the induction or maintenance of a hypoglycemic state.

Besides being useful for human ent, n compounds and formulations disclosed herein may also be useful for nary treatment of companion animals, exotic animals and farm animals, including mammals, rodents, and the like. More preferred animals include horses, dogs, and cats.

Methods General Synthetic Methods for Preparing Compounds The following s can be used to practice the present invention.

Scheme 1 0 o p-F—PhCOCI HOJH W —%NazCos OH 1.Me28-BH3/THF u"\/\/OH m‘ OH dioxane/H2 HOJHwW (\N N o o /N\} HN o NH2 O 63 /°0 2. ylpiperazine DEPBT imidazole % for 2 steps F F Dess Martin O "\x H oxidation; (\NJS W J><©_F v HN’ N\2 HN ‘ \/\/ ’J><©’F —> O H2 (\N / 0 N )S CHZCI2 /N\} HN o Na(OAC)3BH CHZCI2\ F 25% Scheme 1 depicts an example of a synthesis wherein R3 and R5 are each para— fluorophenyl in the final product. However, by tuting reagents wherein the fluorine is ed by another substituent such as methoxy or chlorine, or wherein additional substituents on the phenyl are present, or wherein the phenyl is replaced by another aryl or a heteroaryl in either step 1 or step 4, additional compounds of Formula I can be made. The 2-phenyl-aminocyclopropane substituent can exist in two distinct steric forms that are prepared from the (+) and the (-) forms of the starting material trans-Z-phenyl aminocyclopropane. Further, compounds where n=3 may be prepared from amic acid rather than L-adipic acid by the same methods. Additional variations can be accomplished h methods known in the art.

The invention is further illustrated by the following examples, which have not been made yet or tested. The methods exemplified below may also be extrapolated to compounds disclosed herein which may not yet have not been made or tested.

Intermediate A: (lR,2 S)—2-(4-fluorophenyl)- l -methylcyclopropanamine O O 0 IPT0V /\ A o H |AK") EC (3 KOH "'BUL'DME F F’ : MeOH/HZO F \AQN8°C DPPA TEA ,t-BuOH HCI , \AgNH2 —> —> Toleune MeOH A solution of ethyl 2-(diethoxyphosphoryl)propanoate (3.45 g, 14.48 mmol, 2.00 equiv) in ne glycol dimethyl ether (20 mL) was treated with n-BuLi (2.5M) (5.8 mL) dropwise with stirring at 0°C. The resulting solution was stirred for 30 min at room temperature. To this was added 2-(4-fluorophenyl)oxirane (1 g, 7.24 mmol, 1.00 equiv). The ing solution was stirred for 12 h while the temperature was maintained at 80°C in an oil bath. The reaction mixture was cooled to RT. The reaction was then quenched by the addition of 20 mL of water. The resulting solution was extracted with ethyl e and the organic layers was dried and concentrated. The residue was chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1:100). This resulted in 1 g (62%) of ethyl (1R)(4- fluorophenyl)—1-methylcyclopropanecarboxylate as yellow oil. A solution of ethyl (1R)—2- (4-fluorophenyl)methylcyclopropanecarboxylate (1 g, 4.50 mmol, 1.00 equiv) in methanol/H2O (10/2 mL) and potassium hydroxide (1.26 g, 22.46 mmol, 4.99 equiv) was stirred for 10 h at room temperature. The resulting on was diluted with H2O. The pH value of the solution was adjusted to 2 with hydrochloric acid (2 mol/L). The resulting solution was extracted with ethyl acetate and the c layers ed and dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in 800 mg (92%) of (1R)(4-fluorophenyl)methylcyclopropanecarboxylic acid as yellow oil. A solution of (1R)(4-fluorophenyl)methylcyclopropanecarboxylic acid (400 mg, 2.06 mmol, 1.00 equiv) in toluene (10 mL) was mixed with diphenoxyphosphoryl azide (680 mg, 2.47 mmol, 1.20 , and triethylamine (312 mg, 3.08 mmol, 1.50 equiv). The resulting on was stirred for 30 min at 90°C in an oil bath. Then, tert-butanol (2 mL) was added.

The resulting solution was allowed to react, with stirring, for an additional 12 h while the temperature was maintained at 90°C in an oil bath. The on mixture was cooled to room temperature and the resulting solution was diluted with ethyl acetate. The resulting mixture was washed with H2O. The mixture was dried over anhydrous sodium e and concentrated under vacuum. The residue was chromatographed on a silica gel column and eluted with ethyl acetate/petroleum ether (1:100). This resulted in 350 mg (64%) of tert—butyl N—[(1R)—2-(4-fluorophenyl)—1-methylcyclopropyl]carbamate as yellow oil. A solution of tertbutyl N—[(1R,2S)—2-(4-fluorophenyl)—1-methylcyclopropyl]carbamate (350 mg, 1.32 mmol, 1.00 equiv) in methanol (HCl) (10 mL) was stirred for 2 h at room temperature. The resulting on was diluted with 10 mL of H20. The pH value of the solution was adjusted to 9 with saturated sodium bicarbonatesolution. The ing solution was extracted with 3x10 mL of ethyl acetate and the organic layers combined and dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in 200 mg (92%) of (lR,2S)—2—(4—fluorophenyl)—l— methylcyclopropan-l-amine as yellow oil.

EXAMPLE 1: N—((S)- l -oxo((( l R,2S)—2-phenylcyclopropyl)amino)- l -(pyrrolidin- l - yl)hexanyl)benzamide HN 0 on Scheme for Alkyl—Linked nds HOJH'MWOH O HN O @H 01%..0W0H NJH'MWO HN O Dess Martin C/ HN O émh/lldaZOIe’. .

DCM ,rt 2 h 0 H2N,,' H C/N)S\"\O/VN/hHN Na(OAc)3BH/DCM/rt (S)—2—benzamido—6—hydroxyhexanoic acid was prepared from (S)-amino- hydroxyhexanoic acid. This material (1 g, 3.98 mmol, 1.00 equiv)1n ydrofuran was reacted with 3-(diethoxyphosphoryloxy)—l,2,3-benzotriazin-4(3H)-one (DEPBT) (2.4 g, 8.03 mmol, 2.00 equiv) and imidazole (542 mg, 7.97 mmol, 2.00 equiv). This was followed by the addition of a solution of pyrrolidine (283 mg, 3.98 mmol, l.00 equiv) in tetrahydrofuran at 0°C in 30 min. The resulting solution was stirred for 16 h at room temperature. The solution was diluted with KH2P04(aq.). The s layer was extracted with ethyl acetate and the organic layers were washed with brine and dried over anhydrous sodium sulfate.

After filtration, solvent was removed under reduced pressure. The residue was purified by preparative HPLC and eluted with MeCN with 0.5% NH4HCO3. This resulted in 640 mg (53%) of (S)—N—(6—hydroxy-l-oxo-l-(pyrrolidin-l-yl)hexanyl)benzamide as a light yellow oil. (S)-N—(6-hydroxy-l-oxo-l-(pyrrolidin-l-yl)hexanyl)benzamide (640 mg, 2.10 mmol, l.00 equiv) in dichloromethane (100 ml) was oxidized with Dess-Martin periodinane (DMP) (893 mg, 2. 11 mmol, 1.00 equiv). The resulting solution was stirred for 30 min at 0°C in a water/ice bath and was then diluted with NazSO3(aq.) and NaHCO3(aq.). The aqueous layers were extracted with ethyl acetate and the organic layers were washed with brine and dried over anhydrous sodium sulfate. After tion, solvent was removed under d pressure.

The residue was chromatographed on silica gel and eluted with ethyl e/petroleum ether (10:1). This gave 150 mg (24%) of (S)-N-(l,6-dioxo-l-(pyrrolidin-l-yl)hexan yl)benzamide as a white solid. (S)-N-(l,6-dioxo-l-(pyrrolidin-l-yl)hexanyl)benzamide (150 mg, 0.50 mmol, 1.00 equiv) was dissolved in dichloromethane (25 mL). )—2— phenylcyclopropanamine (66 mg, 0.50 mmol, 1.00 equiv) was added. After stirring 5 minutes, sodium triacetoxyborohydride (252 mg, 1.19 mmol, 2.40 equiv) was added. The resulting solution was stirred for 30 min at 0°C. After the reaction was completed, the resulting on was diluted with sat.NaHCO3. Then it was ted with dichloromethane.

The organic layers were washed with brine and dried over anhydrous sodium sulfate. Solvent was removed under reduced pressure and the residue was ed by Prep-HPLC (CAN/ H20 with 0.5% NH4HCO3). This resulted in 29 mg (14%) of N—((S)—l-oxo(((lR,2S) phenylcyclopropyl)amino)—l-(pyrrolidin-l-yl)hexanyl)benzamide as colorless oil. 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.85(d, J: 7.5 Hz, 2H), 7.60—7.00( m, 8H), 4.85— 4.75(m, 1H), 3.92—3.80 (m, 1H), 3.70—3.30 (m, 4H), 2.74(t, J: 7.2 Hz, 1H), 2.36—2.28(m, lH), 2.07—l.75(m, 7H), l.74—l.37(m, 4H), 1.10—0.95(m, 2H); MS (ES, m/z): 420 (M + H).

EXAMPLE 2: N—((S)- l -oxo((( l R,2S)—2-phenylcyclopropyl)amino)- l -(piperidin- l - anyl)benzamide NJS"\\\/\/ Nh.H H N O [025 9] N—((S)— l —((( l R,2S)phenylcyclopropyl)amino)- l -(piperidin- l -yl)hexan yl)benzamide was prepared in the same manner as was described for the sis of N—((S)— l-oxo((( l R,2S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexanyl)benzamide.

(S)benzamido—6—hydroxyhexanoic acid was coupled with piperidine using 3— (diethoxyphosphoryloxy)-l,2,3-benzotriazin-4(3H)-one and imidazole. The resultant alcohol (S)-N-(6-hydroxy- l -oxo- l -(piperidin- l -yl)hexanyl)benzamide was ed under Dess— Martin conditions to the aldehyde (S)-N-( l ,6-dioxo- l -(piperidin- l -yl)hexanyl)benzamide.

This was coupled with (1R,2S)—2-phenylcyclopropanamine under reductive amination conditions (Na(OAc)3BH) to yield the desired product N—((S)—l—oxo—6—(((1R,ZS) phenylcyclopropyl)amino)-l-(piperidin-l-yl)hexanyl)benzamide as a colorless oil. ES, m/z = 434 (M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.86(d, J= 7.2Hz, 2H), 7.70— 7.40( m, 3H), 7.30—7.15(m, 2H), 7.15—7.08(m, 1H), 7.06(d, J= 7.2Hz, 2H), 5.15—5.00(m, 1H), 3.80—3.60(m, 2H), 3.60—3.40(m, 2H), 2.34(t, J= 7.2Hz, 2H), 2.40—2.30(m, 1H), 2.10—1.40(m, 4H), 1.15—1.00(m, 2H).

E 3: 4-fluoro-N—((S)((( 1R,2 S)(4-fluorophenyl)cyclopropyl)amino)- l -(4- methylpiperazin- l -yl)- l -oxohexanyl)benzamide NJSMWNH 4—fluoro—N—((S)—6—((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -(4- methylpiperazin-l-yl)-l-oxohexanyl)benzamide was prepared in a manner analogous to the previous example. The alcohol 4—fluoro—N—((S)—6—(((1R,ZS)—2—(4— fluorophenyl)cyclopropyl)amino)- l -(4-methylpiperazin- l -yl)- l -oxohexan-2—yl)benzamide was prepared by reduction of (S)—2—(4—fluorobenzamido)hexanedioic acid with H3.

This type of reduction was used to prepare similar ls (e. g. The alcohol starting material (S)benzamidohydroxyhexanoic acid for the synthesis of -l-oxo(((1R,ZS) phenylcyclopropyl)amino)-l-(pyrrolidin-l-yl)hexanyl)benzamide). Into a 1000-mL 3- necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed a solution of (S)—2—(4—fluorobenzamido)hexanedioic acid (10 g, 35.30 mmol, 1.00 equiv) in tetrahydrofuran (300 ml). Then a on of MezS—BH3 (11 mL, 3.00 equiv) in ydrofuran (50 ml) was added at 0°C. The resulting on was stirred for 3 h at 0°C in an ice/salt bath. The on was then quenched by the addition of 20 ml of methanol. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 300 ml of sat.Na2CO3. The resulting solution was extracted with 3x100 mL of ethyl acetate and the aqueous layers combined. The pH value of the solution was adjusted to 2 with hydrochloric acid(2 mol/L). The resulting solution was extracted with 3x200 ML of ethyl acetate and the c layers combined. The resulting mixture was washed with 1x500 mL of brine. The mixture was dried over anhydrous sodium sulfate. The solids were filtered out.

The resulting mixture was concentrated under vacuum. This resulted in 6 g (63%) of (S)—2— (4-fluorobenzamido)hydroxyhexanoic acid as colorless oil. This material was reacted with yl piperazine followed by Dess-Martin oxidation and coupling via reductive amination with (lR,2S)(4-fluorophenyl)cyclopropanamine in the manner bed for the synthesis of N—((S)— l -oxo(((lR,2 S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexan- 2-yl)benzamide to yield the desired product 4—fluoro—N—((S)—6—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)- l thylpiperazin- l -yl)- l -oxohexanyl)benzamide as colorless oil. ES, m/s =485 *M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.83 (dd, Hz, J2=l.4Hz, 2H), 7.18—7.04 (m, 3H), 7.00—6.87 (m, 4H), .05 (m, 1H), 3.78— 3.50 (m, 4H), 2.71 (t, J=6.9Hz, 2H), 2.30 (s, 3H), 2.28—2.21 (m, 1H), .78 (m, 2H), 1.72—1.31 (m, 9H), 1.07—0.96 (m, 1H), 0.94—0.86 (m, 1H).

EXAMPLE 4: N—((S)- l -(4-methylpiperazin- l -yl)- l -oxo((( l R,2 S)—2- phenylcyclopropyl)amino)hexanyl)benzamide N—((S)- l -(4-methylpiperazin- l -yl)- l -oxo(((lR,2S) cyclopropyl)amino)hexanyl)benzamide was prepared in the same manner as was described for the synthesis of N—((S)-l-oxo—6—(((lR,2S)phenylcyclopropyl)amino)—l— (pyrrolidin-l-yl)hexanyl)benzamide. (S)benzamidohydroxyhexanoic acid was coupled with N—methyl piperidine using 3-(diethoxyphosphoryloxy)-l,2,3-benzotriazin- 4(3H)—one ) and imidazole. The resultant alcohol (S)-N—(6-hydroxy-l-(4- methylpiperazin-l-yl)- l -oxohexanyl)benzamide was oxidized under Dess-Martin conditions to the aldehyde (S)-N-(l-(4-methylpiperazin-l-yl)—l,6-dioxohexan yl)benzamide. This was coupled with (lR,2S)phenylcyclopropanamine under reductive amination conditions (Na(OAc)3BH) to yield the desired N—((S)—l—(4—methylpiperazin—l—yl)— 6-(((lR,2S)phenylcyclopropyl)amino)hexanyl)benzamide as a colorless oil. 1H NMR (300 MHz, CD3OD—d4) 5 ppm: .80 (m, 2H), 7.60—7.42 (m, 3H), 7.26—7.18 (m, 2H), 7.15—7.02 (m, 3H), 5.03 (dd, J: 8.1 Hz, 6.0 Hz, 1H), 3.85—3.48 (m, 4H), 2.73(t, J: 7.2 Hz, 2H), 2.60—2.35 (m, 4H), 2.35—2.25 (m, 4H), 1.95—1.72 (m, 3H), 1.70—1.38 (m, 4H), 1.10— 0.95 (m, 2H); MS (ES, m/z): 449 (M + H).

EXAMPLE 5: 4—fluoro—N—((R)—3 —((2—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropanyl)benzamide ANJSJOWN/hH ) HN o 4—fluoro—N—((R)—3—((2—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropanyl)benzamide was prepared by the method described for N—((S)—l—oxo—6—(((lR,2S)-2— phenylcyclopropyl)amino)— l -(pyrrolidin- l -yl)hexanyl)benzamide. (S)—2-benzamido hydroxyhexanoic acid was reacted with diethyl amine with thoxyphosphoryloxy)-l,2,3- riazin-4(3H)-one (DEPBT)/imidazole to give (S)—N—(l-(diethylamino)hydroxy-l- oxohexan—2—yl)benzamide in 45% yield as a colorless oil. This was ed under Dess Martin ions to give the aldehyde (S)-N—(l-(diethylamino)—l,6-dioxohexan yl)benzamide in 45% yield as a yellow oil. The aldehyde was reacted with (lR,2S) phenylcyclopropanamine under ive amination conditions (Na(OAc)3BH) to give N— ((S)- l -(diethylamino)- l -oxo((( l R,2S)phenylcyclopropyl)amino)hexanyl)benzamide as a light yellow oil (6% yield). ES, m/z = 422 (M+H). 1H NMR (300 MHz, CD3OD-d4) 5 ppm: 7.85 (dd, J1: 5.25 Hz, J2: 1.65Hz, 2H), .40 (m, 3H), 7.22 (t, J: , 2H), 7.15—7.00 (m, 3H), 4.94—5.05 (m, 1H), 3.60—3.45 (m, 3H), 3.30—3.21 (m, 1H), 2.73 (t, J: 7.2Hz, 1H), 2.27—2.35 (m, 1H), 1.79—1.72 (m, 3H), 1.67—1.39 (m, 4H), 1.31 (t, J=7.05Hz, 3H), 1.14 (t, J=7.05Hz, 3H), 1.10—0.95 (m, 2H) EXAMPLE 6: N—((S)- l -morpholino- l -oxo((( l R,2 S)phenylcyclopropyl)amino)hexan- 2-yl)benzamide .u‘\/\/ N "I N—((S)— l —morpholino- l -((( 1R2 S)phenylcyclopropyl)amino)hexan-2— yl)benzamide was prepared by the method that was described r for N—((S)—l—oxo—6— ((( 1R2 S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexan-2—yl)benzamide. (S) benzamidohydroxyhexanoic acid was reacted with morpholine, 3- (diethoxyphosphoryloxy)-l,2,3-benzotriazin-4(3H)-one (DEPBT) and imidazole to give (S)- N—(6—hydroxy—l—morpholino—l—oxohexan-2—yl)benzamide in 37% yield as a colorless oil.

This was oxidized under Dess-Martin conditions to give (S)-N—(l-morpholino-l,6- dioxohexan-2—yl)benzamide in 45% yield as a ess oil. .

This material was reacted with (lR,2S)phenylcyclopropanamine under reductive amination conditions (Na(OAc)3BH) to give, after prep-hplc, a 7% yield of N—((S)-l-morpholino-l-oxo(((lR,2S)-2— phenylcyclopropyl)amino)hexan-2—yl)benzamide as a light yellow oil. ES, m/z = 436 (M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.85(d, J = 6.9Hz, 2H), .50( m, 1H), 7.46(t, J =7.35Hz, 2H), 7.22 (t, J =7.35Hz, 2H), .08(m, 1H), 7.05 (d, J =6.9Hz, 2H), .00(t, J =7.05Hz, 1H), 3.80—3.48(m, 8H), 2.32(t, J =3.0Hz, 2H), 2.36—2.08(m, 1H), 1.95— , 1H), l.86—l.72(m, 2H), l.70—l.54(m, 2H), l.54—l.40(m, 2H), 0.97—1.12(m, 2H) E 7: N—[(2S)[[(1R,2 S)(4-fluorophenyl)cyclopropyl]amino]— l -oxo- l - (piperidin- l -yl)hexanyl]pyridinecarboxamide O NJSN‘WNMH HN O N—[(2S)[[( l R,2S)(4-fluorophenyl)cyclopropyl]amino]— l -oxo- l -(piperidin- l - yl)hexanyl]pyridine—2—carboxamide was prepared by the method that was described for of N—((S)— l —oxo—6—((( l R,2S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexan-2— yl)benzamide. A 240 mg sample of(S)-N—(6-hydroxy-l-oxo-l-(piperidin-l-yl)hexan yl)picolinamide was converted under Dess—Martin conditions to the aldehyde (l,6— dioxo-l-(piperidin-l-yl)hexanyl)picolinamide as a yellow oil. Under reductive amination conditions with (lR,2S)—2-(4-fluorophenyl)cyclopropanamine the aldehyde gave the product N—((S)—6—((( l R,2S)—2—(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l -yl)hexan-2— yl)picolinamide as a light yellow oil. ES, m/z =453 (M+l). 1H NMR (300MHz, DMSO— d6, 5): 8.65 (d, J= 3.3 Hz, 1H), 8.09 (d, J= 7.8 Hz, 1H), 8.03—7.94 (m, 1H), 7.65—7.42 (m, 1H), 7.15—6.89 (m, 4H), 5.05—5.18 (m, 1H), 3.74—3.43 (m, 4H), 2.70 (t, J: 7.4 Hz, 2H), 2.32— 2.17 (m, 1H), 1.99—1.79 (m, 2H), 1.80—1.38 (m, 11H), 1.07—0.89 (m, 2H).

EXAMPLE 8: N-((S)—6-(((1R,2 S)—2-(4-fluorophenyl)- l -methylcyclopropyl)amino)— l -(4- lsulfonyl)piperazin- l -yl)- l -oxohexanyl)benzamide O§,© NJH"\\\/\/Nl"H HN O N—((S)—6—((( l R,2 S)—2-(4-fluorophenyl)- l -methylcyclopropyl)amino)- l -(4- (methylsulfonyl)piperazin-l-yl)-l-oxohexanyl)benzamide was prepared with a modification of the method that used that for N—((S)—l-oxo(((lR,2S) phenylcyclopropyl)amino)-l-(pyrrolidin-l-yl)hexanyl)benzamide. The key l intermediate (S)-N—(6-hydroxy- l -(4-(methylsulfonyl)piperazin- l -yl)- l -oxohexan yl)benzamide was prepared in a slightly different manner than was described earlier. (2S) aminomethoxyoxohexanoic acid was first reacted with l de to give (S)—2— benzamidomethoxyoxohexanoic acid. This was converted to the amide (S)—methyl 5- ido(4-(methylsulfonyl)piperazin- l -yl)oxohexanoate with l-methane sulfonylpiperazine/HATU and DIEA. The ester was hydrolyzed with LiOH in methanol/water to yield the acid (S)benzamido(4-(methylsulfonyl)piperazin-l-yl)—6- oxohexanoic acid in 72% yield as a yellow solid. This was reduced with BH3/THF to give the alcohol (S)—N—(6-hydroxy- l -(4-(methylsulfonyl)piperazin- l -yl)- l -oxohexan yl)benzamide in 59% yield as a yellow oil. This was converted to the mesylate (S)—N—(l—(4— (methylsulfonyl)piperazin- l -yl)-l,6-dioxohexanyl)benzamide using e sulfonyl chloride and triethyl amine. The mesylate was a white solid. Yield was 40%. The mesylate was reacted in sn2 fashion with (S)benzamido(4-(methylsulfonyl)piperazin-l-yl) anoic acid in the ce of DIEA/Kl in acetonitrile to give N—((S)—6-(((lR,2S)(4- fluorophenyl)- l lcyclopropyl)amino)— l -(4-(methylsulfonyl)piperazin- l -yl)- l - oxohexanyl)benzamide as a white solid in 18% yield. ES, m/z = 545 (M+H). 1H NMR (300 MHz, CDCl3, ppm): 7.80—7.83(m, 2H), 7.51—7.53(m, 3H), 7.07—7.l2(m, 2H), 6.92— 7.01(m, lH), 5.13—5.18(m, lH), 3.88—4.05(m, 2H), 3.43—3.92(m, 4H), 3.09—3.2l(m, 2H), 2.72— 2.80(m, 5H), 2.12—2.l7(m, lH), l.50—l.89(m, 6H), 0.81—l.ll(m, 4H) N—((S)— l —((( l R,2S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexan- enzamide: This preparation is similar to that of N-((S)—l-oxo(((lR,2S)—2- phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexanyl)benzamide via Dess-Martin oxidation and reductive amination to form product. However, the synthesis of intermediate (S)-N—(6-hydroxy- l -(4-(methylsulfonyl)piperazin- l -yl)- l -oxohexanyl)benzamide ed in that this method gave superior optical purity.

EXAMPLE 9: N-((S)—6-(((1R,2 S)—2-(4-fluorophenyl)cyclopropyl)amino)- l -(4- (methylsulfonyl)piperazin- l -yl)- l -oxohexanyl)benzamide O /O \\S/ O [N] O HOJSMW\ CH N OH H (\NJS W‘0 NHZOHHCI N EDC|,HOBt,DCM,rt Oe ,Nd N NH20H(aq-)/MeOH/60°C U .9 \ / O O (\NJW- \/\/ O .\\ OH ©__< (\NJH \/\/OH\ O\\ ,Nd NHZ CI 0\ ,Nd HN O artin,DCM fi\ NEt3, THF, rt .\ rt 0 o N—((S)—6—((( l R,2 S)—2—(4-fluorophenyl)cyclopropyl)amino)- l -(4- (methylsulfonyl)piperazin-l-yl)-l-oxohexanyl)benzamide. A solution of (S)—2-(2,5- dimethyl-lH-pyrrol-l-yl)hydroxyhexanoic acid (820 mg, 3.64 mmol, 1.00 equiv) in dichloromethane l-methanesulfonylpiperazine (2.18 g, 13.27 mmol, 3.00 equiv), l—3- (3 -dimethylaminopropyl)carbodiimide "EDCI"(1.7 g, 2.00 equiv) and hydroxybenzatriazole, "HOBT"(1.2 g, 2.00 equiv) was stirred for l h at room temperature. After the reaction was completed, the reaction was quenched with water and extracted with dichloromethane. The organic layers were washed with brine, dried over sodium sulfate, concentrated, and chromatographed on silica gel and eluted with ethyl e/petroleum ether (1:3). This resulted in 340 mg (25%) of (S)(2,5-dimethyl-lH-pyrrol-l-yl)—6-hydroxy-l-(4- (methylsulfonyl)piperazinyl)hexanone as a yellow solid. A solution this material (440 mg, 1.18 mmol, 1.00 equiv) in ethanol was treated with a solution ofNH20H.HCl (430 mg, .20 equiv) in water and NHzOH (1.97 g, 28.70 equiv). The resulting solution was d for 6 days at 80°C. The reaction mixture was concentrated under vacuum and quenched with ice water. The pH was ed to pH 10 with aqueous NaOH and the e was ted with ethyl acetate. The organics were concentrated and the residue tographed on silica gel and eluted with dichloromethane/methanol (10:1). This resulted in 210 mg (60%) of (S)aminohydroxy(4-(methylsulfonyl) piperazin-l-yl)hexanone as a solid. A 322 mg sample of this material (1.1 mmol, 1.0 equiv in tetrahydrofuran and NEt3 (133 mg, 1.32 mmol, 1.20 equiv was reacted with a solution of benzoyl chloride (185 mg, 1.32 mmol, 1.20 equiv) in tetrahydrofuran at 0°C. during addition and stirred for 1h at room temperature. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with brine, dried, and concentrated. The residue was chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1:5). This resulted in 304 mg (70%) of (S)— 2-aminohydroxy(4-(methylsulfonyl)piperazinyl)hexanone as colorless oil. This (S)aminohydroxy(4-(methylsulfonyl)piperazinyl)hexanone was oxidized to the corresponding aldehyde in the manner bed for N-((S)oxo(((1R,2S) phenylcyclopropyl)amino)(pyrrolidinyl)hexanyl)benzamide. Yield 66%. The resulting (S)—N—(1-(4-(methylsulfonyl)piperazinyl)-1,6-dioxohexanyl)benzamide was reacted with (1R,2 4-fluorophenyl)cyclopropanamine under the reductive amination conditions described earlier to yield the desired N—((S)—6—(((1R,2S)—2—(4— fluorophenyl)cyclopropyl)amino)(4-(methylsulfonyl)piperazinyl)oxohexan yl)benzamide as a ess oil, yield 14%. ES, m/z =457 (M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.86 (d, z, 2H), 7.43—7.60 (m, 3H), 6.90—7.12 (m, 4H), 5.03 (t, J=7.05Hz, 1H), 3.89—4.03 (m, 2H), 3.62—3.77 (m, 1H), 3.45—3.58 (m, 1H), 3.33—3.45 (m, 2H), 3.20—3.330 (m, 1H), 3.08—3.20 (m, 1H), 2.87 (s, 3H), 2.73 (t, J=8.2Hz, 2H), 2.25—2.32 (m, 1H), 1.78—1.95 (m, 3H), 1.40—1.68 (m, 4H), 0.92—1.08 (m, 2H).

EXAMPLE 10: N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l - (piperidin- l -yl)hexanyl)benzamide NAB-"\WN/nH HN O N—((S)—6—((( l R,2S)—2—(4—fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l - yl)hexan—2—yl)benzamide was prepared by the same method that was described for —l— oxo(((1R,2 S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexanyl)benzamide. (S)- 2-(2,5-dimethyl-lH-pyrrol-l-yl)hydroxyhexanoic acid was d with piperidine/EDCl/HOBt to yield (S)—2-(2,5-dimethyl- l H-pyrrol- l -yl)hydroxy- l -(piperidinl-yl )hexan-l-one. This was deprotected with hydroxyl amine in methanol to yield (S) aminohydroxy-l-(piperidin-l-yl)hexan-l-one which could be reacted with benzoyl chloride and ylamine to yield (6-hydroxy-l-oxo-l-(piperidin-l-yl)hexan yl)benzamide. This alcohol was oxidized under Dess-Martin conditions to yield the de (S)-N—(l,6-dioxo-l-(piperidin-l-yl)hexanyl)benzamide. The aldehyde was, in turn, coupled with (lR,2S)—2-(4-fluorophenyl)cyclopropanamine under ive amination conditions (Na(OAc)3BH) to yield the product N—((S)—6—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l -yl)hexanyl)benzamide as a light yellow oil. 1H NMR (300 MHz, d4) 5 ppm: 7.92—7.83(m, 2H), 7.60—7.45( m, 3H), 7.12—7.00(m, 2H), 7.00—6.90 (m, 2H), 5.07(dd, J: 8.1 Hz, 5.7 Hz, 1H), 3.75—3.42 (m, 4H), 2.72(t, J: 7.2 Hz, 2H), 2.31—2.25(m, 1H), l.95—l.40(m, 13H), l.lO—0.90(m, 2H); MS (ES, m/z): 452 (M + H).

EXAMPLE 1 l: N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -morpholino- l - oxohexanyl)benzamide CC} "‘\\/\/N’I.

HN o h N—((S)—6—((( l R,2S)—2—(4—fluorophenyl)cyclopropyl)amino)- l -morpholino- l - oxohexan—2—yl)benzamide was prepared by the same method that was described for N—((S)—l— oxo(((1R,2 S)phenylcyclopropyl)amino)- l olidin- l -yl)hexanyl)benzamide. (S)- 2-(2,5-dimethyl-lH-pyrrol-l-yl)hydroxyhexanoic acid was reacted with line/EDCl/HOBt to yield (S)(2,5-dimethyl- lH-pyrrol- l -hydroxy- l - morpholinohexan-l-one with a 69% yield. This was deprotected with hydroxyl amine in methanol to yield (S)—2—aminohydroxy—l—morpholinohexan—l—one which could be reacted with benzoyl chloride and triethylamine to yield (S)-N—(6-hydroxy-l-morpholino- l - oxohexanyl)benzamide with a 63% yield on deprotection and 69% on amide formation.

This alcohol was oxidized under Dess-Martin conditions to yield the aldehyde (S)—N—(l— morpholino-l,6-dioxohexanyl)benzamide with a 70% yield. The aldehyde was, in turn, coupled with (lR,2S)—2-(4-fluorophenyl)cyclopropanamine under reductive amination conditions (Na(OAc)3BH) to yield the product N—((S)—6—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)-l-morpholino-l-oxohexanyl)benzamide with a yield of 21% after purification by prep HPLC. ES, m/z=454 (M+H). 1H NMR (300 MHz, CD3OD— d4) 5 ppm: 7.85 (dd, J1=5.1Hz, J2=l.8Hz 2H), 7.83—7.45 (m, 3H), 7.09—7.04 (m, 2H), 6.98— 6.92 (m, 2H), 5.03 (d, J=4.05Hz, 1H), .67 (m, 8H), 2.72 (d, J=7.2Hz, 2H), 2.29—2.27 (m, 1H), 0.95—1.90 (m, 10H) EXAMPLE l2: N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -(4- methylpiperazin- l -yl)- l -oxohexanyl)benzamide N’I.H N—((S)—6—((( l R,2S)—2—(4-fluorophenyl)cyclopropyl)amino)— l -(4-methylpiperazin- l-yl)—l-oxohexanyl)benzamide was prepared by the same method that was bed for N—((S)— l —oxo—6—((( l R,2S)—2-phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexan yl)benzamide. (S)-N—(6-hydroxy- l thylpiperazin- l -yl)- l -oxohexanyl)benzamide was prepared in the usual way. This l was oxidized under Dess-Martin conditions to yield the aldehyde (S)—N—(l-(4-methylpiperazin- l -yl)-l,6-dioxohexanyl)benzamide as a yellow solid with a 75% yield. The aldehyde was, in turn, coupled with (lR,2S)—2-(4- fluorophenyl)cyclopropanamine under reductive amination conditions (Na(OAc)3BH) to yield the product N—((S)—6—(((lR,2S)(4-fluorophenyl)cyclopropyl)amino)— l -(4- piperazin-l-yl)-l-oxohexanyl)benzamide with a yield of 3% after purification by prep HPLC on a chiral column. ES, m/z=467 (M+l). H—NMR: (CD3OD, ppm): 7.86—7.85 2014/049906 (d, J = 1.8 Hz, 2H), 7.59—7.52 (m, 1H), 7.50—7.41 (m, 2H), 7.12—7.01 (m, 2H), .88 (t, J = 8.7 Hz, 2H), 5.01—5.12 (d, J= 6 Hz, 1H), 3.86—3.69 (m, 2H), 3.67—3.43 (m, 2H), 2.66—2.78 (m, 2H), 2.56—2.39 (m, 4H), .21 (m, 4H), 1.98—1.73 (m, 3H), 1.64—1.38 (m, 4H), 0.91— 1.11 (m, 2H) EXAMPLE l3: 4-fluoro-N—((S)((( l R,2 S)(4-fluorophenyl)cyclopropyl)amino)— l -oxo- l - (piperidin- l xanyl) NJR-MWN’I.H HN O 4—fluoro—N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l - (piperidin- l -yl)hexan-2 -yl) (S)fluoro-N—(6-hydroxy- l -oxo- l -(piperidin- l -yl)hexan yl)benzamide was prepared in a manner similar to that exemplified in the synthesis of N—((S)- l-oxo((( l R,2S)phenylcyclopropyl)amino)- l -(pyrrolidin- l -yl)hexanyl)benzamide.

The alcohol precursor fluoro—N—(6—hydroxy— l —oxo— l -(piperidin— l —yl)hexan—2— yl)benzamide was oxidized under des-Martin conditions and the resultant aldehyde was coupled with (lR,2S)—2-(4-fluorophenyl)cyclopropanamine under the usual reductive amination conditions to yield the desired product 4—fluoro—N—((S)—6—(((lR,2S)—2—(4— fluorophenyl)cyclopropyl)amino)- l -oxo- l ridin- l -yl)hexanyl) as a colorless oil.

ES, m/z = 470 (M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.91—7.74 (m, 2H), 7.20 (m, 2H), 7.01—7.12 (m, 2H), 6.94 (t, 2H), 5.05 (t, J=6.9Hz, 1H), 3.42—3.73 (m, 4H), 2.73 (t, J=7.2Hz, 2H), 2.25—2.33 (m, 1H), .97 (m, 13H), 0.92—1.08 (m, 2H) EXAMPLE l4: N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l - (piperidin- l -yl)hexanyl)(trifluoromethyl)benzamide NJS-MWNMH HN O 2014/049906 N—((S)—6—((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l - yl)hexanyl)(trifluoromethyl)benzamide was prepared in a manner similar to that exemplified in the synthesis of N—((S)-l-oxo(((lR,2S)phenylcyclopropyl)amino)-l- (pyrrolidin- l -yl)hexanyl)benzamide. The alcohol (S)-N—(6-hydroxy- l -oxo- l -(piperidin- l - yl)hexanyl)—4-(trifluoromethyl)benzamide was oxidized under des-Martin conditions and the resultant aldehyde (S)-N—(l,6-dioxo-l-(piperidin-l-yl)hexanyl)—4- (trifluoromethyl)benzamide was coupled with (lR,2S)(4-fluorophenyl)cyclopropanamine under the usual reductive amination conditions to yield the desired product N—((S)—6— ((( 1R2 S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l -yl)hexanyl) (trifluoromethyl)benzamide as an off-white semi-solid. ES, m/z = 520 (M+l). H-NMR (CD3OD,ppm): 8.14—7.89 (m, 2H), 7.88—7.71 (d, J: 7.5 Hz, 2H), 7.26—6.83 (m, 4H), 5.13 (s, 1H), .81 (m, 2H), 3.58—3.38 (m, 2H), 3.02—2.71 (b, 2H), .33 (b, 1H), .95 (b, 1H), 1.91—1.36 (m, 12H), 1.27—1.11 (m, 2H).

EXAMPLE 15: N—((S)((( l R,2S)(4-fluorophenyl)cyclopropyl)amino)- l -oxo- l - (piperidin-l-yl)hexanyl)-[1,1'-biphenyl]carboxamide NJS-‘WN/I.H HN o N—((S)—6—((( l R,2S)—2—(4—fluorophenyl)cyclopropyl)amino)- l -oxo- l -(piperidin- l - yl)hexanyl)-[l, l'-biphenyl]carboxamide was prepared by the same method that was bed for — l —oxo—6—(((lR,2S)—2—phenylcyclopropyl)amino)— l —(pyrrolidin— l - yl)hexanyl)benzamide. (S)aminohydroxy- l -(piperidin- l -yl)hexanone was d with 4-phenylbenzoyl chloride and triethylamine to yield (S)-N—(l,6-dioxo-l- (piperidin-l-yl)hexanyl)-[l,l'-biphenyl]carboxamide. This alcohol was oxidized under Dess—Martin conditions to yield the aldehyde (S)-N-(l,6-dioxo-l-(piperidin-l-yl)hexanyl)- [1,1'-biphenyl]carboxamide. The aldehyde was, in turn, coupled with (lR,2S)(4- henyl)cyclopropanamine under reductive amination conditions (Na(OAc)3BH) to yield the product N—((S)—6—(((lR,2S)(4-fluorophenyl)cyclopropyl)amino)-l-oxo-l- (piperidin—l—yl)hexan—2-yl)—[l,l'—biphenyl]—4—carboxamide as a colorless oil. ES, m/z: 528 (M+1) .

H—NMR: , ppm): 7.94 (d, J: 8.4 Hz, 2H), 7.90—7.65 (m, 4H), 7.60—7.35 (m, 3H), 7.20—7.00 (m, 2H), 7.00-6.90 (m, 2H), 5.12 (t, .1: 7.2 Hz, 1H), 3.85—3.40 (m, 4H), 2.74 (t, .1: 7.2 Hz, 1H), 2.40—2.30 (m, 1H), 2.00—1.45 (m, 13H), 1.15-0.85 (m, 2H).

EXAMPLE l6: N—((S)— l -( l , l -dioxidothiomorpholino)((( l R,2S)(4- fluorophenyl)cyclopropyl)amino)- l -oxohexanyl)benzamide 0:9 N'UH HN o h o F N—((S)- l -(l , l-dioxidothiomorpholino)—6-((( l 2-(4- fluorophenyl)cyclopropyl)amino)-l-oxohexanyl)benzamide was prepared in a manner similar to that described for —l-oxo(((lR,2S)phenylcyclopropyl)amino)-l- (pyrrolidin-l-yl)hexanyl)benzamide. Dess-Martin oxidation resulted in 80 mg (80%) of )-l-(l,l-dioxo-l [6],4-thiomorpholinyl)—l,6-dioxohexanyl]benzamide as a yellow solid. Coupling under reductive amination conditions with (lR,2S)—2—(4— fluorophenyl)cyclopropanamine yielded the desired product in 24% yield as an off white solid. ES, m/z =502 (M+l). H—NMR—:(DMSO—d6, ppm): 8.90 (d, J: 7.2 Hz, 2H), 7.89 (d, J = 7.2 Hz, 2H), 7.46 (d, J: 7.2 Hz, 2H), 7.15—6.90 (m, 4H), .80 (m, 1H), 4.28—4.05 (m, 2H), 3.95—3.80 (m, 1H), 3.75—3.55 (m, 1H), 3.38—3.12 (m, 3H), 3.12—2.95 (m, 1H), 2.70—2.45 (m, 2H), 2.25—2.15 (m, 1H), 1.85—1.60 (m, 3H), 1.55—1.30 (m, 4H),l.00—0.80 (m, 2H).

EXAMPLE l7: 4-fluoro-N—((R)-3 (( l R,2S)(4-fluorophenyl)- l - methylcyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropanyl)benzamide Reaction Scheme for SOz—Linker O O HOJH"\\\SH O HO \/OH HN O Br/\/OH HN O (\NJSJ\\\S/\/OH DEPBT,imidazole 0d HN O K3C03, DMF, 0 °C-rt morpholine, THF,rt,15h 68% 60% F F \\ OTBS l.\\8\ IBVOTBS pN SN N S TBSCI, imidazole m-CPBA —> TBAF DCMJLB" OJ HN O 0d HN O DCMrtGh 60'0/ THF, rt, 10h o HZN, .0‘80? (\N S/\/0H (\N ’NOMS 0d HN O MsCI 0d HN O —> F TEA,THF,rt,2h DIEA,KI, MeCN,5o°c .39 H (\N .x‘ S/\/ ll, 0d HN o 4—fluoro—N—((R)-3 -((2-(((1R,2 S)(4-fluorophenyl)- l - methylcyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropanyl)benzamide.

A solution of (R)(4-fluorobenzamido)mercaptopropanoic acid (5 g, 20.55 mmol) in N,N—dimethylformamide (50 mL) was stirred with potassium methaneperoxoate (5.7 g, 40.94 mmol). This was followed by the addition of a solution of 2-bromoethan-l-ol (2.8 g, 22.41 mmol) in N,N—dimethylformamide (20 mL) dropwise with stirring at 0°C. The resulting solution was stirred for 5 h at room ature. The resulting solution was diluted with 200 mL of H20. The pH value of the on was adjusted to 3 with hydrochloric acid (2 mol/L).

The resulting solution was extracted with ethyl acetate and the organic layers ed and dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was chromatographed on silica gel and eluted with dichloromethane/methanol (20: 1). This ed in 4 g (68%) of (R)(4-fluorobenzamido)-3 -((2-hydroxyethyl)thio)propanoic acid as yellow oil. (R)fluoro-N—(3-((2-hydroxyethyl)thio)morpholinooxopropan yl)benzamide as yellow oil. To a solution of (4-fluorobenzamido)((2- hydroxyethyl)thio)propanoic acid (4 g, 13.92 mmol, 1.00 equiv) in tetrahydrofuran (50 mL), was added 3—(diethoxyphosphoryloxy)—1,2,3—benzotriazin—4(3H)-one (DEPBT) (6.25 g, 20.90 mmol, 1.50 equiv) and imidazole (1.42 g, 20.88 mmol, 1.50 equiv).The mixture was stirred for 30 minutes at 0°C. Then morpholine (1.2 g, 13.77 mmol, 0.99 equiv) was added. The resulting solution was stirred for 12 h at room temperature. The resulting solution was diluted with 200 mL of ethyl acetate. The resulting mixture was washed with brine. The organic layers was dried over ous sodium sulfate and concentrated under vacuum. The residue was chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1: 10). This resulted in 3 g (60%) of the desired product. To a solution of this (R)—4—fluoro—N—(3—((2— hydroxyethyl)thio)morpholinooxopropanyl)benzamide (3 g, 8.42 mmol, 1.00 equiv) in dichloromethane (30 mL) and imidazole (1.14 g, 16.76 mmol, 1.99 equiv) was added tert— imethylsilyl chloride "TBSCl" (1.9 g, 12.58 mmol, 1.49 equiv), dropwise at 0°C. The resulting solution was stirred for 6 h at room ature. The reaction was then quenched by the addition of water. The resulting solution was extracted with romethane and the organic layers combined and dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1 :30). This ed in 2 g (50%) of (R)—N—(3—((2—((tert— butyldimethylsilyl)oxy)ethyl)thio)morpholinooxopropanyl)fluorobenzamide which was white solid. A solution of (R)-N—(3-((2-((tert-butyldimethylsilyl)oxy)ethyl)thio) linooxopropanyl)fluorobenzamide (2 g, 4.25 mmol, 1.00 equiv) and meta— chloroperbenzoic acid, " m-CPBA" (1.84 g, 10.66 mmol, 2.51 equiv)was for 6 h at room temperature. This was d with of DCM. The resulting mixture was washed with of saturated sodium bicarbonate solution. This was washed with of brine. The organic layers was dried over anhydrous sodium sulfate and concentrated under . The residue was chromatographed column with ethyl acetate/petroleum ether (1 :30). This resulted in 1.3 g (61%) of (R)-N—(3 -((2-((tert-butyldimethylsilyl)oxy)ethyl)sulfonyl)morpholino oxopropanyl)fluorobenzamide as a white solid. This was dissolved in THF and treated with tetrabutylammonium fluoride, "TBAF" with stirring for 10h. The on was diluted with ethyl acetate and washed with brine. The mixture was dried over anhydrous sodium sulfate and concentrated under . The residue was chromatographed on a silica gel column and eluted with ethyl acetate/petroleum ether (1:20). This resulted in 300 mg (78%) of (R)fluoro-N—(3 -((2-hydroxyethyl)sulfonyl)- l -morpholino- l -oxopropan-2—yl)benzamide as yellow oil. This alcohol was converted to the mesylate with methanesulfonyl de, "MsCl" and triethyl amine and the mesylate was reacted with )(4-fluorophenyl)-l- cyclopropanamine to yield 4-fluoro-N—((R)((2-(((lR,ZS)(4-fluorophenyl)-l- methylcyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropanyl)benzamide in a 28% yield as a white solid. : [M+H]= 536; H-NMR (400MHz ,CDCl3): 87.87~7.82 (m, 2H), 7.l7~7.07 (m, 4H), .99~6.95 (m, 2H),5.70~5.65 (m, 1H), 3.81~3.64 (m, 10H), 3.42~3.50(m, 1H), 3.39~3.33 (m, 3H), 2.22~2.30 (m, 1H), 1.10~l.20 (m, 1H), 1.00 (s, 3H), 0.90~0.87 (m, 1H).

EXAMPLE 18: 4—fluoro—N—((R)—3 —((2—(((1R,ZS)—2—(4— methoxyphenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropan yl)benzamide (\NJH‘\\\\\\SlI/\/ NI"o O H 0d HN O The method that was described for the synthesis of 4—fluoro—N—((R)—3—((2— ((( 1R2 S)-2—(4-fluorophenyl)- l lcyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l - panyl)benzamide was used in the preparation of 4-fluoro-N—((R)—3-((2-(((1R,ZS) (4-methoxyphenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropan zamide. The mesylate, 4-fluoro-N—((R)—3-((2—(((1R,ZS)(4- methoxyphenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropan yl)benzamide, was prepared by the methods described earlier. The te was reacted with (lR,2S)—2-(4-methoxyphenyl)cyclopropanamine in the presence of DIEA/KI in acetonitrile at 50 degrees to give the desired 4—fluoro—N—((R)—3 —((2—(((1R,ZS)(4— methoxyphenyl)cyclopropyl)amino)ethyl)sulfonyl)- l -morpholino- l -oxopropan yl)benzamide as a white solid. ES, m/z=534 (M+H). H-NMR (300MHz, CDCl3):87.86~7.81 (m, 2H), 7.l2~7.07 (m, 2H), 6.97~6.94(m, 2H), 6.8l~6.78 (m, 2H), .67~5.64 (m, 1H), 3.80~3.77 (m, 4H),3.71~3.58 (m, 11H), 3.42~3.38(m, 2H), 2.48~2.42 (m, 1H), 2.20~2.10 (m, 1H) 2.28~1.23 (m, 1H), 1.03~1.00 (m, 1H).

EXAMPLE 19; 4—flu0r0—N—((R)—3—((2—(((1R,2S)—2—(4— henyl)cyclopropyl)amin0)ethyl)sulf0nyl)- l -m0rpholin0- l -0x0pr0pan-2—yl)benzamide (\NJH"\\\\\SH/\/N/"00 H 0d HN o 4—flu0r0—N—((R)—3—((2—(((1R,2S)—2—(4— fluorophenyl)cyclopropyl)amin0)ethyl)sulf0nyl)- l -m0rpholin0- l -0x0pr0panyl)benzamide was ed by the method used to prepare .4—flu0r0—N—((R)—3—((2—(((1R,ZS)—2-(4— fluorophenyl)- l -methylcyclopropyl)amin0)ethyl)sulf0nyl)- l olin0- l -0x0pr0pan-2— yl)benzamide. The mesylate (R)—2—((2-(4-flu0r0benzamid0)m0rpholin0 0x0pr0pyl)sulf0nyl)ethyl methanesulfonate was prepared as described earlier. This was reacted with (lR,2S)—2—(4-flu0r0phenyl)cyclopropanamine and DIEA/KI in acetonitrile at 50 degrees to yield 4—flu0r0—N—((R)—3—((2-(((1R,ZS)—2—(4— henyl)cyclopropyl)amin0)ethyl)sulf0nyl)- l -m0rpholin0- l 0panyl)benzamide as a white solid. ES,rn/z:=522 (M+H). H-NMR(300MHZ,CDC13): 87.85~7.8l (m, 2H), 7.l4~7.08 (m, 2H), .90 (m, 4H), 5.69~5.62 (m, 1H), 3.79~3.54 (m, 12H), 3.40~3.35 (m, 2H), 2.44~2.42 (m, 1H), 2.18~2.14 (m, 1H), 1.28~1.23 (m, 1H), 1.09~1.03 (m, 1H).

EXAMPLE 20: 4—flu0r0—N—((S)—3 —(2—(((1R,ZS)—2—(4— fluorophenyl)cyclopropyl)amin0)eth0xy)- l -m0rpholin0- l -0x0pr0pan-2—yl)benzamide Reaction Scheme for oxygen—linked compounds: CI HOJS""\OH 0 (\N \\ [j - OH O HN o 0d HN o HOJH" \OH —.F_ N \\ H Nazcoatzoyd'Oxane NH2 imidazole,THF 0 C Br/\n/0v 0 \\o/\lr0\/ (\NJW""\O/\/OH 0d HN o o NaBH4,THF,rt 0d HN o NaH,DMF F F O H \\\ /\/OMs N " o (\NJSIM\o/\/N"' f (V 0d HN o MSCI HN o F TEATHF rt,'2h ’ NaHC03,KI,MeCN,50°C A on of (2S)—2-aminohydroxypropanoic acid (21 g, 199.82 mmol, 1.00 equiv) in H20/dioxane (450/210 mL) was treated with sodium carbonate in water. To this was added a solution of 4—fluorobenzoyl de in dioxane at 0°C. The solution was stirred for 1 h at 0°C. The reaction was extracted with ethyl acetate. The water layers were acidified and extracted with ethyl acetate. The organic layers were washed with brine and dried over anhydrous sodium sulfate and concentrated to yield 45 g (99%) of (S)—2—(4— enzamido)-3 -hydroxypropanoic acid as a white solid. The material (4g) was dissolved in tetrahydrofuran and treated with 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)—one, (DEPBT) (10.54 g, 2.00 equiv) and imidazole (2.4 g, 2.00 equiv), and, after stirring 30 min, morpholine (1.53 g, 17.56 mmol, 1.00 equiv) in tetrahydrofuran at 0°C and then at RT for 16. The reaction was diluted with 150 mL of KH2P04(aq.) and extracted with ethyl acetate.

The organics were washed with brine and dried over anhydrous sodium sulfate. After concentration, the e was chromatographed on silica gel and eluted with 10/1 dichloromethane/methanol (10:1). This resulted in 800 mg (15%) of (S)—4—fluoro—N—(3— hydroxymorpholinooxopropanyl)benzamide as a yellow oil. This was dissolved in DMF and d with sodium hydride (130 mg, 5.42 mmol, 2.00 equiv) at 0°C. The mixture was stirred for 30min at 25°C. This was followed by the on of a solution of ethyl 2- bromoacetate (903 mg, 5.41 mmol, 2.00 equiv) in methylformamide at 0°C. The ing solution was stirred for 16 h at 25°C. The reaction was then ed by the addition of 10 mL of water/ice. The resulting solution was diluted with H20 and extracted with ethyl acetate. After a brine wash, the cs were dried and concentrated and then chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1: 1). This resulted in 500 mg (48%) of (S)—ethyl 2-(2—(4—fluorobenzamido)—3—morpholino oxopropoxy)acetate as a light yellow oil. This was dissolved in THF and treated with NaBH4 (100 mg, 2.64 mmol, 2.00 equiv at 0°C. The resulting solution was stirred for 16 h at °C. The reaction was quenched with water/ice and extracted with ethyl e. The organics were washed with brine, dried over sodium sulfate, concentrated, and chromatographed on silica gel and eluted with ethyl acetate/petroleum ether (1 :0). This resulted in 350 mg (79%) of (S)—4—fluoro—N—(3-(2-hydroxyethoxy)—1—morpholino—1— oxopropan—2—yl)benzamide as colorless oil. This was converted to the mesylate using MsCl/TEA/THF in the usual way (81% as an off white solid) and the mesylate was reacted with (1R,2S)(4—fluorophenyl)cyclopropanamine to yield the desired 4—fluoro—N—((S)—3—(2— (((1R,2 S)(4-fluorophenyl)cyclopropyl)amino)ethoxy)—1-morpholinooxopropan yl)benzamide as a light yellow oil (12%). ES, m/z= 474 (M+H). 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.76 (dd, J1=5.4Hz, J2=8.4Hz, 2H), 7.16 (d, J=7.5Hz, 1H), 7.04 (t, z, 2H), 7.4—6.85 (m, 4H), 5.22 (dd, J1=7.2Hz, J2=12.6Hz, 1H), 3.90—3.48 (m, 12H), 2.84 (t, J=5.1Hz, 2H), 2.40—2.25 (m, 1H), 2.05—1.80 (m, 1H), 1.03—0.99 (m, 1H), 0.95—0.80 (m, 1H) EXAMPLE 21: 4—fluoro—N—((S)—3 —(2—((( l R,2S)—2—(4— fluorophenyl)cyclopropyl)amino)acetamido)- l -morpholino- l -oxopropanyl)benzamide Reaction Scheme for Amine—Linked Compounds O O 0 \_/NH HOJSI"\\NH2 CICHZCOCI,Na2003 \ JK/CI Dioxane, water,rt,6h HOJHH\\NJK/C' (\NJH\" NHBoc NHBoc HOBT EDCI, 0d NHBoc DMFrt 10h O O F \\ JK/CI TFA'DCM (\NJS-ZHHNj\/ N \\\ CI Cl1 C (\NJH —. 0Q HN 5 Cd N32003, H20, dioxane,rt,2h O 0 F H2N,,'RE) (\NJS"\\\NJJ\/H"" wHN 5 F Ag — F K2003, Nal, acetone o—N—((S)—3—(2—(((lR,2S)(4-fluorophenyl)cyclopropyl)amino)acetamido)— l-morpholino-l-oxopropanyl)benzamide: (2 S)amino[[(tertbutoxy nyl]amino]propanoic acid was reacted with chloroacetyl chloride in sodium carbonate/dioxane/water for 6h to give (S)((tert-butoxycarbonyl)amino)(2- chloroacetamido)propanoic acid in 44% yield as a white solid. The acid was d with line in the usual fashion with HOBT, EDCl in DMF to give a 49% yield of (S)—tert— butyl (3-(2-chloroacetamido)-l-morpholino-l-oxopropanyl)carbamate as a white solid.

The amine was deprotected with TFA in methylene chloride to give (S)—N—(2-amino morpholino-3—oxopropyl)—2—chloroacetamide in 25% yield as a white solid. This could be ted with p—fluorobenzoyl chloride to (S)—N—(3—(2-chloroacetamido)-l—morpholino-l- oxopropanyl)fluorobenzamide. Yield was 25% after silica gel chromatography and eluted with ethyl acetate/petroleum ether (1/3). The ketone was reacted with (lR,2S)— 2—(4-fluorophenyl)cyclopropanamine, K2CO3, NaI in acetone to yield the desired 4—fluoro—N— ((S)-3 -(2-(((1R,2 S)(4-fluorophenyl)cyclopropyl)amino)acetamido)- l -morpholino- l - oxopropanyl)benzamide (32% yield) as a white solid. ES, m/z = 487 (M+H). 1H NMR (400 MHz, CD3Cl, ppm): 7.49—7.87(m, 4 H), 7.01—7.26(m, 2 H), 6.89—6.99 (m, 4 H), 5.18— .22 (m, l H), 3.94—3.80 (m, 12 H), 2.46—2.49 (m, l H), 2.02—2.06(m, l H), 0.93—1.16 (m, 2 EXAMPLE 22: 4-f1uoro-N—((S)—6-((( 1R2 4-fluorophenyl)cyclopropyl)amino) morpholino-1,6-dioxohexanyl)benzamide F~©— O OH WOH—> HOO \/\n/ ACI HN o o —.

NH2 0 MeOH, 000, 1 h NaZCO3, dioxane/HZO, 0 00, 2 h F (1) o o o HOJS"‘\\/\[ro\ [j (\N .\\‘\/\[ro\ HN O O H 0x} HN 0 0 LiOH,H20/THF DEPBT, imidazole, DCM F (2) F (3) o H2N,h NAB \ OH H, HATU,D|EA,DMF Step 1. (S)—2—(4—fluorobenzamido)hexanedioic acid (1) In a 1000—mL round-bottom flask, was placed a solution of (2S) aminohexanedioic acid (10 g, 62.05 mmol, 1.00 equiv) in hydrogen chloride (0.5 mol/L) (250 mL). Then dioxane (80 mL) was added. This was followed by the addition of a solution of sodium carbonate (23.1 g, 3.50 equiv) in water (60 mL) and a solution of 4—fluorobenzoyl chloride (11.8 g, 74.42 mmol, 1.20 equiv) in dioxane (20 mL) were added dropwise with stirring at 0°C at the same time. The resulting solution was stirred for 1 h at 0°C in a water/ice bath. After the reaction was completed, the resulting solution was ted with 2x400 mL of ethyl acetate. Then the pH value of the aqueous layers was adjusted to 2 with hydrogen chloride (1 mol/L). The aqueous layers were extracted with 3x400 mL of ethyl acetate and the organic layers combined. The organic layers were washed with 1x1000 mL of brine and dried over ous sodium sulfate. After filtration, solvent was removed under reduced 2014/049906 pressure. The residue was washed with 1x100 mL of DCM. This resulted in 11 g (63%) of (S)(4-fluorobenzamido)hexanedioic acid as a white solid.

Step 2. (S)—2—(4—fluorobenzamido)methoxyoxohexanoic acid (2) Into a 1000-mL round-bottom flask, was placed a solution of (4- fluorobenzamido)hexanedioic acid(10 g, 35.30 mmol, 1.00 equiv) in methanol (3 60 mL).

This was followed by the addition of acetyl de (3.3 g, 42.04 mmol, 1.20 equiv) dropwise with stirring at 0°C in 30 min. The resulting solution was stirred for 60 min at 0°C.

After the reaction was completed, NazCO3(aq.) was added to the reaction. The resulting solution was extracted with 3x300 mL of ethyl acetate and then. Then the pH value of the aqueous layers were adjusted to 2 with hydrogen chloride (1 mol/L). The aqueous layers were extracted with 3x400 mL of ethyl acetate and the organic layers combined. The organic layers were washed with 1x1000 mL of brine and dried over anhydrous sodium sulfate. After filtration, solvent was removed under reduced pressure. This resulted in 6.4 g (61%) of (S)—2— (4—fluorobenzamido)—6-methoxy—6—oxohexanoic acid as colorless oil Step 3. (S)—methyl 5—(4—fluorobenzamido)—6—morpholino—6—oxohexanoate (3) Into a 250-mL 3-necked bottom flask, was placed a solution of (4- fluorobenzamido)—6—methoxy—6—oxohexanoic acid (3.5 g, 11.77 mmol, 1.00 equiv) in tetrahydrofuran (90 mL), DEPBT (7 g, 23.41 mmol, 2.00 equiv) and imidazole (1.6 g, 2.00 equiv). The mixture solution was stirred for 30 min at 0°C. To this was added a solution of morpholine (1 g, 11.48 mmol, 1.00 equiv) in tetrahydrofuran (30 mL) se with stirring at 0°C in 30 min. The resulting solution was stirred for 16 h at room temperature. The resulting solution was diluted with 150 mL of (aq.). The ing solution was extracted with 3x150 mL of ethyl acetate and the organic layers combined. The resulting mixture was washed with 1x300 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1). This resulted in 1.7 g (39%) of (S)—methyl 5—(4— fluorobenzamido)morpholinooxohexanoate as yellow oil Step 4. (S)—5—(4—fluorobenzamido)—6—morpholino—6—oxohexanoic acid (4) Into a 100-mL round-bottom flask, was placed a on of (S)-methyl 5-(4- fluorobenzamido)—6—morpholino—6—oxohexanoate (1.6 g, 4.37 mmol, 1.00 equiv) in tetrahydrofuran (16 mL). This was followed by the addition of a on of LiOH (112 mg, 4.68 mmol, 1.10 equiv) in water (14.4 mL) dropwise with stirring at 0°C in 5 min. The resulting solution was stirred for 1 h at 25°C. The resulting mixture was concentrated under vacuum. The residue was diluted with 20 mL of water. The resulting solution was extracted with 2x20 mL of ethyl e and the aqueous layers combined. The pH value of the on was adjusted to 2 with hydrogen chloride (1 . The resulting solution was extracted with 3x30 mL of ethyl acetate and the organic layers combined. The resulting mixture was washed with 1x40 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. This ed in 1.3 g (84%) of (4—fluorobenzamido)—6— morpholinooxohexanoic acid as light yellow oil Step 5. 4—fluoro—N—((S)—6—((1R,2S)—2—(4-fluorophenyl)cyclopropylamino)morpholino- 1 ,6- dioxohexanyl)benzamide Into a 100-mL round—bottom flask, was placed a solution of (S)—5—(4— fluorobenzamido)—6—morpholino—6—oxohexanoic acid (200 mg, 0.60 mmol, 1.00 equiv) in N,N—dimethylformamide (30 mL), HATU (500 mg, 1.31 mmol, 2.00 equiv), DIEA (170 mg, 1.32 mmol, 2.00 equiv) and (1R,2S)(4-fluorophenyl)cyclopropanamine (94.3 mg, 0.62 mmol, 1.10 equiv). The resulting on was d for 2 h at 25°C. The resulting solution was diluted with 100 mL of H20. The resulting solution was extracted with 3x30 mL of ethyl acetate and the organic layers combined. The c layers were washed with 1x100 mL of Brine. The mixture was dried over anhydrous sodium sulfate. The solids were filtered out.

The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC. This resulted in 196.1 mg (67%) of 4-fluoro—N—((S)—6—((1R,2S)—2—(4— fluorophenyl)cyclopropylamino)morpholino-1,6-dioxohexanyl)benzamide as a white solid. 1H NMR (300 MHz, CD3OD—d4) 5 ppm: 7.95 (dd, J1=6.7Hz, Jz=3.15Hz, 2H), 7.23— 7.25 (m, 4H), 6.99 (t, J=9.3Hz, 2H), 4.87—5.04 (m, 1H), .58 (m, 8H), 2.84—2.79 (m, 1H), 2.29—2.75 (m, 2H), 2.04—1.99 (m, 1H), 1.84—1.72 (m, 4H), 1.19—1.14 (m, 2H). LC/MS: (ES, m/z): 486 [M+H]+ EXAMPLE 23: N—((S)((1R,2S)(4-fluorophenyl)cyclopropylamino)—1-oxo(piperidin- 1-yl)hexanyl)-1H-imidazolecarboxamide N)S_\\\V\/N\H HN O / NH EXAMPLE 24: 4-fluoro-N-((S)((1R,2S)—2-(4-fluorophenyl)cyclopropylamino)- 1-(4- methylpiperazinyl)-1,6-dioxohexanyl)benzamideH /NQJSHN O N HOJH W_‘\\ O\ E j N /NQNJHM\\O/\n/O\HN LiOH, HZO/THF DEPBT, imidazole, DCM F (1) o H /"(> )8 \ OH N H2N/I,%>\ p )S \V\n/N/N "N m /N\) HN O O F AU\F IEA,DMF F (2) F Step 1. thyl 5-(4-fluorobenzamido)(4-methylpiperazinyl)oxohexanoate (1) Into a 500—mL 3—necked round—bottom flask, was placed a solution of (S)—2—(4— fluorobenzamido)—6—methoxy—6—oxohexanoic acid (3.5 g, 11.77 mmol, 1.00 equiv) in tetrahydrofuran (90 mL), DEPBT (7 g, 23.41 mmol, 2.00 equiv) and ole (1.6 g, 23.53 mmol, 2.00 equiv). The mixture solution was stirred for 30 min at 0°C. This was followed by the addition of a solution of 1-methylpiperazine (1.2 g, 11.98 mmol, 1.00 equiv) in tetrahydrofuran (50 mL) dropwise with stirring at 0°C in 40 min. The resulting solution was stirred for 16 h at 25°C. The resulting solution was diluted with 150 mL of KH2P04 (aq.). The resulting solution was extracted with 3x150 mL of ethyl acetate and the organic layers combined. The resulting mixture was washed with 1x300 mL of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1). This ed in 2.4 g (54%) of (S)-methyl 5-(4-fluorobenzamido)(4-methylpiperazinyl)oxohexanoate as light yellow oil.

Step 2. (S)—5—(4—fluorobenzamido)(4-methylpiperazinyl)—6-oxohexanoic acid (2) Into a 100-mL round-bottom flask, was placed a solution of (S)—methyl 5-(4- fluorobenzamido)(4-methylpiperazinyl)oxohexanoate (1.75 g, 4.64 mmol, 1.00 equiv) in tetrahydrofuran (17 mL). This was followed by the addition of a solution of LiOH (118 mg, 4.93 mmol, 1.10 equiv) in H20 (15 mL) se with stirring at 0°C. The resulting solution was stirred for 1 h at 25°C. The resulting e was concentrated under vacuum.

This resulted in 1.3 g(crude) (77%) of (S)—5—(4—fluorobenzamido)—6—(4—methylpiperazinyl)- 6—oxohexanoic acid as yellow oil.

Step 3. 4—fluoro—N—((S)—6—((1R,2S)(4-fluorophenyl)cyclopropylamino)(4- methylpiperazinyl)—1,6-dioxohexanyl)benzamide Into a 100-mL round—bottom flask, was placed a on of (S)—5—(4— fluorobenzamido)(4-methylpiperazinyl)oxohexanoic acid (400 mg, 1.10 mmol, 1.00 equiv) in N,N—dimethylformamide (50 mL), HATU (832 mg, 2.19 mmol, 2.00 , DIEA (284 mg, 2.20 mmol, 2.00 equiv) and (1R,2S)(4-fluorophenyl)cyclopropanamine (182 mg, 1.20 mmol, 1.10 equiv). The resulting solution was stirred for 1 h at 25°C. The resulting mixture was concentrated under vacuum. The crude product was purified by PLC .

This resulted in 63.1 mg (11%) of 4-fluoro-N—((S)((1R,2S)—2-(4- fluorophenyl)cyclopropylamino)(4-methylpiperazinyl)—1,6-dioxohexanyl)benzamide as a white solid. 1H NMR (300 MHz, CD3OD-d4) 5 ppm: 7.95 (dd, J1=6.7Hz, 12:3. 15Hz, 2H), 7.23—7.25 (m, 4H), 6.99 (t, J=9.3Hz, 2H), 4.87—5.04 (m, 1H), 3.73—3.58 (m, 8H), 2.84— 2.79 (m, 1H), 2.29—2.75 (m, 2H), 2.04—1.99 (m, 1H), 1.84—1.72 (m, 4H), .14 (m, 2H).

LC/MS: (ES, m/z): 486 [M+H]+ EXAMPLE 25: 4-fluoro-N—((S)(2-((1R,2S)(4-methoxyphenyl)cyclopropylamino) oxoethoxy)morpholinooxopropanyl)benzamide NJH"‘\\O H 6::J N\ HN 0/81; OCH3 O O O IQ, MeOH WO/ CHzqu Pd(OAC)2 wo/ MeO MeO (1) MeO O OH OH H2Nj\© <1)LN KOH, MeOH — H (3) EDCI, HOBt, DMF MeO H so O didxania, DPPA, TEA, q» t—BuOH "uQ’NHBOC HCL MeOH \\..<’NH2 O" —’ MeOD" M Oe MeO (5) (6) (7) O H N HATU, DMF, rt + 0d —> e Step 1. (E)-methyl 3-(4-methoxyphenyl)acrylate (1) In a 1000—mL 3—necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed a solution of (E)-3 -(4-methoxyphenyl)acrylic acid (40 g, 224.49 mmol, 1.00 equiv) in ol (300 mL). This was followed by the addition of thionyl dichloride (54 g, 453.90 mmol, 2.00 equiv) dropwise with stirring at 0°C in 2 hr. The resulting solution was stirred for 16 h at 65°C in an oil bath. After the reaction was completed, the mixture was concentrated under vacuum. The residue was diluted with 300 mL of ethyl acetate and then washed with 1x400 mL of sat.NaHCO3, 1x300 mL of brine. The mixture was dried over anhydrous sodium sulfate. After filtration, solvent was removed under reduced pressure. This resulted in 41 g (95%) of (E)—methy1 ethoxypheny1)acrylate as an off—white solid.

Step 2. Methyl 2-(4-methoxyphenyl)cyclopropanecarboxylate (2) Into a L 3—necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed a on of (E)-methyl 3-(4-methoxyphenyl)acrylate (41 g, 213.31 mmol, 1.00 equiv) in dichloromethane (500 mL), Pd(OAc)2 (480 mg, 2.14 mmol, 0.01 equiv). This was followed by the addition of a solution of CH2N2 in ether (1500 mL) se with stirring at -5°C. The resulting solution was stirred for 4 h at 0°C. After the reaction was completed, the reaction was quenched by the addition of 4 mL of AcOH. The resulting mixture was washed with 1x400 mL of sat.Na2CO3 and then concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1 :3). The collected fractions were combined and trated under vacuum. This ed in 42 g (97%) of methyl 2-(4-methoxyphenyl)cyclopropanecarboxylate as an off-white solid.

Step 3. 2-(4-methoxyphenyl)cyclopropanecarboxylic acid (3) Into a 1000—mL bottom flask, was placed a solution of methyl 2-(4- methoxyphenyl)cyclopropanecarboxylate (42 g, 203.65 mmol, 1.00 equiv) in methanol (250 mL), then a solution of ium hydroxide ((57 g, 1.02 mol, 5.00 equiv) in methanol (200 mL) was added. The resulting solution was stirred for 5 h at room temperature. After the reaction was completed, it was concentrated under vacuum. The residue was diluted with 1000 mL of H20. The pH value of the solution was adjusted to 2 with hydrogen de (2 mol/L). The resulting solution was extracted with 3x1000 mL of dichloromethane and the organic layers combined. The combined organic layers were washed with 1x1500 mL of brine and dried over anhydrous sodium sulfate. After filtration, solvent was removed under reduced pressure. This resulted in 36 g (90%) of 2-(4- methoxyphenyl)cyclopropanecarboxylic acid as an off-white solid.

Step 4. (1R,2R)-N—((R)hydroxyphenylethyl)(4- methoxyphenyl)cyclopropanecarboxamide (4) Into a 1000-mL bottom flask, was placed a solution of 2-(4- methoxyphenyl)cyclopropanecarboxylic acid (36 g, 187.29 mmol, 1.00 equiv) in N,N— dimethylformamide (500 mL), HOBt (25 g, 185.02 mmol, 1.00 equiv), EDCI (36 g, 187.79 mmol, 1.00 equiv), (2R)aminophenylethanol (26 g, 189.53 mmol, 1.00 equiv). The resulting solution was stirred for 2 h at room temperature. After the reaction was completed, the mixture was poured into 300 mL of ice/water with stirring. The solids were collected by filtration. The e was applied onto a silica gel column with dichloromethane/ethyl acetate (10: 1-1:1). This resulted in 10.0 g (17%) of (1R,2R)-N—((R)hydroxy phenylethyl)(4-methoxyphenyl)cyclopropanecarboxamide as a white solid.

Step 5. (1R,2R)(4-methoxyphenyl)cyclopropanecarboxylic acid (5) Into a 250-mL bottom flask, was placed a solution of )-N—((R)—2- hydroxyphenylethyl)(4-methoxyphenyl)cyclopropanecarboxamide (10 g, 32.12 mmol, 1.00 equiv) in 1, 4—dioxane (70 mL) and sulfuric acid (70 mL, 3 mol/L). The resulting solution was stirred for 16 h at 100°C in an oil bath. After the on was completed, it was cooled to room temperature. The resulting mixture was concentrated under vacuum. The residue was diluted with 300 mL of water, extracted with 3x300 mL of ethyl acetate and the organic layers combined. The ed organic layers were washed with 1x500 mL of brine and dried over anhydrous sodium sulfate. After filtration, solvent was removed under reduced pressure. This resulted in 4.8 g (78%) of (1R,2R)—2-(4— methoxyphenyl)cyclopropanecarboxylic acid as an ite solid.

Step 6. Tert-butyl (1R,2S)(4-methoxyphenyl)cyclopropylcarbamate (6) Into a 250—mL round-bottom flask, was placed a solution of )(4- methoxyphenyl)cyclopropanecarboxylic acid (4.8 g, 24.97 mmol, 1.00 equiv) in tert-Butanol (50 mL), DPPA (6.9 g, 25.07 mmol, 1.00 equiv), TEA (2.5 g, 24.71 mmol, 1.00 equiv). The resulting solution was stirred for 5 h at 90°C in an oil bath. After the reaction was completed, it was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1: 10). The collected fractions were combined and concentrated under . This resulted in 2.5 g (3 8%) of tert—butyl (1R,2S)—2—(4— methoxyphenyl)cyclopropylcarbamate as a light yellow solid.

Step 7. (1R,2S)(4-methoxyphenyl)cyclopropanamine (7) Into a 100-mL bottom flask, was placed a solution of tert—butyl (1R,2S) (4-methoxyphenyl)cyclopropylcarbamate (2.5 g, 9.49 mmol, 1.00 equiv) in HCl/MeOH (40 mL). The resulting solution was stirred for 2 h at room temperature. After the on was completed, it was concentrated under . The resulting solution was diluted with 50 mL of H20, extracted with 2x30 mL of ethyl acetate and the aqueous layers combined.

Sat.NaHCO3 was employed to adjust the pH to 9. The resulting solution was extracted with 3x40 mL of ethyl acetate and the organic layers combined. The ed organic layers were washed with 1x100 mL of brine and dried over anhydrous sodium sulfate. After tion, solvent was removed under reduced pressure. This resulted in 1.4 g (90%) of (1R,2S)—2—(4— methoxyphenyl)cyclopropanamine as light yellow oil. 1H-NMR (300 MHz, CDCl3): 5 ppm: 7.00—6.90(m, 2H), 6.83—6.76(m, 2H), 3.77(s, 3H), 2.52—2.45(m, 1H), 1.85—1.78(m, 1H), 1.72(s, 2H), 1.02—0.86(m 2H).

Step 8. 4—fluoro—N—((S)—6—((1R,2S)(4-methoxyphenyl)cyclopropylamino)morpholino- 1,6-dioxohexanyl)benzamide Into a 100-mL round—bottom flask, was placed a solution of (S)—5—(4— fluorobenzamido)—6—morpholino—6—oxohexanoic acid (200 mg, 0.57 mmol, 1.00 equiv), HATU (500 mg, 1.31 mmol, 2.00 equiv) and DIEA (170 mg, 1.32 mmol, 2.00 equiv) in N, N—dimethylformamide (30 mL). The e was stirring for 5 min at room temperature.

Then (1R,2S)(4-methoxyphenyl)cyclopropanamine (102 mg, 0.62 mmol, 1.10 equiv) was added. The resulting solution was continued to stir for 2 h at room temperature. After the on was completed, it was diluted with 50 mL of H20. The resulting on was extracted with 3x30 mL of ethyl acetate and the organic layers combined. The organic layers were washed with 1X100 mL of brine and dried over anhydrous sodium sulfate. After filtration, solvent was d under reduced pressure. The residue was purified by Prep— HPLC (ACN/ H20 with 0.5% NH4HCO3). This resulted in 138.7 mg (49%) of 4—fluoro—N— ((S)((1R,2 S)(4-methoxyphenyl)cyclopropylamino)morpholino-1,6-dioxohexan yl)benzamide as a white solid. 1H NMR (400 MHz, CD3OD-d4) 5 ppm: 8.00—7.90(m, 2H), 7.25—7.17( m, 2H), 7.15—7.05(m, 2H), 6.88—6.80 (m, 2H), 5.05—5.00(m, 1H), 3.87—3.55(m, 11H), 2.85—2.76(m, 1H), 2.35—2.20(m, 2H), 2.00—1.92(m, 1H), 1.90—1.64(m, 4H), 1.18— 1.05(m, 2H); MS (ES, m/z): 498 (M + H).

EXAMPLE 26: 4-fluoro-N—((S)((1R,2S)(4-methoxyphenyl)cyclopropylamino)(4- piperazinyl)-1,6-dioxohexanyl)benzamide /©W" W.

HN :1? Ag OCH3 HATU, DIEA,DMF Into a 100-mL round—bottom flask, was placed a solution of (S)—5—(4— fluorobenzamido)—6—(4—methylpiperazin—1—yl)oxohexanoic acid (300 mg, 0.83 mmol, 1.00 equiv) in methylformamide (30 ), HATU (624 mg, 1.64 mmol, 2.00 equiv), DIEA (213 mg, 1.65 mmol, 2.00 equiv) and (1R,2S)(4-methoxyphenyl)cyclopropanamine (Example 25 Step 7) (147 mg, 0.90 mmol, 1.10 equiv). The resulting solution was stirred for 1 h at 25°C. The resulting e was trated under vacuum. The crude product was purified by Prep—HPLC. This resulted in 78.3 mg (19%) of 4—fluoro-N—((S)((1R,2S)—2—(4— methoxyphenyl)cyclopropylamino)(4-methylpiperazinyl)-1,6-dioxohexan zamide as a white solid. 1H NMR (300 MHz, CD3OD-d4) 5 ppm: 7.90—8.00 (m, 2H), 7.22 (m, 2H), 7.08 (d, J=6.6Hz, 2H), 6.85 (d, J=6.6Hz, 2H), 5.02—5.08 (m, 1H), 3.78 (s, 3H), 3.53—3.75 (m, 3H), 2.77—2.86 (m, 1H), 2.42—2.56 (m, 4H), 2.33 (s, 3H), 2.23—2.30 (m, 2H), 1.95—2.05 (m, 1H), 1.66—1.87 (m, 4H), 1.07—1.19 (m, 2H). LC/MS (ES, m/z): 511 [M+H]+.

EXAMPLE 27: N—[(2S)[[(1R,2S)(4-fluorophenyl)methylcyclopropyl]amino](4- methanesulfonylpiperazinyl)oxohexanyl]-N-methylbenzamide .x"\/\/N‘ 0" N / [‘0 O O O (\NJW W \o )5 o NaH,CH3I,DMF LiOH 0 IO N \O/jbr /, :fi MeOH,H20,2h msww O gI,N\) /N o o BH3_THF pNJH"‘\V\/OH Bess-Martin periodinane 0\ NJ /N o o 2% O O (\NJH \WO H H2N, NJS \WN, 0; /N\) /N O 0\ IQ /N O I I F O O DIEA,K|,MeCN,50°C Step 1. Methyl (5 S)(4-methanesulfonylpiperazinyl)-5 -(N-methylphenylformamido)- exanoate (1) Into a 100-mL round-bottom flask, was placed a solution of methyl (SS)(4- methanesulfonylpiperazinyl)oxo(phenylformamido)hexanoate (100 mg, 0.24 mmol, 1.00 equiv) in N,N—dimethylformamide (20 mL), sodium hydride (10 mg, 0.42 mmol, 1.77 equiv), Mel (100 mg). The ing solution was stirred for 1 overnight at 25°C. After the reaction was completed, the reaction was then quenched by the addition of water (100 mL).

The resulting mixture was extracted with ethyl acetate (4 x 50 mL) and the organic layers were combined. The resulting mixture was washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The obtained residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3). This resulted in 50 mg (48%) of methyl (5 4— methanesulfonylpiperazinyl)(N—methylphenylformamido)oxohexanoate as a yellow solid.

Step 2. (5S)(4-methanesulfonylpiperazinyl)(N—methylphenylformamido)—6- oxohexanoic acid (2) Into a 100-mL round-bottom flask, was placed a solution of methyl (SS)(4- esulfonylpiperazinyl)(N-methylphenylformamido)oxohexanoate (100 mg, 0.23 mmol, 1.00 equiv) in tetrahydrofuran (30 mL), a solution of LiOH (100 mg) in water(20 mL). The ing solution was stirred for l h at room temperature. After the reaction was ted .The reaction was then ed by the addition of water (100 mL). The resulting mixture was extracted with ethyl acetate (4 x 50 mL) and the organic layers were combined.

The resulting mixture was washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The obtained residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3). This ed in 50 mg (52%) of (5S)(4-methanesulfonylpiperazin-l-yl)—5-(N—methyl-l- phenylformamido)oxohexanoic acid as a white solid.

Step 3. N— [(2 S)—6—hydroxy- l thanesulfonylpiperazin- l -yl)- l -oxohexanyl] -N- methylbenzamide (3) Into a 100-mL round—bottom flask, was placed (5 S)—6—(4— methanesulfonylpiperazin- l -yl)-5 -(N-methyl- l -phenylformamido)oxohexanoic acid (100 mg, 0.24 mmol, 1.00 equiv), BH3/DCM (10 mL). The resulting solution was stirred for l h at room temperature. After the reaction was completed. The reaction was then quenched by the addition of water (100 mL). The resulting mixture was extracted with ethyl acetate (4 x 50 mL) and the organic layers were combined. The resulting mixture was washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The ed residue was ed by silica gel column chromatography using ethyl acetate/petroleum ether (l/3). This resulted in 50 mg (52%) of N—[(2S)hydroxy—l—(4— methanesulfonylpiperazin- l -yl)- l -oxohexanyl] -N-methylbenzamide as yellow oil.

Step 4. N— [(2 S)— l -(4-methanesulfonylpiperazin- l -yl)- l ,6-dioxohexanyl] -N- methylbenzamide (4) Into a 100—mL bottom flask, was placed )hydroxy—l—(4— methanesulfonylpiperazin-l-yl)-l-oxohexanyl]-N-methylbenzamide (200 mg, 0.49 mmol, 1.00 equiv), Dess—Martin (200 mg), dichloromethane (30 mL). The resulting solution was stirred for l h at room temperature after the reaction was completed .The reaction was then quenched by the addition of water (100 mL). The resulting mixture was ted with ethyl acetate (4 x 50 mL) and the organic layers were combined. The resulting mixture was washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The obtained residue was purified by silica gel column chromatography using ethyl e/petroleum ether (1/3). This resulted in 150 mg (75%) of N—[(2S)- l -(4-methanesulfonylpiperazin- l -yl)- l ,6-dioxohexanyl] -N-methylbenzamide as a white solid.

Step 5. N—[(2 S)—6—[[(1R,2S)—2-(4-fluorophenyl)— 1 -methylcyclopropyl]amino](4- methanesulfonylpiperazinyl)oxohexanyl]-N-methylbenzamide Into a 100-mL round-bottom flask, was placed a solution of N—[(2S)— 1 -(4- methanesulfonylpiperazinyl)-1,6-dioxohexanyl]-N-methylbenzamide (150 mg, 0.37 mmol, 1.00 equiv) in dichloromethane (30 mL), Na(OAc)3BH (200 mg), (1R,2S)(4- fluorophenyl)—1-methylcyclopropanamine hydrochloride (150 mg, 0.74 mmol, 2.03 equiv).

The resulting solution was stirred for 1 h at room temperature. After the reaction was complete, the reaction was then quenched by the addition of water (100 mL). The resulting mixture was extracted with ethyl acetate (4 x 50 mL) and the organic layers were combined.

The resulting mixture was washed with brine (3 x 50 mL), dried over ous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The obtained e was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3). This resulted in 22.7 mg (11%) of N—[(2S)—6-[[(1R,2S)—2-(4-fluorophenyl)—1- methylcyclopropyl]amino](4-methanesulfonylpiperazinyl)oxohexanyl]-N— methylbenzamide as a light yellow solid. 1H NMR (300 MHz, MeOD, ppm): 7.48—7.53(m, 5 H), 7.25—7.35(m, 2 H), 7.05—7.15(m, 2 H), 5.56—5.65(m, 1 H), .95(m, 9 H), 2.89— , 6 H), 2.60—2.70(m, 1 H), 1.13—2.08(m, 12 H). LC/MS (ES, m/z): 559 [M+H]+.

EXAMPLE 28: N—[(2S)[[(1R,2S)(4-fluorophenyl)cyclopropyl]amino]— 1 -(4- methanesulfonylpiperazinyl)oxohexanyl]-N-methylbenzamide pN ""V\/N ‘ Q\,N\) /N O o 0 MAS-"\WO H Han. NJS‘MWN’I' o\ ,0 /N o 0‘ ,0 /N 0 /IO /(.3 F DIEA,K|,MeCN,50°C Into a 100-mL round-bottom flask, was placed a solution of N—[(2S)—1-(4- esulfonylpiperazinyl)-1,6-dioxohexanyl]-N-methylbenzamide (150 mg, 0.37 mmol, 1.00 equiv) in dichloromethane (60 mL), (1R,2S)(4-fluorophenyl)cyclopropan amine hydrochloride (100 mg, 0.53 mmol, 1.45 , Na(OAc)3BH (100 mg). The resulting solution was stirred for l h at room temperature. After the reaction was complete, the reaction was then ed by the addition of water (100 mL). The resulting mixture was extracted with ethyl acetate (4 x 50 mL) and the organic layers were combined. The resulting mixture was washed with brine (3 x 50 mL), dried over anhydrous sodium e and filtered. The filtrate was concentrated under vacuum. The obtained e was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3). This resulted in 67 mg (34%) of N—[(ZS)—6—[[(lR,2S)(4-fluorophenyl)cyclopropyl]amino]-l-(4- methanesulfonylpiperazin-l-yl)-l-oxohexanyl]-N-methylbenzamide as an off-white solid. 1H NMR (400 MHz, CDCL3, ppm): 7.30—7.45(m, 5H), 6.90—7.10(m, 4H), 5.55—5.59(m, 1H), 3.75—3.85(m, 4H), 3.25—3.35(m, 4H), 2.83—2.94(m, 8H), 2.40—2.48(m, 1H), l.7l—2.25(m, 5H), l.25—l.51(m, 3H), l.02—l.l4(m, 1H). LC/MS (ES, m/z): 545 .

EXAMPLE 29: 4-fluoro-N—[(ZS)[[(lR,2S)(4-fluorophenyl)cyclopropyl]amino]— l -(4- methanesulfonylpiperazin- l -yl)- l -oxopentan-2—yl]benzamide O 0 o o F_< >_< HOJS\\)J\ / O HN O \_/N—é— HOJHNQkO/ O O DEPBT,imidazole,THF NH NaZCOa dioxane/Hzo, 2 Mom 0°C-rt O NaBH(AcO)3,DCM 0 Step 1. (2S)—2-[(4-fluorophenyl)formamido]methoxyoxopentanoic acid (1) Into a 500—mL round—bottom flask, was placed a solution of (2S)—2—amino—5— methoxyoxopentanoic acid (20 g, 124.10 mmol, 1.00 equiv) in dioxane (200 mL), a solution of sodium carbonate (20 g, 188.70 mmol, 1.52 equiv) in water(100 mL), 4— fluorobenzoyl chloride (20 g, 126.14 mmol, 1.02 equiv). The resulting on was stirred for 2 h at 0°C in a water bath. The resulting solution was extracted with 2x100 mL of ethyl acetate and the c layers combined and dried over anhydrous magnesium e and concentrated under vacuum. This resulted in 15 g (43%) of (2S)—2—[(4— fluorophenyl)formamido]methoxyoxopentanoic acid as an off-white solid.

Step 2. Methyl (4S)[(4-fluorophenyl)formamido](4-methanesulfonylpiperazinyl) oxopentanoate (2) Into a 250-mL bottom flask, was placed a solution of (2S)[(4- fluorophenyl)formamido]-5—methoxy—5-oxopentanoic acid (8 g, 28.30 mmol, 1.00 equiv) in tetrahydrofuran (200 mL), DEPBT (15 g), ole (10 g), 1-methanesulfonylpiperazine (8 g, 48.71 mmol, 1.38 equiv). The resulting solution was stirred for 1 overnight at room temperature. After the reaction was ted, The mixture was diluted with 300 mL of H20.

The aqueous phase was extracted with 3x200 mL of dichloromethane and the organic layers were combined. The organic layers were washed with 1x500 mL of brine and then dried with 2014/049906 anhydrous sodium sulphate. After filtration, solvent was removed under reduced pressure.

The residue was concentrated under vacuum and then applied onto a silica gel column with romethane/methanol (10:1). This resulted in 6 g (50%) of methyl (4S)[(4- fluorophenyl)formamido](4-methanesulfonylpiperazinyl)oxopentanoate as an off- white solid.

Step 3. (S)—4—(4—fluorobenzamido)(4-(methylsulfonyl)piperazinyl)oxopentanoic acid(3) Into a 100—mL round-bottom flask, was placed methyl (4S)[(4- fluorophenyl)formamido](4-methanesulfonylpiperazinyl)oxopentanoate (1 g, 2.3 3 mmol, 1.00 equiv), methanol (30 mL), water (30 mL), and lithium ide (1.5 g, 41.76 mmol, 24.88 equiv). The resulting solution was stirred for 3 h at room ature. The pH value of the solution was adjusted to 6 with hydrogen chloride (12 mol/L). The resulting solution was ted with 3x50 mL of ethyl acetate and the organic layers combined and concentrated under vacuum. This resulted in 700 mg (72%) of (S)—4—(4—fluorobenzamido)—5— (4-(methylsulfonyl)piperazinyl)oxopentanoic acid as a white solid.

Step 4. 4—fluoro—N—[(2S)hydroxy(4-methanesulfonylpiperazinyl)oxopentan yl]benzamide (4) Into a 100—mL round—bottom flask, was placed a solution of (S)—4—(4— fluorobenzamido)(4-(methylsulfonyl)piperazinyl)oxopentanoic acid (700 mg, 1.68 mmol, 1.00 equiv) in tetrahydrofuran (60 mL), BH3 (1 mL). The resulting solution was stirred for 3 h at room temperature. After the reaction was completed, The e was diluted with 300 mL of H20. The aqueous phase was extracted with 3x200 mL of romethane and the organic layers were combined. The organic layers were washed with 1x500 mL of brine and then dried with anhydrous sodium sulphate. After filtration, solvent was removed under reduced pressure. The residue was concentrated under vacuum and then applied onto a silica gel column with dichloromethane/methanol (10:1). This resulted in 500 mg (73%) of 4— fluoro-N— [(2 S)-5 -hydroxy(4-methanesulfonylpiperazinyl)oxopentanyl]benzamide as a white solid.

Step 5. 4—fluoro—N—[(2S)(4-methanesulfonylpiperazinyl)-1,5-dioxopentan yl]benzamide (5) Into a 100—mL round—bottom flask, was placed o—N—[(2S)—5—hydroxy—1—(4— methanesulfonylpiperazinyl)oxopentanyl]benzamide (200 mg, 0.50 mmol, 1.00 equiv), dichloromethane (20 mL), D-M (800 mg). The ing solution was stirred for 2 h at room temperature. After the reaction was completed, The mixture was diluted with 300 mL of H20. The s phase was extracted with 3x200 mL of dichloromethane and the organic layers were combined. The organic layers were washed with 1x500 mL of brine and then dried with anhydrous sodium te. After filtration, solvent was removed under reduced pressure. The residue was concentrated under vacuum and then applied onto a silica gel column with dichloromethane/methanol (10:1). This ed in 100 mg (50%) of 4—fluoro—N— l-(4-methanesulfonylpiperazin-l-yl)-l,5-dioxopentanyl]benzamide as yellow oil.

Step 6. 4—fluoro—N—[(2S)—5—[[(lR,2S)(4-fluorophenyl)cyclopropyl]amino]-l-(4- methanesulfonylpiperazin- l -yl)- l -oxopentanyl]benzamide Into a 100—mL round—bottom flask, was placed a solution of 4-fluoro-N—[(2S)—l— (4-methanesulfonylpiperazin-l-yl)-l,5-dioxopentanyl]benzamide (20 mg, 0.05 mmol, 1.00 equiv) in dichloromethane ( mL), (lR,2S)(4-fluorophenyl)cyclopropan-l-amine (40 mg, 0.26 mmol, 5.28 equiv), NaBH(AcO)3 (50 mg). The resulting solution was stirred for 30 min at room temperature. After the on was ted, The e was diluted with 300 mL of H20. The aqueous phase was extracted with 3x200 mL of romethane and the organic layers were combined. The organic layers were washed with 1x500 mL of brine and then dried with anhydrous sodium sulphate. After filtration, solvent was removed under reduced pressure. The residue was concentrated under vacuum and then applied onto a silica gel column with dichloromethane/methanol (10:1). This resulted in 3.4 mg (13%) of 4—fluoro—N— [(2S)—5- [ [( l R,2S)(4-fluorophenyl)cyclopropyl]amino] - l -(4-methanesulfonylpiperazin- l - yl)-l-oxopentanyl]benzamide as yellow oil. 1H NMR (MeOD, 400MHz, ppm): 7.91— 7.92(m, 2H), 7.19—7.42(m, 4H), 7.02—7.10(m, 2H), 5.10—5.22(m, lH), 3.88—3.98(m, 2H), 3.50— , 2H), 3.18—3.33(m, 4H), 2.95—2.96(m, lH), l.86—2.01(m, 4H), l.22—l.59(m, 5H), 0.88— 0.97(m, lH). LC/MS (ES, m/z): 535 [M+H]+.

EXAMPLE 30: l -((S)((1R,2 S)—2-(4-fluorophenyl)cyclopropylamino)- l -oxo- l -(piperidin- l-yl)hexanyl)-3 -phenylurea -"‘V\/N \ HN O HN\© NAS"\—>\/\/OH ;foWoH MsCI, NEt3, DCM O DIEA DCM rt (1) HNO OKNWOMS H2NrSh we 0NASNifWm*0 (2) \© DIEA KI MeCN HMO Step 1. (S)(6-hydroxyoxo(piperidinyl)hexanyl)-3 -phenylurea (1) Into a 50—mL round—bottom flask, was placed (S)—2-amino-6—hydroxy (piperidin-l-yl)hexan-l-one (200 mg, 0.93 mmol, 1.00 equiv), romethane (25 mL), phenyl isocyanate (111 mg, 0.93 mmol, 1.00 equiv) at 0°C in a water/ice bath. To this was added DIEA (3 62 mg, 2.80 mmol, 3.00 equiv) at the same temperature. The resulting solution was stirred for 1 h at room ature. After the reaction was completed, it was diluted with 100 mL of DCM and then washed with 1x75 mL of H20, 1X75 mL of brine. The ed organic layers were dried over anhydrous sodium sulfate. After filtration, solvent was removed under d pressure. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether . This resulted in 220 mg (71%) of (S)-l-(6-hydroxyoxo-l- (piperidin-l-yl)hexanyl)-3 -phenylurea as light yellow oil.

Step 2. (S)—6—oxo—5—(3 —phenylureido)—6—(piperidin-l-yl)hexyl methanesulfonate (2) Into a 50—mL round-bottom flask, was placed (S)(6-hydroxyoxo (piperidinyl)hexanyl)-3 -phenylurea (220 mg, 0.66 mmol, 1.00 equiv), tetrahydrofuran (25 mL), NEt3 (132 mg, 2.00 equiv). The reaction was cooled to 0°C in a water/ice bath.

MsCl (117 mg, 1.50 equiv) was added se at that temperature. The resulting solution was stirred for 3 h at room temperature. After the reaction was completed, it was diluted with 100 mL of H20. The resulting solution was extracted with 3x100 mL of ethyl acetate and the organic layers combined. The combined organic layers were washed with 1x200 mL of brine and dried over anhydrous sodium sulfate. After filtration, solvent was removed under reduced pressure. The residue was applied onto a silica gel column with ethyl e/hexane (1 :3).

This resulted in 230 mg (85%) of (S)—6—oxo—5—(3—phenylureido)(piperidin—l—yl)hexyl methanesulfonate as light yellow oil.

Step 3. 1—((S)—6—((1R,2S)—2-(4-fluorophenyl)cyclopropylamino)oxo(piperidin yl)hexanyl)-3 -phenylurea Into a 50-mL round-bottom flask, was placed (1R,2S)(4- fluorophenyl)cyclopropanamine (85 mg, 0.56 mmol, 1.00 equiv), MeCN (25 mL), (S)—6—oxo— -(3 -phenylureido)(piperidinyl)hexyl methanesulfonate (230 mg, 0.56 mmol, 1.00 equiv), DIEA (145 mg, 1.12 mmol, 2.00 equiv), K1 (9 mg, 0.05 mmol, 0.10 equiv). The resulting solution was stirred for 36 h at 60°C in an oil bath. After the reaction was completed, the solution was diluted with 150 mL of DCM, and then washed with 1x100 mL of H20, 1x100 mL of brine. The combined organic layers were dried over ous sodium sulfate. After filtration, solvent was removed under reduced pressure. The residue was purified by Prep—HPLC (ACN/ H20 with 0.5% NH4HCO3). This resulted in 5.8 mg (2%) of 1—((S)((1R,2S)—2-(4-fluorophenyl)cyclopropylamino)oxo(piperidinyl)hexanyl)- 3-phenylurea as a white solid. 1H NMR (300 MHz, CD3OD-d4) 5 ppm: 7.40—7.15(m, 4H), 7.15—6.90( m, 5H), 4.76—4.52(m, 1H), .42 (m, 4H), 2.72(t, J: 7.2 Hz, 2H), 2.32- 2.25(m, 1H), .85(m, 1H), .35(m, 12H), 1.10—0.85(m, 2H) ; MS (ES, m/z): 467 (M + H).

EXAMPLE 3 1: 1-((S)—6-((1R,2 S)(4-fluorophenyl)cyclopropylamino)oxo(piperidin- 1-yl)hexanyl)-3 -methylurea -"‘V\/N ‘ HN O The title compound may be made in a manner analogous to the method set forth in e 30 and by methods known in the art.

EXAMPLE 32: 3 ,4-dichloro-N—((S)-5 -((1S,2R)(4-fluoropheny1)cyclopropylamino)(4- lsulfony1)piperaziny1)oxopentany1)benzamide The title compound may be made by the method below and by methods known in the art. o o O O ‘\\\\)J\ / \jk / HO O HO 0 CI 0 O HO o HN O /—\ " HN N—fi— 80an NH2 0 —> —> CI NazCO3V dioxane/HzO, DEPBT,imidazole,THF 0' 0 °C, 2 h 0°C-rt CI CI 0 O O O .\\‘\)J\ / (\N "fikaH N O o ,N\2 HN o IN "N L'OH,THF,HO' 0‘ \2 /\| 2 , /| BH3,THF NJHMWOH (\NJWW Han.

HN o 0 IO HN o DMDCM AU —, (DE/Nd / F / —> (I) NaBH(AcO)3,DCM (\NJ‘S \\\/\HNI" o\ ,Nd HN o O F E 3 3: 4-fluoro-N—((S)((1R,2S)(4-fluoropheny1)cyclopropylamino)(4- (methylsulfonyl)piperaziny1)oxopentany1)-N—methy1benzamide 038/0 H /N O The title compound may be made by the method below and by methods known in the art.

O O O O (\N -"‘\)J\o/ (\NJR""\)J\0/ 0‘ ’Nd HN O Me'vNaHDMF 0\ fix} /N 0 LiOH,THF,H20 /b —>/|O —, F F O CO)3,DCM 0 EXAMPLE 34: 4-fluoro-N—((S)-5 -((1 S,2R)(4-fluoropheny1)cyclopropylamino)-1 -(4- methylpiperazin-l-y1)—1-oxopentan-Z-y1)benzamide "\\V\ \" N N The title compound may be made by the method below and by methods known in the art.

O O O O O O Hob"HA"N/ ‘N— mfiw‘vko/ mJH-"VRDH HN O HN\—/ /N\) HN O HF,H20 de HN O —> —> DEPBT,imidazole,THF o°c—n F F 0 0 H2Nn.

(\NJWNWOH .\‘ \AO ;> / \2BH,THF N HN O D-MDCM 0\ ,NO HN O F —>/| —> O NaBH(AcO)3,DCM The ing compounds may be sized using methods analogous to those bed herein and known in the art, using appropriate starting materials and reagents. In the following structures, it should be understood that mixtures of or single isomers, such as racemic es and alternate enantiomers, zwitterions, and the like may be prepared, e. g. by using appropriate L- or D- isomer, or chiral or achiral compound, as a staring material or reagent, or by employing a separation step.

Therefore, in n embodiments in the compounds below, the configuration of the substituents off the cyclopropylamine is trans to the phenyl. In certain embodiments, the trans configuration is R, S; in others, it is S, R. Furthermore, in certain embodiments, the core contains a L-isomer, for example as shown in Formula 11. Additional Examples include: WO 21128 VAQF QN NE :0 W \\\N N 5 LYQW fN¢ H e WO 21128 ¢ HN N o :9 F F \\ \ N\ H N O I HNYQ- CI V‘P CI \ HM o O VIQF 0Me F \N HM N\ HN 0 VIQF Y? i 2014/049906 Q m N .

H N 88 o / N 61' O VIQF HN \ COHKIHn \ NN o N o WO 21128 H O pkflwww N)S_n\\S/\/N\H Ozsd HN o C/ HN O OCF37 C.

O o (\NJWA \O/\"h\\ .x"\ A I I 0% S " /N\} HN 0 HN O OMG 0M6 F CF3 7 7 o o mN‘mu'AQ mN‘mw 02x} HN 0 HN CI OZS,N\} 0 CI ' CF3 CF3 7 7 o o NJK‘ N/H (\NJS'NWN’H HN o 0 8d2 HN o OMe CI Ph Ph WO 21128 0N o»m HN c\ "mu W2 ». HM 0N HN W3 / /\ NI /\ NI WO 21128 Me F 3 N O HN N H 0Ove/NI a , WO 21128 Biological ty The activity of the Examples above may be illustrated in the following assays.

Compounds listed above, which may not yet have been made and/or tested, are predicted to have activity in these assays.

Assaying the inhibition of KDMlA can be determined in vitro, in ed cells, and in animals. There are a variety of spectrophotometric methods to detect the results of demethylation of methylated lysines, viz., ing the products of KDMlA demethylase oxidative activity on a peptide fragment of at least18 amino acid representing the N—terminus of the e H3 substrate that contains a monomethyl at the fourth lysine residue.

Hydrogen peroxide, one product of the KDMlA demethylase reaction, reacts with horseradish peroxidase and dihydroxyphenoxazine (ADHP) to produce the scent compound resorufin (excitation= 530—560nm:emission= 590nm). The KDMlA demethylase enzyme activity can obtained from mammalian cells or tissues sing KDMlA from an endogenous or recombinant gene and purified or assayed from a whole cell t. These methods can be used to determine the concentration of the disclosed compounds can inhibit fifty percent of the enzyme activity (ICso). In one aspect, the disclosed compounds exhibit inhibition fifty percent of the KDMlA enzyme activity at a concentration of less than 500 nM, less than 100 nM, less than 50 nM or less than 10 nM.

The association of KDMlA with other proteins can be determined by a variety of both in vitro and in viva methods known to one skilled in the art. For example, the tion of KDMlA with associated proteins can be determined in an electromobility shift assay (EMSA). In various aspects, the disruption of the al association of KDMlA with CoRest by the disclosed compounds can be observed using EMSA. In another example, the disruption of KDMlA with associated proteins can be determined by immunoprecipitation followed by separation of the co—precipitated ns by mass spectroscopy or by get electrophoresis. In another example, the disruption of KDMlA association with CoRest can be determined by the ability of KDMlA to act on a nucleosomal substrate ning K4 or K9 methylated histone H3, a substrate that requires the ce of both KDMlA and CoRest. The disclosed compounds could be used to assay inhibition of CoRest association with KDMlA using nucleosomal substrate; such compounds may not inhibit KDMlA tic activity as determined by the use of the histone H3 K4 methylated peptide substrate.

The inhibition of KDMlA can be determined in a cell-based assay. For example, KDMlA is an essential enzyme and ged inhibition of KDMlA will result in cell death, thus cell growth inhibition, arrest of cell growth or cell death can be assayed. In another aspect, genes induced by androgens and estrogens require KDMlA activity; inhibition by the disclosed compounds of KDMlA will abrogate the induction of gene expression in cells d with androgens or estrogens. These s can be measured, e. g., using quantitative PCR ofmRNA to measure the magnitude of gene expression for androgen— and estrogen— dependent genes. KDMlA activity is required for the repression of transcription of specific genes. Inhibition of KDMlA by the sed compounds could de—repress the expression such genes in cell. These genes include Meisl, VEG—A, AIMl, HMOXl, VIM, SKAPl, BMP, EOMES, FOXA2, HNF4, SOX17, GH, PSA, p82, GREBl, GR—lb, PRL, TSHB, SYNl, HBG, SCNlA, SCN2a, and SCN3A the expression of which can be assayed using tative PCR of mRNA before and at various time following the treatment of cells with the disclosed compounds. In another aspect, KDMlA is a regulator of leukemic stem cell potential and is required for oncogenic transformation of myeloid cells to acute d leukemia (AML) by MLL-AF9. Inhibition ofKDMlA in MLL-AF9-transformed cells grown in culture mes the arrest in differentiation to resulting in a more mature cell expressing the CD1 lb surface antigen, a monocytic cell antigen. Thus, inhibition of KDMlA can be assayed using an AML cell line such as THP-l grown in culture quantifying the proportion of cells newly expressing the CD1 lb antigen using cence activated cell sorting (FACS).

A similar assay using FACS to count cells displaying the CD14 or CD86 can be also used, each of which are characteristic of more mature cells along the macrophage/monocytic lineage. Other cells lines derived from patients with acute myeloid leukemia such as MV4;ll or 3 cells can be used for this assay. Other markers of entiation along the macrophage/monocyte lineage can be similarly assayed by FACS such as CD14 and CD86.

Other AML cell lines such as MPLM-l3 or MV4;ll can be assayed for the ion of either specific genes mentioned above or the differentiation markers as well as cell growth or apoptosis by Annexin V staining and FACS enumeration.

The selectivity of the disclosed compounds for KDMlA can be determined by assaying the IC50 of the disclosed compounds for other FAD—dependent aminoxidases such as monoamine oxidase A (MAO-A), monoamine oxidase B (MAO-B), IL4Il, KDMlB, or SMOX. As such, a disclosed compound would inhibit KDMlA with an IC50 that is 50—fold, or ld or 250—fold or 500—fold less than for MAO—A or MAO—B.

Additional Demethylase Assays The e demethylase assay can be performed essentially as described in Shi, Y et al. Cell 199, 941— 953 (2004). Briefly, bulk histones, histone peptides or somes are incubated with purified human recombinant KDMlA, in the histone demethylase activity (HDM) assay buffer 1 (50 mM Tris pH 8.5, 50 mM KCl, 5 mM MgCl, 0.5% BSA, and 5% glycerol) from 30 minutes to 4 hours at 37°C. A typical reaction is conducted in 100 microliters in which either 20 rams of purified bulk histones or 3 rams of modified e peptides are used as substrates. Different s of KDMlA g from 1—20 micrograms are used in the on along with, as necessary, other co—factors such as FAD or CoREST, depending on the chosen substrate. The reaction e is analyzed by SDS-PAGE and Western blotting using histone methyl-speciflc antibodies or by formaldehyde formation assay to examine the removal and conversion of the methyl group to formaldehyde, or by mass spectrometry in the case of peptide substrates to fy the demethylated histone peptide.

Bulk histones (e. g., 4 mg) are incubated with the indicated amounts of recombinant proteins or complexes in histone demethylase (HDM) assay buffer A (50mM Tris pH8.5, 50mM KCl, 5mM MgCl, 5% glycerol, 0.2mM phenylmethylsulphonyl fluoride and lmM dithiothreitol) in a final volume of 10 ml for 12—16 h at 37 8C. For nucleosomes (0.3 mg) or mononucleosome (0.3 mg), HDM buffer A containing 0.1% NP40 can be used.

The reaction mixture can then be ed by SDS—PAGE followed by Western blotting.

Antibodies t mono- or di-methyl K4 in e H3 and acetyl-K9/ K14 of histone H3 are used to detect the degree of methylation and acetylation, respectively. Western blots are then quantified by densitometry or by intensity of luminescence.

Alternatively, a standard flurogenic assay can be used in which the methylated histone substrate is tethered to the bottom of a 96 well plate (or to beads resting in the plate) using biotin conjugated to the histone methylated substrate and strepavidin (SA) on beads or SA attached to the plate to secure the biotinylated substrate. After incubation of the KDMlA enzyme in histone demethylase buffer A, the demethylated e substrate can be detected using antibodies c for demethylated H3K4 substrate conjugated to a fluor or some other agent that can be detected. A variation on that assay method would employ an antibody directed against the methylated version of the histone in which the amount of substrate is quantified before and after incubation with the enzyme. Yet another version of a similar assay would employ a fluorescence resonance energy transfer (FRET) system of detection in which the antibody recognizing the methylated version is conjugated or otherwise linked to an entity, e.g., a bead or a large carrier molecule on which a fluorophore ) is attached and the fluorophore tor) is bound to an entity linked to the substrate.

Alternatively, the production of H202 during the KDMlA reaction can be detected fluometrically. In this system, the production of H202 is detected in the HDM assay buffer after exposure to ate, co-factor and enzyme using ADHP (10-Acetyl-3, 7- oxyphenoxazine) as a fluorogenic ate for horse radish peroxidase (HRP). ADHP (also known as Amplex Red Reagent) is the most stable and sensitive fluorogenic ate for HRP. The florescent product is resorufln. Sensitivity can be as low as 10'15 M of target protein. The signal is read using a fluorescence microplate reader at excitation and emission wavelengths of 530—560 nm and 590 nm, respectively.

Additionally, the KDMlA reaction can e other factors which may influence the ty of KDMlA. Such factors might include CoREST, NuRD complexes, DNMTl, HDACl, HDAC2, and HDAC3, for example, as proteins known to associate with KDMlA or KDMlA-containing complexes. ctions that influence any aspect of the KDMlA activity including specificity for template, substrate, Km, Kcat, or sensitivity to FAD concentrations can be assayed. For example, an in vitro interaction assay between KDMlA and CoREST can be performed adding recombinant KDMlA (e. g., 10 mg) and CoREST (e. g., 5 mg) mixed and incubated for 1 h at 4-80C, fractionated by Superdex 200 gel filtration column in a buffer containing 20mM Tris-HCl pH 7.9, 500mM KCl, 10% glycerol, 0.2mM EDTA, lmM dithiothreitol, 0.1% Nonidet P40 and 0.2mM phenylmethylsulphonyl fluoride, and then ed by silver staining.

For co—immunoprecipitation of mononucleosomes with KDMlA and CoREST, somes (1.5 mg) can be digested with micrococcal nuclease and incubated with recombinant KDMlA (e. g., 1 mg), CoREST (e. g., 500 ng) or both proteins in HDM buffer A containing 0.1% NP40 for 1 h at 4-80C. Antibodies directed against KDMlA or CoREST attached to an affinity resin are added and after extensive washing with HDM buffer A containing 0.1% NP40, the bound proteins are eluted with a wash buffer. KDMlA activity can be assayed in the eluate or the concentration of KDMlA can be determined by quantitative Western blotting.

Compounds were tested in a 10—dose ICso mode fluorescence coupling enzyme assay with 3-fold serial dilution in duplicate ng at 100 uM. The production of FAD- dependent H2O2 as a result of demethylase ty of LSDl on 10 uM histone H3(1— 21)K4me2 peptide substrate was ed by coupling with HRP and Amplex Red to yield resorufin (fluorescence measured at Ex/Em=53 5/590 nm on EnVision, Perkin Elmer).

Results are given below in Table 1.

Table l. % at 30 min, MLM: KDMlA ICso Example N0. + is 2 20% + is S 1 uM —is<20% —is>luM ._i O ._. [\J ._. 4; 3 U NNNNNNNl—‘l—‘l—‘l— omngi—toooouo Z [\J \] N 00 [\J \0 ZO Ex vivo differentiation of purified human CD34+ cells into the erythroid lineage Human CD34+ cells isolated from the venous blood of healthy donors after mobilization by granulocyte colony stimulating factor (G-CSF) are grown and differentiated ex vivo for a 14 day incubation using a two—phase culture method described in Cui, S., et al.

Mol Cell Biol 31, 311 (2011). Cells are counted using a hemocytometer and viability determined by trypan blue exclusion. Test article date compounds) dissolved in an appropriate t compatible with logic conditions is added daily to fresh culture medium beginning on Day 4 through Day 14 at a range of test concentrations. Cell morphology and stage of entiation is ined by Wright-Giemsa staining.

Flow cytometry to determine differentiation surface markers and HbF content Cultured erythroid cells are stained with phycoerythrin (PE)—Cy7—conjugated anti— CD34, PE-conjugated anti-CD71, and PECyS-conjugated anti-glycophorin A antibodies. To determine the concentration of cytoplasmic HbF, cells are fixed in 0.05% glutaraldehyde for minutes, bilized with 0.1%Triton X-100 for 5 minutes and stained with allophycocyanin—conjugated bF antibody. Stained cells are sorted and counted using a FACS analyzer.

Western blots to determine presence and tration of KDMlA and histone H3 and H3 modifications.

Cells are lysed in Laemmli sample buffer and subjected to SDS-PAGE. Proteins are transferred from the gel to nitrocellulose and probed with antibodies against KDMlA, and/or histone H3, mono-methyl (H3K4mel) and/or dimethyl histone H3K4 (H3K4me2) and then probed with fluorescence—conjugated secondary antibodies. Proteins concentrations are fied with an imaging system.

Chromatin immunoprecipitation (ChIP) assays to determine protein occupancy at genome— specific sites.

ChIP assays are carried out in an immunoprecipitation (1P) buffer with or without SDS depending on the ivity of the KDMlA antibody to SDS. Briefly, typically 3X107cells are used per KMDlA ChIP and 3X106 cells per 2 ChIP. After 10 minutes of 0.75% formaldehyde treatment, cells are harvested and sonicated in the ChIP lysis buffer (1% Triton X-100, 10 mM EDTA, 50mM Tris-HCl and protease inhibitors) to produce soluble chromatin with average sizes between 300 and 1000 bp. The tin samples are then diluted d in the dilution buffer (5mM EDTA, 25mM Tris-HCl, l67mM NaCl, and cocktails of protease inhibitors) and pre-cleaned for 1 hour using salmon sperm DNA/protein- A agarose beads. Ten micrograms of rabbit anti-KDMlA dy, 3 microliters of anti- H3K4me2 or control antibodies are then added to each sample and incubated overnight at 40C. To collect the immunocomplexes, 40 iters of salmon sperm DNA/protein-A agarose beads are added to the samples for 1 hour at 4°C. The beads are washed three times in wash buffer (0.1% Triton X-100, 5mM EDTA, 30mM Tris-HCl, 150mM NaCl and the washed once in wash buffer 2 (1% Triton X-100, 5mM EDTA, 30mM Tris—HCl, 150mM NaCl). The bound protein—DNA complexes are eluted with 100 microliters of elution buffer (1% SDS, 0.1 M NaHCO3, 250mM NaCl, and 0.2 micrograms protease K) and de—cross— linked at 65°C for 4 hr. The de-crosslinked chromatin DNA is further purified by QIAquick rase chain reaction (PCR) ation Kit (Qiagen) and eluted in 100 microliters of TE buffer. Four microliters of eluted DNA sample is used for each PCR reaction. Thirty-six PCR cycles can be used for KDMlA ChIP and 32 PCR cycles for H3K4mme2 ChIP.

Appropriate primers for loci of interest, e. g., the gamma globin gene, are used.

For globin—specific ChIP analysis, the assays are performed as described in Cui, S., et al. Mol Cell Biol 31, 3298-3311 (2011). For example, ethylene glycol bis(succinimidyl succinate) or formaldehyde can be used as a cross—linker. Antibodies against target proteins such as KDMlA and histone H3 with or t methyl modifications can be used for immunoprecipitation. DNA contained in the immunoprecipitate can be quantified by real- time tative PCR (RT-qPCR) assay using primer for human embryonic and , gamma, adult beta—globin promoter sequences; s for intergenic s between the embryonic and gammaG —globin genes can be used as a negative control.

Hemoglobin analysis by HPLC Cells are lysed and can be analyzed for hemoglobin composition using the Bio— Rad Variant II Hemoglobin Testing System equipped with an ion-exchange HPLC column (Hercules).

Mouse Models for testing induction of gamma globin gene expression Test e can be dissolved in a physiologically compatible t for injection into normal mice or mice transgenic for the yeast ial chromosome (YAC) containing the entirety of the human beta-globin locus as described in Tanabe, O., et al. EMBO J 26, 2295—2306 (2007) or portions of the human beta—globin locus. Test article can be administered daily intraperitoneally or subcutaneously or by gavage at appropriate test doses for up to 26 weeks. At intervals, eral whole blood and bone marrow cells are harvested WO 21128 to determine gene expression by RT—qPCR of the mouse embryonic beta—like globin genes or the beta-like globin composition of red cell lysates or in the case transgenic mice carrying human beta-like globin genes both the human and mouse fetal y- and adult B—globin genes.

Testing for ion of human gamma globin gene sion or HbF.

Patients with hemoglobinopathies including sickle cell disease and beta- thalassemia might benefit from ent with an inhibitor of KDMlA. After appropriate dosing, the measure of HbF can be determined as described above. Gamma globin gene expression can be assayed in bone marrow cells using qPCR. Further, the clinical benefit of an agent inducing HbF can be measured as an increase in total hemoglobin, a reduction in sickle cell crises, a decrease in transfusion dependence, a se in ineffective hematopoiesis, and decrease in inflammatory biomarkers such as plasma levels of GDFlS, etc.

Pharmacokinetics The pharmacokinetic properties of the Examples above, including absorption, distribution, metabolism, and excretion, may be rated in the following assays.

Compounds listed above, which may not yet have been made and/or tested, are predicted to have activity in these assays.

Metabolic Stability in Human and Murine Liver Microsomes The metabolic stability of compounds disclosed herein in pooled human liver microsomes (HLM) and pooled male mouse liver microsomes (MMLM) was determined according to the following protocol, in which the concentrations of compounds in reaction systems were evaluated by LC/MS/MS for estimating the stability in liver microsomes.

Study Design Pooled human liver microsomes (HMMCPL; PL050B) and pooled male mouse liver omes (MSMCPL; M803 3) were purchased from CellzDirect (Invitrogen).

Microsomes were stored at -80 C prior to use.

A master solution was prepared containing microsome (stock concentration 5 mg/mL, volume 50 uL, final concentration 0.5 mg/mL), MgClz solution (stock concentration 50 mM, volume 50 uL, final concentration 5 mM), phosphate buffer (stock concentration 200 mM, volume 250 uL, final tration 100 mM), and water (volume 95 uL. Five uL of 200 uM test nds or control solution (control nd: verapamil) was then added.

The final concentration of test compounds or verapamil in the reaction system was 2 uM.

The mixture was pre-warmed at 37 C for 5 min.

The on was d with the addition of 50 uL of 10 mM NADPH solution at the final concentration of 1 mM and carried out at 37 C. 50 uL of ultra-pure H20 was used instead ofNADPH solution in the negative control.

Aliquots of 50 uL were taken from the reaction solution at 0 and 30 min. The reaction was d by the addition of 3 volumes of cold ol with IS (200 nM imipramine, 200 nM labetalol and 2uM ketoprofen) at the designated time points. Samples were centrifuged at 16,000 g for 10 minutes to precipitate protein. Aliquot of 100 uL of the supernatant was diluted by 100 uL ultra-pure H20, and the mixture was used for LC/MS/MS is. All experiments were performed in duplicate.

Bioanalytical Method Samples were analyzed using liquid chromatography-mass spectrometry. The LC system comprised a Shimadzu liquid chromatograph separation system equipped with degasser DGU-20A3, solvent delivery unit LC-20AD, system ller CBM-20A, column oven CTO-10ASVP and CTC ics HTC PAL System. Chromatographic conditions included a Phenomenex column, 5.0 u C18 (2.0><50 mm); a mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water; an elution rate of 500 uL/min; column temperature 25 C; injection volume 10 uL. Mass spectrometric analysis was performed using an API 4000 instrument from AB Inc. (Canada) with an ESI interface. The data acquisition and control system were created using Analyst 1.5.1 software from ABI Inc. A turbo spray ion source and electrospray tion were employed in a multiple reaction monitoring (MRM) scan. Additional parameters included: collision gas, 6 L/min; curtain gas, 30L/min; nebulize gas, 50 L/min; auxiliary gas, 50 L/min; temperature, 500 C; ionspray voltage, +5500 v (positive MRM). Quadripoles Q1 and Q3 were set to 456.2 and 200.2, respectively; declustering potential (DP), entrance potential (EP), and collision cell entrance potential (CE) were set to 120, 10, and 55 v, tively; ion cell exit ial (CXP) was 12 v.

Analysis All calculations were carried out using Microsoft Excel. Peak areas were determined from extracted ion chromatograms. The control compounds were included in the assay. Any value of the compounds that was not within the specified limits was rejected and the experiment was repeated. The reaction system without the cofactors was used to exclude the misleading factor that ed from instability of chemical .

Results Results are shown above in Table l and below in Table 2. Without wishing to be bound by theory, comparative data indicates that methylation of the cyclopropyl group results in an increase in metabolic stability while at least maintaining efficacy.

Table 2. Metabolic ity in Murine Liver Microsomes % at 30 Example min, MLM: KDMlA Structure N0. + is > 20% IC50 - is < 20% (\NJS..\\\/\/ 1,, 9 o\\ ,NQ HN o 54% 5.1 a F 2014/049906 Compositions The following are examples of compositions which may be used to deliver compounds disclosed herein. These may be encapsulated or wet granulated using methods known in the art. ition Example 1 Ingredients Concentration (w/w %) Compound of Formula130% ium Stearate 2% Composition Example 2 Ingredients Concentration (w/w %) Compound of Formula I Lactose Magnesium Stearate 2% From the foregoing description, one skilled in the art can easily ascertain the essential teristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Other Embodiments The detailed description set—forth above is ed to aid those skilled in the art in practicing the present disclosure. However, the disclosure described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed e these embodiments are ed as illustration of several aspects of the disclosure. Any equivalent embodiments are intended to be within the scope of this disclosure. Indeed, various modifications of the sure in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description, which do not depart from the spirit or scope of the present ive discovery. Such modifications are also intended to fall within the scope of the appended claims.

All references cited in this specification are hereby incorporated by reference. The discussion of the references herein is ed merely to summarize the assertions made by their authors and no admission is made that any reference constitutes prior art relevant to patentability. Applicant reserves the right to challenge the accuracy and pertinence of the cited references.

Claims (51)

1. A compound of Formula I: O Y Z R5 ( )m ( )n R1 N R4 or a salt thereof, wherein: Y is chosen from a bond, NR4a, O, , NHC(O), S, SO2, and CH2; Z is chosen from a bond, NR4b, O, C(O)NH, NHC(O), S, SO2, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, aryl, kyl, cycloalkyl, cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl or R1 and R2, together with the nitrogen to which they , form a nitrogen-containing heterocycloalkyl ring, which is optionally substituted with between 0 and 3 R 6 groups; R3 is chosen from alkylamino, cycloalkylamino, arylamino, heteroarylamino, heterocycloalkylamino, cycloalkyl, lkylalkyl, aryl, arylalkyl, heteroaryl, arylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which is optionally substituted with between 0 and 3 R6 groups; R4, R4a, and R4b are independently chosen from hydrogen, alkyl, alkenyl, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which is ally tuted with between 0 and 3 R6 groups; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, heteroarylalkyl, cyano, alkoxy, amino, alkylamino, dialkylamino, COR7, SO2R7, NHSO2R7, NHSO2NHR7, NHCOR7, NHCONHR7, CONHR7, and CONR7R8; and R7 and R8 are independently chosen from en, and lower alkyl; or R7 and R8 may be taken together to form a nitrogen-containing heterocycloalkyl or aryl ring, which is optionally substituted with lower alkyl.

2. The nd as recited in claim 1, wherein the compound has Formula IIa or IIb: R3 R3 O R4 R5 NH NH O Y Z O Y Z ( )m ( )n ( )m ( )n R1 N R4 R5 R1 N R2 R2 (IIa) (IIb) or a salt thereof, wherein: Y is chosen from a bond, NR4a, O, C(O)NH, , S, SO2, and CH2; Z is chosen from a bond, NR4b, O, C(O)NH, NHC(O), S, SO2, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each independently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkoxyalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, phenyl, yl, heteroaryl, arylalkyl, heterocycloalkyl, and heterocycloalkylalkyl or R1 and R2, together with the nitrogen to which they attach, form a nitrogen-containing heterocycloalkyl ring, which is optionally substituted with between 0 and 3 R 6 groups; R3 is chosen from alkylamino, cycloalkylamino, ino, heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which is optionally substituted with between 0 and 3 R6 groups; R4, R4a, and R4b are independently chosen from hydrogen, alkyl, alkenyl, alkynyl, and cycloalkyl; R5 is chosen from aryl and heteroaryl, any of which is optionally substituted with between 0 and 3 R6 groups; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, aralkyl, heterocycloalkyl, heteroaryl, heteroarylalkyl, cyano, alkoxy, amino, alkylamino, dialkylamino, COR7, SO2R7, NHSO2R7, NHSO2NHR7, NHCOR7, NHCONHR7, CONHR7, and CONR7R8; and R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a en-containing heterocycloalkyl or heteroaryl ring, which is optionally substituted with lower alkyl.

3. The compound as recited in claim 2, wherein the compound has Formula IIIa or IIIb: R3 R3 O O R4 R5 NH NH O Y Z O Y Z ( )m ( )n ( )m ( )n R1 N R4 R5 R1 N R2 R2 (IIIa) (IIIb) or a salt thereof, wherein: Y is chosen from a bond, NR4a, O, C(O)NH, NHC(O), S, SO2, and CH2; Z is chosen from a bond, NR4b, O, C(O)NH, NHC(O), S, SO2, and CH2; m is an integer from 0 to 5; n is an integer from 0 to 3; R1 and R2 are each ndently chosen from, alkyl, aminoalkyl, alkylsulfonylalkyl, alkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, phenyl, biphenyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, and heterocycloalkylalkyl or R1 and R2, together with the en to which they attach, form a nitrogen-containing cycloalkyl ring, which is optionally substituted with between 0 and 3 R 6 groups; R3 is chosen from alkylamino, cycloalkylamino, arylamino, heteroarylamino, heterocycloalkylamino, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, arylalkyl, heterocycloalkyl, and heterocycloalkylalkyl any of which is optionally substituted with n 0 and 3 R6 groups; R4, R4a, and R4b are ndently chosen from hydrogen, alkyl, alkenyl, alkynyl, and lkyl; R5 is chosen from aryl and heteroaryl, any of which is optionally substituted with between 0 and 3 R6 groups; each R6 is independently chosen from hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkoxy, aryl, l, heterocycloalkyl, heteroaryl, arylalkyl, cyano, alkoxy, amino, alkylamino, dialkylamino, COR7, SO2R7, NHSO 7, NHSO 7, NHCOR7, R7, CONHR7, and CONR7R8; and 2R 2NHR R7 and R8 are independently chosen from hydrogen, and lower alkyl; or R7 and R8 may be taken together to form a nitrogen-containing heterocycloalkyl or heteroaryl ring, which is optionally substituted with lower alkyl.

4. The compound as recited in claim 1, wherein Z is NR4b .

5. The compound as d in claim 4, wherein R4b is chosen from methyl and hydrogen.

6. The compound as d in claim 5, wherein the alkyl, whether by itself or as a named part of another non-cyclic substituent, is C1-C8 alkyl.

7. The compound as recited in claim 6, wherein R3 is chosen from aryl, arylalkyl, heteroaryl, and heteroarylalkyl, any of which is optionally substituted with between 0 and 3 R6 groups.

8. The compound as recited in claim 7, wherein R3 is chosen from aryl and heteroaryl, any of which is optionally substituted with between 0 and 3 R6 groups.

9. The compound as recited in claim 8, wherein m is an integer from 0 to 1; Y is chosen from NR4a , O, S, SO2, and CH2; n is an integer from 1 to 3; and R4a is chosen from hydrogen and alkyl.

10. The compound as recited in claim 9, wherein m is 0; Y is CH2; and n is an r from 1 to 3.

11. The compound as recited in claim 10, wherein R3 is 5-6 membered monocyclic or 8-12 membered bicyclic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which is optionally tuted with n 0 and 3 R6 groups.

12. The compound as recited in claim 11, wherein R3 is 5-6 membered monocyclic heteroaryl, in which between one and four ring members may be atoms chosen from N, O, and S, and which is optionally substituted with between 0 and 3 R6 groups.

13. The compound as recited in claim 12, wherein each R6 is chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

14. The compound as recited in claim 13, wherein R3 is chosen from , , , , , , , , , , , and .

15. The nd as recited in claim 14, wherein R4 is hydrogen.

16. The compound as recited in claim 15, wherein the nitrogen-containing heterocycloalkyl ring formed by R1 and R2 together with the nitrogen to which they are attached contains 3 to eight atoms.

17. The compound as recited in claim 16, wherein the nitrogen-containing heterocycloalkyl is chosen from: , , , , , , , , , , and .

18. The compound as recited in claim 17, wherein the nitrogen-containing heterocycloalkyl is chosen from: , , , , and .

19. The compound as recited in claim 18, wherein n is 2 or 3.

20. The compound as recited in claim 19, wherein R5 is aryl, which is optionally tuted with between 0 and 3 R6 groups.

21. The compound as recited in claim 20, wherein R5 is phenyl, which is optionally substituted with between 0 and 3 R6 groups.

22. The compound as recited in claim 19, n R5 is heteroaryl, which is optionally tuted with n 0 and 3 R6 groups.

23. The compound as recited in claim 22, wherein R5 is a 5-6 membered monocyclic or 8-12 membered bicyclic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which is optionally tuted with between 0 and 3 R6 groups.

24. The compound as recited in claim 23, wherein R5 is a 5-6 membered monocyclic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which is optionally substituted with 1 or 2 R6 groups.

25. The compound as recited in claim 24, wherein R5 is chosen from: , , , , , , , , , , , and .

26. The nd as recited in claim 6, wherein R3 is aryl, optionally substituted with between 0 and 3 R6 groups.

27. The nd as d in claim 26, wherein R3 is chosen from phenyl and biphenyl, either of which is optionally substituted with n 0 and 3 R6 groups.

28. The compound as recited in claim 27, wherein m is an integer from 0 to 1; Y is chosen from NR4a, O, S, SO2, and CH2; n is an integer from 1 to 3; and R4a is chosen from hydrogen and alkyl.

29. The compound as recited in claim 28, wherein m is 0; Y is CH2; and n is an integer from 1 to 3.

30. The compound as recited in claim 28, wherein each R6 is chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

31. The compound as recited in claim 30, wherein R4 is hydrogen.

32. The compound as recited in claim 29, wherein n is 2 or 3.

33. The compound as recited in claim 32, n the en-containing heterocycloalkyl ring formed by R1 and R2 together with the nitrogen to which they are attached contains 3 to eight atoms.

34. The compound as recited in claim 33, wherein the nitrogen-containing heterocycloalkyl is chosen from: , , , , , , , , , , and .

35. The compound as recited in claim 34, wherein the nitrogen-containing heterocycloalkyl is chosen from: , , , , and .

36. The compound as recited in claim 35, wherein R5 is aryl, which is optionally substituted with between 0 and 3 R6 groups, each of which is independently chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

37. The compound as recited in claim 36, wherein R5 is phenyl, which is optionally substituted with between 0 and 3 R6 groups, each of which is independently chosen from lower alkyl, halogen, lower alkoxy, OCF3 and CF3.

38. The compound as recited in claim 35, wherein R5 is heteroaryl, which is optionally substituted with between 0 and 3 R6 groups.

39. The compound as recited in claim 38, wherein R5 is a 5-6 membered monocyclic or 8-12 membered ic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which is optionally substituted with n 0 and 3 R6 , each of which is independently chosen from lower alkyl, n, lower alkoxy, OCF3 and CF3.

40. The compound as d in claim 39, wherein R5 is a 5-6 membered clic heteroaryl, in which between one and five ring members may be heteroatoms chosen from N, O, and S, and which is optionally substituted with 1 or 2 R6 groups, each of which is independently, if t, a lower alkyl groups.

41. The compound as recited in claim 40, n R5 is chosen from: , , , , , , , , , , , and .

42. Use of a compound as recited in claim 1, or a salt thereof, in the manufacture of a medicament for the prevention or treatment of a disease or condition chosen from sickle cell disease, thalassemia major, and other beta-hemoglobinopathies.

43. A pharmaceutical composition comprising a compound as recited in claim 1, or a salt thereof, together with a pharmaceutically acceptable carrier.

44. The pharmaceutical composition as recited in claim 43, formulated for oral administration.

45. The pharmaceutical composition as recited in claim 44, additionally sing another therapeutic agent.

46. Use of a compound as recited in claim 1, or a salt thereof, in the manufacturing of a ment for inhibition of KDM1A.

47. Use of a compound as recited in any one of claims 1 to 41, or a salt thereof, in the manufacturing of a ment for the treatment of a globin-mediated disease to a patient in need thereof.

48. The use as recited in claim 47, wherein the disease is chosen from Myelodysplastic Syndrome (MDS), Acute Myelogenous Leukemia (AML), and Chronic Myelogenous Leukemia (CML).

49. Use of a compound as claimed in any one of claims 1 to 41, or a salt thereof, in the manufacturing of a medicament for achieving an effect in a patient, wherein the effect is chosen from an ion of red blood cell count, an elevation of the red blood cell count of red cells containing fetal hemoglobin, an elevation in the total tration of fetal hemoglobin in red cells, an elevation in the total concentration of fetal hemoglobin in reticulocytes, an increase in the transcription of the gamma globin gene in bone marrowderived red cell precursors, e.g., ythroblasts, a reduction in the number of sickle cell crises a patient experiences over a unit period of time, a halt to or prevention of tissue damage e.g., in the heart, spleen, brain or kidney caused by sickling cells, a reduction in the proportion of red cells that undergo sickling under physiological conditions of relative hypoxia as measured using patient blood in an in vitro assay, an increase in the amount of histone 3 lysine methylation at lysine position 4 (H3K4me1 and 2), and/or a decrease in the amount of histone 3 methylation at lysine position 9 (H3K9me1 or 2) near or at the gamma globin promoter as assayed by ChIP using a cell sample derived from a treated patient.

50. Use of a compound as recited in claim 1, or a salt thereof in the manufacturing of a medicament for inhibiting at least one KDM1A on, wherein KDM1A is to be contacted with the compound or salt, and wherein the tion is measured by phenotype of red cells or their precursors cultured in vitro for humans, mice or transgenic mice or cultured in vivo in mice or transgenic mice, the ability of cancer cells to proliferate, become differentiated, or d to undergo apoptosis, the sion of specific genes known to be regulated by KDM1A activity such as gamma globin or HOXA9, a change in the histone methylation states, a change in the methylation state of proteins known to be demethylated by KDM1A such as G9a , p53, DNMT1 or SUV39H1, expression of KDM1A-regulated genes, or g of KDM1A with a natural binding partner such as CoREST, NuRD, DNMT1 or HDACs.

51. The compound as recited in claim 1, chosen from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and , or a salt f.