Small molecules for degrading DOT1L and uses thereof

Abstract

The present invention relates to bifunctional compounds, compositions, and methods for treating diseases or conditions mediated by aberrant disruptors of telomere silencing 1-like (DOT1L) activity.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 256,899, filed October 18, 2021, which is incorporated by reference in its entirety.

[0002] Government Licensing Rights This invention was made with Government support under Grant No. R01 CA176745 awarded by the National Institutes of Health. The Government has certain rights in this invention. [Background technology]

[0003] E3 ubiquitin ligases are proteins that, in combination with E2 ubiquitin-conjugating enzymes, facilitate the attachment of ubiquitin to lysines on target proteins via isopeptide bonds (e.g., amide bonds that are not present in the protein backbone). Ubiquitination of target proteins leads to degradation of the target protein by the proteasome.

[0004] There remains a need to identify compounds that effectively promote the degradation of target proteins (e.g., disruptor of telomeric silencing 1-like (DOT1L)) that have been found to be associated with certain pathological conditions, including proliferative diseases and cancer. DOT1L has been found to be associated with certain pathological conditions, including proliferative diseases and cancer. In particular, compounds that can exploit cellular mechanisms involved in protein homeostasis (e.g., ubiquitination and proteasomal degradation) to target the degradation of certain proteins can find use as therapeutic agents. There is also a need for compounds that target both the target protein DOT1L and E3 ubiquitin ligases, thereby inducing proteasomal degradation of DOT1L. Summary of the Invention [Means for solving the problem]

[0005] The first aspect of the present invention is an uninterrupted divalent C4-C 20 1. A bifunctional compound comprising moieties that bind to a telomere silencing 1-like disruptor (DOT1L) and a degron, covalently linked to each other by a linker comprising an alkylene chain or a polyethylene glycol (PEG) chain comprising 2 to 8 PEG units, the compound having formula (I): [ka] [In formula: R1 represents H, halogen, CH3, CH2F, CF2H, CF3, CN, or NH2; R2 is [ka] wherein said degron represents a ligand that binds to cereblon (CRBN), or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0006] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bifunctional compound of formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof, and a pharma- ceutically acceptable carrier.

[0007] Another aspect of the invention relates to a method for preparing a bifunctional compound.

[0008] A further aspect of the present invention is directed to a method of treating a disease or disorder characterized by or mediated by aberrant DOT1L activity, comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0009] In some embodiments, the disease or disorder is cancer, hi some embodiments, the cancer is a hematological cancer.

[0010] In some embodiments, the hematological cancer is multiple myeloma, lymphoma, or leukemia.

[0011] In some embodiments, the leukemia is acute myeloid leukemia, mixed lineage leukemia (MLL)-rearranged acute myeloid leukemia, acute myeloid leukemia with a mutation in nucleophosmin 1 (NPM1), acute myeloid leukemia with a mutation in DNA methyltransferase 3A (DNMT3A), or acute myeloid leukemia.

[0012] The bifunctional compounds of the present invention may serve as a new set of chemical tools for PRMT5 knockdown and provide effective treatment of PRMT5-mediated diseases and disorders, such as cancer (e.g., multiple myeloma, lymphoma, and leukemia). [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a plot of a fluorescence polarization (FP) displacement assay of telomere silencing 1-like disruptor (DOT1L) with bifunctional compounds 2, 3, and 6 of the present invention, as well as EPZ5676 and pomalidomide (Pom).

[0014] [Diagram 2] FIG. 2 is a plot of a cellular CRBN dimerization assay using bifunctional compounds 2, 3, and 6 of the invention, as well as EPZ5676 and pomalidomide (Pom).

[0015] [Diagram 3] 3A-3E are a series of plots of cell proliferation assays in EOL1 cells using bifunctional compounds of the present invention 2 (FIG. 3A), 3 (FIG. 3B), 4 (FIG. 3C), 5 (FIG. 3D), and 6 (FIG. 3E).

[0016] [Figure 4]4A-4B are Western blots (FIG. 4A) and cell proliferation assay plots (FIG. 4B) showing DOT1L degradation in EOL1 cells using bifunctional compound 2 of the present invention.

[0017] [Diagram 5] 5A-5B are Western blots (FIG. 5A) and cell proliferation assay plots (FIG. 5B) showing DOT1L degradation in MV411 cells using bifunctional compound 2 of the present invention.

[0018] [Figure 6] 6A-6B are Western blots (FIG. 6A) and cell proliferation assay plots (FIG. 6B) showing DOT1L degradation in EOL1 cells using bifunctional compound 3 of the present invention.

[0019] [Figure 7] FIG. 7 is a plot of a cell proliferation assay in EOL1 cells using bifunctional compound 4.

[0020] [Figure 8] 8A-8B are Western blots (FIG. 8A) and cell proliferation assay plots (FIG. 8B) showing DOT1L degradation in EOL1 cells using bifunctional compound 5 of the present invention.

[0021] [Figure 9] 9A-9B are Western blots (FIG. 9A) and cell proliferation assay plots (FIG. 9B) showing DOT1L degradation in EOL1 cells using bifunctional compound 6 of the present invention.

[0022] [Figure 10] 10A-10B are Western blots (FIG. 10A) and cell proliferation assay plots (FIG. 10B) showing DOT1L degradation in MV411 cells using bifunctional compound 6 of the present invention.

[0023] [Figure 11] FIG. 11 is a plot of a cell proliferation assay in SEMK2 cells using bifunctional compound 6 of the present invention.

[0024] [Figure 12] FIG. 12 is a plot of a cell proliferation assay in THP1 cells using bifunctional compound 6 of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of this specification belongs. As used in this specification and the appended claims, unless specified to the contrary, the following terms have the meanings set forth to facilitate understanding of the invention.

[0026] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "composition" includes mixtures of two or more such compositions, reference to an "inhibitor" includes mixtures of two or more such inhibitors, and so forth.

[0027] Unless otherwise specified, the term "about" means within 10% (eg, within 5%, 2%, or 1%) of the particular value modified by the term "about."

[0028] The transitional term "comprising" is synonymous with "including," "containing," or "characterized by" and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes elements, steps, or ingredients not specified in the claim. The transitional phrase "consisting essentially of" limits the claim to certain materials or steps, "and which do not materially affect the basic and novel characteristics" of the claimed invention.

[0029] To the extent that the following terms are used herein with respect to and to further describe the compounds of the present invention, the following definitions apply.

[0030] As used herein, the term "alkyl" refers to a saturated linear or branched monovalent hydrocarbon radical. In one embodiment, the alkyl radical is a C1-C 18 In other embodiments, the alkyl radical is a C0-C6, C0-C5, C0-C3, C1-C 12 , C1-C8, C1-C6, C1-C5, C1-C4 or C1-C3 groups (CO alkyl refers to a bond). Examples of alkyl groups include methyl, ethyl, 1-propyl, 2-propyl, i-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, heptyl, octyl, nonyl, decyl, undecyl and dodecyl. In some embodiments, the alkyl group is a C1 to C3 alkyl group.

[0031] As used herein, the term "alkylene" refers to a straight or branched divalent hydrocarbon chain that links the remainder of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and having 1 to 12 carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, etc. The alkylene chain may be attached to the remainder of the molecule through a single bond or to the radical group through a single bond. In some embodiments, the alkylene group contains 1 to 8 carbon atoms (C1-C8 alkylene). In other embodiments, the alkylene group contains 1 to 5 carbon atoms (C1-C5 alkylene). In other embodiments, the alkylene group contains 1 to 4 carbon atoms (C1-C4 alkylene). In other embodiments, the alkylene group contains 1 to 3 carbon atoms (C1-C3 alkylene). In other embodiments, the alkylene group contains 1 to 2 carbon atoms (C1-C2 alkylene). In other embodiments, the alkylene group contains 1 carbon atom (C1 alkylene).

[0032] As used herein, the term "alkenyl" refers to a straight or branched chain monovalent hydrocarbon radical having at least one carbon-carbon double bond. Alkenyl includes radicals having "cis" and "trans" orientations, or alternatively, "E" and "Z" orientations. In one example, an alkenyl radical is a C2-C 18 In another embodiment, the alkenyl radical is a C2-C 12 , C2~C 10 , C2-C8, C2-C6 or C2-C3 groups. Examples include ethenyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl and hex-1,3-dienyl.

[0033] The term "alkoxyl" or "alkoxy" as used herein refers to an alkyl group, as defined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy, and the like. An "ether" is two hydrocarbyl groups covalently linked by an oxygen. Thus, the substituent of an alkyl that makes it an ether is an alkoxyl or resembles an alkoxyl, and can be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-alkynyl.

[0034] As used herein, the term "alkoxylenes" refers to compounds having the general formula (-OC), where n represents an integer (e.g., 1, 2, 3, 4, 5, 6, or 7). n H 2n "alkoxylene" refers to a saturated monovalent aliphatic radical of the formula (-), including both straight and branched chain radicals. The alkoxylene chain may be attached to the remainder of the molecule through a single bond or to the radical group through a single bond. In some embodiments, the alkoxylene group contains 1 to 3 carbon atoms (-O-C1-C3 alkoxylene). In other embodiments, the alkoxylene group contains 1 to 5 carbon atoms (-O-C1-C5 alkoxylene).

[0035] As used herein, the term "cyclic group," used alone or as part of a larger moiety, broadly refers to any group including saturated, partially saturated, or aromatic ring systems, such as carbocyclic (cycloalkyl, cycloalkenyl), heterocyclic (heterocycloalkyl, heterocycloalkenyl), aryl, and heteroaryl groups. A cyclic group can have one or more (e.g., fused) ring systems. Thus, for example, a cyclic group can contain one or more carbocyclic, heterocyclic, aryl, or heteroaryl groups.

[0036] As used herein, the term "carbocycle" (also "carbocyclyl"), used alone or as part of a larger moiety, refers to a group that is alone or part of a larger moiety (e.g., an alkyl carbocyclic group) containing saturated, partially unsaturated, or aromatic ring systems having from 3 to 20 carbon atoms. The term carbocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged, and spiro ring systems, and combinations thereof. In one embodiment, carbocyclyl refers to a ring system containing from 3 to 15 carbon atoms (C3-C4). 15 In one embodiment, the carbocyclyl contains 3 to 12 carbon atoms (C 12 In another embodiment, carbocyclyl is C3-C8, C3-C 10 Or C5~C 10 In another embodiment, the carbocyclyl, as a monocyclic ring, includes C3-C8, C3-C6, or C5-C6. In some embodiments, the carbocyclyl, as a bicyclic ring, includes C7-C 12 In another embodiment, the carbocyclyl, as a spiro system, is 12Representative examples of monocyclic carbocyclyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, perdeuteriocyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, phenyl, and cyclododecyl. Bicyclic carbocyclyls having 7 to 12 ring atoms include [4,3], [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems, such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, naphthalene, and bicyclo[3.2.2]nonane. Representative examples of spirocarbocyclyl include spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane and spiro[4.5]decane. The term carbocyclyl includes aryl ring systems as defined herein. The term carbocyclyl also includes cycloalkyl rings (e.g., saturated or partially unsaturated monocyclic, bicyclic, or spirocarbocycles). The term carbocyclic group also includes carbocyclic rings fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., aryl or heterocycles), where the radical or point of attachment is on the carbocyclic ring.

[0037] Thus, the term carbocycle as used herein means R c is an alkylene chain; c The term carbocyclyl, as used herein, also encompasses carbocyclylalkyl groups, which refer to the group R c is an alkylene chain of the formula --O--R c Also included are carbocyclylalkoxy groups, which refer to groups attached through the oxygen atom of a -carbocyclyl.

[0038] As used herein, the terms "aryl" (e.g., "aralkyl," where a terminal carbon atom on the alkyl group is the point of attachment, e.g., a benzyl group), "aralkoxy" where an oxygen atom is the point of attachment, or "aroxyalkyl" where the point of attachment is on an aryl group, used alone or as part of a larger moiety, refer to groups that contain monocyclic, bicyclic, or tricyclic, carbocyclic ring systems, including fused rings, in which at least one ring in the system is aromatic. In some embodiments, an aralkoxy group is a benzoxy group. The term "aryl" may be used interchangeably with the term "aryl ring." In one embodiment, aryl includes groups having 6 to 18 carbon atoms. In another embodiment, aryl includes groups having 6 to 10 carbon atoms. Examples of aryl groups include phenyl, naphthyl, anthracyl, biphenyl, phenanthrenyl, naphthacenyl, 1,2,3,4-tetrahydronaphthalenyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, naphthyridinyl, and the like, which may be substituted or independently substituted with one or more substituents described herein. A particular aryl is phenyl. In some embodiments, an aryl group comprises an aryl ring fused to one or more (e.g., one, two, or three) different cyclic groups (e.g., carbocyclic or heterocyclic), where the radical or point of attachment is on the aryl ring. The structure of any aryl group that may have differently positioned double bonds is considered to encompass any and all such resonance structures.

[0039] Thus, the term aryl refers to R c is an alkylene chain such as methylene or ethylene; c -aryl, which refers to a group of aryl, includes aralkyl groups (e.g., benzyl). In some embodiments, the aralkyl group is an optionally substituted benzyl group. The term aryl, as used herein, refers to a group of aryl, such as aryl, which is an optionally substituted benzyl group. c is an alkylene such as methylene or ethylene; c Also included is aralkoxy, which refers to a group bonded through the oxygen atom of an --aryl.

[0040] As used herein, the term "heterocyclyl" refers to "carbocyclyl" used alone or as part of a larger moiety and includes saturated, partially unsaturated, or aromatic ring systems in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced with a heteroatom (e.g., O, N, N(O), S, S(O), or S(O)2). The term heterocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged, and spiro ring systems, and combinations thereof. In some embodiments, heterocyclyl refers to a 3- to 15-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a 3- to 12-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a saturated ring system, such as a 3- to 12-membered saturated heterocyclyl ring system. In some embodiments, heterocyclyl refers to a heteroaryl ring system, such as a 5- to 14-membered heteroaryl ring system. The term heterocyclyl also includes C3-C8 heterocycloalkyl, which are saturated or partially unsaturated monocyclic, bicyclic or spirocyclic ring systems containing 3 to 8 carbons and one or more (1, 2, 3 or 4) heteroatoms.

[0041] In some embodiments, the heterocyclyl group contains 3-12 ring atoms, including monocyclic, bicyclic, tricyclic, and spirocyclic systems, where the ring atom is carbon and 1-5 ring atoms are heteroatoms such as nitrogen, sulfur, or oxygen. In some embodiments, the heterocyclyl contains a 3- to 7-membered monocyclic ring having one or more heteroatoms selected from nitrogen, sulfur, and oxygen. In some embodiments, the heterocyclyl contains a 4- to 6-membered monocyclic ring having one or more heteroatoms selected from nitrogen, sulfur, and oxygen. In some embodiments, the heterocyclyl contains a 3-membered monocyclic ring. In some embodiments, the heterocyclyl contains a 4-membered monocyclic ring. In some embodiments, the heterocyclyl contains a 5- to 6-membered monocyclic ring. In some embodiments, the heterocyclyl group contains 0-3 double bonds. In any of the foregoing embodiments, the heterocyclyl contains 1, 2, 3, or 4 heteroatoms. Any nitrogen or sulfur heteroatom may be optionally oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatom may be optionally quaternized (e.g., [NR4] + Cl - , [NR4] + OH -Representative examples of heterocyclyl include oxiranyl, aziridinyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydropyranyl, dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, and the like. thioxanyl, thiazinyl, thioxanyl, homopiperazinyl, homopiperidinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinyl Dinonyl, oxazolidinonyl, imidazolidinonyl, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazole[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, thiophenyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl yl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiapyranyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinonyl, pyrimidinedionyl, pyrimidine-2,4-dionyl, piperazinonyl, piperazinedionyl, pyrazolidinylimidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.2.1]octanyl, 8-azabicyclo[3.2.1]octanyl, 2-azabicyclo[2.2.2]octanyl, 8-azabicyclo[2.2.2]octanyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl, 1-azaspiro[4.5]decane-2-only, azaspiro[5.5]undecanyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocyclyls containing a sulfur or oxygen atom and 1 to 3 nitrogen atoms are thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl, such as oxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocyclyls containing 2 to 4 nitrogen atoms include imidazolyls such as imidazol-2-yl; triazolyls such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and tetrazolyls such as 1H-tetrazol-5-yl. Representative examples of benzo-fused 5-membered heterocyclyls are benzoxazol-2-yl, benzthiazol-2-yl and benzimidazol-2-yl. Examples of 6-membered heterocyclyls contain 1 to 3 nitrogen atoms and optionally sulfur or oxygen atoms, for example pyridyl, such as pyrid-2-yl, pyrid-3-yl and pyrid-4-yl; pyrimidyl, such as pyrimid-2-yl and pyrimid-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, especially pyridazin-3-yl, and pyrazinyl. Pyridine N-oxide and pyridazine N-oxide and pyridyl, pyrimid-2-yl, pyrimid-4-yl, pyridazinyl and 1,3,4-triazin-2-yl groups are further examples of heterocyclyl groups. In some embodiments, a heterocyclic group comprises a heterocycle fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocycles or heterocycles), where the radical or point of attachment is on the heterocycle, and in some embodiments, the point of attachment is a heteroatom contained in the heterocycle.

[0042] Thus, the term heterocycle, as used herein, includes N-heterocyclyl groups, which refer to heterocyclyl groups containing at least one nitrogen, where the attachment point of the heterocyclyl group to the rest of the molecule is through a nitrogen atom in the heterocyclyl group. Representative examples of N-heterocyclyl groups include 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl and imidazolidinyl. The term heterocycle, as used herein, also includes C-heterocyclyl groups, which refer to heterocyclyl groups containing at least one heteroatom, where the attachment point of the heterocyclyl group to the rest of the molecule is through a carbon atom in the heterocyclyl group. Representative examples of C-heterocyclyl radicals include 2-morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, and 2- or 3-pyrrolidinyl. The term heterocycle, as disclosed above, refers to R c is an alkylene chain; c The term heterocycle as used herein also encompasses heterocyclylalkyl groups, which refer to the group R -heterocyclyl. c is an alkylene chain of the formula --O--R c Also included are heterocyclylalkoxy groups, which refer to radicals attached through the oxygen atom of a -heterocyclyl.

[0043] As used herein, the term "heteroaryl," used alone or as part of a larger moiety (e.g., "heteroarylalkyl" (also "heteroaralkyl") or "heteroarylalkoxy" (also "heteroaralkoxy"), refers to a monocyclic, bicyclic, or tricyclic ring system having 5 to 14 ring atoms, in which at least one ring is aromatic and contains at least one heteroatom. In one embodiment, heteroaryl includes 5-6 membered monocyclic aromatic groups in which one or more ring atoms are nitrogen, sulfur, or oxygen. Representative examples of heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, imidazopyridine, cyclohexyl ... The heteroaryl groups include diphenyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolo[1,5-b]pyridazinyl, purinyl, deazapurinyl, benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, indolyl, 1,3-thiazol-2-yl, 1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl, 1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl, 1,2,3-triazol-5-yl, and pyrid-2-yl N-oxide. The term "heteroaryl" also includes groups in which a heteroaryl is fused to one or more cyclic (e.g., carbocyclyl or heterocyclyl) rings, where the radical or point of attachment is on the heteroaryl ring.Non-limiting examples include indolyl, indolizinyl, isoindolyl, benzothienyl, benzothiophenyl, methylenedioxyphenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzodioxazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or tricyclic. In some embodiments, a heteroaryl group comprises a heteroaryl ring fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocyclic or heterocyclic rings), where the radical or point of attachment is on the heteroaryl ring, and in some embodiments, the point of attachment is a heteroatom contained in the heterocyclic ring. The structure of any heteroaryl group that can have differently positioned double bonds is considered to encompass all such resonance structures.

[0044] Thus, the term heteroaryl, as used herein, encompasses N-heteroaryl groups, which refer to heteroaryl groups as defined above that contain at least one nitrogen, and the point of attachment of the heteroaryl group to the remainder of the molecule is through a nitrogen atom in the heteroaryl group. The term heteroaryl, as used herein, also encompasses C-heteroaryl groups, which refer to heteroaryl groups as defined above that the point of attachment of the heteroaryl group to the remainder of the molecule is through a carbon atom in the heteroaryl group. The term heteroaryl also encompasses R, as disclosed above. c is an alkylene chain as defined above; c The term heteroaryl as used herein also includes heteroarylalkyl groups, which refer to the group R c is an alkylene group as defined above;c Further included are heteroaralkoxy (or heteroarylalkoxy) groups, which refer to groups attached through the oxygen atom of a -heteroaryl.

[0045] Unless otherwise stated and unless further defined for a particular group, any of the groups described herein may be substituted or unsubstituted. As used herein, the term "substituted" refers broadly to all permissible substituents, with the implicit proviso that such substitution is consistent with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, and the like. Representative substituents include halogens, hydroxyl groups, and any other organic group containing any number of carbon atoms, e.g., 1-14 carbon atoms, which may include one or more (e.g., 1, 2, 3, or 4) heteroatoms, such as oxygen, sulfur, and nitrogen, grouped in the form of a straight chain, branched chain, or cyclic structure.

[0046] Thus, unless otherwise disclosed for any particular group, representative examples of substituents include alkyl, substituted alkyl (e.g., C1-C6, C1-C 5、 C1~C 4、 C1~C 3、 C1~C 2、 C1), alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), substituted alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), haloalkyl (e.g., CF3), alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), cyclic (e.g., C3-C 12 , C5-C6), substituted cyclic (e.g., C3-C 12 , C5-C6), carbocyclic (e.g., C3-C12 , C5-C6), substituted carbocyclic rings (e.g., C3-C 12 , C5-C6), heterocycles (e.g., C3-C 12 , C5-C6), substituted heterocycles (e.g., C3-C 12 , C5-C6), aryl (e.g., benzyl and phenyl), substituted aryl (e.g., substituted benzyl or phenyl), heteroaryl (e.g., pyridyl or pyrimidyl), substituted heteroaryl (e.g., substituted pyridyl or pyrimidyl), aralkyl (e.g., benzyl), substituted aralkyl (e.g., substituted benzyl), halo, hydroxyl, aryloxy (e.g., C6-C 12 , C6), substituted aryloxy (e.g., C6-C 12 , C6), alkylthio (e.g., C1-C6), substituted alkylthio (e.g., C1-C6), arylthio (e.g., C6-C 12 , C6), substituted arylthio (e.g., C6-C 12 , C6), cyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, thio, substituted thio, sulfinyl, substituted sulfinyl, sulfonyl, substituted sulfonyl, sulfinamide, substituted sulfinamide, sulfonamide, substituted sulfonamide, urea, substituted urea, carbamate, substituted carbamate, amino acid, and peptide groups.

[0047] The term "binding", when referring to the interaction between a targeting ligand and a targeted protein, which in the present invention is disruptor of telomere silencing 1-like (DOT1L), typically refers to a preferential (also referred to herein as "selective") intermolecular interaction in that binding of the targeting ligand to other proteins present in the cell is substantially less and may not be functionally significant. The terms "selective" and "selectivity" refer to the ability of a bifunctional compound to discriminate between and among molecular targets.

[0048] The term "binding" with respect to the interaction between a degron and an E3 ubiquitin ligase typically refers to an intermolecular interaction that may or may not exhibit an affinity level equal to or exceeding the affinity between the targeting ligand and the target protein, but is nevertheless sufficient to achieve recruitment of the ligase to the target protein, which in this disclosure is DOT1L, for selective degradation.

[0049] Generally, bifunctional compounds are uninterrupted divalent C4-C 20 The compound comprises a moiety that binds to DOT1L and a degron covalently linked to each other by a linker comprising an alkylene chain or a polyethylene glycol (PEG) chain containing 2 to 8 PEG units, wherein the compound has formula (I): [ka] [In formula: R1 represents H, halogen, CH3, CH2F, CF2H, CF3, CN, or NH2; R2 is [ka] represents; and degron represents a ligand that binds to cereblon (CRBN)], or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0050] In some embodiments, R1 is H or CN.

[0051] In some embodiments, the bifunctional compound of the present invention has the structure: [ka] or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0052] Linker The linker ("L") provides a covalent bond between the DOT1L binding moiety and the degron. The structure of the linker may not be important so long as it does not substantially interfere with the activity of the DOT1L binding moiety or the degron. In some embodiments, the linker comprises an alkylene chain (e.g., having 2-20 alkylene units). In other embodiments, the linker is -O-, -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, - N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, - S(O)2O-, -N(R')S(O)2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, C3~C 12 It comprises an alkylene chain which may be interrupted and / or terminated (at either or both termini) by at least one of a carbocyclene, a 3- to 12-membered heterocyclene, a 5- to 12-membered heteroarylene, or any combination thereof, R' is H or a C1-C6 alkyl, and the interrupting group and either or both terminating groups may be the same or different.

[0053] In some embodiments, the linker is a C 20 It may contain an alkylene chain.

[0054] In some embodiments, the linker is a C1-C 12 It may contain an alkylene chain, and the nitrogen is also attached to the degron.

[0055] In some embodiments, the linker is [ka] and / or [ka] The alkylene chain includes an alkylene chain having 1 to 10 alkylene units that terminates with.

[0056] In some embodiments, the alkylene linker contains 11 to 15 uninterrupted alkylene units.

[0057] "Carbocyclene" refers to an optionally substituted divalent carbocycle radical.

[0058] "Heterocyclene" refers to a divalent heterocyclyl radical which may be optionally substituted.

[0059] "Heteroarylene" refers to a divalent heteroaryl radical which may be optionally substituted.

[0060] Representative examples of alkylene linkers that may be suitable for use in the present invention include the following: [ka] [In the formula, n is an integer of 1 to 12 ("of" means inclusive), for example, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, 9-10 and 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10], examples include: [ka] Alkylene chains terminated with various functional groups (as above), examples of which are: [ka] Alkylene chains interrupted by various functional groups (as above), examples of which are: [ka] An alkylene chain interrupted or terminated by a heterocyclene group, e.g. [ka] [wherein m and n are independently integers from 0 to 10], examples of which include the following: [ka] Alkylene chains interrupted by amide, heterocyclene and / or aryl groups, examples of which include: [ka] Heterocyclenes, aryl groups, and alkylene chains interrupted by heteroatoms, examples of which include: [ka] and Alkylene chains interrupted by heteroatoms such as N, O or B, e.g. [ka] wherein each n is independently an integer from 1 to 10, e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, 9-10, and 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and R is H or C1-C4 alkyl, examples of which are: [ka] It is.

[0061] In some embodiments, the linker is -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2- , -S(O)2O-, -N(R')S(O)2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, C 3-12 It may comprise a polyethylene glycol chain which may be terminated (at one or both termini) with at least one of a carbocyclene, a 3- to 12-membered heterocyclene, a 5- to 12-membered heteroarylene, or any combination thereof, where R' is H or a C1-C6 alkyl, and one or both termini may be the same or different.

[0062] In some embodiments, the linker comprises a polyethylene glycol chain having 2-8 PEG units. [ka] One or both ends are terminated with

[0063] In some embodiments, the linker comprises three uninterrupted PEG units.

[0064] Representative examples of linkers comprising polyethylene glycol chains include: [ka] [wherein n is an integer from 2 to 10], examples of which include the following: [ka]

[0065] In some embodiments, the polyethylene glycol linker may terminate in a functional group, examples of which are as follows: [ka] .

[0066] In some embodiments, the linker further comprises: [ka] [ka] or [ka] In some embodiments, the linker comprises a [ka] Contains a group.

[0067] In some embodiments, the linker has the following structure: [ka] It is represented by one of the following:

[0068] Thus, in some embodiments, the bifunctional compounds of the present invention have the structures (I-5) to (I-28): [ka] [ka] [ka] [ka] [ka] [ka] or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0069] Degron The ubiquitin-proteasome pathway (UPP) is a crucial cellular pathway that controls key regulator proteins and degrades misfolded or abnormal proteins. The UPP is central to multiple cellular processes. Covalent attachment of ubiquitin to specific protein substrates is achieved by the action of E3 ubiquitin ligases. These ligases include over 500 different proteins and are divided into multiple classes defined by the structural elements of their E3 functional activity.

[0070] Degrons bind to the E3 ligase cereblon (CRBN).

[0071] Representative examples of such degrons include structures (D1a)-(D1d): [ka] [ka] [wherein X1 is CH2 or C(O), and X2 is a bond, CH2, NH, or O].

[0072] Still other degrons that bind cereblon and may be suitable for use in the present invention are disclosed in U.S. Pat. No. 9,770,512, as well as U.S. Patent Application Publication Nos. 2018 / 0015087, 2018 / 0009779, 2016 / 0243247, 2016 / 0235731, 2016 / 0235730, and 2016 / 0176916, and International Patent Publication Nos. WO2017 / 197055, WO2017 / 197051, WO2017 / 197036, WO2017 / 197056, and WO2017 / 197046.

[0073] Thus, in some embodiments, the bifunctional compounds of the present invention have the structures (I-29) to (I-44): [ka] [ka] [ka] [ka] [ka] or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0074] In some embodiments, the bifunctional compound of the present invention has the following structure: [ka] [ka] [ka] or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0075] The bifunctional compounds of formula (I) may be in the form of a free acid or free base, or a pharma- ceutically acceptable salt. As used herein, the term "pharma-ceutically acceptable" in the context of a salt refers to a relatively non-toxic salt of a compound that does not abolish the biological activity or properties of the compound. That is, the salt form of the compound may be administered to a subject without causing undesirable biological effects (such as dizziness or stomach upset) or interacting in a harmful manner with any of the other components of the composition in which it is included. The term "pharma-ceutically acceptable salt" refers to the product obtained by reacting a compound of the present invention with a suitable acid or base. Examples of pharma-ceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic bases, such as Li, Na, K, Ca, Mg, Fe, Cu, Al, Zn, and Mn salts. Examples of pharma- ceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, 4-methylbenzenesulfonate or p-toluenesulfonate, etc. Certain compounds of the present invention can form pharma- ceutically acceptable salts with various organic bases, such as lysine, arginine, guanidine, diethanolamine or metformin.

[0076] The bifunctional compounds of formula (I) may have at least one chiral center and therefore, as used herein, may be in the form of stereoisomers, encompassing all isomers of the individual compounds that differ only in the orientation of their atoms in space. The term stereoisomer includes enantiomers (enantiomers containing the (R-) or (S-) configuration of the compound), mixtures of enantiomers of the compounds (physical mixtures of enantiomers, and racemates or racemic mixtures), geometric (cis / trans or E / Z, R / S) isomers of the compounds, and isomers of the compounds with multiple chiral centers that are not mirror images of each other (diastereoisomers). The chiral centers of the compounds may undergo epimerization in vivo. Thus, in these compounds, administration of the compound in the (R-) form is considered equivalent to administration of the compound in the (S-) form. Thus, the compounds of the present invention may be made and used in the form of a single isomer, substantially free of other isomers, or in the form of mixtures of various isomers, such as racemic mixtures of stereoisomers.

[0077] In some embodiments, the bifunctional compound of formula (I) is an isotopic derivative in that it has at least one desired isotopic substitution of an atom at a level greater than the natural abundance of the isotope, i.e., enriched. In one embodiment, the compound contains deuterium or multiple deuterium atoms. Deuterium, i.e. 2 Substitution with heavier isotopes, such as H, may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be advantageous in some circumstances.

[0078] Additionally, the bifunctional compounds of formula (I) encompass N-oxides of the compounds, crystalline forms (also known as polymorphs), active metabolites of the compounds having the same type of activity, tautomers, and unsolvated and solvated forms with pharma- ceutically acceptable solvents such as water, ethanol, etc. Solvated forms of the bifunctional compounds are encompassed by this term.

[0079] Synthesis method In some embodiments, the present invention relates to a method for preparing a bifunctional compound of formula (I) or its pharma- ceutically acceptable salt or stereoisomer. Generally, the compounds of the present invention or its pharma- ceutically acceptable salt or stereoisomer can be prepared by any process known to be applicable to the preparation of chemically related compounds. The compounds of the present invention will be better understood in relation to the synthetic schemes described in the various examples, which show non-limiting methods by which the compounds of the present invention can be prepared.

[0080] Pharmaceutical Compositions Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bifunctional compound of formula I, or a pharma- ceutically acceptable salt or stereoisomer thereof, and a pharma- ceutically acceptable carrier. The term "pharma- ceutically acceptable carrier" as known in the art refers to a pharma- ceutically acceptable material, composition, or vehicle suitable for administering the compounds of the present invention to a mammal. Suitable carriers may include, for example, liquids (both aqueous and non-aqueous, and combinations thereof), solids, encapsulating materials, gases, and combinations thereof (e.g., semi-solids), and gases, which function to carry or transport the compound from one organ or part of the body to another organ or part of the body. A carrier is "acceptable" in the sense of being physiologically inert, compatible with the other ingredients of the formulation, and not harmful to the subject or patient. Depending on the type of formulation, the composition may include one or more pharma- ceutically acceptable excipients.

[0081] In general, the bifunctional compounds of formula (I) and their pharma- ceutically acceptable salts and stereoisomers can be formulated into a given type of composition according to conventional pharmaceutical practices, such as conventional mixing, dissolving, granulating, dragee-making, wet-milling, emulsifying, encapsulating, entrapping and compression processes (see, for example, Remington: The Science and Practice of Pharmacy (20th ed.), ed. A. R. Gennaro, Lippincott Williams & Wilkins, 2000, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York). The type of formulation depends on the mode of administration, which may include enteral (e.g., oral, buccal, sublingual and rectal), parenteral (e.g., subcutaneous (sc), intravenous (iv), intramuscular (im)), and intrasternal injection, or infusion techniques, intraocular, intraarterial, intramedullary, intrathecal, intracerebroventricular, transdermal, intradermal, intravaginal, intraperitoneal, mucosal, nasal, intratracheal instillation, bronchial instillation, and inhalation), and topical (e.g., transdermal). In general, the most appropriate administration route will depend on various factors, including, for example, the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration). For example, parenteral (e.g., intravenous) administration may also be advantageous in that the compound may be administered relatively quickly, such as in the case of single-dose treatments and / or acute conditions.

[0082] In some embodiments, the bifunctional compounds are formulated for oral or intravenous administration (eg, systemic intravenous injection).

[0083] Thus, the bifunctional compounds of the present invention can be formulated into solid compositions (e.g., powders, tablets, dispersible granules, capsules, cachets, and suppositories), liquid compositions (e.g., solutions in which the compound is dissolved, suspensions in which solid particles of the compound are dispersed, emulsions, and solutions containing liposomes, micelles, or nanoparticles, syrups, and elixirs), semi-solid compositions (e.g., gels, suspensions, and creams), and gases (e.g., propellants for aerosol compositions). The compounds can also be formulated for rapid, intermediate, or sustained release.

[0084] Oral solid dosage forms include capsules, tablets, pills, powders and granules.In such solid dosage forms, the active compound is mixed with a carrier such as sodium citrate or dicalcium phosphate, and additional carriers or excipients, such as a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) wetting agents such as glycerol, d) crosslinked polymers (e.g., crosslinked polyvinylpyrrolidone (crospovidone), crosslinked carbo Disintegrating agents such as sodium hydroxymethylcellulose (croscarmellose sodium), sodium starch glycolate, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarders such as paraffin, f) absorption promoters such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents. Solid compositions of a similar type may also be used as fillers for soft and hard filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings. They may further contain opacifying agents.

[0085] In some embodiments, the bifunctional compound of the present invention can be formulated into hard or soft gelatin capsules.Representative excipients that can be used include pregelatinized starch, magnesium stearate, mannitol, sodium stearyl fumarate, anhydrous lactose, microcrystalline cellulose and croscarmellose sodium.Gelatin shells can include gelatin, titanium dioxide, iron oxide and coloring agents.

[0086] Liquid dosage forms for oral administration include solutions, suspensions, emulsions, microemulsions, syrups and elixirs.In addition to the compound, liquid dosage forms can contain aqueous or non-aqueous carriers (depending on the solubility of the compound) commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.Oral compositions can also contain excipients, such as wetting agents, suspending agents, coloring agents, sweeteners, flavoring agents and aromatic agents.

[0087] Injectable preparations may include sterile aqueous or oily suspensions. They may be formulated according to standard techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions or emulsions in non-toxic parenterally acceptable diluents or solvents, for example, solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspending medium. For this purpose, any mild fixed oil may be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. Injectable preparations may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. The effect of the compound may be prolonged by delaying its absorption, which may be achieved by the use of liquid suspensions with poor water solubility or crystalline or amorphous materials. Prolonged absorption of a compound from a parenterally administered formulation may also be accomplished by suspending the compound in an oil vehicle.

[0088] In certain embodiments, the bifunctional compounds of formula (I) can be administered locally rather than systemically, for example, by injecting the conjugate directly into an organ, often in a depot or sustained release preparation. In certain embodiments, long-acting formulations are administered by infusion (e.g., subcutaneously or intramuscularly) or intramuscular injection. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). The release rate of the compound can be controlled by varying the ratio of compound to polymer and the nature of the particular polymer used. Depot injectable formulations are also prepared by encapsulating the compound in liposomes or microemulsions that are compatible with body tissues. Furthermore, in other embodiments, the compound is delivered in a targeted drug delivery system, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to and taken up selectively by the organ.

[0089] The bifunctional compounds can be formulated for buccal or sublingual administration and examples include tablets, lozenges and gels.

[0090] The bifunctional compound may be formulated for administration by inhalation. Various forms suitable for administration by inhalation include aerosol, mist or powder. The pharmaceutical composition may be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In some embodiments, the dosage unit of the pressurized aerosol may be determined by providing a valve to deliver a metered amount. In some embodiments, capsules and cartridges, for example, comprising gelatin for use in an inhaler or insufflator, may be formulated to contain a powder mixture of the compound and a suitable powder base, for example, lactose or starch.

[0091] The bifunctional compound of formula (I) can be formulated for topical administration, which refers to intradermal administration by applying the formulation to the epidermis as used herein.These types of compositions are typically in the form of ointments, pastes, creams, lotions, gels, solutions and sprays.

[0092] Representative examples of carriers useful for formulating compositions for topical application include solvents (e.g., alcohol, polyalcohol, water), creams, lotions, ointments, oils, plasters, liposomes, powders, emulsions, microemulsions, and buffer solutions (e.g., hypotonic or buffered saline).For example, creams can be formulated using saturated or unsaturated fatty acids, such as stearic acid, palmitic acid, oleic acid, palmitoleic acid, cetyl or oleyl alcohol.Cream may also contain non-ionic surfactants, such as polyoxy-40-stearate.

[0093] In some embodiments, topical formulations can also include excipients, examples of which are penetration enhancers. These agents can transport pharmacologically active compounds through the stratum corneum, preferably to the epidermis or dermis with little or no systemic absorption. A wide variety of compounds have been evaluated for their effectiveness in enhancing the penetration rate of drugs through the skin. For example, see Percutaneous Penetration Enhancers, Maibach HI and Smith HE (eds.), CRC Press, Inc., Boca Raton, Fla. (1995), which reviews the use and testing of various skin penetration enhancers, and Buyuktimkin et al., Chemical Means of Transdermal Drug Permeation Enhancement in Transdermal and Topical Drug Delivery Systems, Gosh TK, Pfister WR, Yum SI (Eds.), Interpharm Press Inc., Buffalo Grove, Ill. (1997). Representative examples of penetration enhancers include triglycerides (e.g., soybean oil), aloe compositions (e.g., aloe vera gel), ethyl alcohol, isopropyl alcohol, octriphenyl polyethylene glycol, oleic acid, polyethylene glycol 400, propylene glycol, N-decyl methyl sulfoxide, fatty acid esters (e.g., isopropyl myristate, methyl laurate, glycerol monooleate and propylene glycol monooleate), and N-methylpyrrolidone.

[0094] Representative examples of further excipients that may be included in topical and other types of formulations (to the extent that they are compatible) include preservatives, antioxidants, moisturizers, emollients, buffers, solubilizers, skin protectants, and surfactants. Suitable preservatives include alcohols, quaternary amines, organic acids, parabens, and phenols. Suitable antioxidants include ascorbic acid and its esters, sodium bisulfite, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, and chelating agents such as EDTA and citric acid. Suitable moisturizers include glycerin, sorbitol, polyethylene glycol, urea, and propylene glycol. Suitable buffers include citrate buffer, hydrochloric acid buffer, and lactate buffer. Suitable solubilizers include quaternary ammonium chloride, cyclodextrin, benzyl benzoate, lecithin, and polysorbates. Suitable skin protectants include vitamin E oil, allatoin, dimethicone, glycerin, petrolatum, and zinc oxide.

[0095] Transdermal formulations typically use transdermal delivery devices and transdermal delivery patches, where the compound is formulated in a lipophilic emulsion or buffered aqueous solution, dissolved and / or dispersed in a polymer or adhesive. Patches can be constructed for continuous, pulsatile, or on-demand delivery of pharmaceuticals. Transdermal delivery of compounds can be achieved by iontophoretic patches. Transdermal patches can provide controlled delivery of compounds, where the absorption rate is slowed by using a rate-controlling membrane or by trapping the compound within a polymer matrix or gel. Absorption enhancers can be used to increase absorption, examples of which include pharma- ceutically acceptable absorbent solvents that aid passage through the skin.

[0096] Ophthalmic preparations include eye drops.

[0097] Formulations for rectal administration include enemas, rectal gels, rectal foams, rectal aerosols, and retention enemas, which may contain conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinylpyrrolidone, PEG, etc. Compositions for rectal or vaginal administration may also be formulated as suppositories, which can be prepared by mixing the compound with suitable non-irritating carriers and excipients, such as cocoa butter, mixtures of fatty acid glycerides, polyethylene glycol, suppository waxes, and combinations thereof, all of which are solid at ambient temperature but liquid at body temperature, and therefore will melt in the rectum or vaginal cavity and release the compound.

[0098] Dosage As used herein, the term "therapeutically effective amount" refers to an amount of a bifunctional compound of formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof; or a composition comprising a bifunctional compound of formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof, effective to bring about a desired therapeutic response in a particular patient suffering from a disease or disorder characterized by or mediated by abnormal DOT1L activity. Thus, the term "therapeutically effective amount" includes an amount of a bifunctional compound of the present invention or a pharma- ceutically acceptable salt or stereoisomer thereof, when administered, that is sufficient to induce a positive degeneration of the disease or disorder being treated, or inhibit or even prevent the onset or progression of the disease or disorder, or to alleviate to some extent one or more symptoms of the disease or disorder being treated in a subject, or simply kill or inhibit the growth of diseased (e.g., cancer (e.g., blood cancer)) cells, or reduce the amount of DOT1L in diseased cells.

[0099] The total daily dosage of the bifunctional compound and its usage can be determined in accordance with standard medical practice, for example by the attending physician using sound medical judgment. The specific therapeutically effective dose for any particular subject may depend on a variety of factors, including the disease or disorder being treated and its severity (e.g., its current condition); the age, weight, general health, sex and diet of the subject; the time of administration, route of administration and excretion rate of the particular compound used; duration of treatment; drugs used in combination or simultaneously with the bifunctional compound; and similar factors known in the medical arts (see, e.g., Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 10th Edition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press, 155-173, 2001).

[0100] The bifunctional compounds of formula (I) and their pharma- ceutically acceptable salts and stereoisomers may be effective over a wide dosage range. In some embodiments, the total daily dosage (e.g., for adults) may range from about 0.001 to about 1600 mg, 0.01 to about 1600 mg, 0.01 to about 500 mg, about 0.01 to about 100 mg, about 0.5 to about 100 mg, 1 to about 100 to about 400 mg per day, about 1 to about 50 mg per day, and about 5 to about 40 mg per day, and in other embodiments, may range from about 10 to about 30 mg per day. Individual doses may be formulated to contain the desired dosage depending on the number of times the compound is administered per day. By way of example, capsules may be formulated with about 1 to about 200 mg of the bifunctional compound (e.g., 1, 2, 2.5, 3, 4, 5, 10, 15, 20, 25, 50, 100, 150, and 200 mg). In some embodiments, individual doses may be formulated to contain a desired dosage depending on the number of times the compound is to be administered per day.

[0101] How to use In some aspects, the present invention relates to a method for treating a disease or disorder associated with abnormal (e.g., dysfunctional or dysregulated) DOT1L activity, the method involving administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof.

[0102] A disease or disorder is characterized by or mediated by abnormal DOT1L activity (e.g., DOT1L that is elevated or otherwise functionally abnormal compared to non-pathological conditions). A "disease" is generally considered to be a state of health of a subject in which the subject is unable to maintain homeostasis and the subject's health will continue to deteriorate unless the disease is ameliorated. In contrast, a "disorder" in a subject is a state of health in which the subject is able to maintain homeostasis, but the subject's health is less favorable than if the disorder does not exist. If left untreated, the disorder does not necessarily cause further deterioration of the animal's health.

[0103] The term "subject" (or "patient") as used herein includes all members of the animal kingdom susceptible to or afflicted with the indicated disease or disorder. In some embodiments, the subject is a mammal, such as a human or a non-human mammal. The method is also applicable to companion animals, such as dogs and cats, as well as domestic animals, such as cows, horses, sheep, goats, pigs, and other farm animals, and wild animals. A subject "in need" of treatment according to the present invention may be "suffering or suspected of suffering from" a particular disease or disorder, may have been positively diagnosed, or otherwise exhibits a sufficient number of risk factors, or a sufficient number or combination of signs or symptoms, such that a medical professional may diagnose or suspect that the subject suffers from the disease or disorder. Thus, subjects suffering from a particular disease or disorder and subjects suspected of suffering from it are not necessarily two distinct populations.

[0104] In some embodiments, the bifunctional compounds of formula (I) may be useful in the treatment of cell proliferative diseases and disorders (e.g., cancer or benign neoplasms). As used herein, the term "cell proliferative disease or disorder" refers to conditions characterized by unregulated or abnormal cell proliferation or both, including non-cancerous conditions such as neoplasms, pre-cancerous conditions, benign tumors, and cancers.

[0105] In other embodiments, the method relates to treating a subject with cancer.In general, the bifunctional compounds of the present invention can be effective in treating carcinomas (solid tumors, including both primary and metastatic tumors), sarcomas, melanomas, and hematological cancers (cancers that affect the blood, including lymphocytes, bone marrow, and / or lymph nodes), such as leukemia, lymphoma, and multiple myeloma.Included are adult tumors / cancers and pediatric tumors / cancers.Cancers can be vascularized or not yet substantially vascularized or non-vascularized tumors.

[0106] Representative examples of cancer include adrenal cortical carcinoma, AIDS-related cancers (e.g., Kaposi's and AIDS-related lymphomas), appendix cancer, childhood cancers (e.g., childhood cerebellar astrocytoma, childhood cerebral astrocytoma), basal cell carcinoma, skin cancer (non-melanoma), bile duct cancer, extrahepatic bile duct cancer, intrahepatic bile duct cancer, bladder cancer, urinary bladder cancer, brain cancer (e.g., gliomas and glioblastomas, e.g., brain stem glioma, gestational trophoblastic tumor glioma, cerebellar astrocytoma, cerebral astrocytoma / malignant tumor glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma), breast cancer, bronchial adenoma / carcinoid, carcinoid tumor, nervous system cancer (e.g., central nervous system cancer, central nervous system lymphoma), cervical cancer, chronic bone marrow cancer, Myeloproliferative disorders, colorectal cancer (e.g., colon cancer, rectal cancer), lymphoid neoplasms, mycosis fungoides, Sezary syndrome, endometrial cancer, esophageal cancer, extracranial germ cell tumors, extragonadal germ cell tumors, extrahepatic bile duct cancer, eye cancer, intraocular melanoma, retinoblastoma, gallbladder cancer, gastrointestinal cancer (e.g., gastric cancer, small intestine cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST)), bile duct cancer, germ cell tumors, ovarian germ cell tumors, head and neck cancer, neuroendocrine tumors, Hodgkin's lymphoma, AnnArbor Class III and Class IV pediatric non-Hodgkin's lymphoma, ROS1-positive refractory non-Hodgkin's lymphoma, leukemia, lymphoma, multiple myeloma, hypopharyngeal carcinoma, intraocular melanoma, eye cancer, pancreatic islet cell tumors (endocrine pancreas), kidney cancer (e.g., Wilms' tumor, renal cell carcinoma), liver cancer, lung cancer (e.g., non-small cell lung cancer and large cell lung cancer), ALK-positive anaplastic large cell lymphoma, ALK-positive advanced malignant solid neoplasms, Waldenström's macroglobulinemia, melanoma, intraocular (eye) melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck of unknown primary site, multiple endocrine neoplasia (MEN), myelodysplastic syndromes, myelodysplastic / myeloproliferative disorders, nasopharyngeal carcinoma, neuroblastoma, oral cancer (e.g., oral cancer, lip cancer, oral cancer) , tongue cancer, oropharyngeal cancer, throat cancer, pharyngeal cancer), ovarian cancer (e.g., ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor), pancreatic cancer, islet cell pancreatic cancer, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineoblastoma, metastatic anaplastic thyroid cancer, anaplastic thyroid cancer, papillary thyroid cancer, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleural embryonal tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, uterine cancer (e.g., endometrial cancer, uterine sarcoma, endometrial cancer), squamous cell carcinoma, testicular cancer, thymoma, thymic carcinoma, thyroid cancer, juvenile xanthogranuloma, transitional cell carcinoma of the renal pelvis and ureter and other urinary organs, urethral cancer, gestational trophoblastic tumor, vaginal cancer, vulvar cancer, hepatoblastoma, rod body tumor, and Wilms' tumor.

[0107] Sarcomas that may be treatable with the bifunctional compounds of the invention include both soft tissue and bone cancers as well, representative examples of which include osteosarcoma or osteogenic sarcoma (bone) (e.g., Ewing's sarcoma), chondrosarcoma (cartilage), leiomyosarcoma (smooth muscle), rhabdomyosarcoma (skeletal muscle), mesothelioma or mesothelioma (membranous lining of body cavities), fibrosarcoma (fibrous tissue), angiosarcoma or hemangioendothelioma (blood vessels), liposarcoma (fatty tissue), glioma or astrocytoma (neurogenic connective tissue found in the brain), myxosarcoma (primitive embryonic connective tissue), mesenchymal or mixed mesodermal tumor (mixed connective tissue types), and histiocytic sarcoma (immune cancer).

[0108] In some embodiments, the methods of the invention involve treatment of a subject having a cell proliferative disease or disorder of the blood system, liver, brain, lung, colon, pancreas, prostate, ovaries, breast, skin, and endometrium.

[0109] As used herein, "cell proliferative diseases or disorders of the blood system" includes lymphoma, leukemia, myeloid neoplasms, mast cell neoplasms, myelodysplasia, benign monoclonal gammopathy, lymphomatoid papulosis, polycythemia vera, primary myelofibrosis, and essential thrombocythemia. Thus, representative examples of hematological cancers include multiple myeloma, lymphomas (T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma (diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL) and ALK+ anaplastic large cell lymphoma (e.g., B-cell non-Hodgkin's lymphoma selected from diffuse large B-cell lymphoma (e.g., germinal center B-cell-like diffuse large B-cell lymphoma or activated B-cell-like diffuse large B-cell lymphoma), Burkitt's lymphoma / leukemia, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, follicular lymphoma, ALK+ anaplastic large cell lymphoma (ALK+ anaplastic large cell lymphoma), ... and relapsed B-cell non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas of lymphocytic and cutaneous origin, e.g., small lymphocytic lymphoma, childhood leukemia, hairy cell leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute myeloid leukemia (e.g., acute monocytic leukemia), chronic lymphocytic leukemia, small lymphocytic leukemia, chronic myeloid leukemia, chronic myelogenous leukemia, and leukemias, myeloid neoplasms and mast cell neoplasms, including leukemia, marginal zone lymphoma, lymphoplasmacytic lymphoma / Waldenstrom's macroglobulinemia, metastatic pancreatic adenocarcinoma, refractory B-cell non-Hodgkin's lymphoma, and relapsed B-cell non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas of lymphocytic and cutaneous origin, e.g., small lymphocytic lymphoma, childhood leukemia, hairy cell leukemia, acute lymphocytic leukemia, acute myeloid leukemia (e.g., acute monocytic leukemia), chronic lymphocytic leukemia, small lymphocytic leukemia, chronic myeloid leukemia, and mast cell leukemia.

[0110] In some embodiments, the hematological cancer is multiple myeloma, lymphoma, or leukemia. In some embodiments, the leukemia is acute myeloid leukemia, mixed lineage leukemia (MLL) rearrangement acute myeloid leukemia, acute myeloid leukemia with a mutation in nucleophosmin 1 (NPM1), acute myeloid leukemia with a mutation in DNA methyltransferase 3A (DNMT3A), or acute myeloid eosinophilic leukemia.

[0111] As used herein, "hepatic cell proliferation disease or disorder" includes all forms of cell proliferation disorder that affect the liver.Hepatic cell proliferation disorder can include liver cancer (e.g., hepatocellular carcinoma, intrahepatic cholangiocarcinoma and hepatoblastoma), precancerous or precancerous conditions of the liver, benign growth or lesion of the liver, and malignant growth or lesion of the liver, as well as metastatic lesions in tissues and organs in the body other than the liver.Hepatic cell proliferation disorder can include liver hyperplasia, metaplasia, and dysplasia.

[0112] As used herein, "brain cell proliferation disease or disorder" includes all forms of cell proliferation disorder that affect the brain.Brain cell proliferation disorder can include brain cancer (e.g., glioma, glioblastoma, meningioma, pituitary adenoma, vestibular schwannoma, and primitive neuroectodermal tumor (medulloblastoma)), brain precancerous or precancerous condition, brain benign growth or lesion, and brain malignant growth or lesion, and metastatic lesion in body tissue and organ other than brain.Brain cell proliferation disorder can include brain hyperplasia, metaplasia, and dysplasia.

[0113] As used herein, "cell proliferative disease or disorder of the lung" includes all forms of cell proliferative disorders that affect germ cells. Cell proliferative disorders of the lung include lung cancer, precancerous and precancerous conditions of the lung, benign growths or lesions of the lung, hyperplasia, metaplasia, and dysplasia of the lung, and metastatic lesions in tissues and organs of the body other than the lung. Lung cancer includes all forms of cancer of the lung, such as malignant lung neoplasms, carcinoma in situ, typical carcinoid tumors, and atypical carcinoid tumors. Lung cancer includes small cell lung cancer ("SLCL"), non-small cell lung cancer ("NSCLC"), adenocarcinoma, small cell carcinoma, large cell carcinoma, squamous cell carcinoma, and mesothelioma. Lung cancer can include "scar carcinoma", bronchioveolar carcinoma, giant cell carcinoma, spindle cell carcinoma, and large cell neuroendocrine carcinoma. Lung cancer also includes lung neoplasms with histological and ultrastructural heterogeneity (e.g., mixed cell types).In some embodiments, the compounds of the present invention can be used to treat non-metastatic or metastatic lung cancer (e.g., NSCLC, ALK-positive NSCLC, NSCLC with ROS1 fusion gene, lung adenocarcinoma, and squamous cell lung cancer).

[0114] As used herein, "cell proliferative diseases or disorders of the colon" include all forms of cell proliferative disorders affecting colon cells, including colon cancer, precancerous or precancerous conditions of the colon, adenomatous polyps of the colon, and metachronous lesions of the colon. Colon cancer includes sporadic and hereditary colon cancer, malignant colon neoplasms, carcinoma in situ, typical carcinoid tumors, and atypical carcinoid tumors, adenocarcinoma, squamous cell carcinoma, and squamous cell carcinoma. Colon cancer may be associated with genetic syndromes, such as hereditary nonpolyposis colorectal cancer, familial adenomatous polyposis, MYH-associated polyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndrome, and juvenile polyposis. Cell proliferative disorders of the colon may also be characterized by colonic hyperplasia, metaplasia, or dysplasia.

[0115] As used herein, "cell proliferation disease or disorder of pancreas" includes all forms of cell proliferation disorder that affect pancreatic cells.Cell proliferation disorder of pancreas can include pancreatic cancer, precancerous or precancerous conditions of pancreas, pancreatic hyperplasia, pancreatic dysplasia, benign growth or lesion of pancreas, and malignant growth or lesion of pancreas, as well as metastatic lesions in tissues and organs of the body other than pancreas.Pancreatic cancer includes all forms of cancer of pancreas, including ductal adenocarcinoma, adenosquamous carcinoma, pleomorphic giant cell carcinoma, mucinous adenocarcinoma, osteoclast-like giant cell carcinoma, mucinous cystadenocarcinoma, acinar carcinoma, unclassified large cell carcinoma, small cell carcinoma, pancreatic blastoma, papillary neoplasm, mucinous cystadenoma, papillary cystic neoplasm, and serous cystadenoma, as well as pancreatic neoplasms with histological and ultrastructural heterogeneity (e.g., mixed cell).

[0116] As used herein, "prostate cell proliferation disease or disorder" includes all forms of cell proliferation disorder that affect the prostate.Prostate cell proliferation disorder can include prostate cancer, prostate precancerous or precancerous conditions, prostate benign growth or lesion, prostate malignant growth or lesion, and metastatic lesion in body tissues and organs other than the prostate.Prostate cell proliferation disorder can include prostate hyperplasia, metaplasia, and dysplasia.

[0117] As used herein, "cell proliferative disease or disorder of the ovary" includes all forms of cell proliferative disorders that affect cells of the ovary. Cell proliferative disorders of the ovary may include precancerous or precancerous conditions of the ovary, benign growths or lesions of the ovary, ovarian cancer, and metastatic lesions in tissues and organs of the body other than the ovary. Cell proliferative disorders of the ovary may include ovarian hyperplasia, metaplasia, and dysplasia.

[0118] As used herein, "cell proliferative disease or disorder of the breast" includes all forms of cell proliferative disorders affecting breast cells. Cell proliferative disorders of the breast can include breast cancer, precancerous or precancerous conditions of the breast, benign growths or lesions of the breast, and metastatic lesions in body tissues and organs other than the breast. Cell proliferative disorders of the breast can include hyperplasia, metaplasia, and dysplasia of the breast.

[0119] As used herein, "cell proliferative diseases or disorders of the skin" include all forms of cell proliferative disorders affecting skin cells. Cell proliferative disorders of the skin may include precancerous or precancerous conditions of the skin, benign growths or lesions of the skin, melanoma, malignant melanoma or other malignant growths or lesions of the skin, and metastatic lesions in tissues and organs of the body other than the skin. Cell proliferative disorders of the skin may include hyperplasia, metaplasia, and dysplasia of the skin.

[0120] As used herein, "endometrial cell proliferative disease or disorder" includes all forms of cell proliferative disorder that affect endometrial cells.Endometrial cell proliferative disorder can include endometrial precancerous or precancerous conditions, endometrial benign growths or lesions, endometrial cancer, and metastatic lesions in body tissues and organs other than endometrium.Endometrial cell proliferative disorder can include endometrial hyperplasia, metaplasia, and dysplasia.

[0121] The bifunctional compounds of formula (I) may be administered to patients, such as cancer patients, as monotherapy or in combination therapy. The therapy may be "front / first line" as an initial treatment for patients who have not previously undergone an anticancer treatment regimen, either alone or in combination with other treatments; or "second line" as a treatment for patients who have previously undergone an anticancer treatment regimen, either alone or in combination with other treatments; or "third line", "fourth line", etc., treatment, either alone or in combination with other treatments. The therapy may also be administered to patients who have previously undergone unsuccessful or partially successful treatments, except for patients who have become intolerant to a particular treatment. The therapy may also be administered as an adjuvant treatment, i.e., to prevent recurrence of cancer in patients who currently have no detectable disease or after surgical removal of a tumor. Thus, in some embodiments, the bifunctional compounds may be administered to patients who have undergone another therapy, such as chemotherapy, radioimmunotherapy, surgical therapy, immunotherapy, radiation therapy, targeted therapy, or any combination thereof.

[0122] The method of the present invention can involve administration of a bifunctional compound of formula I or a pharmaceutical composition thereof to a patient in a single dose or multiple doses (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 10, 15, 20 or more doses). For example, the frequency of administration can range from once a day to approximately once every 8 weeks. In some embodiments, the frequency of administration varies from about once a day for 1, 2, 3, 4, 5, or 6 weeks, and in other embodiments involves a 28-day cycle that includes daily administration for 3 weeks (21 days) followed by a 7-day "off" period. In other embodiments, the bifunctional compound can be administered twice a day (BID) over a period of 2 1 / 2 days (a total of 5 doses), or once a day (QD) over a period of 2 days (a total of 2 doses). In other embodiments, the bifunctional compound can be administered once a day (QD) over a period of 5 days.

[0123] Combination therapy The bifunctional compounds of formula (I) and their pharma- ceutically acceptable salts and stereoisomers can be used in combination or simultaneously with at least one other active agent, such as an anti-cancer agent or regimen, in the treatment of diseases and disorders. In this context, the terms "in combination" and "concurrently" mean that the agents are administered simultaneously, including substantially simultaneous administration, by the same or separate dosage forms, by the same or separate modes of administration, or sequentially, such as as part of the same treatment regimen, or by consecutive treatment regimens. Thus, when given sequentially, at the start of administration of the second compound, the first of the two compounds is still detectable at effective concentrations at the treatment site, as the case may be. The order and time intervals can be determined so that they can act together (e.g., synergistically) to provide a greater benefit than if administered otherwise. For example, the therapeutic agents can be administered simultaneously or at different times in any order sequentially. However, if not administered simultaneously, they can be administered close enough in time to provide the desired therapeutic effect, which may be in a synergistic form. Thus, these terms are not limited to administering the active agents exactly at the same time.

[0124] In some embodiments, the treatment regimen may include administration of the bifunctional compound of formula (I) in combination with one or more additional therapeutic agents known for use in treating a disease or condition (e.g., cancer). The dosage of the additional anti-cancer therapeutic agent may be the same as or lower than the known or recommended dose. See Hardman et al., eds., Goodman & Gilman's The Pharmacological Basis Of Basis Of Therapeutics, 10th ed., McGraw-Hill, New York, 2001; Physician's Desk Reference 60th ed., 2006. For example, anti-cancer agents that may be suitable for use in combination with the bifunctional compounds of the present invention are known in the art. See, for example, U.S. Pat. No. 9,101,622 (section 5.2 therein) and U.S. Pat. No. 9,345,705 B2 (columns 12-18 therein). Representative examples of additional active agents and treatment regimens include radiation therapy, chemotherapeutic agents (e.g., antimitotic agents, angiogenesis inhibitors, antihormones, autophagy inhibitors, alkylating agents, intercalating antibiotics, growth factor inhibitors, antiandrogens, signal transduction pathway inhibitors, microtubule inhibitors, platinum coordination complexes, HDAC inhibitors, proteasome inhibitors, and topoisomerase inhibitors), immunomodulatory agents, therapeutic antibodies (e.g., monospecific and bifunctional antibodies), and CAR-T therapy.

[0125] In some embodiments, the bifunctional compound of Formula (I) and the additional (e.g., anticancer) therapeutic agent are administered at intervals of less than 5 minutes, less than 30 minutes, less than 1 hour, about 1 hour, about 1 to about 2 hours, about 2 to about 3 hours, about 3 to about 4 hours, about 4 to about 5 hours, about 5 to about 6 hours, about 6 to about 7 hours, about 7 to about 8 hours, about 8 to about 9 hours, about 9 hours, or about 10 hours. The therapeutic agents may be administered at intervals of about 10 to about 10 hours, at intervals of about 10 to about 11 hours, at intervals of about 11 to about 12 hours, at intervals of about 12 to 18 hours, at intervals of 18 to 24 hours, at intervals of 24 to 36 hours, at intervals of 36 to 48 hours, at intervals of 48 to 52 hours, at intervals of 52 to 60 hours, at intervals of 60 to 72 hours, at intervals of 72 to 84 hours, at intervals of 84 to 96 hours, or at intervals of 96 to 120 hours. Two or more (e.g., anti-cancer) therapeutic agents may be administered during a single patient visit.

[0126] In some embodiments involving cancer therapy, the bifunctional compound of formula (I) and the additional anti-cancer or therapeutic agent are administered cyclically. Cyclic therapy includes administering one anti-cancer therapeutic agent for a period of time, followed by administering a second anti-cancer therapeutic agent for a period of time, and repeating this sequential administration, i.e., cycle, to reduce the occurrence of resistance to one or both of the anti-cancer therapeutic agents, to avoid or reduce the side effects of one or both of the anti-cancer therapeutic agents, and / or to improve the efficacy of the therapy. In one example, cyclic therapy includes administering a first anti-cancer therapeutic agent for a period of time, followed by administering a second anti-cancer therapeutic agent for a period of time, optionally followed by administering a third anti-cancer therapeutic agent for a period of time, etc., and repeating this sequential administration, i.e., cycle, to reduce the occurrence of resistance to one anti-cancer therapeutic agent, to avoid or reduce the side effects of one anti-cancer therapeutic agent, and / or to improve the efficacy of the anti-cancer therapeutic agent.

[0127] Medicine Kit The bifunctional compounds of the present invention and / or compositions comprising them can be assembled into a kit or pharmaceutical system.The kit or pharmaceutical system according to this aspect of the present invention includes a carrier or package such as a box, carton, tube, etc., in which one or more containers, such as a vial, tube, ampoule, or bottle, containing the bifunctional compound of formula (I) or its pharmaceutical composition, are tightly enclosed.The kit or pharmaceutical system of the present invention can also include printed instructions for using the compounds and compositions.

[0128] These and other aspects of the present invention will be further understood in light of the following examples, which are intended to illustrate particular embodiments of the invention, but are not intended to limit its scope, which is defined by the claims. EXAMPLES

[0129] These and other aspects of the present invention will be further understood in light of the following examples, which are intended to illustrate particular embodiments of the invention, but are not intended to limit its scope, which is defined by the claims.

[0130] Example 1: Synthesis of intermediates int-1 to int-5 [ka] (2R,3R,4S,5R)-2-(6-((4-aminobutyl)amino)-9H-purin-9-yl)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)tetrahydrofuran-3,4-diol (int-1)

[0131] To a solution of compound A (20.0 mg, 25.8 μmol, 1.00 equiv) in THF (1 mL) was added 4.0 M HCl in dioxane (1.00 mL) dropwise. The mixture was stirred at room temperature for 30 min, and then methanol (300 μL) was added. The resulting reaction mixture was stirred for 3 h until the starting material was consumed by MS. The crude mixture was concentrated under reduced pressure to give int-1 (16.4 mg, 25.8 μmol, quantitative).

[0132] MS(ESI):C 34 H 51 Calculated N9O3: 633.84, measured: 634.84.

[0133] UPLC-MS RT: 1.32 min (method A), mass m / z: 389.87 [M-tBu+H] + . [ka] 5-((4-((9-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)amino)-5-oxopentanoic acid (int-2)

[0134] To a 0.3 μM solution of int-1 (16.4 mg, 25.8 μmol, 1.00 equiv.) and glutaric anhydride (2.94 mg, 25.8 μmol, 1.00 equiv.) in MeCN was added triethylamine (7.18 μL, 51.6 μmol, 2 equiv.) dropwise. The reaction was stirred at room temperature for 30 min until the starting material was depleted by MS. The resulting crude mixture was concentrated under reduced pressure to give int-2 (16.4 mg, 25.8 μmol, quantitative).

[0135] MS(ESI):C 39 H 57 Calculated value of N9O6: 747.44, measured value: 748.47. [ka] 1-(3-((((2R,3S,4R,5R)-5-(6-((4-aminobutyl)amino)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)(isopropyl)amino)propyl)-3-(4-tertbutyl)phenyl)urea (int-3)

[0136] To a solution of compound B (20.0 mg, 26.6 μmol, 1.00 equiv) in THF (1 mL) was added 4.0 M HCl in dioxane (1.00 mL) dropwise. The mixture was stirred at room temperature for 30 min, and then methanol (300 μL) was added. The resulting reaction mixture was stirred for 3 h until the starting material was consumed by MS. The resulting crude mixture was concentrated under reduced pressure to give int-3 (16.4 mg, 25.8 μmol, quantitative).

[0137] MS(ESI):C 34 H 51 Calculated value of N9O3: 611.39, measured value: 612.40. [ka] 1-(3-((((2R,3R,4S,5R)-5-(4-((4-aminobutyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)(isopropyl)amino)propyl)-3-(4-(tert-butyl)phenyl)urea (int-4)

[0138] Int-4 was prepared in a similar manner to Int-3 above. [ka] 4-((4-aminobutyl)amino)-7-((2R,3S,4R,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (int-5)

[0139] Int-5 was prepared in a similar manner to Int-1 above.

[0140] Example 2: Synthesis of N-(4-((9-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-12-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide)dodecanamide (1). [ka]

[0141] To a 0.3 M solution of int-1 (16.4 mg, 25.8 μmol) and IMiD acid 1 (13.6 mg, 25.8 μmol) in anhydrous dimethylformamide (DMF), triethylamine was added (77.4 μmol, 3.0 equiv.). After stirring for 5 min, hexafluorophosphate azabenzotriazole tetramethyluronium (HATU) (25.8 μmol, 1.0 equiv.) was added. The resulting reaction mixture was stirred at 23° C. for 1 h. Upon completion of the reaction, the resulting crude product was purified by preparative high performance liquid chromatography (prep-HPLC) to give compound 1 as a white powder (5.05 mg, 17.0% yield).

[0142] MS(ESI):C 61 H 84 N 12 O 10Calculated value: 1144.64, actual value: 1145.66.

[0143] Example 3: N-(4-((9-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexadecanamide (2). [ka]

[0144] Compound 2 was synthesized in a manner similar to compound 1 in Example 2 from int-1 (32.8 mg, 51.6 μmol) and the appropriate IMiD acid (27.2 mg, 51.6 μmol) and was isolated as a light yellow powder (25 mg, 42.3%).

[0145] MS(ESI):C 63 H 90 N 12 Calculated value of O8: 1142.70, measured value: 1143.70.

[0146] Example 4: Synthesis of N-(4-((9-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexadecanamide (3). [ka]

[0147] Compound 3 was synthesized in a similar manner to compound 1 in Example 2 from int-1 (8.15 mg, 12.9 μmol) and the appropriate IMiD acid (6.45 mg, 12.9 μmol) and was isolated as a light yellow powder (3.80 mg, 22.2%).

[0148] MS(ESI):C 61 H 86 N 12 Calculated value of O8: 1114.67, measured value: 1115.67.

[0149] Example 5: Synthesis of N1-(4-((9-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-N5-(8-(((R)-1-((2R,4S)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-8-oxooctyl)glutaramide (4). [ka]

[0150] Compound 4 was synthesized from int-2 (9.6 mg, 12.9 μmol) and compound VHL-C7-NH2 (6.45 mg, 12.9 μmol) in a manner similar to compound 1 in Example 2, and was isolated as a white powder (2.95 mg, 17.6%).

[0151] MS(ESI):C 69 H 100 N 14 Calculated O9S value: 1300.75, measured value: 1301.74.

[0152] Example 6: Synthesis of N-(4-((7-((2R,3R,4S,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexadecanamide (5). [ka]

[0153] Compound 5 was synthesized from int-4 (5.00 mg, 8.08 μmol) and the appropriate IMiD acid (4.2 mg, 8.08 μmol) in a manner similar to compound 1 in Example 2 and was isolated as a light yellow powder (1.52 mg, 16.7%).

[0154] MS(ESI):C 63 H 89 N 11 Calculated value of O8: 1127.69, measured value: 1128.70.

[0155] Example 7: Synthesis of N-(4-((9-((2S,3R,4S,5R)-5-(((3-(3-(4-(tert-butyl)phenyl)ureido)propyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexadecanamide (6). [ka]

[0156] Compound 6 was synthesized in a similar manner to compound 1 in Example 2 from int-3 (5.00 mg, 8.08 μmol) and the appropriate IMiD acid (4.2 mg, 8.08 μmol) and was isolated as a light yellow powder (1.52 mg, 16.7%).

[0157] MS(ESI):C 60 H 88 N 12 Calculated value for O9: 1120.68, observed value: 1121.69.

[0158] Example 8: Synthesis of N-(4-((9-((2R,3S,4R,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexadecanamide (7). [ka]

[0159] Compound 7 was synthesized from int-1 and the appropriate IMiD acid in a manner similar to compound 1 in Example 2, and was isolated as a white powder.

[0160] MS(ESI):C 63 H 89 N 11 Calculated value of O9: 1144.47, observed value: 1145.47.

[0161] Example 9: Synthesis of N-(4-((9-((2R,3S,4R,5R)-5-(((3-(3-(4-(tert-butyl)phenyl)ureido)propyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-9H-purin-6-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexadecanamide (8). [ka]

[0162] Compound 8 was synthesized from int-3 and the appropriate IMiD acid in a manner similar to compound 1 in Example 2, and was isolated as a white powder.

[0163] MS(ESI):C 60 H 87 N 11 O 10 Calculated value: 1122.42, actual value: 1123.42.

[0164] Example 10: Synthesis of N-(4-((7-((2R,3S,4R,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexadecanamide (9). [ka]

[0165] Compound 9 was synthesized from int-5 and the appropriate IMiD acid in a manner similar to compound 1 in Example 2, and was isolated as a bright yellow powder.

[0166] MS(ESI):C 65 H 89 N 11 Calculated value for O9: 1167.51, measured value: 1168.52.

[0167] Example 11: Synthesis of N-(4-((7-((2R,3S,4R,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexadecanamide (10). [ka]

[0168] Compound 10 was synthesized from int-5 and the appropriate IMiD acid in a manner similar to compound 1 in Example 2, and was isolated as a white powder.

[0169] MS(ESI):C 65 H 89 N 11 Calculated value for O9: 1168.50, observed value: 1169.51.

[0170] Example 12: Synthesis of N-(4-((7-((2R,3S,4R,5R)-5-(((3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-5-cyano-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)butyl)-16-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexadecanamide (11). [ka]

[0171] Compound 11 was synthesized from int-1 and the appropriate IMiD acid in a manner similar to compound 1 in Example 2, and was isolated as a light yellow powder.

[0172] MS(ESI):C 63 H 90 N 14 Calculated value of O8: 1171.50, measured value: 1172.51.

[0173] Example 13: Fluorescence polarization (FP) displacement assay of cellular telomere silencing 1-like disruptor (DOT1L).

[0174] DOT1L FP displacement assays were performed with bifunctional compounds 2, 3, and 6 of the invention, as well as pomalidomide and the positive control EPZ5676, at different concentrations [M] and measured in [M] per mP.

[0175] The results are shown in Figure 1. They show that bifunctional compounds 2, 3, and 6 of the present invention bound to DOT1L via displacement of the fluorescein isothiocyanate (FITC)-probe in the FP assay.

[0176] Example 14: DOT1L: Cereblon (CRBN)-DNA damage-binding protein 1 (DDB1) / DOT1L dimerization assay.

[0177] Both GST-CRBN-DDB1 and DOT1L (6xHIS tagged) were diluted to 125 nM in assay buffer (50 mM 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethane-1-sulfonic acid (HEPES) pH 7.4, 150 mM NaCl, and 0.01% Tween®2, 0.1% bovine serum albumin (BSA)) and 20 μL of the protein mixture was added to each well of a 384-well AlphaPlate™ (PerkinElmer®). Compounds were then added at 100 nL per well from DMSO stock plates using a Janus® Workstation (PerkinElmer®). After 1 hour incubation at room temperature, 20 μL of Nickel Chelate AlphaLISA® Acceptor and Glutathione AlphaLISA® Donor beads (PerkinElmer®) diluted to 20 ng / μl in assay buffer were added to each well. After 1 hour incubation at room temperature, luminescence was measured on an Envision® 2104 plate reader (PerkinElmer®). Data were analyzed and plotted using GraphPad PRISM v6, and the "Gaussian" analysis module was used to determine the amplitude, mean, and sd values ​​of dimerization.

[0178] Dimerization of DOT1L:CRBN by bifunctional compounds 2, 3, and 6 of the invention, as well as pomalidomide and EPZ5676, was evaluated by the AlphaScreen® assay.

[0179] The results are shown in Figure 2. They show that the bifunctional compounds 2, 3, and 6 of the present invention induced the formation of a ternary complex between DOT1L and CRBN.

[0180] Example 15: Cell proliferation assay in EOL1 (human eosinophilic leukemia) cells with bifunctional compounds of the invention.

[0181] EOL1 cells, 0.25 x 106 Cells were seeded in 6-well plates at a density of cells / well, then treated with compounds at the indicated concentrations and incubated for the indicated times. After staining with trypan blue, cell counts were performed using a Countess® II FL automated cell counter (ThermoFisher Scientific). [ka]

[0182] The results are shown in Figures 3A-3E, which show that bifunctional compounds 2-6 of the present invention reduced EOL1 cell viability in a dose-dependent manner and at early time points.

[0183] The results are shown in Figures 4A-4B. Figure 4A shows that the bifunctional compound 2 of the present invention degraded DOT1L in EOL1 cells in a dose-dependent manner up to 2.5 μM. Figure 4B shows that the bifunctional compound 2 of the present invention reduced EOL1 cell viability in a dose-dependent manner and at early time points.

[0184] The results are shown in Figures 6A-6B. Figure 6A shows that the bifunctional compound 3 of the present invention degraded DOT1L in EOL1 cells in a dose-dependent manner up to 5.0 μM. Figure 6B shows that the bifunctional compound 3 of the present invention reduced EOL1 cell viability in a dose-dependent manner and at early time points.

[0185] The results are shown in Figure 7. They show that the bifunctional compound 4 of the present invention did not decrease EOL1 cell viability in a dose-dependent manner.

[0186] The results are shown in Figures 8A-8B, which show that bifunctional compound 5 of the present invention did not decrease EOL1 cell viability in a dose-dependent manner and at early time points.

[0187] The results are shown in Figures 9A-9B. Figure 9A shows that the bifunctional compound 6 of the present invention degraded DOT1L in EOL1 cells up to 5.0 μM in a dose-dependent manner. Figure 9B shows that the bifunctional compound 6 of the present invention reduced EOL1 cell viability in a dose-dependent manner and at early time points.

[0188] Example 16: Cell proliferation assay in MV411 (human myelomonocytic leukemia) cells with bifunctional compounds of the invention.

[0189] MV411 cells were cultured at 0.25 x 10 in a 6-well plate. 6 Cells were seeded at a density of 1000 cells / well, then treated with compounds at the indicated concentrations and incubated for the indicated times. After staining with trypan blue, cell counts were performed using a Countess® II FL automated cell counter (ThermoFisher Scientific).

[0190] The results are shown in Figures 5A and 5B. Figure 5A shows that bifunctional compound 2 of the present invention degraded DOT1L in MV411 cells in a dose-dependent manner up to 10.0 μM. Figure 5B shows that bifunctional compound 2 of the present invention reduced MV411 cell viability in a dose-dependent manner and at early time points.

[0191] The results are shown in Figures 10A-10B. Figure 10A shows that bifunctional compound 6 of the present invention degraded DOT1L in MV411 cells up to 5.0 μM in a dose-dependent manner. Figure 10B shows that bifunctional compound 6 of the present invention reduced MV411 cell viability in a dose-dependent manner and at early time points.

[0192] Example 17: Cell proliferation assay in SEMK2 (B acute lymphoblastic leukemia) cells with bifunctional compounds of the present invention.

[0193] SEMK2 cells were cultured at 0.25 x 10 in a 6-well plate. 6Cells were seeded at a density of 100 cells / well, then treated with compounds at the indicated concentrations and incubated for the indicated times. After staining with trypan blue, cell counts were performed using a Countess® II FL automated cell counter (ThermoFisher Scientific).

[0194] The results are shown in Figure 11. They show that bifunctional compound 6 of the present invention reduced SEMK2 cell viability in a dose-dependent manner and at early time points.

[0195] Example 18: Cell proliferation assay in THP1 (acute monocytic leukemia) cells with bifunctional compounds of the invention.

[0196] THP1 cells were cultured at 0.25 × 10 in a 6-well plate. 6 Cells were seeded at a density of 100 cells / well, then treated with compounds at the indicated concentrations and incubated for the indicated times. After staining with trypan blue, cell counts were performed using a Countess® II FL automated cell counter (ThermoFisher Scientific).

[0197] The results are shown in Figure 12. They show that bifunctional compound 6 of the invention reduced THP1 cell viability in a dose-dependent manner and at early time points.

[0198] All patent and non-patent publications are indicative of the level of skill of those skilled in the art to which this invention pertains. All such publications, including the specific portions referred to, are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

[0199] Although the invention herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention. It is therefore to be understood that many modifications can be made to the illustrative embodiments and other arrangements can be devised without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. Uninterrupted C ４ ~C ２０ A bifunctional compound comprising a telomere silencing 1-like disruptor (DOT1L) and a deglon, covalently bonded to each other by an alkylene chain or a linker containing 2 to 8 PEG units of polyethylene glycol (PEG), wherein the compound is of formula (I): 【Chemistry 1】 [In the formula: R １ H, halogen, CH ３ ,CH ２ F, CF ２ H, CF ３ , CN, or NH ２ It represents; R ２ is 【Chemistry 2】 It represents; Degron is a bifunctional compound having a structure represented by [a ligand that binds to cereblon (CRBN)], or a pharmaceutically acceptable salt or stereoisomer thereof.

2. R １ A bifunctional compound according to claim 1, wherein is H or CN, or a pharmaceutically acceptable salt or stereoisomer thereof.

3. structure: 【Transformation 3】 【Chemistry 4】 The difunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, having any one of the above.

4. The difunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the alkylene chain comprises 11 to 15 consecutive alkylene units.

5. The bifunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the linker comprises three PEG units.

6. The linker is -O-, -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O- , -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N (R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O) ２ -, -OS(O)-, -S(O)O-, -S(O)-, -OS(O) ２ -, -S(O) ２ O-, -N(R')S(O) ２ -, -S(O) ２ N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O) ２ N(R')-, -N(R')S(O)N(R')-, C ３ ~C １２ The material further comprises at least one group selected from carbocyclene, 3- to 12-membered heterocyclene, and 5- to 12-membered heteroarylene, where R' is H or C １ ~C ６ It is alkylene, The difunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the aforementioned groups may be the same or different.

7. The aforementioned linker, 【Transformation 5】 or 【Transformation 6】 A bifunctional compound according to claim 1, further comprising a group, or a pharmaceutically acceptable salt or stereoisomer thereof.

8. The aforementioned linker, 【Transformation 7】 A bifunctional compound according to claim 7, comprising a group, or a pharmaceutically acceptable salt or stereoisomer thereof.

9. The aforementioned linker has the following structure: 【Transformation 8】 A bifunctional compound according to claim 1, which is any one of the above, or a pharmaceutically acceptable salt or stereoisomer thereof.

10. Structures (I-5) to (I-28): 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 The difunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, which is represented by any one of the above, or a pharmaceutically acceptable salt or stereoisomer thereof.

11. The aforementioned degron has structures (D1a) to (D1d): 【Chemistry 15】 【Chemistry 16】 [In the formula, X １ CH ２ or C(O), X ２ CH ２ A bifunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, represented by one of the following: , NH, or O.

12. Structures (I-29) to (I-44): 【Chemistry 17】 [Chemistry 18] 【Chemistry 19】 【Chemistry 20】 The difunctional compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, which is represented by any one of the above, or a pharmaceutically acceptable salt or stereoisomer thereof.

13. Structure (1) to (11): 【Chemistry 21】 【Chemistry 22】 【Chemistry 23】 A bifunctional compound, or a pharmaceutically acceptable salt or stereoisomer thereof, which is one of the following.

14. A pharmaceutical composition comprising a therapeutically effective amount of a bifunctional compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition comprising a bifunctional compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt or stereoisomer thereof, for use in a method of treating a disease or disorder characterized by or mediated by the abnormal activity of DOT1L, If you choose to have cancer as your disease or disorder, If, at the discretion of the party, the cancer is a blood cancer, At the discretion of the patient, the blood cancer may be multiple myeloma, lymphoma, or leukemia. Optionally, the leukemia may be acute myeloid leukemia, mixed lineage leukemia (MLL) rearranged acute myeloid leukemia, acute myeloid leukemia with a mutation in nucleophosmin 1 (NPM1), acute myeloid leukemia with a mutation in DNA methyltransferase 3A (DNMT3A), or acute myeloid eosinophilic leukemia. Pharmaceutical composition.

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