Methods and compositions for the treatment of cancer

Abstract

The present disclosure describes compounds and pharmaceutical compositions for inhibiting the ENL YEATS domain for the treatment of leukemia in a subject in need thereof. The present disclosure further provides methods for use of such compounds and pharmaceutical compositions in the treatment of leukemia.

Description

TITLE OF THE INVENTIONMETHODS AND COMPOSITIONS FOR THE TREATMENT OF CANCERCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority of U. S. Provisional Appl. Ser. No. 63 / 716,616, filed November 5, 2024, the entire disclosure of which is incorporated herein by reference.STATEMENT OF GOVERNMENT RIGHTS

[0002] This invention was made with government support under grant numbers R21CA267512 and R01CA291968 awarded by the National Institutes of Health. The government has certain rights in the invention. This invention was also made with the support of the Welch Foundation under grant number A- 1715.FIELD OF THE INVENTION

[0003] This present disclosure relates to the field of cancer therapeutics, and more specifically to methods and compositions for inhibiting the growth of ENL-dependent leukemia cells.BACKGROUND OF THE INVENTION

[0004] The evolutionally conserved histone-reading function of the YEATS domain is essential for the functionality of all the YEATS domain proteins in both yeast and humans. Dysregulated interactions between YEATS domains and the posttranslational modifications they recognize have been found to be associated with various human diseases, including cancers. For example, the YEATS domain-containing protein Eleven-Nineteen Leukemia (ENL) has been shown to be essential for disease maintenance and progression of acute leukemias. As such, inhibition of the ENL YEATS domain is considered to have high therapeutic potential. Chemical compounds and peptide-mimic probes have been developed as competitive inhibitors of the ENL YEATS domain. Target selectivity has been a challenge in inhibitor development due to the high structural similarity between the YEATS domains of YEATS-containing proteins, in particular between ENL and its close homologue AF9. While peptide-mimic probes showed slightly higher potency to the ENL YEATS domain than to the YEATS domains of other proteins, the small molecule ENL inhibitor compounds reported so far failed to distinguish ENL from AF9. In addition, none of these small molecule compoundsshowed significant impact on ENL-dependent leukemia cell growth. There exists a great need for potent, selective ENL YEATS domain inhibitors. The present disclosure describes small molecule inhibitor compounds that have high selectivity, metabolic stability, and antiproliferative effects against ENL-dependent leukemia cells. This provides a significant advance in the art compared to the currently known small molecule ENL inhibitor compounds.SUMMARY OF THE INVENTION

[0005] In one aspect, the present disclosure provides a compound of Formula I or a derivative thereofor compound 13 or a derivative thereof:wherein X is a carbon or a nitrogen, andRi is a 4-, 5-, or 6-membered ring, which may be cycloalkyl, heterocyclic, heteroaryl, or aryl, wherein said ring is optionally substituted with one or more groups selected from hydrogen, C1-C12 alkyl, C1-C12 haloalkyl, C1-C12 heteroalkyl, aralkyl, and aryl sulfamide. In certain embodiments, the compound may have a formula selected from the group consisting of:,a derivative of any thereof. In some embodiments, the compounds of the present disclosure selectively target the ENL YEATS domain.

[0006] Another aspect of the present disclosure provides a pharmaceutical composition comprising an effective amount of the compounds provided herein. In some embodiments, thepharmaceutical composition further comprises one or more pharmaceutically acceptable carriers, buffers, or diluents. In other embodiments, the compound comprised in the pharmaceutical composition selectively targets the ENL YEATS domain. In some embodiments, the compound comprised in the pharmaceutical composition is effective for treating leukemia. In further embodiments, the leukemia is selected from the group consisting of acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, chronic myelomonocytic leukemia, and myeloproliferative neoplasms.

[0007] Yet another aspect of the present disclosure provides a method of treating a subject in need thereof, the method comprising administering a therapeutically effective amount of a compound of the present disclosure to the subject. In some embodiments, the subject is afflicted with leukemia. In other embodiments, the subject is afflicted with acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, chronic myelomonocytic leukemia, or myeloproliferative neoplasms. In some embodiments, said administering comprises local, regional, systemic, or continual administration. In other embodiments, said administering comprises oral administration, intravenous administration, buccal administration, rectal administration, parenteral administration, intraperitoneal administration, topical administration, intradermal administration, intratracheal administration, intramuscular administration, subcutaneous administration, or inhalation. In some embodiments, the method further comprises administering a second therapy to said subject. In further embodiments, the second therapy is selected from the group consisting of a chemotherapy, a radiotherapy, a targeted therapy, a small molecule inhibitor, an immunotherapy, and surgery. In some embodiments, the subject is a mammalian subject. In yet further embodiments, the subject is a human subject.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0009] FIG. 1 demonstrates in vitro activity of newly synthesized ENL inhibitor compounds.FIG. 1A shows dose response curve generated from AlphaScreen assays. FIG. 1B shows thenormalized NanoBRET signal response of ENL inhibitor compounds in HEK293T cells expressing NLuc-ENL YEATS.

[0010] FIG. 2 demonstrates the metabolic stability of newly synthesized ENL inhibitor compounds. FIG. 2A shows a graphical representation of the metabolic stability of each inhibitor in human plasma. FIG.2B shows a graphical representation of the metabolic stability of each inhibitor in human liver microsome was determined by plotting the incubation time (x-axis) against the natural logarithm (In) of the remaining inhibitor concentration.

[0011] FIG. 3 shows results of in vitro cytotoxicity assay using ENL inhibitor compounds in various leukemia cell lines. FIG. 3A shows cell viability data from MOLM-13 cells post 8-day treatment with inhibitor compound. FIG. 3B shows cell viability data from MV4-11 cells post 8-day treatment with inhibitor compound. FIG. 3C shows cell viability data from Jurkat cells post 8-day treatment with inhibitor compound. FIG. 3D shows cell viability data from RS4;11 cells post 8-day treatment with inhibitor compound. FIG. 3E shows cell viability data from K-562 cells post 8-day treatment with inhibitor compound.

[0012] FIG.4 shows results of cell proliferation assays of various leukemia cell lines when treated with 1 uM ENL inhibitor compounds. FIG.4A shows growth curve of treated MOLM-13 cells over 14 days. FIG.4B shows growth curve of treated MV4-11 cells over 14 days. FIG.4C shows growth curve of treated Jurkat cells over 14 days.

[0013] FIG. 5 demonstrates in vivo activity of compound 13. FIG. 5A shows the time-course of the plasma concentration of compound 13 after intravenous (IV) and oral (PO) administration in CD-I mice (n=3). FIG. 5B shows the quantification of bioluminescence levels as a measure of tumor burden (total flux = photons / s) in M0M13-luc xenograft model mice. FIG. 5C shows body weights on the indicated day post-treatment in M0M13-luc xenograft model mice. Treatment started after confirmation of AML engraftment (Day 0). FIG.5D shows Kaplan-Meier survival curves (n=5) of M0M13-luc xenograft model mice. All data represent the mean ± SD.

[0014] FIG. 6 demonstrates in vivo activity of SR-C-107 (R). FIG.6A shows the time-course of the plasma concentration of SR-C-107 (R) after intravenous (IV) and oral (PO) administration in CD-I mice (n=3). FIG. 6B shows the quantification of bioluminescence levels as a measure of tumor burden (total flux = photons / s) in M0M13-luc xenograft model mice. FIG. 6C shows body weights on the indicated day post-treatment in M0M13-lucxenograft model mice. Treatment started after confirmation of AML engraftment (Day 0). FIG.6D shows Kaplan-Meier survival curves (n=5) of MOM13-luc xenograft model mice.DETAILED DESCRIPTION OF THE INVENTION

[0015] Chromatin modifications play crucial roles in various biological processes. Chromatinmodifying complexes typically contain signature domains that either have catalytic activity or recognize and bind to specific histone modifications. A number of conserved protein domains have been identified as involved in establishing and recognizing different chromatin modifications. The YEATS domains are one such example. YEATS domains are a class of histone acetylation readers which are present in four human proteins: eleven-nineteen leukemia (ENL), YEATS domain-containing protein 2 (YEATS2), ALL1-fused gene from chromosome 9 (AF9), and glioma amplified sequence 41 (GAS41). The YEATS domain constitutes 120-140 amino acids and is evolutionarily conserved from yeast to human. The YEATS domain is an acetyllysine-binding domain that binds strongly to histone H3K9 acetylation and, to a lesser extent, H3K27 and H3K18 acetylation. It therefore serves as a histone acylated reader of various chromatin remodeling complexes and plays a role in regulation of chromatin structure, histone acetylation and deposition, gene transcription, and DNA damage response. Given their critical role in numerous processes, the YEATS domain family members from humans have been found to be associated with numerous diseases.

[0016] Dysregulated interactions between YEATS domains and the posttranslational modifications they recognize have been found to be associated with various cancers. For example, GAS41 is amplified in glioblastoma and astrocytoma and both ENL and AF9 have been found to be implicated in acute leukemias. Acute myeloid leukemia (AML) is the second most frequently diagnosed leukemia in children and adults. AML is characterized by the uncontrolled proliferation of abnormal myeloblasts, which impairs the production of normal blood components. Genetic anomalies, such as gene mutations, chromosomal rearrangements, and altered gene and microRNA expression patterns, are often implicated in AML. Among these, translocations involving the mixed-lineage leukemia (MLL) gene on chromosome 11 band q23 are common, occurring in roughly 5 to 10% of AML cases. The MLL gene encodes a histone 3 lysine 4 methyltransferase that positively regulates gene expression during development. Chromosomal translocations of the MLL gene lead to in-frame chimeric proteins consisting of an N-terminal fragment of the MLL protein fused to a C-terminal domain of a fusion partner. These MLL fusion proteins are known to function as “drivers” of the disease.Over 80 genes have been identified as fusion partners with MLL, however the most common are ENL and AF9. It has recently been shown that the YEATS domain-containing protein ENL is required for disease maintenance in AML (Wan et al.. Nature 543:265-269, 2017). Depletion of ENL led to anti-leukemic effects, including increased terminal myeloid differentiation and suppression of leukemia growth. Disrupting the interaction between the YEATS domain and histone acetylation via structure-based mutagenesis reduced RNA polymerase II recruitment to ENL target genes, leading to suppression of oncogenic gene expression. Thus, inhibition of ENL represents promising therapy for AML. The acetyllysine binding pocket of the YEATS domain of ENL has emerged as a target for developing small-molecule inhibitors due to its long, narrow, hydrophobic structures. However, designing molecules with high selectivity for the ENL YEATS domain over its close homologue, the AF9 YEATS domain, has emerged as a considerable challenge in developing these inhibitors. The present disclosure describes small molecule inhibitor compounds that have high selectivity for the ENL YEATS domain. These compounds also demonstrate high metabolic stability and robust anti-proliferative effects against AML in vitro and in vivo. The compounds and methods for their use described herein provide a significant advance in the art compared to the currently known small molecule ENL inhibitor compounds.A. Small Molecule Inhibitors of the ENL YEATS Domain

[0017] In certain aspects, the present disclosure provides ENL YEATS domain inhibitors useful as therapeutics for the treatment of a variety of leukemias, including but not limited to acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, or chronic myelogenous leukemia.

[0018] In some embodiments, the present disclosure provides inhibitor compounds of general Formula I or pharmaceutically acceptable salts thereof:

[0019] In particular embodiments, X is a carbon or a nitrogen, and Ri is a 4-, 5-, or 6-membered ring, which may be cycloalkyl, heterocyclic, heteroaryl, or aryl, wherein said ring is optionally substituted with one or more groups selected from hydrogen, C1-C12 alkyl, C1-C12 haloalkyl, C1-C12 heteroalkyl, aralkyl, and aryl sulfamide.

[0020] In some embodiments, the present disclosure provides an inhibitor compound of compound 13, shown below, or pharmaceutically acceptable salts thereof:

[0021] Additional exemplary compounds of the present disclosure are provided in Table 1.Table 1: Exemplary ENL Inhibitors

[0022] As used herein, the term “amino" refers to the -NH2 radical, “nitro" refers to the -NO2 radical, “cyano" or “nitrile" refers to the -CN radical, “hydroxy" or “hydroxyl" refers to the -OH radical, “oxo” refers to the =0 substituent, “thioxo” refers to the =S substituent, “sulfinyl” refers to bilvalent radical -S (=0)-, “sulfonyl” refers to the bilvalent radical -S(=O)2-, “sulfonamide” refers to -NH-S (=0)2-, and “sulfamide” refers to diradical group -NH-S(=0)2-NH-.

[0023] In some embodiments, “alkyl” refers to a straight or branched hydrocarbon chain radical, which is fully saturated or comprises unsaturations, has from one to thirty carbon atoms, and is attached to the rest of the molecule by a single bond. Alkyls comprising any number of carbon atoms, from 1 to 30, are included. An alkyl comprising up to 30 carbon atoms is referred to as a C1-C30 alkyl, likewise, for example, an alkyl comprising up to 12 carbon atoms is a C1-C12 alkyl. Alkyls (and other moieties defined herein) comprising other numbers of carbon atoms are represented similarly. Alkyl groups include, but are not limited to, C1-C30 alkyl, C1-C20 alkyl, Cl -C15 alkyl, Cl -CIO alkyl, C1-C8 alkyl, C1-C6 alkyl, C1-C4 alkyl, Cl-C3 alkyl, C1-C2 alkyl, C2-C8 alkyl, C3-C8 alkyl, and C4-C8 alkyl. Representative alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1 -methyl ethyl (isopropyl), n-butyl, / -butyl, s-butyl, n-pentyl, 1,1 -dimethylethyl (tert-butyl), 3 -methylhexyl, 2-methylhexyl, vinyl, allyl, propynyl, and the like. Alkyl comprising unsaturations include alkenyl and alkynyl groups. Unless stated otherwise specifically in the specification, an alkyl group may be optionally substituted as described below.

[0024] As used herein “alkylene” or “alkylene chain” refers to a straight or branched divalent hydrocarbon chain, as described for alkyl above. Unless stated otherwise specifically in the specification, an alkylene group may be optionally substituted as described below.

[0025] As used herein, “alkoxy” refers to a radical of the formula -ORawhere Rais an alkyl radical as defined. Unless stated otherwise specifically in the specification, an alkoxy group may be optionally substituted as described below.

[0026] As used herein, “cycloalkyl” or “carbocycle” refers to a stable, non-aromatic, monocyclic or polycyclic carbocyclic ring, which may include fused or bridged ring systems, which is saturated or unsaturated. Representative cycloalkyls or carbocycles include, but are not limited to, cycloalkyls having from three to fifteen carbon atoms, from three to ten carbon atoms, from three to eight carbon atoms, from three to six carbon atoms, from three to five carbon atoms, or three to four carbon atoms. Monocyclic cycloalkyls or carbocycles include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyls or carbocycles include, for example, adamantyl, norbomyl, decalinyl, bicyclo [3.3.0] octane, bicyclo[4.3.0]nonane, cis-decalin, trans-decalin, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, and bicyclo[3.3.2]decane, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Unless otherwise stated specifically in the specification, a cycloalkyl or carbocycle group may be optionally substituted.

[0027] As used herein, “fused” refers to any ring structure described herein which is fused to an existing ring structure. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure which becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be replaced with a nitrogen atom.

[0028] As used herein, “halo” or “halogen” refers to bromo, chloro, fluoro, or iodo.

[0029] [As used herein, “haloalkyl” refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2- fluoropropyl, 1,2-dibromoethyl, and the like. Unless stated otherwise specifically in the specification, a haloalkyl group may be optionally substituted.

[0030] As used herein, “haloalkoxy” similarly refers to a radical of the formula -ORawhere Rais a haloalkyl radical as defined. Unless stated otherwise specifically in the specification, a haloalkoxy group may be optionally substituted as described below.

[0031] As used herein, “heterocycloalkyl” or “heterocyclyl” or “heterocyclic ring” or “heterocycle” refers to a stable 3- to 24-membered non-aromatic ring radical comprising 2 to 23 carbon atoms and from one to 8 heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorous and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; the nitrogen atom may be optionally quatemized; and the heterocyclyl radical may be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, azetidinyl, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolyl, imidazolinyl. imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tctrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1 -oxo-thiomorpholinyl, 1, 1-dioxo-thiomorpholinyl, 12-crown-4, 15-crown-5, 18-crown-6, 21-crown-7, aza-18-crown-6, diaza-18-crown-6, aza-21 -crown-7, and diaza-21 -crown-7. Unless stated otherwise specifically in the specification, a heterocyclyl group may be optionally substituted.

[0032] The term heterocycloalkyl also includes all ring forms of the carbohydrates, including but not limited to the monosaccharides, the disaccharides and the oligosaccharides. Unless otherwise noted, hctcrocycloalkyls have from 2 to 10 carbons in the ring. It is understood that when referring to the number of carbon atoms in a heterocycloalkyl, the number of carbon atoms in the heterocycloalkyl is not the same as the total number of atoms (including the heteroatoms) that make up the heterocycloalkyl (i.e. skeletal atoms of the heterocycloalkyl ring). Unless stated otherwise specifically in the specification, a heterocycloalkyl group may be optionally substituted.

[0033] As used herein, “hctcroaryl” refers to a 5- to 14-mcmbcrcd ring system radical comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorous, and sulfur, and at least one aromatic ring. For purposes of this disclosure, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quatemized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl,benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2- a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1 -oxidopyridinyl, 1-oxidopyrimidinyl, 1- oxidopyrazinyl, 1 -oxidopyridazinyl, 1 -phenyl, IH-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise specifically in the specification, a heteroaryl group may be optionally substituted.

[0034] All the above groups may be either substituted or unsubstituted. The term “substituted” as used herein means any of the above groups (e.g, alkyl, alkylene, alkoxy, aryl, cycloalkyl, haloalkyl, heterocyclyl and / or heteroaryl) may be further functionalized wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atom substituent. Unless stated specifically in the specification, a substituted group may include one or more substituents selected from: oxo, amino, -CO2H, nitrile, nitro, hydroxyl, thiooxy, alkyl, alkylene, alkoxy, aryl, cycloalkyl, heterocyclyl, hctcroaryl, dialkylamincs, arylamincs, alkylarylamincs, diarylamines, trialkylammonium, N-oxides, imides, sulfamide and enamines; a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, triarylsilyl groups, perfluoroalkyl or perfluoroalkoxy, for example, trifmoromethyl or trifiuoromeihoxy.

[0035] As used herein, “substituted” also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g. a double- or triple-bond) to a hctcroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, “substituted' includes any of the above groups in which one or more hydrogen atoms are replaced with -NH2, -NRaC(=O)NRaRb, -NRaC(=O)ORb, - NRaSO2Rb, -OC(=O)NRaRb, -ORa, -SRa, -SORa, -SO2Ra, -OSO2Ra, -SO2ORa, =NSO2Ra, and -SO2NRaRb. In the foregoing, Raand Rb are the same or different and independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl. In addition, each of the foregoing substituents may also beoptionally substituted with one or more of the above substituents. Furthermore, any of the above groups may be substituted to include one or more internal oxygen, sulfur, or nitrogen atoms. For example, an alkyl group may be substituted with one or more internal oxygen atoms to form an ether or polyether group. Similarly, an alkyl group may be substituted with one or more internal sulfur atoms to form a thioether, disulfide, etc.

[0036] As used herein, “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, “optionally substituted alkyl” means either “alkyl” or “substituted alkyl” as defined above. Further, an optionally substituted group may be un- substituted (e.g., -CH2CH3), fully substituted (e.g., -CF2CF3), mono-substituted (e.g., -CH2CH2F) or substituted at a level anywhere in between fully substituted and mono-substituted (e.g., -CH2CHF2, - CH2CF3, -CF2CH3, -CFHCHF2, etc). It will be understood by those skilled in the art with respect to any group containing one or more substituents that such groups are not intended to introduce any substitution or substitution patterns (e.g., substituted alkyl includes optionally substituted cycloalkyl groups, which in turn are defined as including optionally substituted alkyl groups, potentially ad infinitum) that are sterically impractical and / or synthetically non- feasible.

[0037] In some embodiments, the compounds described herein exist as geometric or stereoisomeric forms. They may have one or more chiral centers where each center exists in the R configuration or S configuration. As used herein, the term "stereoisomers" refers to compounds made up of the same atoms having the same bond order but having different three-dimensional arrangements of atoms which are not interchangeable. Such stereoisomers, as encompassed by the present disclosure, can exist as a single enantiomer, a mixture of diastereomers, or a racemic mixture.

[0038] Enantiomers are stereoisomers that are non-superimposable mirror images. As used herein, the term "optical isomer" is equivalent to the term "enantiomer." As used herein the term "diastereomer" refers to two stereoisomers which are not mirror images but also not superimposable. The terms "racemate," "racemic mixture," or "racemic modification" refer to a mixture of equal parts of enantiomers. The term "chiral center" refers to a carbon atom to which four different groups are attached. Choice of the appropriate chiral column, eluent, and conditions necessary to effect separation of pairs of enantiomers is well known to one of ordinary skill in the art using standard techniques (sec, e.g., Jacques et al., "Enantiomers, Racemates, and Resolutions," John Wiley and Sons, Inc., 1981). In some embodiments, thecompounds described herein possess one or more double bonds. The compounds presented herein include all cis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers as well as the corresponding mixtures thereof. In some situations, compounds exist as tautomers. The compounds described herein include all possible tautomers within the formulas described herein. The present disclosure contemplates all such compounds, including cis-and transisomers, R- and S-enantiomers, diastereomers, (D) -and (L) -isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention.

[0039] In some embodiments, the compounds described herein exist in their isotopically-labeled forms. In other embodiments, the methods disclosed herein include methods of treating diseases by administering such isotopically labeled compounds. In certain embodiments, the methods disclosed herein include methods of treating diseases by administering such isotopically labeled compounds as pharmaceutical compositions. The present invention therefore includes all pharmaceutically acceptable isotopically labeled compounds, which are identical to those recited herein, wherein one or more atoms may be replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number which predominates in nature. Non-limiting examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as2H,3H,13C,14C,15N,18O,17O,35S,18F, and36C1. Compounds described herein, and the metabolites, pharmaceutically acceptable salts, prodrugs, solvates, or derivatives thereof which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of the present disclosure. Certain isotopically labeled compounds, for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drag and / or substrate tissue distribution assays. In some embodiments, substitution with heavier isotopes such as deuterium (2H) can afford certain therapeutic advantages such as greater metabolic stability, increased in vivo halflife, or reduced dosage requirements. Isotopically labeled compounds and the pharmaceutically acceptable salts, esters, prodrags, solvates, hydrates, or derivatives thereof can generally be prepared by carrying out the methods provided in the present disclosure and by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. In some embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

[0040] In some embodiments, the compounds described herein exist as their pharmaceutically acceptable salts. In other embodiments, the methods disclosed herein include methods of treating diseases by administering such pharmaceutically acceptable salts. In certain embodiments, the methods disclosed herein include methods of treating diseases by administering such pharmaceutically acceptable salts as pharmaceutical compositions. Examples of pharmaceutically acceptable salts include those salts prepared by reaction of the compounds described herein with a mineral, organic acid, or inorganic base. In some embodiments, the compounds described herein possess acidic or basic groups and therefore react with any of a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt. In some embodiments, these salts are prepared in situ during the final isolation and purification of the compounds of the disclosure, or by separately reacting a purified compound in its free form with a suitable acid or base, and isolating the salt thus formed.

[0041] The degree of ionization in the resulting salt may vary from completely ionized to almost non-ionized. Suitable non-toxic, acid-addition pharmaceutically acceptable salts include, but are not limited to, the acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulphate / sulphate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonatc, mandelates mesylate, mcthylsulphatc, naphthylatc, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, salicylate, saccharate, stearate, succinate, sulfonate, stannate, tartrate, tosylate, trifluoroacetate, and xinofoate salts.

[0042] Suitable non-toxic, base-addition pharmaceutically acceptable salts include, but are not limited to the aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olaminc, potassium, sodium, tromethamine and zinc salts. For a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002), which is incorporated herein by reference.

[0043] In some embodiments, the compounds of the present disclosure may form prodrugs. Prodrugs are generally drug precursors that, following administration to an individual and subsequent absorption, arc converted to an active, or a more active species via some process, such as conversion by a metabolic pathway. Some prodrugs have a chemical group present onthe prodrug that renders it less active and / or confers solubility or some other property to the drug. Once the chemical group has been cleaved and / or modified from the prodrug the active drug is generated. Prodrugs are often useful because, in some situations, they are easier to administer than the parent drug. They are, for instance, bioavailable by oral administration whereas the parent is not. In certain instances, the prodrug also has improved solubility in pharmaceutical compositions over the parent drug. An example, without limitation, of a prodrug would be a compound as described herein which is administered as an ester to facilitate transmittal across a cell membrane where water solubility is detrimental to mobility but which then is metabolically hydrolyzed to the carboxylic acid, the active entity, once inside the cell where water-solubility is beneficial. A further example of a prodrug might be a short peptide bonded to an acid group where the peptide is metabolized to reveal the active moiety. Preparations of such prodrug derivatives are discussed in various literature sources and any such method known in the art may be used to prepare prodrags of the compounds of the present disclosure.

[0044] In some embodiments, the compounds of the present disclosure can be present as a solvate. Solvates may contain either stoichiometric or non-stoichiometric amounts of a solvent and, in certain embodiments, are formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. In some embodiments, the solvent used to prepare the solvate is an aqueous solution and may be referred to as a hydrate. The compounds of the present disclosure, in certain embodiments, can be present as a hydrate. In certain embodiments, a hydrate may be obtained by crystallization from a solvent or from aqueous solution. In particular embodiments, one, two, three or any arbitrary number of solvent or water molecules can combine with the compounds of the present disclosure to form solvates and hydrates. Unless otherwise specified, the present disclosure includes all such possible solvates. For the purposes of the compounds and methods described herein, the solvated forms should be considered equivalent to the unsolvated forms.

[0045] As used herein the term “derivative” when used in reference to the compounds of the present disclosure refers to pharmaceutically acceptable salts, prodrags, isotopically labeled forms, deuterated forms, radioactively labeled forms, isomers, solvates, and any combination thereof of the compounds described herein.B. Methods of Treatment and Pharmaceutical Compositions

[0046] In certain aspects, the present disclosure provides methods, pharmaceutical compositions, and therapeutic compositions for the treatment of cancer. In one embodiment, the pharmaceutical and therapeutic compositions of the present disclosure comprise a compound described herein, or a pharmaceutically acceptable salt, prodrug, solvate, or isomer thereof.

[0047] In some embodiments, a compound of the present disclosure may be combined with a pharmaceutically acceptable carrier. As used herein, a “pharmaceutically acceptable carrier,’’ “pharmaceutically acceptable adjuvant,” or “adjuvant” refers to reagents, cells, compounds, materials, compositions, and / or dosage forms that are not only compatible with a therapeutic agent, or other agents to be administered therapeutically, but also are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other complication commensurate with a reasonable benefit / risk ratio. Also included may be an agent that modifies the effect of other agents and is useful in preparing a therapeutic compound or pharmaceutical compound or composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable. Such an agent may be added to a therapeutic composition or pharmaceutical composition to modify for example the cellular target, cellular localization, or cellular uptake of a therapeutic agent as described herein. Such an agent may include any excipient, diluent, carrier, or adjuvant that is acceptable for pharmaceutical use. Such an agent may be non-naturally occurring, or may be naturally occurring, but not naturally found in combination with other agents in the therapeutic or pharmaceutical composition.

[0048] As used herein, a “therapeutic compound” or “therapeutic composition” refers to a composition comprising a therapeutic agent of the present disclosure. In one embodiment, the composition is capable of reducing, stabilizing, or eliminating tumor growth or tumor progression in a subject. In another embodiment, the composition is capable of reducing, stabilizing, or eliminating tumor size in a subject.

[0049] A compound or composition of the present disclosure is meant to encompass a composition suitable for administration to a subject, such as a mammal, particularly a human subject. In general, a therapeutic composition is sterile, and preferably free of contaminants that are capable of eliciting an undesirable response within the subject (e.g., the compound(s) in the composition arc pharmaceutical grade). Therapeutic compositions may be designed for administration to subjects in need thereof via a number of different routes of administrationincluding, but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular, intra-articular), intranasal, buccal, topical, rectal, or transdermal administration routes. The appropriate dosage of a composition, as described herein, may be determined based on the type of disease to be treated, the severity and course of the disease, the clinical condition of the individual, clinical history, response to the treatment, and the discretion of the attending physician.

[0050] In some embodiments, therapeutic compositions provided by the present disclosure may include various "unit doses." A unit dose is defined as containing a predetermined quantity of the therapeutic composition. The quantity to be administered, and the particular route and formulation, is within the skill of determination of those in the clinical arts. A unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time. In some respects, a unit dose comprises a single administrable dose.

[0051] As used herein, an “effective amount’’ or therapeutically effective amount” refers to an amount of a compound administered to a mammalian subject, either as a single dose or as part of a series of doses, which is effective to produce a desired therapeutic effect.

[0052] Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are specific to each individual. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability and toxicity of the particular therapeutic substance or other therapies a subject may be undergoing.

[0053] As used herein, “treatment” of an individual (e.g. a mammal such as a human) or a cell is any type of intervention used in an attempt to alter the natural course of the individual or cell. In some embodiments, treatment includes administration of a pharmaceutical composition, subsequent to the initiation of a pathologic event or contact with an etiologic agent and includes stabilization of the condition (e.g., condition does not worsen) or alleviation of the condition.

[0054] As used herein, “subject” or “patient” refers to animals, including humans, who are treated with the inhibitors, therapeutic compounds, or compositions or in accordance with the methods described herein. For diagnostic or research applications, a wide variety of mammals may be suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and swine, such as inbred pigs and the like. In particular embodiments, a subject in need of therapy may be any subject who comprises a cancer cell as described herein. In another embodiment,the subject may be afflicted with leukemia as described herein. Non-limiting examples of such diseases or conditions include acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, chronic myelomonocytic leukemia, or myeloproliferative neoplasms.

[0055] A composition, as described herein, may include, in particular embodiments, a combination of therapeutic agents. In some embodiments, a composition as described here may be administered as a single composition or as more than one composition. Different compositions as provided herein, in certain embodiments, may be administered by the same route of administration or by different routes of administration.

[0056] A pharmaceutical composition of the present disclosure may comprise, in some embodiments, a targeting molecule. In one embodiment, the targeting molecule may be cellspecific or tissue-specific. Numerous such targeting molecules are known in the art and any such targeting molecule may be used according to the present disclosure. In certain embodiments, a composition of the present disclosure may be modified with or conjugated to a peptide, a protein, a colloidal molecule, or a polymer to facilitate delivery or adsorption. The pharmaceutical composition of the present disclosure, in some embodiments, may be serum-free, endotoxin-free, or sterile.

[0057] In certain embodiments, the compositions and methods for treating an individual described herein may be combined with any other composition or method of treatment known in the art. The compositions and methods may be administered in any suitable manner known in the art. For example, a first and a second therapeutic agent or inhibitor may be administered sequentially (at different times) or concurrently (at the same time). In some aspects, a first and a therapeutic agent or inhibitor may be administered in separate compositions. In certain embodiments, a first and a second cancer treatment or inhibitor may be administered in the same composition.

[0058] Non-limiting examples of additional treatment modalities that may be included in combination with the compositions and methods provided herein include a therapeutic agent or surgery. In specific embodiments, the methods and compositions of the present disclosure may be combined with other therapies directed towards the treatment of cancer as described herein.

[0059] In certain embodiments, the methods of the present disclosure may comprise administering a second therapy selected from the group consisting of a chemotherapy, a radiotherapy, a targeted therapy, a small molecule inhibitor, an immunotherapy, and surgery.

[0060] The term "about" is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term "or" in the claims is used to mean "and / or" unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word "comprising" or other open language in the claims, the words "a" and "an" denote "one or more," unless specifically noted otherwise. The terms "comprise," "have," and "include" are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as "comprises," "comprising," "has," "having," "includes," and "including," are also open-ended. For example, any method that "comprises," "has," or "includes" one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any system or method that "comprises," "has," or "includes" one or more components is not limited to possessing only those components and covers other unlisted components.

[0061] Other objects, features, and advantages of the present disclosure are apparent from detailed description provided herein. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. Any embodiment of the present disclosure may be used in combination with any other embodiment described herein.

[0062] All references herein are incorporated herein by reference in their entirety.EXAMPLES

[0063] The following examples are included to illustrate embodiments of the present disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventor to function well in the practice of the invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which areboth chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

[0064] The compounds of the present disclosure may be prepared by any suitable method known in the art. In some embodiments, the compounds of the present disclosure may be prepared by methods described in the Examples, together with synthetic methods known in the art of organic chemistry, or modifications and derivatizations that are familiar to those of ordinary skill in the art.

[0065] In certain embodiments, preferred methods for preparation of the compounds of the present disclosure include, but are not limited to, those described in the Examples. During any of the synthetic sequences described herein, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules concerned. This can be achieved by means of conventional protecting groups, such as those described in T. W. Greene, Protective Groups in Organic Chemistry, John Wiley & Sons, 1981; and T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, 1991, which are hereby incorporated by reference.

[0066] Embodiments of the present disclosure are further described in the following examples. The examples are merely illustrative and do not in any way limit the scope of the invention as claimed.EXAMPLE 1: Synthesis of Compound 13

[0067] Compound 13 was initially developed based on results obtained from a structureactivity relationship study of small molecule compounds identified through a high-throughput screen of inhibitors of the ENL YEATS domain. Scheme 1 shows synthesis of compound 13.Scheme 1.

[0068] Synthesis of intermediate 3: methyl 3-hydroxy-4-methylpentanoate.

[0069] Intermediate 2 was dissolved in 100 mL absolute MeOH and cooled to -10°C under inert atmosphere conditions. NaBH₄ (2.62 g, 69 mmol) was added to the resultant solution over several minutes and the starting material was consumed (TLC analysis). The reaction was then quenched with acetone and after several minutes concentrated on the rotary evaporator. The residue was dissolved in EtOAc and washed with 1.2 M HC1 followed by saturated NaHCCh and brine. The organic phase was dried over Na2SO4, filtered, and concentrated on the rotary evaporator. The residue was purified by filtration to obtain intermediate 3 (yield: 22.7 g, 90%). ’ll NMR (400 MHz, CDC13) 5 3.78 (ddd, J= 9.6, 5.7, 2.9 Hz, 1H), 2.56 - 2.35 (m, 2H), 1.70 (pd, J = 6.9, 5.9 Hz, 1H), 0.93 (dd, J= 11.2, 6.8 Hz, 6H).

[0070] Synthesis of intermediate 4: N-(benzyloxy)-3-hydroxy-4-methylpentanamide.

[0071] In Scheme 1, AftCHs (2.0 M in heptane, 2 eq.) was slowly added to an ice bath cooled suspension of OBHA’HCl (2 eq.) in dry CH2CI2 via syringe. After the evolved gas subsided, the remaining solution was warmed to room temperature and stirred for 1 hour. The resulting clear solution was cooled to 0°C and intermediate 3 (22 g 150 mmol) in CH2O2 (88 mL) was added via cannula. The heterogenous mixture was stirred for 12-15 hours at room temperature and subsequently quenched by the addition of 1.2 M HC1. The two layers were separated, and the aqueous layer was extracted (4x 50 mL) with CH2O2. The extract and the original organic layer were combined and washed with 1.2 M HC1, saturated NaHCOj, and brine. The organic phase was then dried over Na2SC>4, filtered, and concentrated on a rotary evaporator to obtaina crude product. The crude product was purified by recrystallization using CH2Q2 and hexane to obtain intermediate 4 (yield: 28.8 g, 81%). ¹H NMR (400 MHz, DMSO-d6) 5 10.93 (s, 1H), 7.45 - 7.29 (m, 5H), 4.78 (s, 2H), 4.59 (d, J = 5.2 Hz, 1H), 3.65 (dq, J = 9.2, 4.7 Hz, 1H), 2.12 - 1.89 (m, 2H), 1.54 (pd, J = 6.8, 4.7 Hz, 1H), 0.82 (dd, J = 8.1, 6.8 Hz, 6H).13C NMR (101 MHz, DMS0-&) 5 168.37, 136.14, 128.74, 128.27, 128.17, 76.78, 71.52, 37.72, 33.06, 18.77, 17.13.

[0072] Synthesis of intermediate 5: l-(benzyloxy)-4-isopropylazetidin-2-one.

[0073] In Scheme 1, intermediate 4 (28.4 g, 120 mmol) was added to round-bottom flask with triphenylphosphine (37.77 g, 144 mmol). The flask was purged with argon and freshly distilled acetonitrile was added by syringe. The resulting slurry was cooled in an ice bath to 0°C. Freshly distilled EtjN (41.8 mL, 300 mmol) was added to the cooled solution, followed by dropwise addition of CCI4 (13.96 mL, 144 mmol). After 30 min, flask was removed from the ice bath and the reaction was warmed to room temperature and stirred for 20-26 hours. The reaction mixture was then concentrated on a rotary evaporator and the residue was redissolved in EtOAc or Et2O and filtered through a pad of silica by vacuum filtration to remove the hydrochloride salts. The filtrate was concentrated and chromatographed to obtain intermediate 5 (yield: 21.8 g, 83%). ’ll NMR (400 MHz, DMSO-ifc) 57.47 - 7.34 (m, 5H), 4.99 - 4.85 (m, 2H), 3.60 (ddd, J = 6.2, 5.3, 2.5 Hz, 1H), 2.61 (dd, 7= 13.6, 5.4 Hz, 1H), 2.37 (dd, 7= 13.7, 2.5 Hz, 1H), 1.81 (dq, 7 = 13.4, 6.7 Hz, 1H), 0.92 (d, 7 = 6.8 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H).13C NMR (101 MHz, DMSO-rfe) 6 163.58, 135.27, 129.04, 128.60, 128.39, 76.58, 62.02, 34.32, 29.78, 18.43, 17.40.

[0074] The racemic mixture of intermediate 5 was separated using a chiral column to obtain optically pure enantiomers, intermediate 5a and intermediate 5b. The configuration of intermediate 5a and intermediate 5b were confirmed by small molecule X-ray crystallography analysis.

[0075] Synthesis of intermediate 6a: (S)-4-isopropylazetidin-2-one. Freshly prepared W-2 Raney nickel was added to a solution of intermediate 5a (10.0 g, 45.63 mmol) and methanol and the mixture was stirred for 16 hours under hydrogen atmospheric conditions. The reaction mixture was filtered through a pad of celite and washed with methanol (2 x 30 mL). The combined organic layers were concentrated under reduced pressure, and the residue was purified by column chromatography using 5-10% methanol and dichloromethane to obtain intermediate 6a (4.9 g, 95%). ¹H NMR (400 MHz, CDCl₃) 56.40 (s, 1H), 3.30 (ddd, 7 = 7.8,5.0, 2.4 Hz, 1H), 3.02 - 2.90 (m, 1H), 2.58 (ddd, 1H), 1.78 - 1.60 (m, 1H), 0.91 (dd, J = 17.1, 6.7 Hz. 6H).13C NMR (101 MHz, CDCh) 5 168.74, 54.03, 41.23. 32.69, 18.53, 17.80.

[0076] Synthesis of intermediate 6b: (Z?)-4-isopropylazetidin-2-one. ¹H NMR (400 MHz, CDCl₃) δ 6.12 (s, 1H), 3.31 (ddd, J = 7.7, 5.0, 2.4 Hz, 1H), 3.02 - 2.92 (m, 1H), 2.60 (ddd, J = 14.9, 2.5, 1.2 Hz, 1H), 1.77 - 1.64 (m, 1H), 0.93 (dd, J = 15.9, 6.7 Hz, 6H).13C NMR (101 MHz, CDCh) 6 168.57, 54.14, 41.40, 32.78, 18.65, 17.90.

[0077] Synthesis of intermediate 7a: (S)-2-isopropylazetidine hydrochloride.

[0078] A 2 M L1AIH4 solution in tetrahydrofuran (THF; 65 mL, 130 mmol) was suspended in dry THF (130 mL) and the resulting solution was cooled to 0°C. Trimethylsilyl chloride (17.0 mL, 134.4 mmol) was added dropwise, and the solution was stirred for 2 hours at the same temperature. The mixture was cooled to -20°C and intermediate 6a (4.9 g, 43.36 mmol) was added in small portions. The resulting mixture was stirred for 2 days at room temperature, and then the excess of LiAlH4 was quenched with 40% aqueous NaOH solution. Inorganic precipitates were filtered out. The filtrate was cooled to 0°C and 2 M HC1 (43.3 mL, 86.7 mmol) was added to the filtrate. The mixture was stirred for 5-10 min at the same temperature. The THF was evaporated from the reaction mass and the aqueous layer was evaporated under lyophilization to obtain intermediate 7a (5.41g, 39.88 mmol). ’ll NMR (400 MHz, DMSO-r / e) 89.39 (s, 2H), 3.99 - 3.87 (m, 1H), 3.87 - 3.74 (m, 1H), 3.65 - 3.53 (m, 1H), 2.37 - 2.26 (m, 1H), 2.26 - 2.16 (m, 1H), 2.16 - 2.04 (m, 1H), 0.88 (d, J = 6.5 Hz, 3H), 0.80 (d, J = 6.8 Hz, 3H).13C NMR (101 MHz, DMSO-tZ6) 665.30, 40.49, 31.14, 23.36, 17.78, 16.42.

[0079] (R)-2-isopropylazetidine hydrochloride (intermediate 7b) was synthesized through the same method as intermediate 7a. ¹H NMR (400 MHz, DMSO-d₆) δ 9.41 (s, 2H), 3.99 - 3.87 (m, III), 3.86 - 3.73 (m, III), 3.66 - 3.53 (m, III), 2.36 - 2.26 (m, III), 2.26 - 2.16 (m, III), 2.16 - 2.06 (m, 1H), 0.87 (d, J = 6.5 Hz, 3H), 0.80 (d, J= 6.8 Hz, 3H).13C NMR (101 MHz, DMSO-rfc) 665.41, 40.61, 31.21, 23.41, 17.82, 16.47.

[0080] Synthesis of racemate compound 13: N-(4-((2-isopropylazetidin-l-yl)methyI)benzyI)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0081] In Scheme 1, intermediate 7a or intermediate 7b (54 mg, 0.4 mmol) and Ti(OiPr)4 (0.6 mmol, 170 mg) were added to a solution of compound N-(4-formylbenzyl)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide (as described previously in Ma et al.,. J. Med. Chem. 64(15): 10997-11013, 2021) in DMF (1 mL). The mixture was stirred at room temperature for 10 min. Then NaBH(OAc)3 (0.6 mmol, 127 mg) was added and the mixture was heated to 70-75 °C for 24 h under N2. The reaction mixture was diluted with saturated Nal ICO solution (20 mL) and the precipitate was filtered. The filtrate was then extracted by EtOAc (2 x 30 mL). The combinedorganic layers were dried with anhydrous Na2SC>4 and concentrated in vacuo. The residue was then purified by column chromatography (silica gel, 20% methanol / EtOAc as eluent) to yield Compound 13 (R) or Compound 13 (S).

[0082] Compound 13 (R): (R)-N-(4-((2-isopropylazetidin-l-yl)methyI)benzyI)- [l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.!H NMR (400 MHz, CD3OD) 5 9.38 - 9.28 (m, 1H), 9.13 (d, J = 1.2 Hz, 1H), 8.00 - 7.80 (m, 2H), 7.59 - 7.43 (m, 4H), 4.68 (d, J = 4.0 Hz, 2H), 4.47 (d, J = 13.1 Hz, 1H), 4.36 (d, J = 13.1 Hz, 1H), 4.22 - 3.97 (m, 2H), 3.92 - 3.78 (m. 1H), 2.65 - 2.51 (m. 1H), 2.37 - 2.21 (m, 1H), 2.19 - 2.03 (m, 1H), 1.00 - 0.89 (m, 6H).13C NMR (101 MHz, CD3OD) 5 164.86, 149.00, 139.59, 137.52, 129.97, 127.85, 127.40, 126.44, 122.37, 114.23, 74.49, 61.57, 49.58, 42.95, 32.88, 21.14, 17.75, 16.12. HRMS (ESI+) was calculated for C21H25N5O [M+H]+364.2000, found 364.2124.

[0083] Compound 13 (S): (S)-N-(4-((2-isopropylazetidin-l-yl)methyl)benzyl)- [l,2,4]triazolo[4,3-a]pyridine-6-carboxamide. ¹H NMR (400 MHz, CD₃OD) δ 9.30 (s, 1H), 9.08 (t, J = 1.3 Hz, 1H), 7.95 - 7.66 (m, 2H), 7.49 - 7.26 (m, 4H), 4.63 (s, 2H), 4.61 (s, 1H), 4.08 (d, J = 12.7 Hz, 1H), 3.73 (d, J = 12.6 Hz, 1H), 3.32 - 3.16 (m, 2H), 2.30 - 2.12 (m,lH), 2.02 - 1.93 (m, 1H), 1.90 - 1.80 (m, 1H), 0.97 (d, J = 6.6 Hz, 3H), 0.86 (d, J = 6.7 Hz, 3H).13C NMR (101 MHz, CD3OD) 5 164.76, 148.98, 137.50, 136.39, 129.21, 127.39, 127.36, 126.39. 122.45, 114.20, 73.27, 63.12, 49.79, 48.06, 47.85, 47.64, 47.42, 47.21, 47.00, 43.10, 34.48, 21.64, 18.38, 16.73. HRMS (ESI+) was calculated for C21H25N5O [M+H]+364.2000, found 364.2122.EXAMPLE 2: Synthesis of Novel ENL Inhibitors.

[0084] Based on a previous ENL-specific NanoBRET assay, compound 13 demonstrated strong cellular permeability and high potency and was therefore determined to be a good candidate for structure-guided optimization. These efforts resulted in the development of the racemate compound SR-C-107, which was generated by removing one of the benzimidazole nitrogen atoms and introducing a new nitrogen atom into the ring system to predispose the molecule to adopt the conformation in which it binds to the ENL YEATS domain. These modifications were designed to enhance hydrogen bond interactions and predispose the molecule to adopt the conformation needed for binding to the ENL YEATS domain. Furthermore, the nitrogen in the azetidine ring is believed to be positioned to potentially form a salt bridge with the Glu75 side chain. To further investigate the role of the 2-isopropyl azetidine group in binding affinity, YR-D-120 and YR-D-121 were developed. Scheme 2exemplifies the synthesis of the compounds YR-D-120, YR-D-121, SR-C-107, SR-C-107 (R) and SR-C-107 (S).Scheme 2.

[0085] General procedure for the synthesis of intermediates 8a, 8b, 8c, 8d, and 8e.

[0086] In Scheme 2, a stirred solution of 6-chloro-lH-pyrrolo[3,2-c]pyridine-2-carbaldehyde (2.0 g, 11.1 mmol) and (S)-2-methylpyrrolidine (2.25 g, 16.6 mmol) in anhydrous DCE (40 mL) was added to acetic acid (0.66 g, 11.1 mmol) at 0°C. The mixture was stirred at room temperature for 3 hours. NaBH(OAc)3 was added and the mixture was stirred at same temperature for 12 hours. After completion of the reaction, the reaction was quenched with a saturated NaHCCh solution (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic layer was washed with brine, dried over MgSC, and concentrated in a vacuum. The residue was then purified with flash chromatography (0-10% MeOH in DCM as the eluent) to obtain the desired compound.

[0087] intermediate 8a: (S)-6-chloro-2-((2-methylpyrrolidin-l-yl)methyl)-lH-pyrrolo[3,2-c]pyridine. Yield: 1.5 g. 60%. ’ll NMR (400 MHz, DMSO-6) 5 11.67 (s. 1H),8.52 (d, J = 0.8 Hz, 1H), 7.33 (t, J = 0.9 Hz, 1H), 6.47 (s, 1H), 4.11 - 4.05 (m, 1H), 3.48 (d, J = 14.0 Hz, 1H), 2.88 (ddd, J = 9.3, 7.1, 4.0 Hz, 1H), 2.24 (q, J = 8.8 Hz, 1H), 1.99 - 1.92 (m, 1H), 1.65 (tdd, J = 8.4, 6.4, 3.4 Hz, 2H), 1.45 - 1.31 (m, 1H), 1.11 (d, J = 6.1 Hz, 3H).

[0088] intermediate 8b: 6-chloro-2-((2-cyclopropylazetidin-l-yl)methyl)-lH-pyrrolo[3,2-c]pyridine. ¹H NMR (400 MHz, DMSO-d₆) δ 11.50 (s, 1H), 8.44 (d, J = 0.9 Hz, 1H), 7.24 (t, J = 0.9 Hz, 1H), 6.35 (s, 1H), 3.97 (q, J = 7.1 Hz, 1H), 3.75 (d, J = 14.0 Hz, 1H), 3.49 (d, J = 13.9 Hz, 1H), 3.08 (ddd, J = 8.6, 6.6, 2.3 Hz, 1H), 2.68 (ddd, J = 9.3, 7.9, 6.5 Hz, 1H), 2.58 (q, J = 7.8 Hz, 1H), 1.99 - 1.87 (m, 1H), 1.82 - 1.68 (m, 1H), 0.81 (qt, J = 8.0, 4.9 Hz, 1H), 0.31 - 0.13 (m, 2H), 0.06 - -0.06 (m, 2H). HRMS (ESI+) was calculated for C21H22N7O [M+H]+388.1880, found 388.1874.

[0089] racemic intermediate 8c (comprising enantiomers 8d and 8e): 6-chloro-2-((2-isopropylazetidin-l-yl)methyI)-lH-pyrrolo[3,2-c] pyridine, ’ll NMR (400 MHz, DMSO-db) 8 11.53 (s, 1H), 8.51 (d, J = 9.1 Hz, 1H), 7.32 (d, J = 3.4 Hz, 1H), 6.42 (s, 1H), 3.94 - 3.45 (m. 2H), 3.16 (t, J = 5.6 Hz. 1H), 3.02 - 2.66 (m, 2H), 1.95 (dd, J = 20.1, 11.6 Hz, 1H), 1.81 - 1.69 (m, 1H), 1.60 (dd, J = 13.3, 6.5 Hz, lH), 0.81 (dd, 7 = 12.3, 6.7 Hz, 6H).

[0090] Synthesis of intermediate 9a: (S)-6-chloro-2-((2-methylpyrrolidin-l-yl)methyl)-l-((2-(trimethylsilyl)ethoxy)methyl)-177-pyrrolo[3,2-c]pyridine.

[0091] In Scheme 2, NaH (4.21 mmol, 0.101 g, 60%) was added to a stirred solution of intermediate 8a (350 mg, 1.40 mmol) in anhydrous THF (10 mL) at 0°C. SEM-C1 (234 mg, 1.40 mmol) was subsequently added to the reaction dropwise at 0°C. The temperature was raised slowly to room temperature and the reaction mixture was stirred for 2 hours. The mixture was then poured into water (10 mL) and extracted with EtOAc (2x20 mL). The organic layer was dried over anhydrous Na2SC>4 and concentrated in vacuo. The residue was then purified with flash chromatography (0-50% EtOAc in hexane as the eluent) to obtain intermediate 9a (yield: 300 mg. 56%). ¹H NMR (400 MHz, CDCl₃) δ 8.56 (d, J = 0.9 Hz, 1H), 7.37 (t, J = 0.9 Hz. 1H), 6.44 (s, 1H), 5.78 (d, / = 10.9 Hz, 1H), 5.49 (d, J = 10.8 Hz, 1H), 4.16 (dd, J = 13.4, 1.0 Hz, 1H), 3.49 (dtd, J = 24.1, 9.1, 7.3 Hz, 2H), 3.32 (d, J = 13.5 Hz, 1H), 2.86 - 2.75 (m, 1H), 2.42 (ddt, J = 13.8, 8.0, 6.1 Hz, 1H), 2.15 (q, J = 8.9 Hz, 1H), 2.07 - 1.91 (m, 1H), 1.66 (tt, J = 8.8, 5.8 Hz, 2H), 1.50 - 1.35 (m, 1H), 1.16 (d, J = 6.0 Hz, 3H), 0.99 - 0.81 (m, 3H), -0.05 (s, 9H).

[0092] Synthesis of intermediate 9b: 6-chloro-2-((2-cyclopropylazetidin-l-yl)methyl)-l-((2(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridine.

[0093] In Scheme 2, NaH (4.21 mmol, 0.101 g, 60%) was added to a slirred solution of intermediate 8b (350 mg, 1.40 mmol) in anhydrous THF (10 mL) at 0°C. SEM-C1 (234 mg, 1.40 mmol) was subsequently added to the reaction dropwise at 0°C. The temperature was raised slowly to room temperature and the reaction mixture was stirred for 2 hours. The mixture was then poured into water (10 mL) and extracted with EtOAc (2x20 mL). The organic layer was dried over anhydrous Na2SC>4 and concentrated in vacuo. The residue was then purified with flash chromatography (0-50% EtOAc in hexane as the eluent) to obtain intermediate 9b. ¹H NMR (400 MHz, CDCl3) 88.65 - 8.57 (m, 1H), 7.45 - 7.35 (m, 1H), 6.47 (s, 1H), 5.71 (q, 7 = 11.0 Hz, 2H), 4.01 (t, J = 13.8 Hz, 1H), 3.63 - 3.47 (m, 3H), 3.22 (s, 1H), 2.84 - 2.77 (m, 1H), 2.67 (d, 7 = 9.7 Hz, 1H), 2.19 - 2.00 (m, 1H), 2.00 - 1.91 (m, 1H), 1.64 (s, 2H), 1.05 -0.86 (m, 1H), 0.45 - 0.38 (m, 2H), 0.12 (tq, 7= 13.3, 4.9 Hz, 2H), -0.00 (s, 9H).

[0094] Synthesis of racemic intermediate 9c (comprising enantiomers 9d and 9e): 6-chloro-2-((2-isopropylazetidin-l-yl)methyl)-l-((2 (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridine.

[0095] In Scheme 2, NaH (4.21 mmol, 0.101 g, 60%) was added to a stirred solution of intermediate 8c (350 mg, 1.40 mmol) in anhydrous THF (10 mL) at 0°C. SEM-C1 (234 mg, I.40 mmol) was subsequently added to the reaction dropwise at 0°C. The temperature was raised slowly to room temperature and the reaction mixture was stirred for 2 hours. The mixture was then poured into water (10 mL) and extracted with EtOAc (2x20 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was then purified with flash chromatography (0-50% EtOAc in hexane as the eluent) to obtain intermediate 9c. NMR (400 MHz, CDCI3) 88.61 (d. 7 = 0.6 Hz, 1H), 7.41 (s, 1H), 6.49 (s, 1H), 5.81 (d, J = II.0 Hz, 1H), 5.58 (d, 7 = 11.0 Hz, 1H), 4.08 (d, 7 = 13.3 Hz, 1H), 3.60 - 3.48 (m, 3H), 3.22 - 3.04 (m, 1H), 3.01 - 2.87 (m, 7 = 15.8, 7.6 Hz, 1H), 2.88 - 2.68 (m, 7 = 16.5, 8.4 Hz, 1H), 2.10 - 1.95 (m, 1H), 1.96 - 1.79 (m, 7 = 17.5, 8.7 Hz, 1H), 1.79 - 1.66 (m, 1H), 0.96 - 0.88 (m, 8H), 0.04 - -0.06 (m, 9H).

[0096] General procedure for the synthesis of intermediates 10a ((S)-N-(2-((2-methyIpyrrolidin-l-yl)methyI)-l-((2-(trimethyIsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)-l,l-diphenylmethanimine), 10b ( 1 / / -pyrrolol 3.2-c|pyridin-6-y 11- 1.1-diphenylmethanimine). and the racemic mixture of 10c, comprising enantiomers lOd and lOe (N-(2-((2-isopropylazetidin-l-yI)methyl)-l-((2-(trimethylsilyI)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)-l,l-diphenylmethanimine).

[0097] In Scheme 2, a stirred solution of intermediate 9a, intermediate 9b, or intermediate 9c (300 mg, 0.787 mmol), benzophenone imine (213 mg, 1.18 mmol), sodium tert-butoxide (150 mg, 1.57 mmol), tris(dibenzylideneacetone) dipalladium(O) (0.017 g, 0.064 mmol) and 2,2'-bis(diphenylphosphino)- 1,1 '-binaphthyl (0.02 g, 0.064 mmol) in toluene (5 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 110°C for 4 hours. The reaction mixture was filtered and concentrated in vacuo and used in the next step without further purification.

[0098] General procedure for the synthesis of intermediates Ila, 11b, 11c, lid, and lie.

[0099] In Scheme 2, 1 M HC1 (5 mL) was added to a stirred solution of crude intermediates 10a, 10b, or 10c (300 mg) in THF: H2O (1:1, 10 mL) and the reaction solution was stirred at room temperature for 16 hours. The mixture was then poured into water (10 mL) and extracted with EtOAc. The aqueous phase was basified using 1 N NaOH to adjust the pH to 8. The aqueous layer was extracted with EtOAc (2x30 mL). The organic layer was dried over anhydrous Na2SO4 and then concentrated in vacuo and used in the next step without further purification.

[0100] Intermediate Ila: (S)-2-((2-methylpyrrolidin-l-yl)methyl)-l-((2-(trimethylsilyl) ethoxy) methyl)-! W-pyrrolo[3,2-c]pyridin-6-amine. ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 (d, J = 0.9 Hz, 1H), 7.88 (t, J = 0.9 Hz, 1H), 6.50 (s, 1H), 5.76 (d, 7 = 11.1 Hz, 1H), 5.58 (d, J = 11.0 Hz, 1H), 4.21 (d, J = 13.5 Hz, 1H), 3.71 - 3.53 (m, 2H), 3.38 (d, 7 = 13.5 Hz, 1H), 2.89 (ddd, 7 = 9.7, 6.4, 4.0 Hz, 1H), 2.50 (td, 7 = 7.6, 5.8 Hz, 1H), 2.25 (q, J = 8.8 Hz, 1H), 2.12 - 1.99 (m, 1H), 1.71 (q, 7 = 8.3 Hz, 2H), 1.52 - 1.38 (m, 1H), 1.22 (d, 7 = 5.9 Hz, 3H), 1.04 - 0.86 (m, 2H), 0.00 (s, 9H).

[0101] Intermediate 11b: 2-((2-cyclopropylazetidin-l-yl)methyl)-l-((2-(trimethylsilyl) ethoxy)methyl)-LH-pyrroIo[3,2-c]pyridin-6-amine. ’ll NMR (400 MHz, DMSO-de) 3 8.50 (s, 1H), 7.87 (s, 1H), 6.48 (s. 1H), 5.75 - 5.61 (m, 2H), 3.99 (d, 7 = 13.5 Hz, 1H), 3.70 - 3.54 (m, 3H), 3.17 (d, 7 = 7.8 Hz, 1H), 2.80 (dt, 7 = 15.5, 8.2 Hz, 2H), 2.10 (d, 7 = 9.9 Hz, 1H), 1.89 (p, 7 = 9.0 Hz, 1H), 0.96 (t, 7 = 7.9 Hz, 3H), 0.42 (td, 7 = 6.4, 2.4 Hz, 2H), 0.26 - 0.15 (m, 2H), 0.00 (s, 9H).

[0102] Racemic intermediate 11c (and enantiomers lid and lie): 2-((2-isopropylazetidin-l-yl)methyI)-l-((2-(trimethylsilyl) ethoxy )methyl)-l / / -pyrrolo|3.2-c]pyridin-6-amine. ’ll NMR (400 MHz, DMSO-cfe) 8 8.20 (d, J = 0.7 Hz, 1H), 6.51 (s, 1H), 6.33 (s, 1H), 5.64 (d, 7 = 11.1 Hz, 1H), 5.47 (d, 7 = 11.1 Hz, 1H), 5.43 (s. 2H), 4.29 - 4.10 (m,J = 28.9, 5.9 Hz, 1H), 3.97 (d, J = 13.2 Hz, 1H), 3.60 - 3.51 (m, 2H), 3.15 - 3.01 (m, 1H), 3.00 - 2.88 (m, J = 15.5, 7.5 Hz, 1H), 2.86 - 2.72 (m, J = 16.2, 8.5 Hz, 1H), 2.08 - 1.93 (m, 1H), 1.86 - 1.73 (m, 1H), 1.73 - 1.62 (m, J = 13.6, 6.8 Hz, 1H), 0.99 - 0.82 (m, 8H), 0.04 - -0.03 (m, 9H).

[0103] Synthesis of intermediate 12a: (S’)-N-(2-((2-methylpyrrolidin-l-yl)methyl)-l-((2-(trimethylsilyl)ethoxy)methyl)- l / / -pyrrolo|3.2-c|pyridin-6-yl)-| 1,2,4 ]triazolo[4,3-a]pyridine-6-carboxamide.

[0104] In Scheme 2, EDCI was added (280 mg, 1.25 mmol) to a stirred solution of crude intermediate 1 la (300 mg) and [l,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid (174 mg, 0.916 mmol) in anhydrous pyridine (5 mL) at 0°C. The reaction mixture was stirred at 50°C for 12 hours. After completion of the reaction, the reaction mixture was added to H2O (10 mL), extracted with EtOAc (2x20 mL), and washed with saturated brine solution (2x10 mL) sequentially. The organic layer was dried over anhydrous Na2SC>4 and then concentrated in vacuo. The residue was then purified with flash chromatography (0-10% MeOH in CH2Q2 as the eluent) to obtain intermediate 12a (yield: 170 mg, 41%). ’ll NMR (400 MHz, DMSO-t / fi) 6 11.00 (s, 1H), 9.49 (d, J = 0.8 Hz, 1H), 9.43 (t, 7 = 1.4 Hz, 1H), 8.67 (d, J = 1.0 Hz, 1H), 8.46 (s, 1H), 8.03 - 7.90 (m, 2H), 6.63 (s, 1H), 5.82 (d, 7= 11.1 Hz, 1H), 5.69 (d, J = 11.1 HZ, 1H), 4.25 (d, J = 13.6 Hz, 1H), 3.61 (did, J = 30.0, 9.2. 6.8 Hz. 2H), 2.92 - 2.82 (m, 1H), 2.50 (p, J = 6.7 Hz, 1H). 2.26 (q, J = 8.8 Hz, 1H), 2.04 (dt. J = 14.6, 7.2 Hz, 1H). 1.71 (q, J = 7.8 Hz, 2H), 1.45 (dq, J = 12.2, 8.3 Hz, 1H), 1.22 (d, J = 5.9 Hz, 3H), 1.04 - 0.86 (m, 2H), 0.00 (s, 9H).

[0105] Synthesis of intermediate 12b: N-(2-((2-cyclopropylazetidin-l-yI)methyl)-l-((2-(trimethyIsilyl)ethoxy)methyl)-LH-pyrrolo[3,2-c]pyridin-6-yl)-[l,2,4]triazoIo[4,3-a]pyridine-6-carboxamide.

[0106] In Scheme 2, EDCI was added (280 mg, 1.25 mmol) to a stirred solution of crude intermediate 1 lb (300 mg) and [l,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid (174 mg, 0.916 mmol) in anhydrous pyridine (5 mL) at 0°C. The reaction mixture was stirred at 50°C for 12 hours. After completion of the reaction, the reaction mixture was added to H2O (10 mL), extracted with EtOAc (2x20 mL), and washed with saturated brine solution (2x10 mL) sequentially. The organic layer was dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was then purified with flash chromatography (0-10% MeOH in CH2Q2 as the eluent) to obtain intermediate 12b. ’ll NMR (400 MHz, DMSO-cfc) 5 10.99 (s, 1H), 9.48(d, J = 0.8 Hz, 1H), 9.42 (t, J = 1.4 Hz, 1H), 8.66 (d, J = 0.9 Hz, 1H), 8.44 (s, 1H), 8.00 - 7.89 (m. 2H), 6.60 (s, 1H), 5.83 - 5.71 (m, 2H), 4.03 (d, J = 13.7 Hz, 1H), 3.76 - 3.50 (m, 3H), 3.21 - 3.13 (m, 1H), 2.81 (dq, J = 20.0, 7.9 Hz, 2H), 2.15 - 2.04 (m, 1H), 1.89 (p, 7 = 9.0 Hz, 1H), 1.02 - 0.88 (m, 3H), 0.41 (ddd, J = 9.2, 4.6, 2.9 Hz, 2H), 0.25 - 0.14 (m, 2H), 0.00 (s, 9H).

[0107] Synthesis of racemic intermediate 12c (and enantiomers 12d and 12e): N-(2-((2-isopropylazetidin- l-yl)methyI)-l-((2-(trimethylsilyl)ethoxy)methyI)-l / 7-pyrrolo[3,2-c]pyridin-6-yl)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0108] In Scheme 2, EDCI was added (280 mg, 1.25 mmol) to a stirred solution of crude intermediate 11c (300 mg) and [l,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid (174 mg, 0.916 mmol) in anhydrous pyridine (5 mL) at 0°C. The reaction mixture was stirred at 50°C for 12 hours. After completion of the reaction, the reaction mixture was added to H2O (10 ml,), extracted with EtOAc (2x20 mL), and washed with saturated brine solution (2x10 mL) sequentially. The organic layer was dried over anhydrous Na2SC>4 and then concentrated in vacuo. The residue was then purified with flash chromatography (0-10% MeOH in CH2Q2 as the eluent) to obtain intermediate 12c. 'll NMR (400 MHz, DMSO-rL) 6 11.01 (s, 1H), 9.50 (s, 1H), 9.43 (s, 1H), 8.68 (s, 1H), 8.46 (s, 1H), 8.05 - 7.86 (m, 2H), 6.63 (s, 1H), 5.84 (d, 7 = II.3 Hz, 1H), 5.71 (d, 7= 11.2 Hz, 1H), 4.11 (d, 7 = 13.5 Hz, 1H), 3.73 - 3.55 (m, 3H), 3.14 (s, 1H), 3.02 (s, 1H), 2.82 (d, 7 = 36.5 Hz, 1H), 2.06 (d, 7 = 7.7 Hz, 1H), 1.92 - 1.65 (m, J = 19.9, 15.7, 7.8 Hz, 2H), 0.99 - 0.88 (m, 8H), -0.00 (s, 9H).

[0109] Synthesis of Compound YR-D-120: (S)-N-(2-((2-methylpyrrolidin-l-yl)methyl)-l / / -pyrrolo|3.2-c|pyridin-6-yl)-| l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0110] In Scheme 2, TFA (1 mL) was added to a stirred solution of intermediate 12a (120 mg, 0.237 mmol) in anhydrous BCM (5 mL) at 0 °C. The reaction mixture was stirred at room temperature for 12 hours. After completion of reaction, the solvent was removed in vacuo. The residue was then purified with reverse phase flash chromatography (0-100% ACN in H2O as the eluent) to obtain compound YR-D-120 (yield: 30 mg, 34%). 01 NMR (400 MHz, CD3OD) 59.39 (s, 1H), 9.34 (s, 1H), 8.93 (s, 1H), 8.09 - 8.01 (m, 2H), 7.94 (d, 7 = 9.6 Hz, 1H), 7.16 (s, 1H), 4.54 (d, 7 = 14.2 Hz, 1H), 3.77 - 3.56 (m, 2H), 3.47 - 3.38 (m, 2H), 2.48 - 2.39 (m, 1H), 2.23 - 2.01 (m, 2H), 1.90 - 1.76 (m, 1H), 1.54 (d, 7 = 6.6 Hz, 3H).13C NMR (126 MHz, CD3OD) 5 164.60. 161.95, 161.68, 144.55, 141.57, 135.45, 134.44, 127.96, 127.36, 122.60, 121.45, 114.55, 106.81, 97.91, 89.73, 64.61, 53.90, 31.04, 20.92, 14.98. HRMS (ESI+) was calculated for C22H26N5O [M+H]+376.1880, found 376.1874.

[0111] Synthesis of Compound YR-D-121: N-(2-(2-cyclopropylazetidin-l-yl) methyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0112] In Scheme 2, TFA (1 niL) was added to a stirred solution of intermediate 12b (120 mg, 0.237 mmol) in anhydrous DCM (5 mF) at 0 °C. The reaction mixture was stirred at room temperature for 12 hours. After completion of reaction, the solvent was removed in vacuo. The residue was then purified with reverse phase flash chromatography (0-100% ACN in H2O as the eluent) to obtain Compound YR-D-121. 'll NMR (400 MHz, CD3OD) 6 9.12 (d, J = 0.9 Hz, 1H), 9.01 (t, J = 1.4 Hz, 1H), 8.44 (d, J = 1.0 Hz, 1H), 8.30 (s, 1H), 8.05 (s, 1H), 7.77 (dd, 7 = 9.6, 1.7 Hz, 1H), 7.66 (dt, J = 9.7, 1.0 Hz, 1H), 6.54 (d, J = 0.9 Hz, lH), 4.10 (s, 1H), 2.95 (t, J = 7.1 Hz, 2H), 2.80 (td, J = 8.5, 3.9 Hz, 1H), 1.86 - 1.61 (m, 2H), 0.75 - 0.62 (m, 1H), 0.38 - 0.23 (m, 2H), 0.19 - 0.04 (m, 1H), 0.04 - -0.04 (m, 1H).13C NMR (126 MHz, CD3OD) 5 163.34, 149.29, 142.36, 141.01, 137.79, 127.92, 126.88, 123.79, 123.18, 114.54, 101.68, 97.78, 74.80, 45.82, 44.70, 34.31, 17.16, 2.35, 1.43.

[0113] Synthesis of Compound SR-C-107: N-(2-(2-isopropylazetidin-l-yl) methyl)- 1 / / -pyrrolo[3,2-c]pyridin-6-yl)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0114] In Scheme 2, TFA ( 1 mL) was added to a stirred solution of intermediate 12c ( 120 mg, 0.237 mmol) in anhydrous DCM (5 mF) at 0 °C. The reaction mixture was stirred at room temperature for 12 hours. After completion of reaction, the solvent was removed in vacuo. The residue was then purified with reverse phase flash chromatography (0-100% ACN in H2O as the eluent) to obtain Compound SR-C-107. ’ll NMR (400 MHz, CD3OD) 5 9.22 (d, J = 0.6 Hz, 1H), 9.14 - 9.06 (m, 1H), 8.45 (t, J = 2.5 Hz, 1H), 8.06 (s, 1H), 7.92 - 7.83 (m, J = 9.6, 4.6, 1.7 Hz, 1H), 7.76 (d, J = 9.6 Hz, 1H), 6.40 (s, 1H), 3.91 (d, J = 13.7 Hz, 1H), 3.59 (t, J = 13.0 Hz, 1H), 3.19 (s, 1H), 2.96 - 2.80 (m, 2H), 2.06 - 1.94 (m, 1H), 1.82 - 1.58 (m, 2H), 0.89 - 0.80 (m, 3H), 0.75 (t, J = 6.1 Hz, 3H).13C NMR (101 MHz, DMSO-6) 5 162.98, 148.84, 145.00, 141.88, 140.21, 139.20, 138.07, 127.80, 127.57, 123.39, 121.91, 114.75, 99.24, 97.13, 72.49, 56.17, 50.75, 49.06, 40.60, 40.39, 40.18, 39.97, 39.76, 39.55, 39.34, 33.68, 20.92, 19.12, 17.95. HRMS (ESI+) was calculated for C21H24N7O [M+H]+390.2037, found 390.2029.

[0115] Synthesis of Compound SR-C-107 (R): (R)-N-(2-((2-isopropylazetidin-l-yl)methyI)-lH-pyrrolo[3,2-c]pyridin-6-yl)-[l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0116] TFA (1 mF) was added to a stirred solution of intermediate 12d in anhydrous DCM (5 mF) at 0°C. The reaction mixture was stirred at room temperature for 12 hours. After completion of reaction, the solvent was removed in vacuo. The residue was then purified withreverse phase flash chromatography (0-100% ACN in H2O as the eluent) to obtain Compound SR-C-107 (R). ’ll NMR (400 MHz, CD3OD) 59.32 (s, 1H), 9.19 (s, 1H), 8.54 (s, 1H), 8.16 (s, 1H), 7.96 (d, J = 9.7 Hz, 1H), 7.84 (d, J = 9.7 Hz, 1H), 6.49 (s, 1H), 4.02 (d, J = 13.7 Hz, 1H), 3.68 (d, J = 13.7 Hz, 1H), 3.30 - 3.28 (m, 1H), 3.10 - 2.89 (m, 2H), 2.17 - 2.06 (m, 1H), 1.94 - 1.81 (m, 1H), 1.80 - 1.66 (m, 1H), 0.95 (d, J = 6.6 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).13C NMR (101 MHz, DMSO-rfc) 5162.34, 148.19, 144.36, 141.23, 139.59, 138.41, 137.42, 127.14, 126.92, 122.73, 121.26, 114.10, 98.67, 96.48. 71.86, 55.45, 50.09, 33.01, 20.27, 18.46, 17.29. HRMS (ESI+) was calculated for C21H23N7O [M+H]+390.2000, found 390.2029.

[0117] Synthesis of Compound SR-C-107 (S): (S)-N-(2-((2-isopropylazetidin-l-yl)methyl)- l / / -pyrrolo|3,2-c|pyridin-6-yl)-| l,2,4]triazolo[4,3-a]pyridine-6-carboxamide.

[0118] TFA (1 ml,) was added to a stirred solution of intermediate 12e in anhydrous BCM (5 mL) at 0°C. The reaction mixture was stirred at room temperature for 12 hours. After completion of reaction, the solvent was removed in vacuo. The residue was then purified with reverse phase flash chromatography (0-100% ACN in H2O as the eluent) to obtain compound SR-C-107 (S). 'll NMR (400 MHz, CD3OD) 5 9.32 (d, J = 0.8 Hz, 1H), 9.19 (t, J = 1.4 Hz, 1H), 8.54 (d, 7= 1.1 Hz, 1H), 8.15 (t, 7= 1.0 Hz, 1H), 7.97 (dd, 7= 9.7, 1.6 Hz, 1H), 7.85 (dt, 7= 9.6, 1.0 Hz, 1H), 6.48 (d, 7= 0.9 Hz, 1H), 3.99 (d, 7= 13.6 Hz, 1H), 3.65 (d, 7= 13.7 Hz, 1H), 3.30 - 3.26 (m, 1H), 3.04 - 2.88 (m, 2H), 2.16 - 2.03 (m, 1H), 1.92 - 1.80 (m, 1H), 1.79 - 1.65 (m, 1H), 0.95 (d, 7= 6.7 Hz, 3H), 0.83 (d, 7 = 6.7 Hz, 3H).13C NMR (101 MHz, DMSO-6) 8 163.47, 149.32, 145.50, 142.36, 140.72, 139.54, 138.55, 128.28, 128.05, 123.86, 122.39, 115.23, 99.80, 97.61, 72.99, 56.58, 51.22, 34.13, 21.40, 19.59, 18.42. HRMS (ESI+) was calculated for C21H23N7O [M+H]+390.2000, found 390.2027.EXAMPLE 3: In vitro Characterization of Novel ENL Inhibitors

[0119] The inhibitory effect of the newly synthesized compounds on ENL YEATS was evaluated using an AlphaScreen assay. Instead of a commonly used biotinylated H3K27cr peptide binding to ENL YEATS, a 7-mer peptide ENL-S1 developed by Chen et al. (ACS Cent. Sci. 10(4):782-792, 2024) was used as it possesses a stronger binding affinity to the ENL YEATS domain. Samples were prepared using the inhibitor compounds, biotinylated ENL-S 1 peptide, his-tagged ENL YEATS protein, Ni -chelate acceptor beads, and streptavidin donor beads. AlphaScreen signals (Rubrene emission at 520 - 620 nM) were measured using a multimode microplate reader equipped with an Alpha laser. (FIG. 1A). In addition, IC50 values for each compound were derived from the collected data.

[0120] A NanoBRET assay was performed to evaluate the ability of the inhibitory compounds to permeate into cells and engage the ENL YEATS domain in the cellular environment. HEK293T cells stably expressing a NLuc-ENL YEATS fusion protein were coincubated with a cell-permeable fluorescent tracer and various concentrations of each inhibitor compound. For BRET detection, the donor emission was measured at 450 nm and the acceptor emission at 610 nm. The BRET ratio was calculated according to the following formula: BRET Ratio = [(Acceptorsample / Donorsample) - (AcCeptorno-tracercontrol / DonOrno-tracer control)] X 1000. (FIG. 1B)

[0121] The partition coefficient between 1 -octanol and aqueous buffer pH 7.4 (logD7.4) of the newly synthesized compounds was measured using a chromatographic method as previously described in Nakashima et al. (Chem Pharm Bulletin 61:1228-38, 2013). The samples were analyzed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS / MS).

[0122] The calculated results of all three assays are summarized in Table 2 below.Table 2. Mean AlphaScreen and NanoBRET IC50 values and logD values obtained from assays.

[0123] Among the tested compounds, SR-C-107 (R) showed the best potency, with an IC50 value of 0.59 pM, indicating its excellent cellular permeability and engagement with the ENL YEATS active site. In contrast, the racemate SR-C-107 and its S-isomer showed reduced cellular engagement, with IC50 values of 1.48 and 5.67 pM, respectively. The comparable cellular permeability of SR-C-107 (R) and SR-C-107 (S) is supported by their similar partitioncoefficients (logD values of 0.97 and 0.89, respectively). Although YR-D-120 and YR-D-121 displayed strong in vitro ENL YEATS binding affinity, their cellular inhibition potencies detected by NanoBRET were moderately lower, with ICso values of 2.02 and 2.36 pM. respectively, this reduction in potency could be attributed to their relatively low lipophilicity (log / ) values of 0.08 and 0.13, respectively).EXAMPLE 4: Metabolic Stability of Novel Inhibitor Compounds in Human Plasma and Liver Microsomes

[0124] Plasma stability testing is a key component of in vitro ADME (Absorption, Distribution, Metabolism, and Excretion) screening assays used to gauge the stability of drug candidates in plasma. The in vitro stability analysis of compound 13, YR-D-120, YR-D-121, SR-C-107, SR-C-107 (R), and SR-C-107 (S) was initiated by the addition of the test compound to 90 pL of pre- warmed human plasma with a final concentration of 5 LIM. All assays were performed in a plate shaker at 37°C and conducted in triplicate. At 0, 5, 15, 30, 60, and 120 min, 400 pL acetonitrile was added to deproteinize the plasma and terminate the reaction, the remaining compound was then analyzed by HPLC-MS / MS (FIG. 2A). The results are presented in Table 3 below.Table 3. Mean values for compound stability in human plasma.

[0125] As shown in Table 3 above, compound 13 and SR-C-107 (R) exhibited minimal degradation after the 2-hour incubation in human plasma at 37°C. Both compounds retained approximately 95% of their original concentration in plasma. SR-C-107(S) showed lower plasma stability compared to SR-C-107 (R). Due to this, SR-C-107 showed quicker elimination than SR-C-107 (R), with 90.77% of SR-C-107 remaining following 2-h incubation in humanplasma. In contrast, YR-D-120 and YR-D-121 displayed reduced stability in human plasma, with approximately 15% degradation after 2-hour incubation.

[0126] Hepatic metabolism is a primary pathway for drug elimination. In vitro studies of hepatic metabolism are often conducted using the microsomal incubation technique to determine the intrinsic clearance rate (CI ant). This assay is essential in drug development as it allows for the screening of lead compounds from a broad array of candidates with similar in vitro potency. To prioritize novel ENL inhibitors, this assay was conducted by incubating each inhibitor with human liver microsomes, either with or without nicotinamide adenine dinucleotide phosphate (NADPH), the cofactor for flavin monooxygenase-dependent oxidation, at 37°C. Degradation levels were quantified using liquid chromatography-tandem mass spectrometry (LC-MS / MS). As shown in FIG. 2B, the metabolic stability of each inhibitor was determined by plotting the natural logarithm (Ln) of the remaining inhibitor concentration (y-axis) against incubation time (x-axis). The slope of this plot was used to calculate the metabolic rate of each inhibitor, which was then used to determine the in vitro half-life (ri / 2) and intrinsic clearance rate. The results are presented in Table 4 below.Table 4. Calculated in vitro half-life (ti / 2) and intrinsic clearance rate values for tested compounds in human liver microsome.

[0127] All compounds were stable in human liver microsome in the absence of NAPDH (data not shown). In the presence of NADPH, compounds 13, YR-D-120, and YR-D-121 displayed excellent microsomal stability, with over 90% of parent compounds remaining post 1-hour incubation. Their determined intrinsic clearance rates were 4.09, 4.76, and 3.51 mL / min / kg, respectively, highlighting their promising metabolic stability profiles. For two enantiomers ofSR-C-107, the more potent SR-C-107(R), with a half-life of 418 minutes and low clearance rate of 4.18 mL / min / kg, demonstrating superior metabolic durability than SR-C-107 (S) (ri / 2 = 165 min, CLint = 10.95 mL / min / kg), further supporting its potential for AML treatment.EXAMPLE 5: In Vitro Anti-Tumor Efficacy of Novel ENL Inhibitors

[0128] After evaluating the in vitro metabolic stability of the inhibitor compounds, their efficacy in inhibiting ENL-dependent tumor cell growth was evaluated. The ENL-dependent leukemia cell lines MV4-11 and MOLM-13, ENL-independent Jurkat leukemia cell line, a chronic myelogenous leukemia cell line K-562, an acute lymphoblastic leukemia cell line RS4;11, and non-tumorigenic HEK293T cell line were utilized for cytotoxicity experiments. Approximately IxlO4cells / well of each line were added to a 96- well plate and treated with DMSO or inhibitory compound at indicated concentrations for 72 hours or 8 days. Cell viability was measured using a cell counting kit (CCK-8; Abeam) according to the manufacturer’s instructions. The cell viability was assessed by detection of absorbance at 450 nm using a spectrophotometer. The 50% cytotoxic concentration (CC50) for each inhibitory compound was calculated as the concentration of the compound that reduces the number of viable cells by 50% compared to untreated cells. The results are shown in FIGS. 3A-3E and in Table 5 below.Table 5. Half-maximum inhibitory concentration (CC50) values for all tested inhibitory compounds in each cell line (n=3).CC; S;(pMOLM-13 MV4-11 drafcat HEK2938.20 t. 0.58s9.15 * 0.87s> 84.00s136.01 i 0.83 6.72 * 1.00 22.04 < 1.78YR-O-120 361 0.49 328 < 0.85 > 64.00sYR-D-121 6.86 1.21 14.18 2.6484.00SR-C-107 257 < 0.28 1,733. 0.181.25 v 0.18 0.81 0.15Ceil was treated ferB- ays except indicated as » (72-h iemsbaticni.

[0129] Compound SR-C-107 (R) exhibited superior antiproliferative effects in MOLM-13 and MV4-11 cells, with CC50 values of 1.25 and 0.81 pM, respectively. Additionally, compounds YR-D-120 and SR-C-107 exhibited antiproliferative activity comparable to one another, with CC50 values below 4 pM for both cell lines. Unexpectedly, despite having comparable cellular ENL YEATS engagement and metabolic stability to compound YR-D-120, compound YR-D-121 displayed weaker cytotoxicity, with CC50 values of 6.96 and 14.16 pM in both ENL-dependent leukemia cell lines. None of the compounds showed antiproliferative activity in Jurkat or HEK293T cells, even at concentrations up to 64 pM.

[0130] Cell proliferation assays were carried out in a 96-well tissue culture plate at 2xl04cells / well for all cell lines. Culture density was determined every 3-4 days, after which 2x104live cells were reseeded in fresh media and compound. Cell proliferation was monitored over 14 days in the presence of 1 uM of the inhibitory compound. The cumulative cell number was achieved by back calculation.

[0131] Consistent with the results from the cytotoxicity assay described above, compound 13 significantly inhibited the growth of MOLM-13 (85% inhibition) and MV4-11 (75% inhibition) at a concentration of 10 pM (data not shown). However, at a lower concentration of compound 13 (1 pM), the cellular inhibition efficacy was reduced, showing 30% and 41% inhibition for MOLM-13 and MV4-11, respectively (data not shown). In contrast, both SR-C-107 and SR-C-107 (R) at 1 pM demonstrated strong inhibition of cell proliferation in MOLM-13 and MV4-11. Notably, SR-C-107 (R) achieved approximately 90% and 70% inhibition for MOLM-13 and MV4-11 cells, respectively (FIGS. 4A and 4B). Co-incubation with all compounds had no effect on the growth of Jurkat cells (FIG. 4C).EXAMPLE 6: In Vivo Pharmacokinetic and Antitumor Efficacy Analyses of Novel ENL Inhibitors

[0132] Given their superior metabolic stability and promising cellular efficacy, compound 13 and compound SR-C-107 (R) were further tested in vivo in pharmacokinetic and antitumor efficacy experiments. The plasma pharmacokinetics of both inhibitors were characterized in CD-I mice following oral administration at a dose of 20 mg / kg or intravenous administration at a dose of 20 mg / kg (compound 13) or 2 mg / kg (SR-C-107 (R)). Three male CD-I mice were used for each administration. Blood samples were taken from a vein at different time points up to 24 hours after dosing, collected in tubes coated with an anticoagulant, and centrifuged to obtain plasma samples. The total plasma concentrations of the inhibitors were measured using LC-MS / MS.

[0133] After oral administration, compound 13 showed excellent systemic exposure (Cmax — 2,080 ng / mL and AUC(o-inf) = 5,137 ng-h / mL) and a favorable half-life (C1 / 2 = 0.84 h) in treated mice (FIG. 5A). The half-life of SR-C-107 (R) following oral administration at a dosage of 20 mg / kg was 1.41 h (FIG. 6A). Compared to compound 13, the systemic exposure of SR-C-107(R) was lower, with an AU o-inf) of 808 ng-h / mL. The Cmax calculated for SR-C-107 (R), 285 ng / ml, corresponds to an effective concentration of 0.75 pM. which was sufficient to induce antiproliferation effects based on in vitro antiproliferation activity described earlier. These advantageous pharmacokinetic properties suggest that both compounds have the potential to achieve high and sustained drug concentrations when administered orally.

[0134] For the anti-tumor efficacy experiments, an orthotopic AML mouse model was generated. MOLM-13-luciferase cells (1 x 106) were intravenously injected into six-week-old female NSG mice. After confirmation of engraftment (6 days after injection), the mice were randomly divided into groups and administered daily gavage treatments of compound 13 at a dose of 200 mg / kg or 400 mg / kg, compound SR-C-107 (R) at a 200 mg / kg dose, or vehicle. Both dosages of compound 13 were found to be well-tolerated, with no significant body weight loss observed (FIG. 5C). The dosage used for SR-C-107 (R) was similarly well-tolerated (FIG.6C).

[0135] The leukemic burden of MOLM-13-luciferase cells was monitored by bioluminescence imaging. While no noticeable tumor reduction was observed with the 200 mg / kg dose of compound 13, increasing the dose to 400 mg / kg resulted in significant reduction in leukemia burden after 13 days of treatment (corresponding to day 20 post- transplantation). This higher dose resulted in nearly 45% tumor growth inhibition compared to the control group, without significant body weight loss until day 15 post treatment (FIGS. 5B and 5C). Furthermore, most mice in the 400 mg / kg dose-treated group not only survived but also displayed a marked reduction in leukemia burden. The overall survival in this group was significantly better compared to the control group, with a 33.3% increase in median survival (P = 0.00185) (FIG. 5D).

[0136] The in vivo antitumor efficacy of compound SR-C-107 (R) at the 200 mg / kg dose surpassed that of compound 13 at the 400 mg / kg dose. SR-C-107 (R) demonstrated significant tumor growth inhibition, achieving 49.5% tumor growth inhibition after 8 days of treatment (P = 0.0058) and 45% tumor growth inhibition after 12 days of treatment (P = 0.0206) (FIG. 6B). Additionally, the median survival time was extended from 10 days post-treatment to 15 days post-treatment (FIG. 6D). Collectively, these findings suggest that compound SR-C-107 (R) has a favorable safety profile and a lower efficacious dose compared to compound 13.* *

[0137] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this disclosure have been described in terms of preferred embodiments or aspects, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit, and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

CLAIMSWhat is claimed is:

1. A compound of Formula I or a derivative thereof:or compound 13 or a derivative thereof:whereinX is a carbon or a nitrogen, andRi is a 4-, 5-, or 6-membered ring, which may be cycloalkyl, heterocyclic, heteroaryl, or aryl, wherein said ring is optionally substituted with one or more groups selected from hydrogen, C1-C12 alkyl, C1-C12 haloalkyl, C1-C12 heteroalkyl, aralkyl, and aryl sulfamide.

2. The compound of claim 1, having a formula selected from the group consisting of:a derivative of any thereof.

3. The compound of any one of claim 1, wherein the compound selectively targets the ENL YEATS domain.

4. A pharmaceutical composition comprising an effective amount of the compound of claim 1.

5. The pharmaceutical composition of claim 4, further comprising one or more pharmaceutically acceptable carriers, buffers, or diluents.

6. The pharmaceutical composition of claim 4, wherein the compound selectively targets the ENL YEATS domain.

7. The pharmaceutical composition of claim 4, wherein the compound is effective for treating leukemia.

8. The pharmaceutical composition of claim 7, wherein the leukemia is selected from the group consisting of acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, chronic myelomonocytic leukemia, and myeloproliferative neoplasms.

9. A method of treating a subject in need thereof, the method comprising administering a therapeutically effective amount of the compound of claim 1 to the subject.

10. The method of claim 9, wherein said subject is afflicted with leukemia.

11. The method of claim 10, wherein said subject is afflicted with acute myeloid leukemia, acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, chronic myelomonocytic leukemia, or myeloproliferative neoplasms.

12. The method of claim 9, wherein said administering comprises local, regional, systemic, or continual administration.

13. The method of claim 9, wherein said administering comprises oral administration, intravenous administration, buccal administration, rectal administration, parenteral administration, intraperitoneal administration, topical administration, intradermal administration, intratracheal administration, intramuscular administration, subcutaneous administration, or inhalation.

14. The method of claim 9, the method further comprising administering a second therapy to said subject.

15. The method of claim 14, wherein the second therapy is selected from the group consisting of a chemotherapy, a radiotherapy, a targeted therapy, a small molecule inhibitor, an immunotherapy, and surgery.

16. The method of claim 9, wherein the subject is a mammalian subject.

17. The method of claim 16, wherein the subject is a human subject.

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