WO2017039318A1

WO2017039318A1 - Benzimidazole derivatives for dna methylation inhibitors

Info

Publication number: WO2017039318A1
Application number: PCT/KR2016/009722
Authority: WO
Inventors: MinJeong CHOI; Nakcheol Jeong; Hak Joong Kim; Jiaojie LI; Sunmi SHIN
Original assignee: Kainos Medicine Inc
Current assignee: Kainos Medicine Inc
Priority date: 2015-09-01
Filing date: 2016-08-31
Publication date: 2017-03-09
Anticipated expiration: 2018-03-01

Abstract

The invention provides for DNA methylation inhibitors, and also methods and use of the compounds of the invention, by themselves or in combination with other therapies, for treating a disease in which DNA hypermethylation is found.

Description

DESCRIPTION

Title of Invention

BENZIMIDAZOLE DERIVATIVES FOR DNA METHYLATION INHIBITORS FIELD OF THE INVENTION

The present invention relates to certain compounds which can modulate the hypermethylation of DNA sequences containing CpG sites that are associated with disease; and relates to processes for their preparation, as well as pharmaceutical compositions containing them as an active ingredient, and their use as medicaments.

BACKGROUND OF THE INVENTION

DNA methylation is the most stable epigenetic mark and means by which gene expression is modulated inside a cell. It occurs most frequently at CpG islands, used to designate "-C-phosphate-G-" or cytosine and guanine separated by only one phosphate and with cytosine being 5 prime to the guanine base, as distinguished from base-pairing of cytosine and guanine. Cytosine methylation in CpG islands is associated with gene silencing and gene transcription regulation.

DNA hypermethylation is observed at a specific CpG islands and in relation to specific genes in a number of diseases, as compared to the same regions of DNA or in relation to the same gene or genes in healthy subjects (e.g., where no methylation or less methylation is observed). Hypermethylation is also used to refer to a plurality of CpG dinucleotides in a specific or defined region of nucleic acid where DNA sequence is methylated. In that regard, such methylation is known to cause transcriptional inactivation of genes, and this methylation and transcriptional inactivation have been associated with, or thought to contribute to, the disease process. For example, in cancer cells showing hypermethylation at selected CpG islands, there is transcriptional inactivation of genes involved in main cellular pathways including, but not limited to, DNA repair (e.g., hMLHl, MGMT, BRAC1), vitamin response (e.g., CRBP1, RARB2), Ras signaling, cell cycle control (e.g., RASSF1A), p53 network, and apoptosis (e.g., DAPK1). In tumors, hypermethylation also causes an overall miRNA downregulation which is not only linked to development of cancer, but also to development of metastasis. It has been proposed by those skilled in cancer research that hypermethylation in certain CpG islands confers a growth advantage for tumor development. DNA hypermethylation has been observed in several tumor types including, but not limited to, colon cancer, endometrial cancer, stomach cancer, esophageal cancer, head and neck cancer, breast cancer, ovarian cancer, non-Hodgkins lymphoma, lung cancer, thyroid cancer, prostate cancer, renal cancer, brain cancer, leukemia, and lymphoma.

In neurological disorders, CpG hypermethylation is observed in Alzheimer's disease, resulting in transcriptional inactivation of a number of genes. For example, it has been reported that the frontal cortex of subjects with Alzheimer's disease showed disease-specific hypermethylation in the promoter region of cAMP response element-binding protein (CREB). The result may be exacerbation of reduced levels of brain-derived neurotrophic factor, along with in reduced neuronal plasticity and long-term memory formation, as observed in brains of subjects with Alzheimer's disease. Hypermethylation in the promoter region of the gene for synaptophysin has also been observed, and the reduced level of that protein is associated with altered synaptic plasticity in subjects with Alzheimer's disease.

In autoimmune disease, CpG hypermethylation is observed in rheumatoid arthritis, resulting in transcriptional inactivation of a number of genes. For example, in rheumatoid arthritis synovial cells, the CpG island containing the death receptor (DR3) gene promoter was specifically methylated. The result is reduced expression of DR3 protein, which may provide resistance to apoptosis. Increased proliferation and decreased apoptosis of synovial cells results in hyperplasia of the lining layer of the synovial tissue, an important feature of rheumatoid arthritis. Hypermethylation was also observed for genes including Transforming growth factor β receptor 2 (TGFBR2) and FOXOl. TGFBR2 affects the function of B cells, and changes in B cells in rheumatoid arthritis are evident by the pathogenic autoantibodies produced in the disease process. FOXOl transcript levels appear to be down-regulated in peripheral blood mononuclear cells in subjects with rheumatoid arthritis, suggesting the possibility that the down-regulation of FOXOl expression may contribute to the development of lymphocyte over-activation in rheumatoid arthritis.

Thus, modulation of the hypermethylation of genes involved in the control of cellular processes such as the control of cell proliferation, apoptosis, and immune activation is one strategy for the use of DNA methylation inhibitors. The DNA methylation inhibitors may be used to reactivate antiproliferative, apoptotic, and regulating genes in cells involved in diseases such as cancer, neurological diseases, and autoimmune diseases, with a resultant decrease in disease severity.

DNA methylation occurs at position 5 of cytosine mainly in a CpG dinucleotide context and it is catalyzed by the C5-DNA methyltransferases (DNMTs). Three families of DNMT have been characterized: DNMTl, DNMT2 and DNMT3 (comprising DNMT 3 A, 3B and 3L). DNMTl is mainly involved in the maintenance of the methylation pattern and is active on hemimethylated DNA, and DNMT 3 A and 3B are active both on unmethylated and hemimetnylated substrates, are responsible of de novo DNA methylation, and include DNMT3A, DNMT3B, and the catalytically inactive DNMT3L (Jurkowska, R. Z., et al, Chembiochem 2011, 12:206-222). If promoter CpG islands are methylated, the corresponding gene is repressed due to poor recognitions by transcription factors and recruitment of proteins involved in the chromatin remodeling such as methyl-binding proteins. By specifically inhibiting DNMTl, the enzyme required for maintenance methylation, the aberrant methylation of the tumor suppressor genes can be prevented.

Known cytidine analogs, such as azacitidine(vidaza®) and decitabine (Dacogen®), antimetabolites that have been found to inhibit the activity of DNA methyltransferases by incorporating into newly synthesized DNA, in the place of cytidine, causing covalent complex formation between DNA methyltransferases and azacitidine or decitabine containing DNA. Covalent complex formation results in the trapping of the enzyme (suicide enzyme inhibitor), preventing turnover even at other sites, and thus leads to DNA demethylation. Both compounds are now used clinically for the treatment of myelodysplasia syndromes (MDS) and lymphoproliferative diseases.

For all these reasons, DNA methylation appears to be a particularly interesting target from a therapeutic point of view. However, these two clinically approved DNMT inhibitors, azacitidine and decitabine are not selective toward the different DNMTs, are chemically instable (degradatde by cytidine deaminase: CDA), and have strong secondary effects, for example, renal toxicity or myelotoxicity (Boumber Y, et al., Oncology, 2011, 25(3), 220-6). Therefore, there is a need to identify novel, more specific and selective inhibitors. The non- nucleoside compounds that differ by chemical structure but in general bind to the DNMTs exhibit their action by variable mechanisms (Jacques F., et al., expert opin, Ther. Patents, 2012, 22(12), 1427-1442) . They should be more specific and less toxic than other inhibitors of DNA methylation. Among these, various inhibitors have been characterized, but most of them are nonspecific and /or do not induce DNA demethylation in cell (Gros, C, et al., Biochimie, 2012, 94:2280-2296, Singh, V., Curr. Cancer Drug Targets, 2013, 13:379-399) except for SGI- 1027, a quinolone derivatives that was described by Datt et al. in 2009 (Cancer Res., 2009, 69:4277-4285) for its enzymatic and cellular DNMT inhibition. Initially synthesized as part of a minor groove binders family of quinolinium bisquaternary salts, SGI- 1027 inhibits bacterial DNA methyltranferase M.SssI, human DNMTl, mouse DNMT3A, and mouse DNMT3B.

Thus, there still exists a need to develop effective modulators of DNA methylation which can be used in the prevention or treatment of diseases associated with aberrant DNA methylation such as cancer.

SUMMARY OF THE INVENTION

The present invention includes compounds that selectively inhibit the activity of mammalian DNA methyltransferases DNMTl and DNMT3s (DNMT 3 A, 3B and 3L). The compounds of the present invention, comprising DNA methylation inhibitors, surprisingly may provide improved metabolic stability, drug permeability, absorption, and increase in the DNMT binding when a compound of the invention inhibits the catalytic domain, transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) as compared to deoxycytidine, strong decrease in the dissociation rate, better toxicity profile. The compounds and compositions herein are useful in treatment or modulation of disease (one or more of pathology, symptoms, disorders, or conditions) in a subject, wherein the disease is treated or modulated by inhibition of DNA methylation of one or more genes which is transcriptionally inactivated by DNA methylation. The compounds are useful to inhibit DNA methyltransferase activity (and methods therefor). The compounds, compositions, and methods are useful in treatment or modulation of a disease in which one or more genes whose transcriptional inactivation is due to methylation and contributes to the disease. Diseases in which one or more genes whose transcriptional inactivation contributes to the disease include, but are not limited to, cancer, neurological disease, autoimmune disease, and symptoms or conditions thereof in a subject having the disease. In one aspect of the present invention, provided is a compound of Formula I and Formula II or a pharmaceutic thereof:

Formula I

wherein,

Z is selected from NH, O or S;

X_! and X₂ are each independently -NR.'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')C(S)N(R')-, -N(R')SO₂-, -N(R')SO₂N(R')-, -N(R')N(R')-, -N(R')C(=N(R'))N-, - SO₂N(R , -0-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R , -C(O)N(R')-, -C=NN(R')-, - C=NO-, -C(=N(R'))N(R or -C(R')₂-;

Y is a phenyl group optionally substituted by C^₄ alkyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo C^₄ alkyl, halo C1-4 alkoxy, hydroxyl C_1--4 alkyl, C_M alkoxy C^₄ alkyl, or C_1--4 alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by C^₄ alkyl, CF₃, halogen, hydroxyl, Ci~₄ alkoxy, cyano, nitro, halo C^ alkyl, halo Ci~4 alkoxy, hydroxyl

alkyl, or C 1-4 alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by C1- alkyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo C^₄ alkyl, halo C^₄ alkoxy, hydroxyl C^ alkyl, C^ alkoxyC^ alkyl, or C_1--4 alkoxycarbonyl ; C^₆ alkyl, C₂~6 alkynyl, or C₂~e alkenyl;

Ri and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 ^{03 04} Q5

wherein,

A is a 5-membered fused heteroaryl ring having 2 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 6-membered aryl or fused heteroaryl ring having 2 to 3 nitrogen atoms;

R4 and e are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -

N0₂, -C(O)R', -C(S)R', -CO₂R'₅ -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(0)R\ -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

B] and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃,

-NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -Ν^')^=Ν(^))Ν(^)₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

Ei, E₂, E₃ and E₄ are each independently selected from CH₂, NH, S, SH₂, or O; p is an integer of 1 to 3; and

R' is hydrogen, Ci ₆ aliphatic, Ci_~6 alkoxy, Ci~₆ alkyl, phenyl, CF₃, halogen, hydroxyl, Ci_~4 alkoxy, cyano, nitro, halo C_w alkyl, halo C1-4 alkoxy, hydroxyl C^ alkyl, Ci_^4 alkoxycarbonyl, Ci_~6 alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen.

Also, there is provided a compound of formula (II), or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof:

Formula II

Xi and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')C(S)N(R'K -N(R')SO₂-, -N(R')SO₂N(R , -N(R')N(R')-, -N(R')C(=N(R'))N-, - SO₂N(R')-, -O-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R')-, -C(O)N(R')-, -C=NN(R')-, - C=NO-, -C(=N(R'))N(R')- or -C(R')₂-; Y is a phenyl group optionally substituted by

alkyl, CF₃, halogen, hydroxyl, Ci alkoxy, cyano, nitro, halo C^₄ alkyl, halo C^ alkoxy, hydroxyl alkyl, C_1--4 alkoxy Ci alkyl, or alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by C^ alkyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo alkyl, halo alkoxy, hydroxyl alkyl, C^ alkoxyC^ alkyl, or C^ alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by alkyl, CF₃, halogen, hydroxyl, C_M alkoxy, cyano, nitro, halo C^₄ alkyl, halo alkoxy, hydroxyl C^ alkyl, Ci

alkoxycarbonyl ; Ci alkyl, C₂_₆ alkynyl, or C_2~6 alkenyl;

R₃ is selected from R\ halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R', - C(S)R', -C0₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, - C(S)OR\ -S(0)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

R and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5

wherein,

A is a 5 -membered fused heteroaryl ring having 2 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 6-membered aryl or fused heteroaryl ring having 2 to 3 nitrogen atoms;

R₄ and ¾ are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -

NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

B_t and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N; R₅ and R₅' are each independently R\ halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R\ -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR\ -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')2, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

Ei, E₂, E₃ and E₄ are each independently selected from CH₂, ΝΉ, S, SH₂, or O; p is an integer of 1 to 3; and

R' is hydrogen, C^₆ aliphatic, C^₆ alkoxy, alkyl, phenyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo C^₄ alkyl, halo alkoxy, hydroxyl Ci^ alkyl, C^ alkoxycarbonyl, alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen.

In accordance with another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating precancerous transformation or a cancer, which comprises the compound of formula (I) or a pharmaceutically acceptable salt, a hydrate, a solvate or an isomer thereof as an active ingredient, and a pharmaceutically acceptable carrier.

In accordance with a further aspect of the present invention, there is provided a method for preventing or treating precancerous transformation or a cancer in a subject, which comprises administering the compound of formula (I) or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof to the subject in need thereof.

In accordance with a still further aspect of the present invention, there is provided a use of the compound of formula (I) or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof for the manufacture of a medicament for preventing or treating precancerous transformation or a cancer.

The compounds of the present invention may be more potent, more metabolically stable and more effective in cancer treatment than other DNMT inhibitors that do not have the benzimidazole core derivatives. Further, the compound of the present invention may exhibit excellent enzyme activity and in vitro DMPK properties.

DETAILED DESCRIPTION OF THE INVENTION While the terms used in the description of the invention are believed to be well understood by one of ordinary skill in the art, definitions, where provided herein, are set forth to facilitate description of the invention, and to provide illustrative examples for use of the terms.

The term "alkyl" is used herein to refer to a hydrocarbon containing normal, secondary, tertiary, or cyclic carbon atoms (e.g., linear saturated aliphatic hydrocarbon groups, branched saturated aliphatic hydrocarbon groups, or a saturated or unsaturated non- aromatic hydrocarbon mono or multi-ring system (e.g., cycloalkyl)). When the term "alkyl" is used without reference to a number of carbon atoms, it is to be understood to refer to a _lo alkyl.

The term "aryl" is used herein to refer to cyclic, aromatic hydrocarbon groups which have 1 to 3 aromatic rings. The aryl group may have fused thereto a second or third ring which is a heterocyclo, cycloalkyl, or heteroaryl ring, provided in that case the point of attachment will be to the aryl portion of the ring system. Examples of "aryl" groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, biphenyl, indanyl, anthracyl or phenanthryl, as well as substituted derivatives thereof.

The term "heteroaryl" is used herein to refer to an aryl group in which at least one of the carbon atoms in the aromatic ring has been replaced by a heteroatom selected from oxygen, nitrogen and sulfur. The nitrogen and/or sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quaternized. The heteroaryl group may be a 5- to 6-membered monocyclic, 7- to 11-membered bicyclic, or 10- to 16- membered tricyclic ring system.

The term "alkenyl" is used herein to refer to a straight or branched chain hydrocarbon containing from 2 to 10 carbon atoms and containing at least one carbon-carbon double bond formed by the removal of two hydrogens.

The term "alkynyl" is used herein to refer to a straight or branched chain hydrocarbon containing from 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond.

The term "alkoxy" is used herein to refer to an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

The term "aralkyl" is used herein to refer to an aryl-alkyl group in which the aryl and alkyl are as defined herein. Preferred aralkyls comprise a lower alkyl group. The term "aryloxy" is used herein to refer to an aryl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

The term "carbocyclyl" (alone or in combination with another term(s)) is used herein to refer to a saturated cyclic (i.e., "cycloalkyl"), partially saturated cyclic (i.e., "cycloalkenyl"), or completely unsaturated (i.e., "aryl") hydrocarbyl substituent containing from 3 to 14 carbon ring atoms ("ring atoms" being the atoms bound together to form the ring or rings of a cyclic substituent). A carbocyclyl may be a single ring, which typically contains from 3 to 6 ring atoms.

The term "cycloalkyl" is used herein to refer to monocyclic or multicyclic (e.g., bicyclic, tricyclic, etc.) hydrocarbons containing from 3 to 12 carbon atoms that is completely saturated or has one or more unsaturated bonds but does not amount to an aromatic group.

The term "cyano" is used herein to refer to a -CN group.

The term "halo" or "halogen" is used herein to refer to -CI, -Br, -I, or -F.

The term "haloalkyl" is used herein to refer to an alkyl, as defined herein, wherein at least one hydrogen atom is replaced with halogen atoms.

The term "heterocyclyl" is used herein to include a saturated (e.g., "heterocycloalkyl"), partially unsaturated (e.g., "heterocycloalkenyl" or "hetercycloalkynyl") or completely unsaturated (e.g., "heteroaryl) ring system, which have 3 to 12 atoms including at least one heteroatom, such as nitrogen, oxygen, or sulfur.

The terms "first" and "second" are used herein for the purpose of distinguishing between two compounds, or between two compositions, as will be clearer from the description.

The phrase "medically effective amount" means an amount of a composition or compound that treats the particular disease, condition or disorder; ameliorates, relieves, or decreases one or more symptoms associated with the particular disease, condition or disorder; and/or delays or prevents the onset of symptoms of, or a pathological process associated with the particular disease, condition or disorder described herein in more detail.

The term "pharmaceutically acceptable carrier" is used herein to mean any compound or composition or carrier medium useful in any one or more of administration, delivery, storage, stability of a composition or compound described herein. The pharmaceutically acceptable carriers include, but are not limited to, a diluent, water, saline, a suitable vehicle (e.g., liposome, microparticle, nanoparticle, emulsion, and capsule), a buffer, a medical parenteral vehicle, an excipient, an aqueous solution, a suspension, a solvent, an emulsion, a detergent, a chelating agent, a solubilizing agent, a salt, a colorant, a polymer, a hydrogel, a surfactant, an emulsifier, an adjuvant, a filler, a preservative, a stabilizer, an oil, a binder, a disintegrant, an absorbant, a flavor agent, and the like as broadly known in the art.

Hereinafter, the present invention is described in detail.

The present invention provides a compound of Formula I and Formula II, or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof:

Formula I

wherein,

Z is selected from NH, O or S;

X_! and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')C(S)N(R , -N(R')SO₂-, -N(R')SO₂N(R')-, -N(R')N(R')-, -N(R')C(=N(R'))N-, - SO₂N(R')-, -0-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R , -C(O)N(R , -C= N(R , -

C=NO-, -C(=N(R'))N(R')- or -C(R')₂-;

Y is a phenyl group optionally substituted by C^₄ alkyl, CF₃, halogen, hydroxyl, Ci-4 alkoxy, cyano, nitro, halo C^ alkyl, halo C^ alkoxy, hydroxyl C_w alkyl, C^₄ alkoxy C^4 alkyl, or Ci_4 alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by C_1--4 alkyl, CF₃, halogen, hydroxyl, C_1--4 alkoxy, cyano, nitro, halo C_1--4 alkyl, halo C^ alkoxy, hydroxyl C^ alkyl, C^ alkoxyC^ alkyl, or C_w alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by C^ alkyl, CF₃, halogen, hydroxyl, C^₄ alkoxy, cyano, nitro, halo C_1--4 alkyl, halo C^₄ alkoxy, hydroxyl C^₄ alkyl, Ci^ alkoxyC^ alkyl, or C^ alkoxycarbonyl ; Ci~₆ alkyl, C₂_6 alkynyl, or C_2~6 alkenyl;

Ri and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5

wherein,

R4 and Re are each independently R\ halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, - NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

B_\ and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

-NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

R' is hydrogen, Ci~₆ aliphatic, Ci_₆ alkoxy, C^₆ alkyl, phenyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo Ci^ alkyl, halo Ci~₄ alkoxy, hydroxyl Ci_4 alkyl, Ci~₄ alkoxycarbonyl, C}^ alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen. Also, there is provided a compound of formula (II), or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof:

Formula II

Xi and X₂ are each independently -NR.'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')C(S)N(R')-, -N(R')SO₂-, -N(R')SO₂N(R')-, -N(R')N(R')-, -N(R')C(=N(R'))N-, - SO₂N(R')-₅ -0-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R')-, -C(O)N(R , -C=NN(R')-, - C=NO-, -C(=N(R'))N(R')- or -C(R')₂-;

Y is a phenyl group optionally substituted by C^ alkyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo Ci_4 alkyl, halo C^ alkoxy, hydroxyl C^₄ alkyl, C^₄ alkoxy C_1--4 alkyl, or alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by C_1--4 alkyl, CF₃, halogen, hydroxyl, C₁- alkoxy, cyano, nitro, halo C^₄ alkyl, halo C^ alkoxy, hydroxyl C_1--4 alkyl, C^₄ alkoxyC^ alkyl, or C_1--4 alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by alkyl, CF₃, halogen, hydroxyl, C_1--4 alkoxy, cyano, nitro, halo C_1-- alkyl, halo Cj_4 alkoxy, hydroxyl C_1--4 alkyl, Ci_^4 alkoxyC^ alkyl, or C_1--4 alkoxycarbonyl ; C_1--6 alkyl, C₂~₆ alkynyl, or C_2~6 alkenyl;

R₃ is selected from R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -NO₂, -C(0)R', - C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, - C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Ri and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5

wherein, A is a 5-membered fused heteroaryl ring having 2 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 6-membered aryl or fused heteroaryl ring having 2 to 3 nitrogen atoms;

R4 and Re are each independently R', halogen, -OR', -SR\ -N(R')₂, -CN, -CF₃, - NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R\ -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Bi and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R\ halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -N0₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

Ei, E₂, E3 and E₄ are each independently selected from CH₂, NH, S, SH₂, or O; p is an integer of 1 to 3; and

R' is hydrogen, Ci~₆ aliphatic, Ci_₆ alkoxy, Ci_₆ alkyl, phenyl, CF₃, halogen, hydroxyl, C^₄ alkoxy, cyano, nitro, halo Ci~ alkyl, halo Ci_4 alkoxy, hydroxyl C_1--4 alkyl, C_1-4 alkoxycarbonyl, Ci~₆ alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen.

In another embodiment, the present invention provides compounds of formula I, wherein,

Z is selected from NH or O;

Xi and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')N(R')-, -N(R')C(=N(R'))N-, -O- , -CO-, -OC(O)-, -C(O)R'C(O)R'-, -C(O)O-, - OC(O)N(R')-, -C(O)N(R')-, -C=NN(R , -C=NO-, -C(=N(R'))N(R')- or -C(R')₂-; Y is a 5- to 6-membered heteroaromatic group optionally substituted by C_1--4 alkyl, CF₃, halogen, hydroxyl, C^₄ alkoxy, cyano, nitro, halo alkyl, halo C^alkoxy, hydroxyl C_1--4 alkyl,

alkoxy carbonyl;

R] and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5 wherein,

R4 and Re are each independently R', halogen, -OR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R\ -CO₂R\ -C(O)N(R')₂-C(O)C(O)R\ -C(O)CH₂C(O)R', -N(R')C(O)R', - N(R')C(O)N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, -C=NN(R')₂, -C=NOR', - C(=N(R'))N(R')_2> -OC(O)R', or -OC(O)N(R')₂;

and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

C-i, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R\ halogen, -OR', -N(R')₂, -CN, -CF₃, -NO₂,

-C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)C(O)R', -C(O)CH₂C(O)R', -N(R')C(0)R', - N(R')C(O)N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, -C=NN(R')₂, -C=NOR', - C(=N(R'))N(R')₂, -OC(O)R' or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH; Ei, E₂, E₃ and E₄ are each independently selected from CH₂, NH, or O; p is an integer of 0 to 3; and

R' is hydrogen, or an optionally substituted group selected from C_w aliphatic,

Ci~6 alkoxy, C^ alkyl, phenyl, CF₃, halogen, hydroxyl or C^ alkoxy, cyano, nitro, halo Ci_4 alkyl, halo C^ alkoxy, hydroxyl C1-4 alkyl, C^ alkoxycarbonyl, or a C^ alkyl substituted with amino or hydroxyl. In another embodiment, the present invention provides compounds of formula I, wherein,

Z is NH;

X] and X₂ are each independently -NR' or -O- ;

Y is a 5- to 6-membered heteroaromatic group optionally substituted by alkyl, CF₃, halogen, hydroxy 1, C_1--4 alkoxy, cyano, nitro, halo C_1-4 alkyl, halo C_1--4 alkoxy, hydroxyl Ci~ alkyl, C^ alkoxy, Ci~4 alkyl or C^ alkoxycarbonyl ;

R and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5 wherein,

A is a 6-membered aryl or fused heteroaryl ring having 2 to 3 nitrogen atoms;

4 and R6 are each independently R', -N(R')₂ or -CN;

i and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R' or -N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH or N;

Ei, E₂, E₃ and E₄ are each independently selected from CH₂ or NH; p is an integer of 0 to 3; and

R' is hydrogen, C^₆ alkyl, cyano, nitro, or a C^₆ alkyl substituted with amino.

Preferable examples of the compound according to the present invention are listed below, and a pharmaceutically acceptable salt and a stereoisomer thereof are also included in the scope of the present invention:

Example 1: 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine Example 2: 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine tri-hydrogen chloride

Example 3: 2-amino-4-methyl-6-((2-(4-(quinolin-4-ylamino)phenyl)-lH- benzo [d] imidazol-5 -y l)amino)pyrimidine 1 -oxide

Example 4: N-(4-(5-(2-aminopyrimidin-4-yloxy)- 1 H-benzo[d]imidazole-2-yl) phenyl)quinolin-4-amine

Example 5: N4-(2-(4-(lH-tetrazol-5-ylamino)phenyl)-lH-benzo[d]imidazol-5-yl) pyrimidine-2 ,4-diamine

Example 6: 6-methyl-N4-(2-(4-((2-methylpyridin-4-yl)amino)phenyl)-lH- benzo[d]imidazol-5-yl)pyrimidine-2,4-diamine

Example 7: 6-methyl-N4-(2-(4-( yridin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine

Example 8: N4-(2-(4-(quinolin-4-ylamino)phenyl)- lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine

Example 9: N-(4-(5-(pyridin-2-ylamino)- 1 H-benzo[d]imidazol-2-yl)phenyl)quinolin-

4- amine

Example 10: N4-(2-(4-((2-methylpyridin-4-yl)amino)phenyl)-lH-benzo[d]imidazol-

5- yl)pyrimidine-2,4-diamine trihydrochloride

Example 11 : 2-(4-((2-methylpyridin-4-yl)amino)phenyl)-N-(pyridin-2-yl)- 1H- benzo[d]imidazol-5-amine tris(2,2,2-trifluoroacetate)

Example 12: 4-methyl-N2-(2-(4-(pyridin-4-ylamino)phenyl)-lH-benzo[d]imidazol- 5 -yl)pyridine-2,6-diamine trihydrochloride

Example 13: 6-methyl-N4-(2-(4-( yridin-2-ylamino)phenyl)-lH-benzo[d]imidazol- 5-yl)pyrimidine-2,4-diamine trihydrochloride

Example 14: 6-methyl-N4-(2-(4-((3-methylpyridin-4-yl)amino)phenyl)-lH- benzo[d]imidazol-5-yl)pyrimidine-2,4-diamine trihydrochloride Example 15: N-(4-(5-(quinolin-4-ylamino)-lH-benzo[d]imidazol-2- yl)phenyl)quinolin-4-amine trihydrochloride

Example 16: N-(4-((2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)amino)pyrimidin-2-yl)acetamide dihydrochloride

Example 17: N4-(2-(4-(pyridin-4-ylamino)phenyI)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine trihydrochloride

Example 18: N-(4-(5-((2-aminopyrimidin-4-yl)oxy)-lH-benzo[d]imidazol-2- yl)phenyl)quinolin-4-amine trihydrochloride

Example 19 : N4-(2-(4-(quinolin-4-ylamino)phenyl)- 1 H-benzo [d] imidazol-5- yl)pyridine-2,4-diamine trihydrochloride

Example 20 : 2-amino-4-methy l-6-((2-(4-(quinolin-4-y lamino)pheny 1)- 1 H- benzo[d]imidazol-5-yl)amino)pyrimidine 1 -oxide trihydrochloride

Example 21 : N4-(2-(4-((2,6-dimethylpyridin-4-yl)amino)phenyl)- 1 H- benzo[d]imidazol-5-yl)-6-methylpyrimidine-2,4-diamine trihydrochloride

Example 22: N4-(2-(4-((2,3-dimethylpyridin-4-yl)amino)phenyl)-lH- benzo[d]imidazol-5-yl)-6-methylpyrimidine-2,4-diamine trihydrochloride

Example 23: N4-(4-(5-((2-amino-6-rnethylpyrimidin-4-yl)amino)-lH- benzo[d]imidazol-2-yl)phenyl)-6-methylpyrimidine-2,4-diamine trihydrochloride

Example 24 : (Z)-2-cyano- 1 -(2-(4-(pyridin-4-ylamino)phenyl)- 1 H-benzo [d]imidazol- 5-yl)guanidine trihydrochloride

Example 25: N4-(2-(4-(pyrimidin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine trihydrochloride

Example 26: N-(pyridin-4-yl)-2-(4-(pyridin-4-ylamino)phenyl)-lH- benzo[d]imidazol-5-amine trihydrochloride

Example 27: N4-(2-(4-((lH-tetrazol-5-yl)amino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine tris(2,2,2-trifluoroacetate). While the terms used in the description of the invention are believed to be well understood by one of ordinary skill in the pharmaceutical arts, definitions, where provided herein, are set forth to facilitate description of the invention, and to provide illustrative examples for use of the terms.

The terms "a," "an," and "the" may mean one or more, and may be used to reference both the singular and the plural.

The terms "purified" or "isolated" for a compound according to Formula I refers to the physical state of the compound following isolation from a synthetic process or purification step described herein or well known to those in the art, and in sufficient purity to be characterizable by standard analytical methods described herein or well known in the art.

The terms "treat", "treats", or "treating" , as used herein, embrace one or more of preventative (prophylactically) or therapeutically (palliative).

The compounds of Formula I or Formula II can form salts, and salts of the compounds are included within the scope of the invention. The terms "salt" or pharmaceutically acceptable salt", as used herein, refers to inorganic or organic salts of a compound. These salts can be prepared, for example, by reacting a compound of Formula I with an amount of acid or base, such as an equivalent amount, and in a medium such as one in which the salt formed then precipitates, or in an aqueous medium followed by lyophilization. Representative salts include bisulfate, sulfate, benzene sulfonate, camphorsulfonate, laurylsulphonate, methanesulfonate, toluenesulfonate, naphthalenesulformate, acetate, trifluoracetate, benzoate, borate, butyrate, citrate, formate, fumarate, hydorbromide, hydrochloride, hydroiodide, lactate, laurate, maleate, malonate, mesylate, nitrate, oxalate, phosphate, hexafiuorophosphate, propionate, salicylate, stearate, succinate, tartrate, thiocyanate, and the like. The salts may include base salts based on the alkali and alkaline earth metals, such as calcium, sodium, lithium, magnesium, and potassium; or with organic bases such as with organic amines (e.g., dicyclohexylamine, t- butyl amine, methylamine, dimethylamine, triethylamine, ethylamine, procaine, morpholine, N-methylpiperidine, dibenzylamine, and the like); or as an ammonium salt.

The compounds of Formula I or Formula II may exist in a solvated form or unsolvated form. Solvates of a compound of the invention may be formed in the synthetic process in which the compound becomes physically associated with one or more solvent molecules (e.g., such as by ionic and/or covalent bonding) or, optionally, may be converted to a solvate such as by dissolving the compound in desired amounts of a solvent of choice (e.g., organic solvent, water, or mixtures thereof) in forming a solution, heating the solution to a temperature higher than ambient temperature, and cooling the solution at a rate sufficient to form crystals of the solvate, which may then be further isolated using methods known the art. Examples of suitable solvents include methanolates, ethanolates, hydrates (where the solvent molecule is water), and the like.

The compounds of Formula I may contain asymmetric or chiral centers, and thus exist in different stereoisomeric forms. All stereoisomers (e.g., geometric isomers, optical isomers, and the like), enantiomeric forms, diastereomeric forms, tautomeric forms, positional isomers, of the compounds of the invention are embraced within the scope of the invention. A first conformational form of a compound can be separated from a second and different conformational form of the compound using methods well known in the chemical arts such as by chromatography, crystallization, and methods of synthesis which selectively result in a particular desired conformational form.

The invention further provides a composition, preferably a pharmaceutical composition or medicament, containing a medically effective amount of a compound of Formula I or Formula II (or pharmaceutically acceptable salts, solvates, esters or prodrugs thereof) and a pharmaceutically acceptable carrier. A composition according to the invention may be administered once, or multiple times, as needed, to deliver a medically effective amount of the composition, e.g., an amount effective to mediate modulation of a disease by inhibiting DNA methyltransferase activity in cells in the subject receiving the composition. For example, a medically effective amount of a composition comprising the compound of the invention may be an amount that enters into cells which are contacted with the compound, and which results in inhibiting DNA methylation in the cells. Such a medically effective amount of the composition will depend on such factors as the mode of administration, the formulation for administration, disease to be modulated, the size and health of the subject to receive such a composition, and other factors which can be taken into consideration by a medical practitioner who is skilled in the art of determining appropriate dosages for treatment. An amount of the compound of the invention in a composition to be administered may vary from 0.01 milligrams to about 500 milligrams, and more typically from about 1 milligram per day to about 200 milligram per day. One skilled in the art can apply known principles and models of drug delivery and pharmacokinetics to ascertain a likely range of dosages to be tested in preclinical and clinical studies for determining a medically effective amount of the compound of the invention. A pharmaceutically acceptable carrier, used in a composition of the invention, may facilitate one or more of storage, stability, administration, and delivery, of the composition. The carrier may be particulate, so that the composition may be in, for example, powder or solid form. The carrier may be in a semi-solid, gel, or liquid formula, so that the composition may be ingested, injected, applied, or otherwise administered. The carrier may be gaseous, so that the composition may be inhaled.

For oral administration of a composition containing a compound of the invention, suitable formulations may be presented in the form of tablets, caplets, capsules, and the like, in which typically the compound of the invention may be present in a predetermined amount as a powder, granules, solution, or suspension as the sole active agent, or in combination with an additional one or more pharmaceutical agents. As known in the art, such oral formulations typically involve one or more of a binder (e.g., syrup, sorbitol, gum, corn starch, gelatin, acacia), a filler (e.g., lactose, sugar, starch, calcium phosphate), an excipient (e.g., dicalcium phosphate), a disintegrating agent (e.g., vegetable starch, alginic acid), a lubricant (e.g., magnesium stearate), a flavoring agent (sweetening agent, natural or artificial flavors). Such oral formulations may be coated or uncoated to modify their disintegration and/or absorption. Coating may be performed using conventional coating agents and methods known in the art.

The mode of administration of a compound or composition of the invention to a subject (such as a human) in need of such composition or compound may be any mode known in the art to be suitable for delivering a pharmaceutical composition, and particularly suitable for treating a disease by inhibiting DNA methyltransferase activity in cells, and may include but is not limited to, intravenously, intraperitoneally, orally, subcutaneously, intramuscularly, intranasally, transdermally, by perfusion, and by peristaltic techniques. The compositions of the invention may also be combined with other therapies, such as one or more additional pharmaceutical agents, to treat a disease in which hypermethylation of CpG islands is observd. Such combination therapy may be administered in concurrently, sequentially, or in regimen alternating between the composition of the invention and the other therapy. Such combination therapies may include administering a compound of Formula I with one or more additional therapeutic agents, for treating one or more diseases selected from the group consisting of cancer, neurological disease (e.g., Alzheimer's disease), autoimmune or inflammatory disease (e.g., rheumatoid arthritis), and a myelodysplastic disorder. The structure of the agents, for combination with a compound of Formula I or Formula II, identified herein, and their generic or trademark names, are readily available to those skilled in the art, such as from the standard compendium of drugs (e.g., The Merck Index) or from the applicable pharmaceutical company's web site, as well as dosages applicable for treatment (see also The Physician's Desk Reference). Alternatively, the doses and dosage regimen of an additional pharmaceutical agent, used in conjunction with a compound of the invention in combination therapy, can be determined by a physician, taking into account the medical literature, the health, age and sex of the patient, the disease to be treated, the mode of administration and dosing schedule of the pharmaceutical agent, and other relevant considerations. Generally, dosages of such agents can range from about 0.1 mg to 1000 mg per day, with more specific dosages dependent on the aforementioned factors.

Accordingly, provided herein is a pharmaceutical composition or medicament comprising a medically effective amount of a compound of one or more of Formula I or Formula II, in combination with a medically effective amount of one or more of a chemotherapeutic agent, anti-inflammatory drug, COX-2 inhibitor, immune modulator, cholinesterase inhibitor, NMDA receptor antagonist, or a combination thereof; and optionally further comprising a pharmaceutically acceptable carrier. Also provided herein is a pharmaceutical composition or medicament comprising a medically effective amount of a compound of one or more of Formula I or Formula II, and a pharmaceutically acceptable carrier.

In this Example 1, illustrated is the use of a compound of the invention, a compound of Formula I or Formula II, (or a pharmaceutical composition comprising such compound) in a medically effective amount to treat a disease modulated by inhibiting DNA methyltransferase activity in cells. In one method according to the invention, provided is a method for treating a disease modulated by inhibiting DNA methyltransferase activity in cells of a subject in need of inhibition of DNA methylation, comprising the step of administering to the subject(a subject, such as a human) in need of such treatment a medically effective amount of one or more compounds having Formula I or Formula II, wherein the disease modulated by inhibiting DNA methyltransferase activity in cells is selected from the group consisting of an atherosclerosis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, liver cirrhosis, cancer (e.g., lymphoma; leukemia; and solid, non- lymphoid tumors), rheumatoid arthritis, Alzheimer's disease, and myelodysplastic disorders. Also provided is a method for treating a disease in a subject by inhibiting DNA methyltransferase activity in cells, including those having DNA hypermethylation, the method comprising the step of administering to the subject in need of such treatment a medically effective amount of a pharmaceutical composition comprising one or more compounds having Formula I or Formula II, and a pharmaceutically acceptable carrier, wherein the disease modulated by inhibiting DNA methyltransferase activity is selected from the group consisting of an atherosclerosis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, liver cirrhosis, cancer (e.g., lymphoma; leukemia; and solid, non-lymphoid tumors), rheumatoid arthritis, Alzheimer's disease, and myelodysplastic disorders. In either case, the pharmaceutical composition may further comprise one or more chemotherapeutic agents. Also provided is a compound having Formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disease modulated by inhibiting DNA methyltransferase activity in cells. Such disease, modulated by inhibiting DNA methyltransferase activity in cells may be selected from the group consisting of an atherosclerosis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, liver cirrhosis, cancer (e.g., lymphoma; leukemia; and solid, non-lymphoid tumors), rheumatoid arthritis, Alzheimer's disease, and myelodysplastic disorders. General Synthetic Methods

NMR spectra were recorded in CDC1₃ and OMSO-d₆ solution in 5-mm o.d. tubes (Norell, Inc. 507-HP) at 30 °C and were collected on Varian VNMRS-400 at 400 MHz for 1H. The chemical shifts (δ) are relative to tetramethylsilane (TMS = 0.00 ppm) and expressed in ppm. LC/MS was taken on Ion-trap Mass Spectrometer on FINNIGAN Thermo LCQ Advantage MAX, Agilent LC 1200 series (Column : YMC Hydrosphere (CI 8, 04.6 x 50 mm, 3 μιιι, 120 A, 40 °C) operating in ESI(+) ionization mode; flow rate = 1.0 mL/min; and Mobile phase = 0.01% heptafluorobutyric acid (HFBA) and 1.0% isopropyl alcohol (IP A) in water or CH₃CN).

Scheme 1 shows the synthesis of Example 1 and Example 2. Schemel.

Intermediate 3: 4-(5-nitro-lH-benzo[d]imidazol-2-yl)aniline

A solution of 4-nitrobenzaldehyde (25.5 mmol, 3.9 g) and stannous chloride dihydrate (166 mmol, 37.5 g) in ethanol (250 mL) was refiuxed at 70 °C. The reaction was followed by TLC. The reaction was quenched with saturated NaHCO₃ solution, extracted with ethyl acetate, dried with MgSO₄ and then filtered. The organic extracts containing 4- aminobenzaldehyde 2 were not fully evaporated to avoid self-condensation problem. The residue was dissolved in acetonitrile (250 mL) and 4-nitrobenzene-l,2-diamine(25.5 mmol, 3.9 g), 37% hydrochloric acid (7 mL), 35% H₂O₂ (18 mL) were added at 0 °C. The solution was stirred at room temperature and the reaction was followed by TLC. The solvent was evaporated under reduced pressure and the resulting residue was quenched with 1 N NaOH solution, extracted with ethyl acetate. The combined organic extracts were dried with MgS0₄, filtered, and concentrated under reduced pressure. The residue obtained upon evaporation of solvent was purified by recrystallization using acetone, ethyl acetate and hexane to provide pure 3 (3.3 g). Yield: 50.9 % 1H NMR (DMSO-d₆, 400 MHz) δ 13.14 (br s, 1H), 8.38 (br s, 1H), 8.16 (d, J = 3.2 Hz, 1H), 7.98 (d, J = 2.4 Hz, 2H) 7.68 (br s, 1H), 6.88 (d, J = 5.6 Hz, 2H), 5.87 (br s, 2H)

Intermediate 4: N-(4-(5-nitro-lH-benzo[d]imidazol-2-yl)phenyl)quinolin-4-amine

4-(5-nitro-lH-benzo[d]imidazol-2-yl)aniline (3.93 mmol, 1.0 g), 4-chloroquinoline (4.7 mmol, 772 mg), and a catalytic amount of 37% hydrochloric acid (4 drops) in ethanol(40 mL) were refluxed for 2 h. The reaction was allowed to cool to room temperature, and the precipitated solid was filtered off, and recrystallized by acetonitrile to afford pure N- (4-(5-nitro-lH-benzo[d]imidazol-2-yl)phenyl)qumolin-4-amine 4 (1.25 g).

Yield: 83.2 % 1H NMR (DMSO-d₆, 300 MHz) δ 9.26 (bs, 1H), 8.56 (d, J = 4.9 Hz, 1H), 8.41 (d, 1H), 8.36 (d, 1H) 8.21 (d, J = 8.5 Hz, 2H), 8.06 (dd, 1H), 7.90 (d, 1H), 7.68 (m, 2H), 7.56(d, 1H), 7.52(d, J = 8.5 Hz, 2H), 7.21 (d, J = 4.9 Hz, 1H)

Example 1: 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine

A solution of 4 (2.62 mmol, 1.0 g) and stannous chloride dihydrate (13.1 mmol, 2.96 g) in ethanol (30 mL) was refluxed for 3 h. Afterward, the solvent was evaporated, and the residue was basified with 6 N sodium hydroxide solution, stirred overnight and the mixture was extracted with ethyl acetate, washed with brine, dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain pure 5 (921 mg). To a solution of 5 (2.62 mmol, 921 mg) in ethanol (30 mL), 4-chloro-6-methylpyrimidin-2-amine (2.62 mmol, 376 mg) and a catalytic amount of 37% hydrochloric acid (3 drops) were added, refluxed for 2 h. After the reaction, the solution was allowed to cool to room temperature and then filtered. The resulting solid was washed with ethanol and recrystallized with acetonitrile to afford pure Example 1 (1.1 g).

Yield: 91.6 % 1H NMR (DMSO-d₆, 300 MHz) δ 9.25 (bs, 1H), 8.97 (s, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.39 (d, 1H) 8.19 (d, J = 8.8 Hz, 2H), 8.07 (s, 1H), 7.88 (d, 1H), 7.68 (ddd, 1H), 7.52(ddd, 1H), 7.48(d, J - 8.8 Hz, 2H), 7.45(s, 1H), 7.25(dd, 1H), 7.14(d, J = 5.2 Hz, 1H), 6.12,(bs, 1H), 5.88(s, 1H), 2.06(s, 3H)

Example 2: 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine tri-hydrogen chloride

A solution of 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol- 5-yl)pyrimidine-2,4-diamine in trifluoroacetic acid (Example 1) was dropped concentration HC1. The mixture was froze in dry ice bath for lh. After freezing, lyophilized overnight to provide 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)- 1 H-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine tri-hydrogen chloride (Example 2).

1H NMR (DMSO-d₆, 400 MHz) δ 12.80 (br s, 1H), 11.23 (s, 1H), 11.04 (br s, 1H), 8.87 (d, J = 8.8 Hz, 1H), 8.62 (d, J = 6.8 Hz, 1H), 8.48 (d, J = 8.8 Hz, 2H), 8.17 (s, 1H), 8.10 (d, J = 8.4 Hz, 1H), 8.04 (t, J - 7.2 Hz, 1H) 7.91-7.73 (m, 6H), 7.11 (d, J = 6.4 Hz, 1H), 6.23 (s, 1H), 2.27 (s, 3H).

Scheme 2 shows the synthesis of Example 3.

Scheme 2.

Example 3: 2-amino-4-methyl-6-((2-(4-(quinolin-4-ylamino)phenyl)-lH- benzo[d]imidazol-5-yl)amino)pyrimidine 1 -oxide

To a solution of 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH- benzo[d]imidazol-5-yl)- pyrimidine-2,4-diamine (Example 1) (30.0 mg, 0.0650 mmol) in MeOH (0.33 mL) was added w-CPBA (55wt%, 48.4 mg, 0.196 mmol) at room temperature. The reaction mixture was stirred at room temperature for 3 days. A precipitated solid was collected by filtration and washed with hexanes, The solid was purified by column chromatography on SiO₂ (EtOAc:MeOH = 10:1 to 8:1) to afford 2-amino-4-methyl-6-((2-(4- (quinolin-4-ylamino)phenyl)- lH-benzo[d]imidazol-5-yI)amino)pyrimidine 1 -oxide

(Example 3, 7.00 mg, 22% yield) as a yellow solid.

LC/MS m/z 475.3 [M+H]⁺, Rt = 2.87 min

Scheme 3 shows the synthesis of Example 4. Scheme 3.

Intermediate 3 : 5 -methoxy-2-(4-nitrophenyl)- 1 H- benzo [d] imidazole

To a mixture of 4-nitrobenzoic acid (1) (1.18 g, 7.09 mmol) and 4-methoxybenzene- 1,2-diamine (2) (0.855 mL, 7.24 mmol) was added POC13 (0.675 mL, 7.24 mmol) at room temperature. The reaction mixture was refluxed for 30 min, cooled to room temperature and poured into ice-water. A generated solid was collected by filtration, washed with saturated aq. NaHCO₃ and dried under vacuum to afford 5-methoxy-2-(4-nitrophenyl)-lH- benzo[d]imidazole (3) (1.00 g, 51%) as a brown solid. nu M 201b. / 0 o 9 7 22

Intermediate 4: 4-(5-methoxy-lH-benzo[d]imidazol-2-yl)aniline

A suspension of 5-methoxy-2-(4-nitrophenyl)-lH-benzo[d]imidazole (3) (1.00 g, 3.71 mmol) Pd/C (10wt%, 0.790 g, 0.371 mmol) in MeOH (18 mL) was stirred at room temperature for 1 hour under H2 atmosphere (balloon). After filtration through a Celite pad, the filtrate was concentrated in vacuo to afford 4-(5-methoxy-lH-benzo[d]imidazol-2- yl)aniline (4) (820 mg, 92%) as a brown solid, which was used for the next step without further purification. Intermediate 6: N-(4-(5-methoxy-lH-benzo[d]imidazol-2-yl)phenyl)quinolin-4- amine

A dried round flask was charged with 4-(5-methoxy-lH-benzo[d]imidazol-2- yl)aniline (4) (820 mg, 3.43 mmol), 4-bromoquinoline (5) (713 mg, 3.43 mmol), and Cs2C03 (1.68 g, 5.14 mmol) and dioxane (17 mL) and then evacuated and refilled with nitrogen several times. After addition of Pd2(dba)3 (94.0 mg, 0.103 mmol) and Xantphos (119 mg, 0.206 mmol), the reaction mixture was refluxed overnight, cooled to room temperature and quenched with water (5.0 mL). The mixture was filtered through a Celite pad. The filtrate was concentrated in vacuo. The residue was partitioned between saturated aq. NaHC03 (5.0 mL) and DCM (10 mL). The separated aqueous layer was extracted with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (EtOAc:MeOH = 20:1), affording N-(4-(5-methoxy-lH-benzo[d]imidazol-2- yl)phenyl)quinolin-4-amine (6) (560 mg, 45%) as a yellow solid.

Intermediate 7: 2-(4-(quinolin-4-ylamino)phenyl) -lH-benzo[d]imidazol-5-ol

To a solution of N-(4-(5-methoxy-lH-benzo[d]imidazol-2-yl)phenyl)quinolin-4- amine (6) (535 mg, 1.46 mmol) in DCM (7.3 mL) was added BBr3 (14.6 mL, 14.6 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 days and quenched with ice-water. After concentration in vacuo, the residue was purified by column chromatography on SiO2 (EtOAc:MeOH = 10:1), affording 2-(4-(quinolin-4- ylamino)phenyl) -lH-benzo[d]imidazol-5-ol (7) (200 mg, 39%) as a yellow solid.

Example 4 : N-(4-(5 -(2-aminopyrimidin-4-yloxy)- 1 H-benzo [d]imidazole-2-y 1) phenyl)quinolin-4-amine

A flask containing a solution of 2-(4-(quinolin-4-ylamino)phenyl)-lH- benzo[d]imidazol-5-ol (7) (50.0 mg, 0.142 mmol), 4-chloropyrimidin-2-amine (8) (18.4 mg, 0.142 mmol) and 2 N aq. KOH (0.071 mL, 0142 mmol) in water (0.70 mL) was subjected microwave irradiation at 165 °C for 2 hours. The reaction mixture was cooled to room temperature. A precipitated solid was collected by filtration and washed with hexanes. The solid was purified by column chromatography on SiO2 (EtOAc:MeOH = 10:1) to afford N- (4-(5-(2-aminopyrimidin-4- yloxy)-lH-benzo[d]imidazole-2-yl)phenyl)quinolin-4-amine (Example 4) (13.0 mg, 21%) as a yellow solid.

LC MS m/z 446.3 [M+H]+, Rt = 2.65 min

Scheme 4 shows the synthesis of Example 5.

Scheme 4.

Intermediate 2: N-(4-(5-nitro-lH-benzo [d]imidazole-2-yl)phenyl)cyanamide To a solution of 4-(5-nitro-lH-benzo[d]imidazol-2-yl)aniline (1) (500 mg, 1.97 mmol) in MeOH (9.8 mL) was added cyanic bromide (312 mg, 2.95 mmol) followed by KOAc (772 mg, 7.87 mmol) at 0 °C . The reaction mixture was stirred at room temperature for 4 days. A precipitated solid was collected by filtration and washed with hexanes. The solid was purified by column chromatography on SiO2 (EtOAc:MeOH = 10: 1) to afford N-(4-(5- nitro-lH-benzo [d]imidazole-2-yl)phenyl)cyanamide (2) (400 mg, 73%) as a brown oil.

Intermediate 3 : N-(4-(5 -nitro- 1 H-benzo[d] imidazol-2-yl)phenyl)- 1 H-tetrazol-5 - amine

To a solution of N-(4-(5-nitro-lH-benzo[d]imidazol-2-yl)phenyl)cyanamide (2) (30.0 mg, 0.107 mmol) in DMF (0.54 mL) was added sodium azide (10.5 mg, 0.161 mmol) followed by A1C13 (7.16 mg, 0.0540 mmol) at room temperature. The reaction mixture was heated at 120 °C for 1 hour and cooled to room temperature. A precipitated solid was collected by filtration and washed with hexanes. The solid was purified by column chromatography on Si02 (EtOAc.-MeOH = 10:1) to afford N-(4-(5-nitro-lH-benzo[d]imidazol-2-yl)phenyl)- lH-tetrazol-5-amine (3) (28.0 mg, 81%) as a brown solid.

Intermediate 4 : 2-(4-( 1 H-tetrazol-5 -ylamino)phenyl)- 1 H-benzo[d] imidazo 1-5 -amine

A suspension of of N-(4-(5 -nitro- lH-benzo[d]imidazol-2-yl)phenyl)-l H-tetrazol-5 -amine (3) (142 mg, 0.441 mmol) and Pd/C (10wt%, 46.9 mg, 0.0220 mmol) in MeOH (4.4 mL) was stirred at room temperature for 1 hour under H2 atmosphere (balloon). The reaction mixture was filtered through a Celite pad and the filtrate was concentrated in vacuo to give 2-(4-(lH- tetrazol-5-ylamino)phenyl)-lH-benzo[d]imidazol-5-amine (4) (1 19 mg, 92%) as a brown solid, which was used for the next step without further purification.

Example 5: N4-(2-(4-(lH-tetrazol-5-ylamino)phenyl)-lH-benzo[d]imidazol-5-yl) pyrimidine-2,4-diamine

A solution of 2-(4-(lH-tetrazol-5-ylamino)phenyl)-lH-benzo[d]imidazol-5-amine (4) (54.0 mg, 0.185 mmol) and 4-chloropyrimidin-2-amine (5) (23.9 mg, 0.185 mmol) in water (0.92 mL) was refluxed for 8 hours and cool to room temperature. A precipitated solid was collected by filtration and washed with hexanes. The solid was purified by prep-HPLC to afford N4-(2-(4-( 1 H-tetrazol-5-ylamino)phenyl)- 1 H-benzo [d] imidazol-5 -yl)pyrimidine-2,4- diamine (Example 5) (10.0 mg, 14%) as a yellow solid.

LC/MS m/z 386.1 [M+HJ+, Rt = 2.73 min

Test Example 1: DNMT1 Inhibitory Assay (IC₅₀ measurement)

Reaction buffer was prepared with 50 mM Tris-HCl (pH 7.5), 50 mM NaCl, 5 mM EDTA, 5 mM DTT, 0.1 mM PMSF, 5% glycerol, and 1% DMSO. h-DNMT assay was used Human Full-length DNMT1 (GenBank Accession No. NM_001130823), (aa 2-1632), with N-terminal GST tag, MW=209 kDa, expressed in baculovirus expression system and Substrate was Poly(dl-dC) for DNMTl ; Poly(dI- dC)rPoly(dl-dC) Sigma cat# P4929. The reaction conditions was following proportion; DNMTl : 100 nM DNMTl, 0.001 mg/ml Poly(dl-dC), 1 μΜ SAM.

After preparation of indicated substrate in freshly prepared Reaction Buffer, deliver indicated DNMT into the substrate solution and gently mixture. The compounds was delivered in DMSO into the DNMT reaction mixture by using Acoustic Technology (Echo 550, LabCyte Inc. Sunnyvale, CA), and incubated for 20 min. ³H-SAM was delivered into the reaction mixture to initiate the reaction and incubated for 60 min at 30°C. The reaction mixture was filtered by paper for detection and analyzed data using Excel and GraphPad Prism software for IC50 curve fits.

Test Example 2: Human/ mouse Liver microsomal stability

Human and mouse liver microsomal clearance assays were carried out in CROWN Biosciences (Taicang, China). The human liver microsomes (Cat No. X008067, Lot No. KQB) and mouse liver microsomes (Cat No. Ml 000, Lot No. 1210302) were purchased from Celsis and Xenotech, respectively.

5 uL of test compound stock solution was diluted with 495 uL of 1 : 1 Methanol/Water (final concentration: 100 μΜ, 50% MeOH) and combined with 534 μΕ of the respective liver microsome solution (final concentration: 1.111 μΜ, 0.555% MeOH). The final concentration of the liver microsome solution was 0.7 mg protein /mL.

Incubations of the liver microsome solutions were performed in a 96 well plate at 37°C. 90 μί of the liver microsome solutions were added to Blank, and 90 μί of working solution of the test compound was added to all plates except the Blank.

All plates thus obtained were warmed in water bath at 37°C for 10 min, and 10 L of NADPH co-factor solution comprising 42 mg of β-nicotinamide adenine dinucleotide phosphate (Sigma Cat. No. N0505 Lot 020M7009V), 84 mg of isocitric acid (Sigma Cat. No. 11252 Lot 119K1099) and 0.478 mL of isocitric dehydrogenase (Sigma Cat. No. 12002 Lot 086K7055, 15 units/mg protein, warmed in water bath at 37°C for 5 min was added to the plates. The resulting plates were incubated at 37°C in the following order: T60 (The test compound was incubated with the liver microsomal solution and NADPH for 60 min at 37°C), T30 (such as for 30 min.), and T10 (such as for 10 min.). 300 uL of a cold (4°C) stop solution (acetonitrile (ACN) including 500 nM of tolbutamide as internal standard) and 10 uL of NADPH co-factor solution to starting plate (TO: 100% of the parent compound without any reaction) were added to the plates. The reaction was stopped by adding 300 μL of the cold (4°C) stop solution to the other plates in the following order: T10 first, then T30 and T60.

The sample was centrifuged at 4,000 rpm for 20 min and transferred to Bioanalytical Services for liquid chromatography-mass spectrometry (LC-MS)/MS (Waters UPLC/API 4000, 10 μL injection) analysis. Test Example 3: Caco-2 permeability assay

The Caco-2 cells (Passage 48) were diluted with culture medium into cell concentration of 6.86x105 cells/mL. This cell concentration can be used to seed 2.40x105 cells/cm2 and dispense 50 uL into the filter well of the 96-well HTS Transwell plate. Cells were cultivated in a cell culture incubator set at 37 °C, 5% CO2, 95% relative humidity. Incubate the plate for 14-18 days and replaced the medium every other day, beginning no sooner than 48 hours after initial plating. The standard transport buffer in the study was HBSS at pH 7.4. Before and after the transport studies, the monolayer integrity was evaluated by measuring transepithelial electrical resistance (TEER). The pre-transport and post-transport TEER were required to be no less than 200 Ω-cm². After incubate plate for 14 days, remove the Caco-2 plate from the incubator. Wash the inserts 2 times with pre-warmed HBSS (10 mM HEPES, pH 7.4). Place the inserts into receiver plate. Add 75 uL and 235 buffer to each Transwell inserts and receiver wells, respectively. Incubate the filter inserts under gentle shaking (150 rpm) for 30 min at 37 °C. The stock solutions of test compound and control compounds were prepared in DMSO and dilute with HBSS (10 mM HEPES, pH 7.4). The final concentration of the test compounds and control compounds was 5 μΜ. To determine the rate of drug transport in the apical to basolateral direction, add 75 μΐ_^ of the test compounds to the filter well (apical compartment). Fill each well in the receiver plate (basolateral compartment) with 235 uL HBSS (10 mM HEPES, pH 7.4). To determine the rate of drug transport in the basolateral to apical direction; add 235 μΐ. of the test compounds to each well of the receiver plate. Fill the transwell inserts with 75 μί, HBSS (10 mM HEPES, pH 7.4). Incubate at 37 °C with shaking at 150 rpm on a rotary shaker for 2 hours. At the end of the transport period, remove 50 uL directly from the apical and basolateral wells and transfer to a new plate. Then add 4 volume of cold acetonitrile containing internal standards (IS, 100 nM Alprazolam, 200 nM Labetalol and 2 μΜ Ketoprofen) to terminate the reaction. Vortex for 5 minutes. Samples are centrifuged at 3,220 g for 20 minutes. An aliquot of 200 μΐ, of the supernatant is used for LC/MS/MS analysis. All incubations are performed in duplicate. Data analysis method was below;

The Papp value has the dimension of a rate (10-6cm/s).

Papp = CRecx VRec/ (A x t x C₀) (1)

Where

C_Rec is the compound concentration in the receiver chamber at the measurement time t;

V_Recis the volume of the receiver chamber;

A is the area of the permeability barrier, which corresponds to the surface area of the filter (0.143 cm2 for HTS Transwell-96 Well Permeable Supports);

Co is the initial concentration in the donor chamber (μΜ).

The Efflux ratio is derived using the equation (2):

Efflux Ratio = P_app (B→A) P_apP (A→B) (2)

A compound is considered as likely Pgp substrate when the efflux ratio is more than

2.0.

Recovery is calculated using the equation (3):

Recovery - [(V_rec x C_rec) + (V_d x C_d)] / (V_d x Co) (3)

Where

V_d and V_rec are the volume of donor and receiver chambers;

C_d and C_rec are the final concentrations of test compound in donor and receiver chambers at time t;

Co is the initial concentration in the donor chamber at time Zero. Test Example 4 : Thermodynamic solubility assay

After weighing dry powders of the test compounds about 1 mg, phosphate buffer (pH 7.0) was added to the 4-ml glass sample vials respectively to make a solution with a nominal concentration of lmg/ml working solution. The sample vials were incubated at room temperature (25 °C), 600rpm for 24 hours. All samples were centrifuged at 14000 rpm, 25°C for 4 minutes. A standard solution was prepared by dissolving the weighed powders of the same test compound in pure DMSO (1 mg/ml), then diluted the lmg/ml standard solution to 200 μg/ml, 40 μg/ml and 8 μg/ml standard solutions with 100% DMSO. 20 μΤΛνεΙΙ working and standard solution were transferred to a new plate which contain 580 μΐ/ννεΐΐ ACN:H2O 20:80 (including 40ng/ml Tolbutamide as internal standard), mixed. 20 μΙ,ΛνεΙΙ step 5 solutions were transferred to a new plate which contain 580 μΙ_Λνε11 ACN:H2O 20:80 (including 40ng/ml Tolbutamide as internal standard), mixed. 20 μΐ ννεΐΐ step 6 solutions were transferred to a new plate which contain 580 uL/well ACN:H₂O 20:80 (including 40ng/ml Tolbutamide as internal standard), mixed. This sample of step above was transferred for LC-MS/MS analysis.

The result is shown in Table 1. [Table 1]

Claims

What is claimed is:

1. A compound of Formula I, or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof:

Formula I

wherein,

Z is selected from NH, O or S;

Xj and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(0)N(R')-, - N(R')C(S)N(R')-, -N(R')SO₂-, -N(R')S0₂N(R')-, -N(R')N(R')-, -N(R')C(=N(R'))N-, - S0₂N(R')-, -0-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R')-, -C(O)N(R')-, -C=N (R')-, - C=NO-, -C(=N(R'))N(R')- or -C(R')₂-;

Y is a phenyl group optionally substituted by C_w alkyl, CF₃, halogen, hydroxyl, Ci_4 alkoxy, cyano, nitro, halo C^ alkyl, halo C^ alkoxy, hydroxyl C^ alkyl, C^ alkoxy Ci-4 alkyl, or C_1--4 alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by Ci_4 alkyl, CF₃, halogen, hydroxyl, C_1-- alkoxy, cyano, nitro, halo C^ alkyl, halo Ci-4 alkoxy, hydroxyl

alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by C^₄ alkyl, CF₃, halogen, hydroxyl, C^ alkoxy, cyano, nitro, halo C_1--4 alkyl, halo C^₄ alkoxy, hydroxyl C^ alkyl, C^alko yC^ alkyl, or C_M alkoxycarbonyl ; C^ alkyl, C_2~6 alkynyl, or C₂~6 alkenyl;

R_\ and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5

wherein,

Rt and Rs are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, - NO₂, -C(O)R\ -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R\ -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')S0₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R\ or -OC(O)N(R')₂;

B] and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

-NO₂, -C(O)R', -C(S)R', -CO₂R\ -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR\ -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

R' is hydrogen, Ci^ aliphatic, C^₆ alkoxy, Ci~₆ alkyl, phenyl, CF₃, halogen, hydroxyl, C1-4 alkoxy, cyano, nitro, halo Ci~ alkyl, halo Ci_₄ alkoxy, hydroxyl Ci_~4 alkyl, C^ alkoxycarbonyl, C^₆ alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen.

2. A compound of Formula II, or a pharmaceutically acceptable salt, a hydrate, a solvate, or an isomer thereof:

Formula II

wherein,

Xi and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')C(S)N(R')-, -N(R')SO₂-, -N(R')SO₂N(R , -N(R')N(R')-, -N(R')C(=N(R'))N-, - SO₂N(R')-, -O-, -CO-, -CS-, -C(O)S-, -OC(O)-, -C(O)R'C(O)R'-, -C(S)O-, -C(O)O-, - S(O)n(wherein, n is an integer of 0 to 2), -OC(O)N(R')-, -C(O)N(R')-, -C=NN(R')-, - C=NO-, -C(=N(R'))N(R')- or -C(R')₂-; Y is a phenyl group optionally substituted by C_1--4 alkyl, CF₃, halogen, hydroxyl, Ci-4 alkoxy, cyano, nitro, halo C^₄ alkyl, halo Ci_4 alkoxy, hydroxyl C1-4 alkyl, C^ alkoxy C_1--4 alkyl, or alkoxycarbonyl ; a 5- to 6- membered heteroaromatic group optionally substituted by alkyl, CF₃, halogen, hydroxyl, alkoxy, cyano, nitro, halo alkyl, halo C^ alkoxy, hydroxyl C^₄ alkyl, alkoxyC^ alkyl, or C_1--4 alkoxycarbonyl ; a 5- to 6-membered heterocarbocyclic group optionally substituted by C_1--4 alkyl, CF₃, halogen, hydroxyl, C_1--4 alkoxy, cyano, nitro, halo C_1--4 alkyl, halo C_M alkoxy, hydroxyl Ci_₄ alkyl, Ci-4 alkoxyC^ alkyl, or alkoxycarbonyl ; C^₆ alkyl, C₂~6 alkynyl, or C_2~ alkenyl;

R₃ is selected from R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R', - C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, - C(S)OR', -S(O)R', -SO₂R\ -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Rj and R₂ are each independently selected from the following groups Ql to Q5;

Q1 Q2 Q3 Q4 Q5

wherein,

R4 and R6 are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, - NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR', -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Bi and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N; R₅ and R₅' are each independently R', halogen, -OR', -SR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)SR', -C(O)C(O)R', -C(O)CH₂C(O)R', -C(S)N(R')₂, -C(S)OR\ -S(O)R', -SO₂R', -SO₂N(R')₂, -N(R')C(O)R', -N(R')C(O)N(R')₂, - N(R')C(S)N(R')₂, -N(R')SO₂R', -N(R')SO₂N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, - C=NN(R')₂, -C=NOR', -C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH, N, or SH;

Ei, E₂, E₃ and E are each independently selected from CH₂, NH, S, SH₂, or O; p is an integer of 1 to 3; and

R' is hydrogen, C^₆ aliphatic, C^₆ alkoxy, C_1--6 alkyl, phenyl, CF₃, halogen, hydroxyl, C_1--4 alkoxy, cyano, nitro, halo alkyl, halo alkoxy, hydroxyl C^₄ alkyl, alkoxycarbonyl, C^₆ alkyl substituted with amino or hydroxyl, or 5- to 6-membered heterocyclic having one or more oxygen, sulfur or nitrogen.

3. The compound of claim 1, wherein

Z is selected from NH or O;

Xi and X₂ are each independently -NR'-, N(R')C(O)-, -N(R')C(O)N(R')-, - N(R')N(R')-, -N(R')C(=N(R'))N-, -O- , -CO-, -OC(O)-, -C(O)R'C(O)R'-, -C(O)O-, - OC(O)N(R')-, -C(O)N(R , -C=NN(R , -C=NO-, -C(=N(R'))N(R')- or -C(R')₂-;

Y is a 5- to 6-membered heteroaromatic group optionally substituted by C^ alkyl, CF₃, halogen, hydroxyl, C_1-- alkoxy, cyano, nitro, halo Ci^ alkyl, halo C ^alkoxy, hydroxyl Ci_ alkyl,

or C_M alkoxycarbonyl;

Ri and R₂ are each independently selected from the following groups Ql to Q5;

m(R₄)|¹2¾-(Re)w

Q1 Q2 Q3 Q4 Q5

wherein,

R_t and R_$ are each independently R', halogen, -OR', -N(R')₂, -CN, -CF₃, -NO₂, -C(O)R', -CO₂R', -C(O)N(R')₂-C(O)C(O)R', -C(O)CH₂C(O)R', -N(R')C(O)R', - N(R')C(0)N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, -C=NN(R')₂, -C=NOR', - C(=N(R'))N(R')₂, -OC(O)R', or -OC(O)N(R')₂;

Bi and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R', halogen, -OR', -N(R')₂, -CN, -CF₃, -NO₂,

-C(O)R', -C(S)R', -CO₂R', -C(O)N(R')₂, -C(O)C(O)R', -C(O)CH₂C(O)R', -N(R')C(O)R', - N(R')C(O)N(R')₂, -N(R')N(R'), -N(R')C(=N(R'))N(R')₂, -C=NN(R')₂, -C=NOR\ - C(=N(R'))N(R')₂, -OC(O)R' or -OC(O)N(R')₂;

Dj, D₂, D₃ and D₄ are each independently selected from CH, N, or SH; Ei, E₂, E₃ and E₄ are each independently selected from CH₂, NH, or O; p is an integer of 0 to 3; and

R' is hydrogen, or an optionally substituted group selected from Ci_₆ aliphatic,

C_1--6 alkoxy, C^₆ alkyl, phenyl, CF₃, halogen, hydroxyl or Ci~₄ alkoxy, cyano, nitro, halo Ci_4 alkyl, halo C^ alkoxy, hydroxyl C^₄ alkyl, C_1--4 alkoxycarbonyl, or a C^ alkyl substituted with amino or hydroxyl.

4. The compound of claim 1, wherein

Z is NH;

Xi and X₂ are each independently -NR' or -O- ;

Y is a 5- to 6-membered heteroaromatic group optionally substituted by Ci^ alkyl, CF₃, halogen, hydroxyl, C^₄ alkoxy, cyano, nitro, halo C^ alkyl, halo C^₄ alkoxy, hydroxyl C^ alkyl, C^ alkoxy, C^ alkyl or Ci~₄ alkoxycarbonyl ;

Q1 Q2 Q3 Q4 Q5

wherein,

A is a 6-membered aryl or fused heteroaryl ring having 2 to 3 nitrogen atoms; R4 and R are each independently R', -N(R')₂ or -CN;

B and B₂ are each independently selected from CH or N;

m is 1 or 2;

w is an integer of 1 to 4;

Ci, C₂ and C₃ are each independently selected from CH or N;

R₅ and R₅' are each independently R' or -N(R')₂;

Di, D₂, D₃ and D₄ are each independently selected from CH or N;

R' is hydrogen, C^ alkyl, cyano, nitro, or a C^₆ alkyl substituted with amino.

5. The compound of claim 1 or 2, which is selected from the group consisting of: 6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine;

6-methyl-N4-(2-(4-(quinolin-4-ylamino)phenyl)- 1 H-benzo[d] imidazol-5 - yl)pyrimidine-2,4-diamine tri-hydrogen chloride;

2-ammo-4-methyl-6-((2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5- yl)amino)pyrimidine 1 -oxide;

N-(4-(5-(2-aminopyrimidin-4-yloxy)- 1 H-benzo[d] imidazole-2-yl) phenyl)quinolin- 4-amine;

N4-(2-(4-( 1 H-tetrazol-5 -ylamino)phenyl)- 1 H-benzo [d] imidazol-5 -yl) pyrimidine- 2,4-diamine;

6-methyl-N4-(2-(4-((2-methylpyridin-4-yl)amino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine;

6-methyl-N4-(2-(4-(pyridin-4-ylamino)phenyl)- 1 H-benzo [d] imidazol-5 - yl)pyrimidine-2,4-diamine;

N4-(2-(4-(quinolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5-yl)pyrimidine-2,4- diamine;

N-(4-(5-(pyridin-2-ylamino)-lH-benzo[d]imidazol-2-yl)phenyl)quinolin-4-amine; N4-(2-(4-((2-methylpyridin-4-y l)amino)pheny 1)- 1 H-benzo [d] imidazol-5 - yl)pyrimidine-2,4-diamine trihydrochloride;

2-(4-((2-methylpyridin-4-yl)amino)phenyl)-N-(pyridin-2-yl)-lH-benzo[d]imidazol- 5 -amine tris(2,2,2-trifluoroacetate);

4-methyl-N2-(2-(4-(pyridin-4-ylamino)phenyl)- 1 H-benzo [d]imidazol-5-yl)pyridine- 2,6-diamine trihydrochloride;

6-methyl-N4-(2-(4-(pyridin-2-y lamino)phenyl)- 1 H-benzo [d] imidazol-5- yl)pyrimidine-2,4-diamine trihydrochloride;

6-methyl-N4-(2-(4-((3-methylpyridin-4-yl)amino)phenyl)-lH-benzo[d]imidazol-5- yl)pyrimidine-2,4-diamine trihydrochloride;

N-(4-(5 -(quinolin-4-ylamino)- 1 H-benzo [d] imidazol-2-yl)phenyl)quinolin-4-amine trihydrochloride;

N-(4-((2-(4-(quinolin-4-ylamino)phenyl)- 1 H-benzo [d] imidazol-5 - yl)amino)pyrimidin-2-yl)acetamide dihydrochloride;

N4-(2-(4-(pyridm-4-ylamino)phenyl)-lH-benzo[d]imidazol-5-yl)pyrimidine-2,4- diamine trihydrochloride;

N-(4-(5-((2-ammopyrimidin-4-yl)oxy)-lH-benzo[d]imidazol-2-yl)phenyl)quinolin- 4-amine trihydrochloride;

N4-(2-(4-(qumolin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5-yl)pyridine-2,4- diamine trihydrochloride;

2-amino-4-methyl-6-((2-(4-(quinolin-4-ylamino)phenyl)-l H-benzo [d] imidazol-5 - yl)amino)pyrimidine 1 -oxide trihydrochloride;

N4-(2-(4-((2,6-dimethylpyridin-4-yl)amino)phenyl)-lH-benzo[d]imidazol-5-yl)-6- methylpyrimidine-2,4-diamine trihydrochloride;

N4-(2-(4-((2,3-dimethylpyridin-4-yl)amino)phenyl)-lH-benzo[d]imidazol-5-yl)-6- methylpyrimidine-2,4-diamine trihydrochloride; N4-(4-(5-((2-amino-6-methylpyrimidin-4-yl)amino)-lH-benzo[d]imidazo yl)phenyl)-6-methylpyrimidine-2,4-diamine trihydrochloride;

(Z)-2-cyano- 1 -(2-(4-(pyridin-4-ylamino)phenyl)- 1 H-benzo [d] imidazol-5 - yl)guanidine trihydrochloride;

N4-(2-(4-(pyrimidin-4-ylamino)phenyl)- 1 H-benzo [d] imidazol-5 -yl)pyrimidine-2,4- diamine trihydrochloride;

N-(pyridin-4-yl)-2-(4-(pyridin-4-ylamino)phenyl)-lH-benzo[d]imidazol-5-amine trihydrochloride; and

N4-(2-(4-((l H-tetrazol-5-yl)amino)phenyl)- 1 H-benzo[d] imidazol-5 -yl)pyrimidine- 2,4-diamine tris(2,2,2-trifluoroacetate).

6. A pharmaceutical composition comprising the compound of Formula I as defined in claim 1 or the compound of Formula II as defined in claim 2, and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition of claim 6, which further comprises at least one additional pharmaceutical agent.

8. A method for treating a disease modulated by inhibiting DNA methyltransferase activity in cells in a subject comprising: administering to the subject in need of such treatment a medically effective amount of the compound of Formula I as defined in claim 1 or the compound of Formula II as defined in claim 2.

9. The method of claim 8, wherein the disease is selected from the group consisting of an atherosclerosis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, liver cirrhosis, cancer, rheumatoid arthritis, and Alzheimer's disease.

10. A method for treating a disease modulated by inhibiting DNA methyltransferase activity in cells in a subject comprising: administering to the subject in need of such treatment, a medically effective amount of two pharmaceutical compositions comprising (i) a first composition comprising the compound of Formula I as defined in claim 1 or the compound of Formula II as defined in claim 2, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising at least one additional pharmaceutical agent selected from a chemotherapeutic agent, an anti-inflammatory drug, a COX-2 inhibitor, an immune modulator, a cholinesterase inhibitor, or an NMDA receptor antagonist.

11. The method of claim 10, wherein the first composition and second composition are administered simultaneously.

12. The method of claim 10, wherein the first composition and second composition are administered sequentially and in any order.

13. A use of the compound of Formula I as defined in claim 1 or Formula II as defined in claim 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disease modulated by inhibiting DNA methyltransferase activity in cells.

14. The use of claim 13, wherein the disease is selected from the group consisting of an atherosclerosis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, liver cirrhosis, cancer, rheumatoid arthritis, and Alzheimer's disease.