Translational inhibitors and binding probes

Abstract

Small molecules for targeting the 5'UTR to modulate oncogenic protein expression levels and methods thereof are described. These small molecules are inhibitors of BMI 1 expression and may be useful in the treatment of cancer. Also disclosed are small molecule probes to map binding sites and small molecule binding.

Description

TRANSLATIONAL INHIBITORS AND BINDING PROBESCROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority to U. S. Provisional Patent Application No.63 / 737,848, filed December 23, 2024, which is incorporated herein in its entirety for all purposes.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH & DEVELOPMENT

[0002] This invention was made with government support under Grant No.7R01CA226746 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0003] Fueled by cancer cell oncogenic addiction, the polycomb oncogene BMI1 (for B cell specific Moloney murine leukemia virus integration site 1) enhances cancer cell selfrenewal, drives cell proliferation, and accelerates cancer progression in 35 various cancer types including the most lethal ones like glioblastoma (GBM). Functionally, BMI1 enhances cellular self-renewal in glioma stem-like cells (GSCs) and drives cell proliferation by repressing multiple tumor suppressors at the epigenetic level with Poly comb-repressive complex 1 (PRC1) complex and contributes to oxidative and DNA damage stresses, adding to its powerful oncogenic effects. Despite many studies showing these powerful oncogenic behaviors, there is currently no targeted therapy against BMI1 in the clinic. BMI1 overexpression, accompanied by high quantities of RNA, correlates with poor patient survival. This same pattern repeats across many similar oncogenes.

[0004] There remains a need in the art for small molecule compounds useful for targeting RNA to modulate oncogenic protein expression levels. There is a further need for compounds useful for inhibiting BMI1 expression and thus useful for treatment of cancer.BRIEF SUMMARY

[0005] One embodiment is a compound according to Formula (I), or a pharmaceutically acceptable salt thereof:whereineach R1independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 haloalkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di- FR2,C1-C4 alkylamino, halogen, hydroxy, nitro,orr3' wherein each R2and R3independently is hydrogen Ci-Ce alkyl, Ci-Ce haloalkyl, or OR7wherein R7is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C8 cycloalkyl, C3-C8 cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0);wherein when R1is Ci-Ce alkyl or Ci-Cehaloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or Ci-Cshaloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Cehaloalkyl forms a diazirine ring; each R4independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di- C1-C4 alkylamino, halogen, hydroxy, or nitro;R is Ci-Cs alkyl substituted with R6, cyano, C2-C6 alkenyl, C2-C6 alkynyl, or a heterocycloalkyl, wherein R6is an azido group, cyano, OR7, SR7, SO2R7, NR7R7, or heterocycloalkyl, wherein each R7individually is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl;R8is hydrogen or C1-C4 alkyl;R9is hydrogen, Ci-Ce alkyl, or Ci-Cehaloalkyl;n is 1, 2, 3, or 4;m is 0, 1, 2, or 3;q is 0, 1, 2, or 3; andZ is CH or N.

[0006] Another embodiment is a pharmaceutical formulation comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0007] In yet another embodiment, a method of treating cancer in a subject comprises administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject in need of treatment.

[0008] In another embodiment, a method for inhibiting BMI1 expression in a subject in need thereof, comprises administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0009] In an embodiment, a method comprises providing an mRNA sequence; identifying a secondary structure of the mRNA sequence, wherein the secondary structure corresponds to a pocket within a 5’UTR; generating highly probable models of the pocket secondary structure, wherein the model accounts for a target minimum and maximum free energy; altering at least one functional group of a small molecule, e.g. compound of Formula (I) or a compound in Table 2, to conform the molecule to a subset of the highly probably models of the pocket to create a targeted small molecule; synthesizing the targeted small molecule; validating the 5'UTR binding of the targeted small molecule.

[0010] These and other embodiments are described in detail below.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1 are images of patient derived 3D cultures treated with inhibitors HS-D1 to HS-D5 compared to compound A 15 and media. The top row are Brightfield images showing sphere size, and the lower row are images of cells stained with DAPI for DNA at 48 hours.

[0012] FIG. 2 is a graph of viability of GBM cells in single cell sphere assay upon treatment for 72 hours.

[0013] FIG. 3 is a Western blot (WB) showing BMI1 protein reduction with no change in phospho-Histone H3 (PHH3) a mitosis marker.

[0014] FIG. 4 is a WB of HS-D4 and HS-D5 after 48 hours.

[0015] FIG. 5 illustrates quantitation of proteins in FIG. 4.

[0016] FIG. 6 is a WB showing BMI1 protein reduction with no change in PHH3 at 48 hours.

[0017] FIG. 7 is a WB showing effects on downstream targets of BMI1; datafrom six independent experiments in GBM-PDOs from GBM#50.DETAILED DESCRIPTION

[0018] The present disclosure is directed to novel small molecules for targeting the 5'UTR to modulate oncogenic protein expression levels and methods thereof. In a particular embodiment, the novel small molecule are inhibitors of BMI1 expression and may be useful in the treatment of cancer. Also disclosed are small molecule probes to map binding sites and small molecule binding.

[0019] Prior research using cellular and small animal reporters led to the identity of compounds that modulate BMI1 levels without inducing cell toxicity or developmental changes. Small molecule N-(4-methoxyphenyl)-4-(2-methylimidazo[l,2-a]pyridin-3-yl)thiazol-2-amine (“Al 5”), which is under preclinical development, has demonstrated favorable pharmacokinetic / pharmacodynamic (PK / PD) profiles, oral bioavailability, and potency in multiple preclinical models and organoids. The mechanism of action of Al 5 has been studied and there is evidence it reduces high BMI1 translation, which in turn, releases repressor functions in cancer cells, effectively killing them, but not healthy cells, therefore, providing tumor selectivity and high therapeutic index.

[0020] Studies revealed that Al 5 shows selective binding to the untranslated regions (5'UTR) of human BMI1, therefore interfering with the translation of the BMI1 main open reading frame (mORF).

[0021] Preliminary data shows that inhibiting BMI1 eliminates GSCs, modulates Programmed death ligand 1 (PD-L1) and immune responses within the tumor microenvironment. It has been demonstrated the oncogenic transcriptional activity of a major undruggable oncogene, providing a model for developing oncogene targeting platform, and validating BMI1 as the target of Al 5, which is the first compound to inhibit GSC self-renewal by specific targeting of 5'UTR.

[0022] Compounds of Formula (I) include BMI1 targeting compounds including several that have been examined for the effects of these compounds in GBM sphere and organoid assays. The compounds were found to inhibit BMI1, albeit without the notable cell cycle arrest effects (increased phospho-histone H3).

[0023] Taking BMI1 as protype for small molecule binding, the technology disclosed herein allows for the identification of binding sites suitable for targeting by an engineered small molecule at nucleotide resolution. Targeting oncogenes to affect cellular functions has the potential to transform oncology and medicine.

[0024] Compounds disclosed herein have been developed for targeting of theoncogenic BMI1. The techniques disclosed are suitable for the formulation of similar compounds created to bind to arbitrary oncogenic targets.

[0025] Compounds described herein include small molecule probes used to map binding sites. These compounds include HS-D1 to HS-D4 described herein. HS-D5 (3-(4-(azidomethyl)phenyl)-3-(trifluoromethyl)-3H-diazirine) can be used as a control photoaffinity probe.

[0026] Disclosed are compounds according to Formula (I), or a pharmaceutically acceptable salt thereof. Also provided are pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. Compounds of Formula (I) can be inhibitors BMI1 expression.

[0027] Disclosed herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereofFormula (I),whereineach R1independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 haloalkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di¬C1-C4 alkylamino, halogen, hydroxy, nitro,or wherein each R2and R3independently is hydrogen Ci-Ce alkyl, C1-C6 haloalkyl, or OR7wherein R7is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C8 cycloalkyl, C3-C8 cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =O);wherein when R1is Ci-Ce alkyl or Ci-Ce haloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3alkyl, or C1-C3 haloalkyl, or a carbon of the Ci-Ce alkyl or Ci-C6 haloalkyl forms a diazirine ring; each R4independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di- C1-C4 alkylamino, halogen, hydroxy, or nitro, specifically R4is Ci-Csalkyl, C1-C3 alkoxy or fluoro; R5 is Ci-C8 alkyl substituted with R6, cyano, C2-C6 alkenyl, C2-C6 alkynyl, or a heterocycloalkyl, wherein R6is an azido group (N3), cyano, OR7, SR7, SO2R7, NR7R7, or heterocycloalkyl, wherein each R7individually is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, specifically R5 is C2-C4 alkyl substituted with an azido group (N3), cyano, or alkynyl;R8is hydrogen or C1-C4 alkyl, specifically R8is hydrogen;R9is hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl, specifically R9is hydrogen;n is 1, 2, 3, or 4, specifically n is 1 or 2;m is 0, 1, 2, or 3, specifically m is 0 or 1;q is 0, 1, 2, or 3, specifically q is 0 or 1; andZis CH or N.

[0028] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein Z is CH.

[0029] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein n is 1 or 2, specifically wherein n is 1.

[0030] Where R2and R3combine to form a C3-C8 cycloalkyl, C3-C8 cycloalkenyl, or heterocycloalkyl, such structures include, for example, diazirine, oxirane / epoxide, furan, oxetane, or 1,3-dioxolane.

[0031] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein each R1independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 haloalkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di- C1-C4 alkylamino, halogen, hydroxy, or nitro. In a further embodiment, R1 is Ci-C6 alkyl or Ci-C6 haloalkyl. In a further embodiment, when R1is Ci-Ce alkyl or Ci-Ce haloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or C1-C3 haloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Ce haloalkyl forms a diazirine ring.

[0032] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is Ci-Ce alkyl or Ci-Ce haloalkyl, wherein the Ci-Ce alkyl or Ci-Ce haloalkyl is substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or C1-C3 haloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Ce haloalkyl forms a diazirine ring.

[0033] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is C1-C3 alkyl substituted with one or two substituents each individually selected from hydroxy, Ci-C2alkoxy, C1-C2 haloalkoxy, C1-C2 alkyl, or C1-C2 haloalkyl, or a carbon of the C1-C3 alkyl forms a diazirine ring.

[0034] In an embodiment, a compound of Formula (I), or a pharmaceutically FR2acceptable salt thereof, whereinR1isr3wherein each R2and R3independently is hydrogen Ci-Ce alkyl, Ci-Ce haloalkyl, or OR7wherein R7is hydrogen, Ci-Cealkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C8 cycloalkyl, C3-C8 cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0); and m is 0, 1, 2, or 3, specifically m is 0 or 1. In a further embodiment, m is 0.

[0035] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R5is C1-C4 alkyl substituted with R6, wherein R6is an azido group (N3), cyano, hydroxy, or NHAc; or R5is C2-C4 alkynyl comprising a terminal alkyne.

[0036] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R8 is hydrogen.

[0037] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R9is hydrogen.

[0038] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein q is 0.

[0039] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R8and R9are both hydrogen.

[0040] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R8and R9are both hydrogen and q is 0.

[0041] In an embodiment, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein each R1independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 haloalkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di- C1-C4 alkylamino, halogen, hydroxy, nitro, orwherein each R2and R3independently is hydrogen Ci-Ce alkyl, Ci-Ce haloalkyl, or OR7wherein R7is hydrogen, Ci-Cealkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C8 cycloalkyl, C3-C8 cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0); wherein when R1is Ci-Ce alkyl or Ci-C6 haloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or C1-C3 haloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Ce haloalkyl forms a diazirine ring; R4is C1-C2 alkyl, C1-C2 alkoxy or fluoro; R5is Ci-Cs alkyl substituted with R6, cyano, C2-C6 alkenyl, C2-C6 alkynyl, or a heterocycloalkyl, wherein R6is an azido group (N3), cyano, OR7, SR7, SO2R7, NR7R7, or heterocycloalkyl, wherein each R7individually is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, specifically R5 is C2-C4 alkyl substituted with an azido group (N3), cyano, or alkynyl; R8and R9are both hydrogen; n is 1 or 2; m is 0 or 1; q is 0 or 1; and Z is CH.

[0042] Also included in this disclosure are compounds of Formula (I) havingTable 1.Structure Compound ])N=C3C=CC=CN23)C=C1N3HS-D2u II - - / / \\ft | \ / / n \ s — JJCF3SMILES: OC(C1=CC=C(NC2=NC(=CS2)C2=C(CCN=[N+]=[N- ])N=C3C=CC=CN23)C=C1)C(F)(F)FN3HS-D3u II -N / / \\fi 1 \ / / IT >II JS‘ -CF3SMILES: CCOC(C1=CC=C(NC2=NC(=CS2)C2=C(CCN=LN+J=LN- ])N=C3C=CC=CN23)C=C1)C(F)(F)FTable 1.Structure Compound N3HS-D4H y —nU I \ / / TU Js' —CF3SMILES: COC(C1=CC=C(NC2=NC(=CS2)C2=C(CCN=[N+]=[N- ])N=C3C=CC=CN23)C=C1 )C(F)(F)F

[0043] The compounds are described using standard nomenclature. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs. Unless clearly contraindicated by the context each compound name includes the free acid or free base form of the compound as well hydrates of the compound and all pharmaceutically acceptable salts of the compound.

[0044] The term “Formula (I)”, as used herein, encompasses all compounds that satisfy Formula (I), including any enantiomers, racemates and stereoisomers, as well as all pharmaceutically acceptable salts and radioisotopes of such compounds. The phrase “a compound of Formula (I)” includes all subgeneric groups of Formula (I), and so forth, as well as all forms of such compounds, including salts and hydrates, unless clearly contraindicated by the context in which this phrase is used.

[0045] Formula (I) includes all subformulae thereof. In certain situations, the compounds of Formula (I) may contain one or more asymmetric elements such as stereogenic centers, stereogenic axes and the like, e.g. asymmetric carbon atoms, so that the compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms. For compounds with two or more asymmetric elements, these compounds can additionally be mixtures of diastereomers. For compounds having asymmetric centers, it should be understood that all of the optical isomers and mixtures thereof areencompassed. In these situations, single enantiomers, i.e., optically active forms, can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or by resolution of the racemates. Resolution of the racemates can also be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example, a chiral high performance liquid chromatography (HPLC) column.

[0046] Where a compound exists in various tautomeric forms, the compound is not limited to any one of the specific tautomers, but rather includes all tautomeric forms.

[0047] All isotopes of atoms occurring in the present compounds are contemplated. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example, and without limitation, isotopes of hydrogen include tritium and deuterium; isotopes of carbon include11C,13C, and14C; and an isotope of fluorine includes18F.

[0048] The term “active agent”, as used herein, means a compound of Formula (I) that when administered to a patient, alone or in combination with another compound, element, or mixture, confers, directly or indirectly, a physiological effect on the patient. The indirect physiological effect may occur via a metabolite or other indirect mechanism. When the active agent is a compound, then salts, solvates (including hydrates) of the free compound, crystalline forms, non-crystalline forms, and any polymorphs of the compound are included. All forms are contemplated herein regardless of the methods used to obtain them.

[0049] A dash that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -(CH₂)C₃-C₈cycloalkyl is attached through carbon of the methylene (CH₂) group.

[0050] “Alkanoyl” is an alkyl group as defined herein, covalently bound to the group it substitutes by a keto (-(C=O)-) bridge. Alkanoyl groups have the indicated number of carbon atoms, with the carbon of the keto group being included in the numbered carbon atoms. For example a C2alkanoyl group is an acetyl group having the formula CH3(C=O)-.

[0051] The term “alkyl”, as used herein, means a branched or straight chain saturated aliphatic hydrocarbon group having the specified number of carbon atoms, generally from 1 to about 12 carbon atoms. The term C₁-C₆ alkyl as used herein indicates an alkyl group having from 1, 2, 3, 4, 5, or 6 carbon atoms. Other embodiments include alkyl groups having from 1 to 8 carbon atoms, 1 to 4 carbon atoms or 1 or 2 carbon atoms, e.g. Ci-Ce alkyl, C1-C4 alkyl, and C1-C2 alkyl. When Co-Cnalkyl is used herein in conjunction with another group, for example, (cycloalkyl)Co-C4 alkyl, the indicated group, in this case cycloalkyl, is either directly bound by a single covalent bond (Co), or attached by an alkyl chain having the specified numberof carbon atoms, in this case 1, 2, 3, or 4 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, t-butyl, n-pentyl, and secpentyl.

[0052] The term “alkoxy’’ represents an alkyl group as defined above with the indicated number of carbon atoms attached through an oxygen bridge. Examples of alkoxy include methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2-butoxy, t-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, n-hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy.

[0053] The term “aryl”, as used herein, means aromatic groups containing only carbon in the aromatic ring or rings. Typical aryl groups contain 1 to 3 separate, fused, or pendant rings and from 6 to about 18 ring atoms, without heteroatoms as ring members. When indicated, such aryl groups may be further substituted with carbon or non-carbon atoms or groups. Bicyclic aryl groups may be further substituted with carbon or non-carbon atoms or groups. Bicyclic aryl groups may contain two fused aromatic rings (naphthyl) or an aromatic ring fused to a 5- to 7-membered non-aromatic cyclic group that optionally contains 1 or 2 heteroatoms independently chosen from N, O, and S, for example, a 3,4-methylenedioxy-phenyl group. Aryl groups include, for example, phenyl, naphthyl, including 1 -naphthyl and 2-naphthyl, and bi-phenyl.

[0054] The term “cycloalkyl”, as used herein, indicates a saturated hydrocarbon ring group, having only carbon ring atoms and having the specified number of carbon atoms, usually from 3 to about 8 ring carbon atoms, or from 3 to about 7 carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl as well as bridged or caged saturated ring groups such as norborane or adamantane.

[0055] The term “heterocycloalkyl”, as used herein, indicates a saturated cyclic group containing from 1 to about 3 heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon. Heterocycloalkyl groups have from 3 to about 8 ring atoms, and more typically have from 5 to 7 ring atoms. Examples of heterocycloalkyl groups include morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl groups. A nitrogen in a heterocycloalkyl group may optionally be quaternized.

[0056] The term “alkenyl”, as used herein, means straight and branched hydrocarbon chains comprising one or more unsaturated carbon-carbon bonds, which may occur in any stable point along the chain. Alkenyl groups described herein typically have from 2 to about 12 carbon atoms. Exemplary alkenyl groups are lower alkenyl groups, those alkenyl groups having from 2 to about 8 carbon atoms, e.g. C2-C8, C2-C6, and C2-C4 alkenyl groups. Examples of alkenyl groups include ethenyl, propenyl, and butenyl groups.

[0057] The term “alkynyl”, as used herein, means straight and branched hydrocarbon chains comprising one or more unsaturated carbon-carbon triple bonds (acetylene), which may occur in any stable point along the chain. Alkynyl groups described herein typically have from 2 to about 12 carbon atoms.

[0058] The term “cycloalkenyl”, as used herein, means a saturated hydrocarbon ring group, comprising one or more unsaturated carbon-carbon bonds, which may occur in any stable point of the ring, and having the specified number of carbon atoms. Monocyclic cycloalkenyl groups typically have from 3 to about 8 carbon ring atoms or from 3 to 7 (3, 4, 5, 6, or 7) carbon ring atoms. Cycloalkenyl substituents may be pendant from a substituted nitrogen or carbon atom, or a substituted carbon atom that may have two substituents may have a cycloalkenyl group, which is attached as a spiro group. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, or cyclohexenyl as well as bridged or caged saturated ring groups such as norbornene.

[0059] The term “heteroaryl", as used herein, indicates a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic ring which contains at least 1 aromatic ring that contains from 1 to 4, or specifically from 1 to 3, heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon. When the total number of S and O atoms in the heteroaryl group exceeds 1, theses heteroatoms are not adjacent to one another. Specifically, the total number of S and O atoms in the heteroaryl group is not more than 2, more specifically the total number of S and O atoms in the heteroaryl group is not more than 1. A nitrogen atom in a heteroaryl group may optionally be quatemized. When indicated, such heteroaryl groups may be further substituted with carbon or non-carbon atoms or groups. Such substitution may include fusion to a 5 to 7-membered saturated cyclic group that optionally contains 1 or 2 heteroatoms independently chosen from N, O, and S, to form, for example, a [l,3]dioxolo[4,5-c]pyridyl group. In certain embodiments 5- to 6-membered heteroaryl groups are used. Examples of heteroaryl groups include, but are not limited to, pyridyl, indolyl, pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl, and 5,6,7,8-tetrahydroisoquinoline.

[0060] “Haloalkyl” includes both branched and straight-chain alkyl groups having the specified number of carbon atoms, substituted with 1 or more halogen atoms, up to the maximum allowable number of halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl.

[0061] “Haloalkoxy” is a haloalkyl group as defined herein attached through an oxygen bridge (oxygen of an alcohol radical).

[0062] “Halo" or “halogen" is any of fluoro, chloro, bromo, and iodo.

[0063] “Amino” is -NH2. “Mono- and / or di-alkylamino” is a secondary or tertiary alkyl amino group, wherein the alkyl groups are independently chosen alkyl groups, as defined herein, having the indicated number of carbon atoms. The point of attachment of the alkylamino group is on the nitrogen. Examples of mono- and di-alkylamino groups include ethylamino, dimethylamino, and methyl-propyl-amino.

[0064] The term “substituted”, as used herein, means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom’s normal valence is not exceeded. When the substituent is oxo (i.e., =0) then 2 hydrogens on the atom are replaced. When an oxo group substitutes aromatic moieties, the corresponding partially unsaturated ring replaces the aromatic ring. For example, a pyridyl group substituted by oxo is a pyridone. A stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and subsequent formulation into an effective therapeutic agent.

[0065] Unless otherwise specified substituents are named into the core structure. For example, it is to be understood that when (cycloalkyl)alkyl is listed as a possible substituent the point of attachment of this substituent to the core structure is in the alkyl portion, or when arylalkyl is listed as a possible substituent the point attachment to the core structure is the alkyl portion.

[0066] Suitable groups that may be present on a “substituted” or “optionally substituted” position include, but are not limited to, halogen; cyano; hydroxyl; nitro; azido; alkanoyl (such as a C2-C6 alkanoyl group such as acyl or the like); carboxamido; alkyl groups (including cycloalkyl groups) having 1 to about 8 carbon atoms, or 1 to about 6 carbon atoms; alkenyl and alkynyl groups including groups having one or more unsaturated linkages and from 2 to about 8, or 2 to about 6 carbon atoms; alkoxy groups having one or more oxygen linkages and from 1 to about 8, or from 1 to about 6 carbon atoms; aryloxy such as phenoxy; alkylthio groups including those having one or more thioether linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; alkylsulfinyl groups including those having one or more sulfinyl linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; alkylsulfonyl groups including those having one or more sulfonyl linkages and from 1 to about 8 carbon atoms, or from 1 to about 6 carbon atoms; aminoalkyl groups including groups having one or more N atoms and from 1 to about 8, or from 1 to about 6 carbon atoms; aryl having 6 or more carbons and one or more rings, (e.g., phenyl, biphenyl, naphthyl, or the like, each ring either substituted or unsubstituted aromatic); arylalkyl having 1 to 3 separate or fused rings and from 6 to about 18 ring carbon atoms, with benzyl being an exemplary arylalkyl group;arylalkoxy having 1 to 3 separate or fused rings and from 6 to about 18 ring carbon atoms, with benzyloxy being an exemplary arylalkoxy group; or a saturated, unsaturated, or aromatic heterocyclic group having 1 to 3 separate or fused rings with 3 to about 8 members per ring and one or more N, O or S atoms, e.g. coumarinyl, quinolinyl, isoquinolinyl, quinazolinyl, pyridyl, pyrazinyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, triazinyl, oxazolyl, isoxazolyl, imidazolyl, indolyl, benzofuranyl, benzothienyl, benzothiazolyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, and pyrrolidinyl. Such heterocyclic groups may be further substituted, e.g. with hydroxy, alkyl, alkoxy, halogen and amino.

[0067] The term “pharmaceutically acceptable salt”, as used herein, includes derivatives of the disclosed compounds in which the parent compound is modified by making inorganic and organic, acid or base addition salts thereof. The salts of the present compounds can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used, where practicable. Salts of the present compounds further include solvates of the compounds and of the compound salts.

[0068] Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts and the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, conventional nontoxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, HOOC-(CH2)x-COOH where x is 0, 1, 2, 3, or 4, and the like.

[0069] In an embodiment, a pharmaceutical formulation comprises a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0070] The term “pharmaceutical compositions”, as used herein, are compositionscomprising at least one active agent, such as a compound or salt of Formula (I), and at least one other substance, such as a pharmaceutical excipient. Pharmaceutical compositions can meet the U. S. FDA’s GMP (good manufacturing practice) standards for human or non-human drugs. The pharmaceutical compositions can be formulated into a dosage form.

[0071] The term “subject” or “patient”, as used herein, is a human in need of medical treatment. Medical treatment can include treatment of an existing condition, such as a disease or disorder, prophylactic or preventative treatment, or diagnostic treatment.

[0072] The term “providing”, as used herein, means giving, administering, selling, distributing, transferring (for profit or not), manufacturing, compounding, or dispensing.

[0073] The term “providing a compound of Formula (I) with at least one additional therapeutic agent”, as used herein, means the compound of Formula (I) and the additional active agent(s) are provided simultaneously in a single dosage form, provided concomitantly in separate dosage forms, or provided in separate dosage forms for administration separated by some amount of time that is within the time in which both the compound of Formula (I) and the at least one additional active agent are within the blood stream of a patient. The compound of Formula (I) and the additional active agent need not be prescribed for a patient by the same medical care worker. The additional active agent or agents need not require a prescription. Administration of the compound of Formula (I) or the at least one additional active agent can occur via any appropriate route, for example, oral tablets, oral capsules, oral liquids, inhalation, injection, suppositories or topical contact.

[0074] In an embodiment, a pharmaceutical formulation comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, formulated for oral administration.

[0075] The term “treatment”, as used herein, includes providing a compound of Formula (I), either as the only active agent or together with at least one additional active agent sufficient to: (a) prevent a disease or a symptom of a disease from occurring in a patient who may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e. arresting its development; and (c) relieving the disease, i.e., causing regression of the disease. “Treating” and “treatment” also means providing a therapeutically effective amount of a compound of Formula (I), as the only active agent or together with at least one additional active agent to a patient suffering from a disease, e.g., a cancer, or symptom.

[0076] The term “therapeutically effective amount” of a pharmaceutical composition, as used herein, means an amount effective, when administered to a patient, to provide a therapeutic benefit such as an amelioration of symptoms, e.g., to treat a patient suffering from a cancer.

[0077] The compounds can be administered as the neat chemical or administered as a pharmaceutical composition. Accordingly, an embodiment provides pharmaceutical compositions comprising a compound or pharmaceutically acceptable salt of Formula (I), together with a pharmaceutically acceptable excipient. The pharmaceutical composition may contain a compound or salt of Formula (I) as the only active agent or may contain one or more additional active agents.

[0078] The compounds may be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, via buccal administration, rectally, as an ophthalmic solution, or by other means, in dosage unit formulations containing conventional pharmaceutically acceptable carriers. The pharmaceutical composition may be formulated as any pharmaceutically useful form, e.g., as an aerosol, a cream, a gel, a pill, a capsule, a tablet, a syrup, a transdermal patch, or an ophthalmic solution. Some dosage forms, such as tablets and capsules, are subdivided into suitably sized unit doses containing appropriate quantities of the active components, e.g., an effective amount to achieve the desired purpose.

[0079] Excipients include carriers and diluents and of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the patient being treated. The excipient can be inert or it can possess pharmaceutical benefits of its own. The amount of an excipient employed in conjunction with the compound is sufficient to provide a practical quantity of material for administration per unit dose of the compound.

[0080] Classes of excipients include, for example, buffering agents, coloring agents, diluents, disintegrants, emulsifiers, flavorants, glidants, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some excipients may be listed in more than one class, for example vegetable oil may be used as a lubricant in some formulations and a diluent in others. Exemplary pharmaceutically acceptable excipients include sugars, starches, celluloses, powdered tragacanth, malt, gelatin, talc, and vegetable oils. Optional active agents may be included in a pharmaceutical composition, which do not substantially interfere with the activity of the compound of Formula (I).

[0081] The pharmaceutical compositions can be formulated for oral administration. These compositions contain between 0.1 and 99 weight percent (“wt.%”) of a compound of Formula (I), and usually at least about 5 wt.%. Some embodiments contain from about 25 wt.% to about 50 wt. % or from about 5 wt.% to about 75 wt.% of a compound of Formula (I).

[0082] The pharmaceutical composition can be formulated in a package comprising the pharmaceutical composition containing a compound of Formula (I) or a saltthereof in a container and further comprising instructions for using the composition in order to elicit a therapeutic effect in a subject.

[0083] The pharmaceutical composition can also be formulated in a package comprising the pharmaceutical composition of Formula (I) or a salt thereof in a container and further comprising instructions for using the composition to treat a subject suffering from, for example, a cancer.

[0084] In an embodiment, a method of treating a cancer comprises administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need of treatment. In an embodiment, the cancer is non-small cell lung cancer, breast cancer, prostate cancer, pancreatic cancer, head and neck cancer, medulloblastoma, glioblastoma, bile duct carcinoma, neuroblastoma, colon cancer, myeloma, gastric cancer, liver cancer, ovarian cancer, colorectal cancer, non-Hodgkin lymphoma, small-cell lung cancer, large cell lung cancer, kidney cancer, esophageal cancer, stomach cancer, cervical cancer, sarcomas, leukemia, or a lymphoma tumor. In an embodiment, the cancer is glioblastoma.

[0085] In an embodiment, use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to prepare a medicament useful for the treatment of cancer in subject.

[0086] In a further embodiment, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, useful in the treatment of a cancer is using in combination with an additional active agent, radiation therapy, or a combination thereof.Suitable additional active agents include, for example, an anti-cancer agent, anti-proliferative agent, immunomodulatory agent, anti-angiogenic agent, anti-inflammatory agent, pain reliever, p2-agonist, anticholinergic agent, antihistamine, anti-malarial agent, anti-viral agent, antibiotic, or a combination thereof.

[0087] In an embodiment, a method for inhibiting BMI1 expression in a subject in need thereof comprises administering to the subject an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0088] In an embodiment, use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to prepare a medicament useful for inhibiting BMI1 expression in a subject.

[0089] Several of the compounds described herein can be used as research tools including as small molecule probes to map binding sites and small molecule binding, particularly in oncogenes. These compounds include photo-reactive and “clickable” affinity-based probes.Table 2.

[0090] Further disclosed herein is a method comprising providing an mRNA sequence; identifying a secondary structure of the mRNA sequence, wherein the secondary structure corresponds to a pocket within a 5’UTR; generating highly probable models of thepocket secondary structure, wherein the model accounts for a target minimum and maximum free energy; altering at least one functional group of a small molecule to conform the molecule to a subset of the highly probable models of the pocket to create a targeted small molecule; synthesizing the targeted small molecule; validating the mRNA binding of the targeted small molecule. Within this embodiment, the small molecule is a compound of Formula (I) or a compound in Table 1 or Table 2. In an embodiment, the mRNA sequence can be an oncogene sequence. The altered functional group is at least one selected from the group of azido, alkyne, alkene, amino, hydroxyl, and thiol. The pocket is one selected from the group consisting of groove fold structure, sphere -like, disc-like, and rod-like. Within the method, when generating the highly probable models of the pocket secondary structure, the model accounts for at least one selected from the group consisting of pocket volume, surface area, effective radius, sphericity, and pocket centroid.

[0091] The invention is further illustrated by the following non-limiting examples.EXAMPLES EXAMPLE 1 Synthesis of HS-D1 4-(2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl)-N-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) thiazol-2-amine

[0092] Step 1: l-(4-Azidophenyl)-2,2,2-trifluoroethanone (2A)

[0093] To a solution of 2,2,2-trifluoro-1-(4-fluorophenyl) ethanone (9.2 g, 47.91 mmol) in dry dimethylformamide (DMF) (80 mL) was added NaNs (15.5 g, 239.5 mmol), and the reaction was stirred at room temperature for 16 hours. The reaction mixture was quenched with water and extracted with EtOAc twice. The combined organic layers were washed with water and brine, dried over Na2SC>4. Filtered and the filtrate was concentrated in vacuo to give the crude product (10.0 g, 92% purity, yield 83%) as yellow oil, which was used in next step without further purification.

[0094] ’H NMR (400 MHz, Chlorofonn-d) 58.07 (dd, J = 1.2, 7.6 Hz, 2H), 7.14-7.17 (m, 2H).

[0095] Step 2 & 3; l-(4-Azidophenyl)-2,2,2-trifluoroethanone oxime (4A)

[0096] l-(4-Azidophenyl)-2,2,2-trifluoroethanone (4.32 g, 20.0 mmol) was dissolved in 60 mL of EtOH containing 10 mL of pyridine. Hydroxylamine hydrochloride (1.68 g, 1.2 equiv) was then added, and the mixture was heated at reflux for 16 hours before the solvent was removed under vacuum and the residue was used directly in next step.

[0097] The oximes prepared above were treated with 4-toluenesulfonyl chloride (4.19 g, 1.1 equiv) in acetone (60 mL) and triethylamine (10 mL). After 1 hour the solution wasfiltered, the filtrate was evaporated to dryness, and purified by flash chromatography (hexane / EtOAc=25:l). The product was obtained as a mixture of isomers (6.0 g, 92%) as yellow solid.

[0098] 1H NMR (400 MHz, Chloroform-d) δ 7.89 (dd, J = 8.4, 3.3 Hz, 2H), 7.48 - 7.42 (m, 2H), 7.41 - 7.35 (m, 2H), 7.14 - 7.03 (m, 2H), 2.47 (d. J = 6.4 Hz, 3H).

[0099] Step 4: 3-(4-Azidophenyl)-3-(trifluoromethyl) diaziridine (5 A)[000100] The tosyloxime 4A (1.6 g, 4.15 mmol) was dissolved in diethyl ether (12 mL) and added to 12 mL of NH3 in MeOH at -78 °C. The mixture was stirred for 4 hours before being brought to room temperature overnight. The mixture was then evaporated to dryness and purified by flash chromatography (hexane / EtOAc=90:10), to obtain the diaziridine 5A (0.8 g, 78.6%) as yellow oil.[000101] 1H NMR (400 MHz, Chloroform-d) δ 7.64 – 7.56 (m, 2H), 7.10 - 7.03 (m, 2H), 2.81 (d, J = 8.8 Hz, 1H), 2.26- 2.14 (m, 1H).[000102] Step 5: 4-(3-(trifluoromethyl) diaziridin-3-yl) aniline (6A)[000103] To a solution of 3 -(4-azidophenyl)-3 -(trifluoromethyl) diaziridine (2.3 g, 10 mmol) in tetrahydrofuran (THF) (20 mL) and H2O (2.5 mL) was slowly added Ph3P (5.25 g, 20 mmol), the reaction mixture was stirred at 70 °C for 48 h. After the reaction was completed, the mixture was concentrated under vacuum to give the crude product. The crude product was purified by Flash Chromatography (solvent gradient: 0% - 30% EtOAc in petroleum ether (PE)) to afford the desired product (1.8 g, 90% yield) as a yellow solid.[000104] 1H NMR (400 MHz, CDCl3) δ 7.37 (d, J = 8.4 Hz, 2H), 6.67 (d, J = 8.6 Hz, 2H), 3.83 (s, 2H), 2.71 (d, J = 8.2 Hz, 1H), 2.14 (d, J = 8.4 Hz, 1H).[000105] Step 6: 4-(3-(trifluoromethyl)-3H-diazirin-3-yl) aniline (7A)[000106] To a solution of 4-(3 -(trifluoromethyl) diaziridin-3-yl) aniline (1.3 g, 6.4 mmol) in dichloromethane (DCM) (10 mL) was added triethylamine (TEA) (1.3 g, 12.8 mmol) and I2 (1.6 g, 6.4 mmol) at 0 °C under N2. The reaction mixture was stirred at 0 °C for 1 hour under N2. After the reaction was completed, the mixture was quenched with saturated Na2S20s aqueous (20 mL) and then extracted with DCM (15 mL x 3). The organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, solvent was removed and the residue was purified by Flash Chromatography (solvent gradient: 0% - 20% EtOAc in PE) to afford the desired product (1.1 g, 85% yield) as a yellow solid.[000107] 1H NMR (400 MHz, CDCl3) δ 6.99 (d. J = 8.4 Hz, 2H), 6.64 (d, J = 8.6 Hz, 2H), 3.84 (s, 2H).[000108] Step 7: N-((4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) carbamothioyl) benzamide (9 A)[000109] To a solution of 4-(3-(trifluoromethyl)-3H-diazirin-3-yl) aniline (1.1 g, 5.4 mmol) in MeCN (12 mL) was slowly added benzoyl isothiocyanate (1.0 g, 6 mmol) at room temperature and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under vacuum to give the crude product, the crude product was purified by Flash Chromatography (solvent gradient: 0% - 10% EtOAc in PE) to afford the desired product (1.4 g, 71% yield) as a yellow solid.[000110] LCMS calculated for C16H11F3N4OS (M+H) + m / z = 365.1; found 365.0.[000111] Step 8: l-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) thiourea (10A) [000112] To a solution of N-((4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) carbamothioyl) benzamide (1.4 g, 3.8 mmol) in MeOH (15 mL) and H2O (1.5 mL) was added K2CO3 (1 g, 7.6 mmol), the reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was filtered and the filter cake was washed with MeOH (10 mL), solvent was removed the residue was purified by Flash Chromatography (solvent gradient: 0% - 30% EtOAc in PE) to afford the desired product (0.9 g, 90% yield) as a white solid.[000113] LCMS calculated for C9H7F3N4S (M+H) + m / z = 261.0; found 261.0.[000114]1H NMR (400 MHz, DMSO-d6) δ 9.91 (s, 1H), 7.68 – 7.57 (m, 2H), 7.23 (d, J = 8.4 Hz, 2H), 8.2 – 7.0 (br s, 2H).[000115] Step 9: ethyl 2-(imidazo[l,2-a] pyridin-2-yl) acetate (Int 2)[000116] To a solution of pyridin-2-amine (30 g, 318.8 mmol) in THF (400 mL) was added ethyl 4-chloro-3-oxobutanoate (52.47 g, 318.8 mmol) and the reaction mixture was stirred at reflux for 16 hours. The reaction mixture was concentrated under reduced pressure and purified by Flash Chromatography (eluting with 0% to 10% MeOH in DCM) to give the title product as black oil (18.33 g, 28.2%).[000117] 1H NMR (400 MHz, Chloroform-d) δ 8.05 (dq, J = 6.8, 1.1 Hz, 1H), 7.58 (s, 1H), 7.56 - 7.50 (m, 1H), 7.13 (ddt, J = 9.0, 6.8, 1.0 Hz, 1H), 6.74 (tt, J = 6.8, 1.0 Hz, 1H), 4.20 (qd, J = 7.1, 0.7 Hz, 2H), 3.86 (s, 2H), 1.28 (td, J = 7.1, 0.8 Hz, 3H).[000118] Step 10: 2-(imidazo[l,2-a] pyridin-2-yl) ethan-l-ol (Int 3)[000119] To a solution of ethyl 2-{imidazo[1,2-a] pyridin-2-yl} acetate (10 g, 49.0 mmol) in THF (120 mL) was added LiAlH4 (73.5 mL, 73.5 mmol) and the reaction mixture was stirred at room temperature for 5 hours. The reaction was quenched with sat. NH4CI and concentrated under reduced pressure to give the crude product. The crude product was purified by Flash Chromatography (eluting with 0% to 10% MeOH in DCM) to give the title product as white solid (6.33 g, 79.6%).[000120] ’H NMR (400 MHz, DMSO-J6) 58.45 (dt, J = 6.7, 1.3 Hz, 1H), 7.72 -7.69 (m, 1H), 7.44 (dq, J = 9.1, 1.0 Hz, 1H), 7.15 (ddd, J = 9.0, 6.8, 1.4 Hz, 1H), 6.83 – 6.78 (m, 1H), 4.70 (s, 1H), 3.75 - 3.69 (m, 2H), 2.87 - 2.79 (m, 2H).[000121] Step 11: 2-(3-bromoimidazo[l,2-a] pyridin-2-yl) ethan-l-ol (Int 4)[000122] To a solution of 2-{imidazo[l,2-a] pyridin-2-yl } ethanol (5.4 g, 33.3 mmol) in DMF (50 mL) at 0 °C was added N-bromosuccinimide (7.11 g, 39.9 mmol) in batches. Then the reaction mixture was stirred at 0 °C for further 20 minutes. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Flash Chromatography (eluting with 0% to 5% MeOH in DCM) to give the title product as off-white solid (5.5 g, 68.5%).[000123] 1H NMR (400 MHz, Chloroform-d) δ 8.08 (dt, J = 6.8, 1.2 Hz, 1H). 7.54 (dt, J = 9.0, 1.2 Hz, 1H), 7.26 - 7.21 (m, 1H), 6.92 (td, J = 6.9, 1.2 Hz, 1H), 4.04 (t, J = 5.7 Hz, 2H), 3.68 (s, 1H), 3.02 (t, J = 5.7 Hz, 2H).[000124] Step 12: 3-bromo-2-(2-((tert-butyldiphenylsilyl) oxy) ethyl) imidazo[l,2-a] pyridine (Int 5)[000125] To a solution of 2-{3-bromoimidazo[l,2-a] pyridin-2-yl } ethanol (450 mg, 1.87 mmol) in DCM (10 mL) was added imidazole (191 mg, 2.80 mmol) and the mixture was stirred at room temperature for 10 minutes. Then tert-butyl(chloro)diphenylsilane (TBDPSC1) (564 mg, 2.05 mmol) was added and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Flash Chromatography (eluting with 0% to 20% EtOAc in PE) to give the title product as white solid (847 mg, 94.5%).[000126] ’H NMR (400 MHz, Chloroform-d) 58.03 (dt, J = 6.8, 1.2 Hz, 1H), 7.62 -7.57 (m, 4H), 7.50 (dt, J = 9.1, 1.1 Hz, 1H), 7.39 - 7.34 (m, 2H), 7.33 - 7.27 (m, 4H), 7.19 (ddd, J = 9.0, 6.7, 1.3 Hz, 1H), 6.87 (td, J = 6.8, 1.2 Hz, 1H), 4.06 (t, J = 6.9 Hz, 2H), 3.09 (t, J = 7.0 Hz, 2H), 1.01 (s, 9H).[000127] Step 13: l-(2-(2-((tert-butyldiphenylsilyl) oxy) ethyl) imidazo[l,2-a] pyridin-3-yl) ethan-l-one (Int 6)[000128] To a solution of 3-bromo-2-{2-[(tert-butyldiphenylsilyl) oxy] ethyl} imidazo[1,2-a] pyridine (80 mg, 0.17 mmol) and tributyl(l -ethoxy ethenyl) stannane (66 mg, 0.18 mmol) in 1,4-dioxane (1 mL) was added Tetrakis(triphenylphosphine)Palladium (10 mg, 0.01 mmol) and the reaction mixture was stirred at 150 °C under microwave for 2 hours. The reaction mixture was diluted with H2O and extracted with EtOAc. The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude product was purified by FlashChromatography (eluting with 0% to 25% EtOAc in PE) to give the title product as white solid (61 mg, 82.6%).[000129] 1H NMR (400 MHz, Chloroform-d) δ 9.73 (d, J = 7.0 Hz, 1H), 7.60 (dt, J = 8.9, 1.2 Hz, 1H), 7.54 - 7.50 (m, 4H), 7.44 (ddd, J = 8.7, 6.8, 1.3 Hz, 1H), 7.37 - 7.32 (m, 2H), 7.27 - 7.23 (m, 4H), 7.00 (td, J = 6.9, 1.3 Hz, 1H), 4.15 (t, J = 6.7 Hz, 2H), 3.40 (t, J = 6.7 Hz, 2H), 2.62 (s, 3H), 0.98 (s, 9H).[000130] Step 14: l-(2-(2-hydroxy ethyl) imidazo[l,2-a] pyridin-3-yl) ethan-l-one (Int 7)[000131] To a solution of 1-(2-{2-[(tert-butyldiphenylsilyl) oxy] ethyl} imidazo[1,2-a] pyridin-3-yl) ethanone (4 g, 9.04 mmol) in THF (50 mL) was added tetrabutylammonium fluoride (TBAF) (10 mL, 10.0 mmol) and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Flash Chromatography (eluting with 0% to 10% DCM / MeOH) to give the title product as off-white solid (1.5 g, 81.3%).[000132] 1H NMR (400 MHz, DMSO-d6) δ 9.63 (dt, J = 7.0, 1.2 Hz, 1H), 7.72 (dt, J = 8.9, 1.2 Hz, 1H), 7.58 (ddd, J = 9.0, 6.9, 1.3 Hz, 1H), 7.18 (td, J = 6.9, 1.4 Hz, 1H), 4.81 (t, J = 5.5 Hz, 1H), 3.87 (td, J = 6.9, 5.5 Hz, 2H), 3.24 (t, J = 6.9 Hz, 2H), 2.64 (s, 3H).[000133] Step 15: 2-(3-acetylimidazo[l,2-a]pyridin-2-yl)ethyl 4-methylbenzenesulfonate (Int 8)[000134] To a solution of l-[2-(2-hydroxyethyl)imidazo[l,2-a]pyridin-3-yl]ethanone (1.5 g, 7.34 mmol) and triethylamine (2.23 g, 22.03 mmol) in DCM (20 mL) was added p-toluenesulfonyl chloride (p-TsCl) (2.1 g, 11.02 mmol) and the reaction mixture was stirred at room temperature for 16 hour. The reaction mixture was concentrated under reduced pressure and purified by Flash Chromatography (eluting with 0% to 25% EtOAc / PE) to give the title product as white solid (2.31 g, 87.8%).[000135] 1H NMR (400 MHz, Chloroform-d) δ 9.70 (dt, J = 6.9, 1.2 Hz, 1H), 7.70 – 7.63 (m, 2H), 7.54 (dt, J = 8.9, 1.3 Hz, 1H), 7.46 (ddd, J = 8.9, 6.8, 1.3 Hz, 1H), 7.20 - 7.14 (m, 2H), 7.03 (td, J = 6.9, 1.4 Hz, 1H), 4.61 (t, J = 6.7 Hz, 2H), 3.48 (t, J = 6.8 Hz, 2H), 2.62 (s, 3H), 2.35 (s, 3H).[000136] Step 16: 1 -(2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl) ethan-l-one (Int 9)[000137] To a solution of 2-{3-acetylimidazo[l,2-a]pyridin-2-yl}ethyl 4-methylbenzenesulfonate (2300 mg, 6.42 mmol) in DMF (20 mL) was added NaNs (834 mg, 12.83 mmol) and the reaction mixture was stirred at 80 °C for 2 hours. The reaction mixture was concentrated under reduced pressure and diluted with H2O. The mixture was extracted withEtOAc. The organic layer was dried over Na2SC>4 and concentrated under vacuum. The crude product was purified by Flash Chromatography (eluting with 0% to 50% EtOAc / PE) to give the title product as off-white solid (1.15 g, 78.2%).[000138] 1H NMR (400 MHz, Chloroform-d) δ 9.75 (dt, J = 7.1, 1.2 Hz, 1H), 7.67 (dt, J = 8.9, 1.2 Hz, 1H), 7.48 (ddd, J = 9.0, 6.9, 1.3 Hz, 1H), 7.04 (td. J = 6.9, 1.3 Hz, 1H), 3.93 (t, J = 7.0 Hz, 2H), 3.39 (t, J = 7.0 Hz, 2H), 2.67 (s, 3H).[000139] Step 17: l-(2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl)-2-bromoethan- 1-one (Int 10)[000140] To a stirred solution of 1 -[2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl] ethanone (500 mg, 2.18 mmol) and p-toluenesulfonic acid (PTS A) (75 mg, 0.44 mmol) in CH3CN (10 mL) at 80 °C was added N-bromosuccinimide (466 mg, 2.62 mmol) and the reaction mixture was stirred at 80 °C for 3 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by Flash Chromatography (eluting with 0% to 35% EtOAc / PE) to give the title product as white solid (224 mg, 33.3%).[000141] 1H NMR (400 MHz, DMSO-d6) δ 9.63 – 9.60 (m, 1H), 7.84 – 7.79 (m, 1H), 7.68 (ddd, J = 8.6, 6.9, 1.3 Hz, 1H), 7.29 (td, J = 6.9, 1.3 Hz, 1H), 4.81 (s, 2H), 3.93 (t, J = 6.7 Hz, 2H), 3.43 (t, J = 6.7 Hz, 2H).[000142] Step 18: 4-(2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl)-N-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) thiazol-2-amine (RU-HS-D1)[000143] To a solution of l-(2-(2-azidoethyl) imidazo[l,2-a] pyridin-3-yl)-2-bromoethan-l-one (250 mg, 0.81 mmol) in MeCN (5 mL) was added l-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl) phenyl) thiourea (191 mg, 0.73 mmol) and the reaction mixture was stirred at room temperature for 3 hours. After completion, 5 drops of DMF were added to make the mixture clear. The reaction mixture was filtered and the filtrate was purified by Reverse-phase column (80 g-C18 column, eluting with 0% to 48% MeCN / H2O containing 0.1% TFA, MeCN 0% hold 2 CV, from 0% to 50% over 8 CV, 120 mL / min) to give the title product as yellow solid (193 mg, 51%).[000144] LCMS (ESI) calcd for C20H15F3N9S [M + H] + m / z 470.11, found 469.80.[000145] 1H NMR (400 MHz, DMSO-d6) δ 10.86 (s, 1H), 8.93 (dd, J = 6.9, 1.2 Hz, 1H), 7.95 - 7.89 (m, 1H), 7.83 - 7.73 (m, 3H), 7.52 (s, 1H), 7.40 (td, J = 6.9, 1.2 Hz, 1H), 7.25 (d, J = 8.4 Hz, 2H), 3.83 (d, J = 6.8 Hz, 2H), 3.25 (t, J = 6.7 Hz, 2H).EXAMPLE 2 Synthesis of HS-D2 l-(4-((4-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)thiazol- 2-yl)amino)phenyl)-2,2,2-trifluoroethan-l -ol hydrochloride[000146] A sample of solid 4-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-N-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)thiazol-2-amine hydrobromide hydrolyzed over time to give a 4: 1 mixture of 4-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-N-(4-(3-(trifhioromethyl)-3H-diazirin-3-yl)phenyl)thiazol-2-amine to l-(4-((4-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)thiazol-2-yl)amino)phenyl)-2,2,2-trifluoroethan-l-ol. 193 mg of this sample was purified by prep HPLC (0.1% FA(aq) / MeCN 12 min gradient, Gemini -C1830x100 mm, 42.5 mL / min, UV 214 / 254). Fractions containing the title compound were combined and concentrated. The concentrate was redissolved in a mixture of MeOH / DCM and 6M HCl / IPA(excess) was added. The sample was concentrated to afford the title compound as a yellow solid. 96% purity by HPLC at 254 nm. (MH+) 460.15.[000147] 1H NMR (300 MHz, DMSO) δ 10.69 (s, 1H), 9.02 (d, J = 7.0 Hz, 1H), 8.03 - 7.86 (m, 2H), 7.70 - 7.62 (m, 2H), 7.55 - 7.47 (m, 2H), 7.43 (d, J = 8.4 Hz, 2H), 6.74 (s, 1H), 5.07 (q, J = 7.4 Hz, 1H), 3.85 (t, J = 6.6 Hz, 3H), 3.28 (t, J = 6.6 Hz, 3H).EXAMPLE 3 Synthesis of HS-D34-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-N-(4-(l-ethoxy-2,2,2-trifluoroethyl)phenyl)thiazol-2-amine[000148] To a solution of l-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-2-bromoethan-l-one (20 mg, 0.065 mmol) in EtOH (0.5 mL) was added l-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)thiourea (17 mg, 0.065 mmol) and the reaction mixture was stirred at 70 °C for 3 hours in the dark. The reaction mixture was concentrated in vacuo. The crude product was purified by prep-TLC (eluting with 10% MeOH / DCM) to give the product as yellow solid (3.68 mg, 12%).[000149] LCMS (ESI) calcd for C22H21F3N7OS [M + H] + m / z 488.15, found 487.90.[000150] 1H NMR (400 MHz, Chloroform-d) δ 8.73 (d, J = 6.9 Hz, 1H), 8.03 – 7.81 (m, 1H), 7.60 (d, J = 9.0 Hz, 1H), 7.50 - 7.37 (m, 4H), 7.23 (d, J = 8.6 Hz, 1H), 6.89 (s, 1H), 6.84 (t, J = 6.8 Hz, 1H), 4.58 (q, J = 6.7 Hz, 1H), 3.85 (t, J = 6.9 Hz, 2H), 3.59 (q, J = 7.0 Hz, 2H), 3.23 (t, J = 7.0 Hz, 2H), 1.28 (d, J = 3.5 Hz, 3H).EXAMPLE 4 Synthesis of HS-D44-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-N-(4-(2,2,2-trifluoro- 1 -methoxy ethyl)phenyl)thiazol-2-amine[000151] To a solution of l-(2-(2-azidoethyl)imidazo[l,2-a]pyridin-3-yl)-2-bromoethan-l-one (20 mg, 0.065 mmol) in MeCN / MeOH (2 / 1, 0.5 mL) was added l-(4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)thiourea (17 mg, 0.065 mmol) and the reaction mixture was stirred at 70 °C for 3 hours. The reaction mixture was concentrated in vacuo. Thecrude product was purified by prep-TLC (eluting with 10% MeOH / DCM) to give the product as yellow solid (10.38 mg, 34%).[000152] LCMS (ESI) calcd for C21H19F3N7OS [M + H] + m / z 474.13, found 473.80.[000153] 1H NMR (400 MHz, Chloroform-d) δ 8.73 (d, J = 7.0 Hz, 1H), 8.07 (s, 1H), 7.60 (d, J = 9.2 Hz, 1H), 7.48 (d, J = 8.3 Hz, 2H), 7.40 (d, J = 8.3 Hz, 2H), 7.22 (d, J = 8.5 Hz, 1H), 6.90 (s, 1H), 6.84 (t, J = 6.8 Hz, 1H), 4.48 (q, J = 6.6 Hz, 1H), 3.85 (t, J = 7.0 Hz, 2H), 3.43 (s, 3H), 3.23 (t, J = 7.0 Hz, 2H).EXAMPLE 5 Synthesis of HS-D5 3-(4-(azidomethyl)phenyl)-3-(trifluoromethyl)-3H-diazirine [000154] Prepared according to von Ballmoos, C.; Appoldt, Y.; Brunner, J.;Granier, T.; Vasella, A.; Dimroth, P. Membrane Topography of the Coupling Ion Binding Site in Na+-translocating FIFO ATP Synthase*. Journal of Biological Chemistry 2002, 277 (5), 3504-3510. DOI: https: / / doi.org / 10.1074 / jbc. M110301200.EXAMPLE 6[000155] Compound HS-D1 is a photo-reactive and “clickable” affinity-based probe and structural mimics of A15 (N-(4-methoxyphenyl)-4-(2-methylimidazo[l,2-a]pyridin-3-yl)thiazol-2-amine) to map binding sites and small molecule binding. The HS-D1 photoaffinity probe technology will allow the identification of binding to a small molecule at nucleotide resolution, taking BMI1 as protype for small molecule binding.[000156] Compounds HS-D2 to HS-D4 are novel BMI1 targeting compounds, and HS-D5 is a negative control for the HS-D1 probe. The effects of these novel probes and compounds in GBM sphere and organoid assays (See, Chadwick et al., iScience 23, 101365, August 21, 2020) were examined and found to inhibit BMI1, albeit without the notable cell cycle arrest effects (increased phospho-histone H3), which is a characteristic feature of A15.[000157] To examine the activity of Compounds HS-D1 to HS-D5, the potency of these compounds was tested against GBM patient derived cells (FIG. 1, bottom row “Use same as DAPI (DAPI has an excitation peak at 350 nm and an emission peak at 465 nm)’j. Optimum seeding densities for 3D cultures were first established by determining the clonal efficiencies of deriving 3D cultures from dissociated patient cells from patient #50 (GBM#50) at limiting dilutions to single cells. Following seeding, GBM spheres or PDOs) with extracellular matrix) were allowed to grow for 2 weeks. After 2 weeks, the media was aspirated from each well and replaced with media containing the treatment drug. Growth inhibitory concentration at 50% (IC50) concentration was determined for each drug, compared to untreated and DMSO-treated(media) cells (FIG. 1 and FIG. 2, compound A5 is N-(2,4-dimethoxyphenyl)-4-(2-methylimidazo[1,2-a]pyrimidin-3-yl)thiazol-2-amine). A reduction in 3D culture size indicates antitumor activity (FIG. 1). As previously demonstrated, Al 5 demonstrated antitumor activity and was used as a reference compound to compare relative to control and new HS-D1 to HS-D5 series. HS-D1 is a photoaffinity probe with a structural mimic of A 15. Similarly, HS-D5 is a control photoaffinity probe with a distinct chemical structure and had limited antitumor activity. The compounds HS-D1 to HS-D4 had measurable antitumor activity (FIG. 1 and FIG. 2), and while HS-D1 to HS-D3 had modest antitumor effects with ICso at IpM concentration, and ICeo at lOpM concentration (FIG. 2), HS-D4 had more potent effects with IC50 at ~5pM concentration (FIG. 2). These data demonstrated the ability of HS-D1 to HS-D4 compounds to kill GBM 3D patient derived cells.[000158] Compound HS-D4 had relatively higher potency than the other compounds. To examine whether the antitumor activity of HS-D1 to HS-D4 is associated with reduced translation of oncogenic BMI1 RNA, the levels of translated protein within 72 hours of treatment were examined. HS-D2 and HS-D3 reduced BMI1 protein levels at IpM and 5pM, respectively (FIG. 3). Notably, HS-D4 reduced BMI1 protein levels in a dose-dependent fashion with ~ 50% reduction of BMI1 protein levels when utilizing HS-D4 at ~5pM concentration (FIG.4 and FIG. 5 expression of BMI1 protein after drug treatment for 48 hours), the same IC50 dose that resulted in antitumor activity in these GBP patient derived cells, suggesting that the antitumor activities of HS-D4 could be due to reduced oncogenic BMI1 translation.[000159] Surprisingly, when compared to the effects of A15, HS-D1 to HS-D4 reduced BMI1 protein levels without the increased expression of the mitosis protein phosphohistone H3 (PHH3) associated with the effects of A15 (FIG. 6). Notably, HS-D4 reduced BMI1 protein levels, and this reduction was associated with reduced PDL1, CDK4, a downstream target of BMI1 transcriptional repressor activity though ubiquitinated y-H2A, which was also reduced (FIG. 7). The specificity of these effects was shown to be similar to a genetic knockout of BMI1 in the same cells (FIG. 7).[000160] Regulation of translation is a key cellular response to oncogenic stress. This disclosure establishes physicochemical principles that direct the binding of small molecules to a 5' UTR target with broader implications in showing the effects of upstream sequences on translation and interaction with small molecules to guide developing more molecular probes of oncogenes. Inhibiting BMI1 eliminates tumor cell self-renewal and inhibits tumor cell proliferation. With the novel compounds disclosed herein, we present a previously unknown function in modulating immune related targets (PD-L1), therefore affecting immune responses within the tumor microenvironment. BMI1 regulates the expression of PD-L1 through theepigenetic machinery, and thus the novel inhibitors of BMI1 translation presented herein could in certain embodiments serve as a potential therapeutic agents alone or in combination with anti-PD-L1 immunotherapy.[000161] This written description uses examples to disclose the invention, including the best mode, and also to enable any person skilled in the art to make and use the invention. The patentable scope of the invention is defined by the claims, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal language of the claims.[000162] All cited patents, patent applications, and other references are incorporated herein by reference in their entirety. However, if a term in the present application contradicts or conflicts with a term in the incorporated reference, the term from the present application takes precedence over the conflicting term from the incorporated reference.[000163] The terms “a” and “an” do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The term “or” means “and / or”.Reference throughout the specification to “one embodiment”, “another embodiment”, “an embodiment”, and so forth, means that a particular element (e.g., feature, structure, and / or characteristic) described in connection with the embodiment is included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it is to be understood that the described elements may be combined in any suitable manner in the various embodiments. The modifier “about” used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context (e.g., includes the degree of error associated with measurement of the particular quantity).[000164] The endpoints of all ranges directed to the same component or property are inclusive of the endpoints, are independently combinable, and include all intermediate points and ranges (e.g., ranges of “up to about 25 wt.%, or, more specifically, about 5 wt.% to about 20 wt.%,” is inclusive of the endpoints and all intermediate values of the ranges of “about 5 wt.% to about 25 wt.%,” such as about 10 wt% to about 23 wt%, etc.).

Claims

1. CLAIMS1. A compound according to Formula (I), or a pharmaceutically acceptable salt thereof:whereineach R1independently is Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce alkoxy carbonyl, C2-C6 alkanoyl, C2-C6 haloalkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, amino, mono- or di¬R2,C1-C4 alkylamino, halogen, hydroxy, nitro,or R3wherein each R2and R3independently is hydrogen Ci-Ce alkyl, Ci-Ce haloalkyl, or OR7wherein R7is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C8 cycloalkyl, C3-C8cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0);wherein when R1is Ci-Ce alkyl or Ci-Ce haloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or Ci-Cshaloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Cehaloalkyl forms a diazirine ring; each R4independently is C1-C6alkyl, Ci-Ce alkoxy, Ci-Ce alkoxycarbonyl, C2-C6 alkanoyl, C2-C6 alkanoyloxy, cyano, Ci-Cehaloalkyl, C1-C6 haloalkoxy, amino, mono- or di- C1-C4 alkylamino, halogen, hydroxy, or nitro;R5is Ci-Cs alkyl substituted with R6, cyano, C2-C6 alkenyl, C2-C6 alkynyl, or a heterocycloalkyl, wherein R6is an azido group, cyano, OR7, SR7, SO2R7, NR7R7, or heterocycloalkyl, wherein each R7individually is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl;Rsis hydrogen or C1-C4 alkyl;R9is hydrogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;n is 1, 2, 3, or 4;m is 0, 1, 2, or 3;q is 0, 1, 2, or 3; andZ is CH or N.

2. The compound of claim 1, wherein R4is C1-C3 alkyl, C1-C3 alkoxy or fluoro; R8and R9are each hydrogen; Z is CH; n is 1 or 2; and q is 0 or 1.

3. The compound of claim 1, wherein R4is C1-C3 alkyl, C1-C3alkoxy or fluoro; R8and R9are each hydrogen; Z is CH; n is 1; and q is 0.

4. The compound of claim 1, wherein n is 1 or 2; and each R1independently is C1-C3 alkyl, Ci-C3 alkoxy, C1-C3 alkoxycarbonyl, C2-C4 alkanoyl, C2-C4 haloalkanoyl, C2-C4 alkanoyloxy, cyano, C1-C3 haloalkyl, C1-C3 haloalkoxy, amino, mono- or di- C1-C2 alkylamino, halogen, hydroxy,R2,nitro,or wherein each R2and R3independently is hydrogen C1-C3 alkyl, Ci- C3 haloalkyl, or OR7wherein R7is hydrogen, C1-C3 alkyl optionally with at least one nonhydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C6 cycloalkyl, C3-C6 cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0);wherein when R1is C1-C3 alkyl or C1-C3 haloalkyl, each is optionally substituted with one or two substituents each individually selected from hydroxy, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkyl, or C1-C3 haloalkyl, or a carbon of the Ci-Ce alkyl or Ci-Ce haloalkyl forms a diazirine ring; and m is 0 or 1.R2,5. The compound of claim 1, whereinR1is wherein each R2and R3independently is hydrogen C1-C3 alkyl, C1-C3 haloalkyl, or OR7wherein R7is hydrogen, C1-C3alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl, with the proviso that R2and R3cannot both be hydrogen; or R2and R3combine to form a C3-C6 cycloalkyl, C3-C6cycloalkenyl, or heterocycloalkyl; or R2and R3combine to form an oxo (i.e., =0); and m is 0 or 1.

6. The compound of claim 1, wherein R5is Ci-Cs alkyl substituted with R6, cyano, C2-C6 alkenyl, C2-C6 alkynyl, or a heterocycloalkyl, wherein R6is an azido group, cyano, OR7, SR7, SO2R7, NR7R7, or heterocycloalkyl, wherein each R7individually is hydrogen, Ci-Ce alkyl optionally with at least one non-hydrogen atom, heterocycloalkyl, or a C2-C4 alkanoyl.

7. The compound of claim 1, wherein R5is C1-C4alkyl substituted with R6, wherein R6is an azido group, cyano, hydroxy, or NHAc; or R5is C2-C4alkynyl comprising a terminal alkyne.

8. The compound of claim 1, wherein the compound is9. A pharmaceutical formulation comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

10. A method of treating cancer in a subject, comprising: administering an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, to the subject in need of treatment.

11. The method of claim 10, wherein the cancer is non-small cell lung cancer, breast cancer, prostate cancer, pancreatic cancer, head and neck cancer, medulloblastoma, glioblastoma, bile duct carcinoma, neuroblastoma, colon cancer, myeloma, gastric cancer, liver cancer, ovarian cancer, colorectal cancer, non-Hodgkin lymphoma, small cell lung cancer, large cell lung cancer, kidney cancer, esophageal cancer, stomach cancer, cervical cancer, sarcomas, leukemia, or a lymphoma tumor.

12. The method of claim 10, wherein the cancer is glioblastoma.

13. A method for inhibiting BMI1 expression in a subject in need thereof, comprising: administering to the subject an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof.

14. A method comprising:providing an mRNA sequence;identifying a secondary structure of the mRNA sequence, wherein the secondary structure corresponds to a pocket within a 5’UTR;generating highly probable models of the pocket secondary structure, wherein the model accounts for a target minimum and maximum free energy;altering at least one functional group of a small molecule to conform the molecule to a subset of the highly probably models of the pocket to create a targeted small molecule;synthesizing the targeted small molecule;validating the 5'UTR binding of the targeted small molecule.

15. The method of claim 14, wherein the 5'UTR sequence is an oncogene sequence.

16. The method of claim 14, wherein the small molecule is a compound of claim 1 or a compound in Table 2.

17. The method of claim 14, wherein the altered functional group is at least one selected from the group of azido, alkyne, alkene, amino, hydroxyl, and thiol.

18. The method of claim 14, wherein the pocket is one selected from the group consisting of groove fold structure, sphere -like, disc-like, and rod-like.

19. The method of claim 14, wherein when generating the highly probably models of the pocket secondary structure, the model accounts for at least one selected from the group consisting of pocket volume, surface area, effective radius, sphericity, and pocket centroid.

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