Trisubstituted pyrimidine compounds and compositions for the treatment of cancer, retinal disorders and cardiomyopathy

Abstract

The present invention relates to a compound of formula (I): or a pharmaceutically acceptable salt or solvate thereof. The present invention further discloses pharmaceutical compositions comprising the compounds of formula (I) and, in particular, their use in the treatment of diseases or disorders associated with an increase in expression or function relative to physiological or desired RhoJ / Cdc42 levels. [Selection diagram] None

Description

[Technical Field]

[0001] [Priority claim] This application claims priority from Italian Patent Application No. 102017000047189, filed May 2, 2017, the disclosure of which is incorporated herein by reference.

[0002] The present invention relates to novel RhoJ inhibitors for the treatment of cardiomyopathy, retinopathy and cancer, particularly melanoma.

[0003] The incidence of melanoma, a type of cancer that develops from the malignant transformation of melanocytes, is increasing. Unfortunately, conventional therapies, such as decarbazine and high-dose interleukin-2 (IL-2) chemotherapy, are cytotoxic and poorly effective, creating a need for new therapeutic approaches.

[0004] More recent strategies include the use of BRAF inhibitors, such as vemurafenib. Immunotherapeutic approaches using antibodies that block immune checkpoint molecules, such as ipilimumab, which targets anti-cytotoxic T-lymphocyte antigen 4 (CTLA-4), are currently being used to treat unresectable melanoma. Further studies are using ipilimumab in combination with PD1 inhibitors, such as nivolumab. While these therapies can induce dramatic responses in some patients, only 30% of patients respond to these therapies. Improved therapy is especially needed for early-stage disease, where only interleukin-2 and oncolytic virus therapy have been used with very limited success.

[0005] In this context, an alternative and promising approach is the combination of therapies, which combine multiple drugs and conventional anti-cancer modalities (radiation, chemotherapy or surgery) to increase therapeutic efficacy and reduce side effects.

[0006] However, all these potential therapeutic strategies have not resulted in a significant reduction in the mortality rate of melanoma patients and the progression of metastatic melanoma. Intrinsic and / or acquired resistance to chemotherapy remains a major problem, and there is a strong need to identify new pathways for treating melanoma. Furthermore, identifying agents with limited toxicity that can be used to treat early stage disease (stage III) has important therapeutic applications.

[0007] RhoJ is now proposed as a validated target in complex therapeutic strategies because its expression regulates melanoma development. RhoJ is a member of the Rho family of small GTPases known to bind and activate PAK kinases. Functional validation studies have revealed that RhoJ activates PAK1, which in turn inhibits p53 signaling and apoptotic pathways in melanoma cells in vitro. [Background technology]

[0008] Further studies have shown that RhoJ and PAK1 also regulate melanoma cell migration and invasion in vitro and tumor growth and invasion in melanoma xenograft models. While RhoJ is known to play a role in endothelial cell biology and angiogenesis, it was unknown whether RhoJ has cell-autonomous effects on melanocytic tumor growth. Recent studies have determined that RhoJ regulates melanoma tumor growth and development in spontaneous mouse models and is expressed at higher levels in stage III compared to stage IV melanomas. Overall, these in vitro and in vivo studies have revealed that RhoJ and PAK1 promote the growth of nascent melanoma tumors, inhibit apoptosis, and stimulate angiogenesis. Recent studies have shown that RhoJ functions to inhibit apoptosis in tumor cells by blocking the phosphorylation of BAD, preventing BAD from inducing apoptosis. RhoJ interaction inhibitors blocked PAK kinase-induced BAD phosphorylation, and similar effects were observed with PAK inhibitors.

[0009] Furthermore, RhoJ is also believed to play a central role in the pathophysiology of cardiomyopathies such as dilated cardiomyopathy (DCM) and as a potential therapeutic target for specifically manipulating endothelial filopodial protrusions.

[0010] For these reasons, blocking RhoJ may be considered a useful tool to treat conditions such as cardiomyopathy, retinopathy, stage III or stage IV melanoma, and other cancers that are resistant to other therapies.

[0011] Therefore, there is a need for novel compounds that inhibit RhoJ. Summary of the Invention [Problem to be solved by the invention]

[0012] An object of the present invention is to provide novel compounds that act as RhoJ inhibitors. [Means for solving the problem]

[0013] The above-mentioned object has been achieved according to the compounds according to claim 1, the pharmaceutical compositions according to claim 7 and the uses according to claims 10, 11 and 12. Preferred embodiments are presented in the dependent claims.

[0014] The present invention will now be described in detail with reference to the accompanying drawings. [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 shows apoptosis induced in WM3248 melanoma cells by the RhoJ interaction inhibitor ARN12405 (Compound 1) as measured by flow cytometry using Annexin V and PI. [Figure 2] Blockade of RhoJ PAK interaction in cell lysates from WM3248 cells incubated with compound 1 (ARN12405) as assessed by SDS PAGE and immunoblotting using (Panel A) RhoJ antibody, (Panel B) Cdc42 antibody, and (Panel C) Rac1 antibody. (Panel D) Cells treated with compound 1 (ARN12405) at concentrations of 10 μM or 50 μM. DETAILED DESCRIPTION OF THE INVENTION

[0016] The following paragraphs provide definitions of the various chemical moieties of compounds according to the invention and are intended to apply uniformly throughout the specification and claims, unless a differently stated definition gives a broader definition.

[0017] The term "alkyl," as used herein by itself or as part of another substituent, refers to an aliphatic hydrocarbon group. Such term includes linear (unbranched) or branched chains and may be fully saturated, monounsaturated, or polyunsaturated.

[0018] The term "unsaturated" aliphatic hydrocarbon group includes alkenyl and alkynyl.

[0019] The term "alkenyl," as used herein, refers to alkyl groups preferably containing from 2 to 6 carbon atoms and containing at least one carbon-carbon double bond.

[0020] The term "alkynyl," as used herein, refers to alkyl groups preferably containing from 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond.

[0021] According to the present invention Alkyl, alkenyl and alkynyl groups Non-limiting examples are, for example, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1- or 2-butenyl, ethynyl, 1-propynyl, 2-propynyl, 1- or 2-butynyl, and the like.

[0022] The term "alkoxy," as used herein, refers to an alkyl group that is linked to the remainder of the compound by an oxygen atom.

[0023] The term "halogen" as used herein refers to fluorine, chlorine, bromine and iodine.

[0024] The term "aromatic ring," as used herein, refers to a moiety whose constituent carbon atoms make up an unsaturated ring system, and all atoms in the ring system are sp 2 hybridization, and the total number of pi electrons is equal to 4n+2, where n is an integer.

[0025] The term "aromatic heterocycle," as used herein, refers to an aromatic ring as defined above in which 1 to 4 carbon atoms are replaced by heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Non-limiting examples of aromatic heterocycle groups include pyrrolyl, furyl, thiophenyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, indolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, benzopyrazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, triazolyl, oxadiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinolinyl, isoquinolinyl, quinazolinyl, and quinoxalinyl.

[0026] Unless otherwise indicated, the term "substituted," as used herein, means that one or more hydrogen atoms of the group described above have been replaced with another non-hydrogen atom or functional group, provided that the normal valences are maintained and that the substitution results in a stable compound.

[0027] The term "pharmaceutically acceptable salts" refers to salts of the compounds of Formula (I) identified below that retain the desired biological activity and that are approved by regulatory authorities.

[0028] As used herein, the term "salt" refers to any salt of a compound according to the invention prepared from an inorganic or organic acid or base and internally formed salts. Typically, such salts have a physiologically acceptable anion or cation.

[0029] Furthermore, the compounds of formula (I) may form acid addition salts or salts with bases depending on the types of substituents, and these salts are included in the present invention as long as they are pharmaceutically acceptable salts.

[0030] Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.), as well as salts formed with organic acids such as acetic acid, trifluoroacetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, alginic acid, polyglutamic acid, and naphthalenesulfonic acid.

[0031] Physiologically or pharmaceutically acceptable salts are particularly suitable for medical applications due to their greater aqueous solubility relative to the parent compounds.

[0032] Pharmaceutically acceptable salts may also be prepared from other salts, including other pharmaceutically acceptable salts, of compounds of formula (I) using conventional methods.

[0033] Those skilled in the art of organic chemistry will understand that many organic compounds can form complexes with the solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates." For example, a complex with water is known as a "hydrate." Solvates of the compounds of the present invention are within the scope of the present invention. The compounds of formula (I) can be easily isolated with associated solvent molecules by crystallization or evaporation of the appropriate solvent to obtain the corresponding solvate.

[0034] The compound of formula (I) may exist in crystalline form. In certain embodiments, the crystalline form of the compound of formula (I) is a polymorph.

[0035] The subject invention also encompasses isotopically labeled compounds identical to the compounds of formula (I) and listed below, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes that may be incorporated into the compounds of the invention and their pharmaceutically acceptable salts include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, e.g., 2 H, 3 H, 11 C.13 C. 14 C. 15 N, 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, 125 I can be mentioned.

[0036] Compounds of the present invention and pharmaceutically acceptable salts of said compounds that contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of the present invention. Isotopically labeled compounds of the present invention, e.g., 3 H, 14 Those in which radioactive isotopes such as 3C are incorporated are useful in drug and / or substrate tissue distribution assays. 3 H and carbon-14, i.e. 14 C isotopes are particularly preferred for their ease of preparation and detectability. 11 C and 18 F isotopes are particularly useful in PET (positron emission tomography), 125 I isotopes are particularly useful in SPECT (single photon emission computed tomography), all of which are useful for imaging the brain. 2 Substitution with heavier isotopes such as H can offer certain therapeutic advantages due to greater metabolic stability, e.g., increased half-life in vivo or reduced dosage requirements, and therefore may be preferred in some circumstances. The isotopically labeled compounds of formula (I) of the invention and described below can generally be made by carrying out the procedures disclosed in the schemes and / or examples below, substituting readily available isotopically labeled reagents for non-isotopically labeled reagents.

[0037] Certain groups / substituents encompassed by the present invention may exist as isomers, or in one or more tautomeric forms. Thus, in certain embodiments, compounds of formula (I) may exist in other tautomeric or geometric isomeric forms, depending on the type of substituent. Although a compound may be described herein in only one of such isomeric forms, the present invention encompasses all such isomers, isolated forms of isomers, or mixtures thereof. Furthermore, compounds of formula (I) may optionally have asymmetric carbon atoms or axial asymmetry, and accordingly, compounds of formula (I) may exist in the form of optical isomers, such as (R)- and (S)-forms. The present invention encompasses within its scope all such isomers, including racemates, enantiomers, and mixtures thereof.

[0038] In particular, all stereoisomers, including enantiomers, diastereomers and mixtures thereof, including racemates, are included within the scope of the present invention, and general references to compounds of formula (I) include all stereoisomers unless otherwise stated.

[0039] In general, the compounds or salts of this invention should be construed to exclude compounds (if any) that are highly chemically unstable by themselves or in water and clearly unsuitable for pharmaceutical use by any route of administration, whether oral, parenteral, or otherwise. Such compounds are known to the expert chemist.

[0040] According to a first aspect of the present invention, a compound of formula (I): [ka] or a pharmaceutically acceptable salt or solvate thereof.

[0041] In the compound of formula (I), X1 and X2 are independently selected from the group consisting of CH2, NR2, and O, with the proviso that X1 and X2 are not both NR2, not both O, or not both NR2 and O; Y is selected from the group consisting of CH and N; A and A' are independently a 6-membered aromatic heterocycle containing 1 or 2 nitrogen atoms and, at any position, C 1~6 Alkyl, halogen, halo-C 1~6 Alkyl, hydroxyl, alkoxy-C 1~6 Alkyl, Amino, Amino-C 1~6 Alkyl and Amino-diC 1~6 a 6-membered aromatic ring optionally substituted with substituents selected from the group consisting of alkyl; R1 is hydrogen and C 1~6 alkyl, R2 is hydrogen, C 1~6 Alkyl, C 1~6 Alkyl-alkoxy, CO-C 1~6 Alkyl and CO-C 1~6 It is selected from the group consisting of alkyl-alkoxy.

[0042] According to the first embodiment, A and A' are independently a 6-membered aromatic heterocycle containing one nitrogen atom and, at any position, a halogen, an alkoxy-C 1~6 Alkyl, Amino, Amino-C 1~6 Alkyl, Amino-diC 1~6 a 6-membered aromatic ring optionally substituted with substituents selected from the group consisting of alkyl; R1 is hydrogen, R2 is hydrogen.

[0043] According to the second embodiment, X1 is selected from the group consisting of CH2, NH and O; X2 is selected from the group consisting of CH2 and NH, provided that X1 and X2 are not simultaneously N; Y is CH2, A is a 6-membered aromatic heterocycle containing one nitrogen atom at the 2- or 3-position, and halogen and alkoxy-C 1~3 a 6-membered aromatic ring optionally substituted with substituents selected from the group consisting of alkyl; A' is a halogen atom, an alkoxy group, or a methyl group at the meta or para position. 1~6 Alkyl and Amino-diC 1~6 a 6-membered aromatic ring optionally substituted with substituents selected from the group consisting of alkyl; R1 is hydrogen, R2 is hydrogen.

[0044] In further embodiments, A is selected from the group consisting of a 6-membered aromatic heterocycle containing one nitrogen atom and an unsubstituted 6-membered aromatic ring.

[0045] In a preferred embodiment, A' is a six-membered aromatic ring substituted at the para or meta position with a substituent selected from the group consisting of methoxy or amino-di-C2 alkyl.

[0046] According to a third embodiment of the present invention, the compound of formula (I) may be selected from the group consisting of:

[0047] [Table 1] TIFF0007776068000003.tif241169TIFF0007776068000004.tif223169TIFF0007776068000005.tif221169TIFF0007776068000006.tif114169

[0048] The compounds exemplified in this invention can be made from readily available starting materials using the following general methods and procedures exemplified, for example, in Michael Smith, Jerry March - March's Advanced Organic Chemistry: reactions mechanisms and structure - 6th Edition, John Wiley & Sons Inc., 2007.

[0049] It is known to those skilled in the art that the conversion of a chemical function into another functional group may require that one or more reactive centers in the compound containing this functional group be protected to avoid undesired side reactions. The protection of such reactive centers, and subsequent deprotection at the end of the synthetic transformation, can be achieved according to standard procedures, for example, as described in Theodora W. Green and Peter GM Wuts - Protective Groups in Organic Synthesis, Fourth Edition, John Wiley & Sons Inc., 2006.

[0050] Where typical or preferred experimental conditions (i.e., reaction temperatures, times, moles of reagents, solvents, etc.) are given, it will be understood that other experimental conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by one skilled in the art using routine optimization procedures.

[0051] The synthesis of compounds of formula (I) can be carried out stepwise according to the synthetic process described below, whereby each intermediate is isolated and purified by standard purification techniques, such as column chromatography, before carrying out the subsequent reaction. Alternatively, two or more steps of the synthetic sequence can be carried out in a so-called "one-pot" procedure, as is known in the art, whereby only the compound resulting from two or more steps is isolated and purified.

[0052] The compounds of formula (I) made using the methods described herein below may be treated or purified by conventional techniques or means, for example, by filtration, distillation, chromatography, recrystallization, and combinations thereof.

[0053] Salts of compounds of formula (I) can be made by reacting the basic compound with the desired acid in solution.

[0054] A second aspect of the present invention relates to a pharmaceutical composition comprising a compound of formula (I) as disclosed above and a pharmaceutically acceptable carrier, stabilizer, diluent or excipient thereof.

[0055] Those skilled in the art are aware of a wide variety of such carrier, diluent or excipient compounds suitable to formulate pharmaceutical compositions.

[0056] The compounds of the present invention, together with conventional adjuvants, carriers, diluents or excipients, may be placed into the form of pharmaceutical compositions and unit dosages thereof, which may be used as solids such as tablets or filled capsules, or as liquids such as solutions, suspensions, emulsions, elixirs or capsules filled therewith, or in the form of sterile injectable solutions for parenteral administration (including subcutaneous and intravenous use), all for oral use. Such pharmaceutical compositions and their unit dosage forms may comprise the ingredients in conventional proportions, with or without additional active compounds or active ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range being used.

[0057] The pharmaceutical composition containing the compound of the present invention can be prepared by a method known in the pharmaceutical art, and comprises at least one active compound.Generally, the compound of the present invention is administered in a pharmaceutically effective amount.The amount of compound actually administered is usually determined by a doctor in consideration of the relevant circumstances, including the condition to be treated, the route of administration selected, the actual compound to be administered, the age, weight and response of individual patient, the severity of the patient's symptoms etc.

[0058] The pharmaceutical compositions of the present invention can be administered by a variety of routes including oral, rectal, subcutaneous, intravenous, intramuscular, intranasal, topical, intratumoral injection and pulmonary routes.

[0059] Compositions for oral administration can be in the form of bulk liquid solutions or suspensions, or bulk powders.However, more commonly, compositions are provided in unit dosage forms to facilitate accurate dosing.The term "unit dosage form" refers to a physically discrete unit suitable as a single dosage for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect, together with suitable pharmaceutical excipients.Typical unit dosage forms include pre-filled and pre-measured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc., in the case of solid compositions.

[0060] Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispersing agents, colorants, flavors, etc. Solid forms may contain, for example, the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth, or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetener such as sucrose or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavoring.

[0061] Injectable compositions are typically based upon injectable sterile saline or phosphate buffered saline or other injectable carriers known in the art.

[0062] The pharmaceutical compositions may be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories, and as sustained release formulations.

[0063] If desired, tablets may be coated using standard aqueous or nonaqueous techniques. In certain embodiments, such compositions and preparations may contain at least 0.1 percent of the active compound. The percentage of the active compound in these compositions may, of course, be varied and may conveniently be from about 1 percent to about 60 percent by weight of the unit. The amount of active compound in such therapeutically useful compositions is such that a therapeutically active dosage will be obtained. The active compound may also be administered intranasally, for example, as drops or a spray.

[0064] Tablets, pills, capsules, etc. may also contain binders such as gum tragacanth, gum arabic, corn starch, or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as corn starch, potato starch, or alginic acid; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose, or saccharin. When the dosage unit form is a capsule, in addition to the above-mentioned materials, it may contain a liquid carrier such as a fatty oil. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetener, methylparaben and propylparaben as preservatives, a dye, and a flavoring such as cherry or orange flavor. To prevent degradation during transit through the upper gastrointestinal tract, the composition may be enterically coated.

[0065] Topical administration of pharmaceutical compositions is particularly useful when the desired treatment involves areas or organs easily accessible by topical application. For topical application to the skin, pharmaceutical compositions are formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petroleum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compounds, emulsifying wax, and water. Alternatively, pharmaceutical compositions can be formulated with a suitable lotion or cream containing the active compounds suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. Topically transdermal patches and iontophoretic administration are also described.

[0066] Compositions for pulmonary administration include, but are not limited to, dry powder compositions consisting of a powder of a compound of formula (I) or a salt thereof and a powder of a suitable carrier and / or lubricant. Compositions for pulmonary administration may be inhaled from any suitable dry powder inhaler device known to those skilled in the art.

[0067] The compounds of the present invention may also be administered by intratumoral injection or by direct injection into the tumor vasculature. Local, regional, or systemic administration may also be appropriate. For tumors larger than 4 cm, the administered volume is about 4 ml to 10 ml (preferably 10 ml), while for tumors smaller than 4 cm, a volume of about 1 ml to 3 ml is used (preferably 3 ml). Multiple injections delivered as a single dose include volumes of about 0.1 ml to about 0.5 ml. In the case of surgical intervention, the present invention can be used preoperatively to target inoperable tumors for resection.

[0068] The composition is administered under a protocol and at a dosage sufficient to reduce inflammation and pain in the subject. In some embodiments, the active ingredient(s) in the pharmaceutical composition of the present invention are generally formulated in a dosage unit. The dosage unit may contain 0.1 mg to 1000 mg of the compound of formula (I) per dosage unit for daily administration.

[0069] In some embodiments, the amount useful in a particular formulation will depend on the severity of the disease, disorder, or condition, medical history, individual health status, and response to the drug, hi some embodiments, the dosage ranges from 0.001% to about 60% by weight of the formulation.

[0070] When used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used singly.

[0071] Methods for administering drugs and formulations for formulations for any of the various routes of administration are disclosed in Remington's Pharmaceutical Sciences, 17th Edition, Gennaro et al. Eds., Mack Publishing Co., 1985, and Remington's Pharmaceutical Sciences, Gennaro AR ed. 20th Edition, 2000, Williams & Wilkins PA, USA, and Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins Eds., 2005, and in Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th Edition, Lippincott Williams & Wilkins Eds., 2005.

[0072] The components described above for orally administered or injectable compositions are merely representative.

[0073] The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems.

[0074] The compound of formula (I) can be used as an independent therapeutic agent or in combination with other chemotherapeutic agents having different mechanisms of action.Preferred chemotherapeutic agents to be used in combination are selected from the group consisting of cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, and non-classical alkylating agents such as dacarbazine and temozolamide.The compound of formula (I) can also be used in combination with radiation therapy.

[0075] A third aspect of the present invention relates to the use of a compound of formula (I) as disclosed above or a pharmaceutical composition thereof, or a pharmaceutically acceptable salt or solvate thereof, as a medicament.

[0076] In particular, the compounds of formula (I) can be used in the treatment of diseases or disorders associated with increased levels of RhoJ / Cdc42 expression or function (relative to physiological or desired levels). In particular, the compounds of formula (I) may act as inhibitors of RhoJ-PAK, and more particularly, the compounds of formula (I) can be used in the treatment of primary or metastatic neoplastic diseases, precancerous conditions such as hyperplasia, metaplasia, or dysplasia, cancer, cancer metastasis, benign tumors, hyperproliferative disorders, cardiomyopathies, and retinal disorders. Preferably, the compounds of formula (I) are used in the treatment of melanoma.

[0077] In the following, the present invention is illustrated by means of several examples, which are not to be construed as being deemed to limit the scope of the present invention.

[0078] The following abbreviations are used below in the accompanying examples: acetic acid (AcOH), acetonitrile (ACN), ammonia (NH), deuterated chloroform (CDCl), deuterated dimethyl sulfoxide (DMSO-d), dichloromethane (DCM), diethyl ether (EtO), dimethyl sulfoxide (DMSO), ethanol (EtOH), ethyl acetate (AcOEt), hydrochloric acid (HCl), tert-butyl methyl ether (TBME), methanol (MeOH), room temperature (rt), sodium bicarbonate (NaHCO), sodium hydroxide (NaOH), sodium sulfate (NaSO), dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium dichloromethane complex (PdCl(dppf) dichloromethane complex), water (HO). [Example]

[0079] Chemicals, materials and methods synthesis All commercially available reagents and solvents were used without further purification when purchased from the supplier. Dry solvents were purchased from Sigma-Aldrich. Automated column chromatographic purification was performed using a Teledyne ISCO instrument (CombiFlash™ Rf) with pre-packed silica gel columns of different sizes (4 g up to 120 g) and mixtures of increasing polarity: cyclohexane and ethyl acetate (AcOEt), cyclohexane and tert-butyl methyl ether (TBME), or dichloromethane (DCM) and methanol (MeOH).

[0080] Characterization Bruker Avance III 400 system equipped with a BBI probe and Z-gradient ( 1 NMR experiments were performed at 400.13 MHz for H. Spectra were acquired at 300 K using deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3) as solvent. 1For H-NMR, data are reported as follows: chemical shift, multiplicity (s = singlet, d = doublet, dd = doublet of doublet, t = triplet, q = quartet, m = multiplet), coupling constant (Hz), and integral. UPLC / MS analysis was performed on a Waters ACQUITY UPLC / MS system consisting of an SQD (single quadrupole detector) mass spectrometer equipped with an electrospray ionization interface and a photodiode array detector. The PDA range was 210 nm to 400 nm. Analysis was performed on an ACQUITY UPLC BEH C18 column (100 × 2.1 mm ID, 1.7 μm particle size) with a VanGuard BEH C18 precolumn (5 × 2.1 mm ID, 1.7 μm particle size). The mobile phases were 10 mM NHOAc in H2O adjusted to pH 5 with CH3COOH (A) and 10 mM NHOAc in CH3CN-H2O (95:5) at pH 5.0. For analytical method 1, the proportion of mobile phase B was increased from 5% to 95% in 3 min. For analytical method 2, the proportion of mobile phase B was increased from 50% to 100% in 3 min. Electrospray ionization in positive and negative modes was applied. ESI was applied in positive and negative modes. All tested compounds showed purity of 90% or higher by NMR and UPLC / MS analysis.

[0081] Preparation and Examples [ka]

[0082] Reaction A: Phenylboronic acid (1 equiv.), K2CO3 2M (2 equiv.), PdCl2(dppf).DCM (0.05 equiv.), 1,4-dioxane, 60 °C, microwave, Ar, 1 h, yield = 72%. Reaction B: Aniline derivative (1 equiv.), Pd(OAc)2 (0.05 equiv.), racBINAP (0.05 equiv.), Cs2CO3 (1.2 equiv.), 1,4-dioxane, Ar, 60 °C, microwave, 4 h. Reaction C: 4,4,5,5-tetramethylboronate (1.2 equiv.), K2CO3 2M (2 equiv.), PdCl2(dppf).DCM (0.05 equiv.), 1,4-dioxane, 120 °C, microwave, Ar, 2 h. Reactions D and E': HCOONH4 (4 equiv.), Pd(OH)2 / C (20 wt%), MeOH, reflux, N2, 4 h, or H-Cube apparatus (Pd(OH)2 cartridge, 50 °C, 50 bar). Reactions D' and E: HCl (4 M), 1,4-dioxane, 0 °C to room temperature, 1 h.

[0083] [ka]

[0084] Reaction 1. 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (1.1 equiv.), LDA 2M THF / heptane / ethylbenzene (1.2 equiv.), THF (dry), -78 °C to room temperature, Ar, 16 h, 28% yield. Reaction 2. Bispinacolatodiboron (1.3 equiv.), KOAc (2.8 equiv.), PdCl(dppf).DCM (0.1 equiv.), 1,4-dioxane, 80 °C, Ar, 3 h, 80% yield.

[0085] General Procedure Reaction B. Palladium-Catalyzed Aniline Derivative Coupling A mixture of Pd(OAc) (0.05 mmol) and racBINAP (0.05 mmol) in 1,4-dioxane (3 mL) was stirred under Ar flushing for 10 min. Subsequently, a solution of the corresponding substituted aniline (1 mmol) in 1,4-dioxane (1 mL) and CsCO (1.2 mmol) were added stepwise to a solution of intermediate A (1 mmol) in 1,4-dioxane (1 mL). The reaction mixture was stirred at 60 °C for 4 h in a CEM™ microwave oven, filtered through a coarse patch of Celite, rinsed with DCM, and concentrated to dryness under reduced pressure. Final normal-phase purification afforded intermediate Bn.

[0086] General Procedure Reaction C. Suzuki Coupling Reaction A suspension of the compound from General Procedure B (1 mmol), the corresponding 4,4,5,5-tetramethylboronate (1.2 mmol), PdCl(dppf) dichloromethane complex (0.1 mmol), and 2M KCO solution (2 mmol) in 1,4-dioxane (10 mL) was stirred at 120 °C for 2 h in a CEM™ microwave. The crude product was partitioned between dichloromethane (25 mL) and saturated NaHCO solution (25 mL), and the organic layer was dried over NaSO and concentrated to dryness under reduced pressure. Final normal-phase purification afforded intermediate Cn.

[0087] General Procedure: Reactions D and E'. Reduction of the Double Bond Method 1. Under a N atmosphere, a suspension of the compound to be reduced (1 mmol), ammonium formate (4 mmol), and Pd(OH) / C (20 wt % of the starting material) was stirred at reflux until the reaction was complete. The catalyst was filtered off through a coarse patch of Celite, and the resulting filtrate was concentrated to dryness under reduced pressure. Final normal-phase purification afforded intermediate Dn.

[0088] Method 2. A 0.01 M solution of the starting material in MeOH or THF was eluted through a Pd(OH)-C cartridge in an H-cube apparatus at 50 °C under 50 bar H pressure until the reaction was complete. Final normal phase purification afforded intermediate D'.n.

[0089] General Procedure for Reactions D' and E. Boc Elimination To a solution of the Boc-protected compound (1 mmol) in 1,4-dioxane (2.6 ml) at 0° C., a solution of HCl (4 M) in 1,4-dioxane (2.6 ml, 10 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 1 hour. The crude reaction mixture was then concentrated to dryness under reduced pressure. The resulting crude was partitioned between DCM (20 ml) and NaOH 0.1 M (20 ml), and the organic layer was dried over NaSO and concentrated to dryness under reduced pressure. Final normal-phase purification afforded the compound of the present invention.

[0090] Example 1. N1,N1-dimethyl-N4-[6-phenyl-2-(3-piperidyl)pyrimidin-4-yl]benzene-1,4-diamine (Compound 8) [ka]

[0091] Step 1. Synthesis of 2,4-dichloro-6-phenyl-pyrimidine (Intermediate A) [ka]

[0092] A suspension of 2,4,6-trichloropyrimidine (1000 mg, 5.29 mmol), phenylboronic acid (665 mg, 5.29 mmol), PdCl(dppf) dichloromethane complex (204 mg, 0.26 mmol), and 2M KCO solution (5.3 mL, 10.58 mmol) in 1,4-dioxane (26.4 mL) was stirred at 60 °C for 1 h in a CEM™ microwave. The crude material was partitioned between dichloromethane (150 mL) and saturated NaHCO solution (100 mL), and the organic layer was dried over NaSO and concentrated to dryness under reduced pressure. Final normal-phase purification (cyclohexane / DCM 100 / 0 to 85 / 15) afforded the pure title compound (857 mg, 72% yield). Retention time = 1.38 minutes (Analysis method 2); MS (ESI) m / z: 225.1 [MH] +, [MH] + Calculated value: 225.0. 1 H NMR (400MHz, CDCl3) δ 8.13~8.03(m,2H), 7.68(s,1H), 7.62~7.48(m,3H).

[0093] Step 2. Synthesis of N1-(2-chloro-6-phenyl-pyrimidin-4-yl)-N4,N4-dimethyl-benzene-1,4-diamine (Intermediate B.1) [ka]

[0094] The title compound was obtained by following the general procedure Reaction B described above using Intermediate A (300 mg, 1.33 mmol) and N1,N1-dimethylbenzene-1,4-diamine (191.1 mg, 1.33 mmol). Final normal phase purification (cyclohexane / TBME 100 / 0 to 80 / 20) afforded the pure title compound (272 mg, 63% yield). Retention time = 1.56 min (Analytical Method 2); MS (ESI) m / z: 325.1 [M−H] + , [MH] + Calculated value: 325.1. 1 H NMR (400MHz, DMSO-d6)δ 9.76(s,1H), 7.93(dd,J=6.7, 3.0Hz,2H), 7.52(dd,J=4.6, 2.4Hz,3H), 7.36(s,2H), 7.00(s,1H), 6.87~6.64(m,2H), 2.89(s,6H).

[0095] Step 3. Synthesis of tert-butyl 5-(trifluoromethylsulfonyloxy)-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate 1) [ka]

[0096] To a 2.0 M solution of lithium diisopropylamide in cyclohexane (8.8 mL, 17.53 mmol) in dry tetrahydrofuran (15.6 mL) was added dropwise a solution of 3-oxo-piperidine-1-carboxylic acid tert-butyl ester (3000 mg, 14.60 mmol) in dry tetrahydrofuran (15.6 mL) at −78° C. The mixture was stirred at −78° C. for 1 hour, and a solution of N-phenylbistrifluoromethanesulfonamide (5855.6 mg, 16.07 mmol) in dry tetrahydrofuran (15.8 mL) was added. The mixture was stirred at −78° C. for 2 hours, then allowed to warm to room temperature and stirred at room temperature for an additional 16 hours. The mixture was evaporated to dryness, and the residue was taken up with diethyl ether (50 mL), washed with water (50 mL), a 2 M solution of sodium hydroxide (50 mL), and brine (50 mL), dried over sodium sulfate, and concentrated to dryness under reduced pressure. Final normal phase purification (hexane / DCM 100 / 0 to 50 / 50) afforded the pure title compound (1354 mg, 28% yield). Retention time = 2.66 min (Analytical Method 1). 1 H NMR (400MHz, CDCl3) δ 7.07(s,1H), 3.52(s,2H), 2.43(td,J=6.4, 1.5Hz,2H), 1.93(tt,J=6.3, 5.0Hz,2H), 1.49(s,9H).

[0097] Step 4. Synthesis of tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate 2) [ka]

[0098] To a degassed solution of intermediate 1 (600 mg, 1.81 mmol) in dioxane (10.7 mL) was added bis-(pinacolato)diboron (603.8 mg, 2.35 mmol), potassium acetate (502.7 mg, 5.07 mmol), and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium dichloromethane complex (139.5 mg, 0.18 mmol). The mixture was stirred at 80 °C for 3 h. After cooling, the mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. Final normal-phase purification (hexane / DCM 70 / 30 to 50 / 50) afforded the pure title compound (448 mg, 80% yield). Retention time = 1.85 min (Analytical Method 2). MS (ESI) m / z 310.2 [M−H] + , [MH] + Calculated value: 310.2. 1 H NMR (400MHz, CDCl3) δ 5.29(s,1H), 3.67~3.39(m,2H), 2.15~1.96(m,2H), 1.81~1.73(m,2H), 1.49(s,9H), 1.32~1.17(m,12H).

[0099] Step 5. Synthesis of tert-butyl 5-[4-[4-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate C.1) [ka]

[0100] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.1 (100 mg, 0.35 mmol) and Intermediate 2 (117.8 mg, 0.37 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 85 / 15) afforded the pure title compound (107.4 mg, 74% yield). Retention time = 2.58 min (Analytical Method 2); MS (ESI) m / z 472.4 [M−H] + , [MH] + Calculated value: 472.3. 1H NMR(400MHz,DMSO-d6)δ 9.28(s,1H), 8.12~7.96(m,2H), 7.64~7.42(m,5H), 7.15(s,1H), 6.96(s,1H), 6.86~6.69(m,2H) , 4.11(d,J=4.0Hz,2H), 3.55(t,J=5.7Hz,2H), 2.87(s,6H), 2.67(d,J=7.0Hz,2H), 1.44(s,9H).

[0101] Step 6. Synthesis of tert-butyl 3-[4-[4-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.1) [ka]

[0102] The title compound was obtained using intermediate C.1 (60.5 mg, 0.12 mmol) according to method 2 of general procedure D described above. Final normal phase purification (hexane / TBME 90 / 10 to 70 / 30) afforded the pure title compound (60 mg, 99% yield). Retention time = 2.22 min (analytical method 2); MS (ESI) m / z: 474.4 [M−H] + , [MH] + Calculated value: 474.3. 1 H NMR(400MHz,DMSO-d6)δ 9.38(s,1H), 8.24~7.80(m,2H), 7.59~7.34(m,5H), 6.98(s,1H), 6.91~6.59(m,2H), 3.37(d,J=9.8Hz,1H), 3.16~3.02(m,1H), 3.02~ 2.91(m,2H), 2.87(s,6H), 2.68(td,J=11.9, 3.0Hz,1H), 2.23~2.05(m,1H), 1.89~1.70(m,2H), 1.63(q,J=12.8Hz,1H), 1.45(s,9H).

[0103] Step 7. Synthesis of N1,N1-dimethyl-N4-[6-phenyl-2-(3-piperidyl)pyrimidin-4-yl]benzene-1,4-diamine (Compound 8) [ka]

[0104] The title compound was obtained using intermediate D.1 (61 mg, 0.13 mmol) following the general procedure Reaction E described above. Final normal phase purification (DCM / DCM:NH3 1M MeOH 4:1 80 / 20 to 60 / 40) afforded the pure title compound (46 mg, 95% yield). Retention time = 1.96 min (Analytical Method 1); MS (ESI) m / z: 374.6 [M−H] + , [MH] + Calculated value: 374.2. 1 H NMR(400MHz,DMSO-d6)δ 9.38(s,1H), 8.24~7.80(m,2H), 7.59~7.34(m,5H), 6.98(s,1H), 6.91~6.59(m,2H), 3.37(d,J=9.8Hz,1H), 3.16~3.02(m,1H) , 3.02~2.91(m,2H), 2.87(s,6H), 2.68(td,J=11.9, 3.0Hz,1H), 2.23~2.05(m,1H), 1.89~1.70(m,2H), 1.63(q,J=12.8Hz,1H).

[0105] Example 2. N-(4-Methoxyphenyl)-6-phenyl-2-(3-piperidyl)pyrimidin-4-amine (Compound 9) [ka]

[0106] Step 1. Synthesis of 2-chloro-N-(4-methoxyphenyl)-6-phenyl-pyrimidin-4-amine (Intermediate B.2) [ka]

[0107] The title compound was obtained by following the general procedure Reaction B described above using Intermediate A (100 mg, 0.44 mmol) and p-methoxyaniline (55.2 mg, 0.44 mmol). Final normal phase purification (cyclohexane / TBME 95 / 5 to 75 / 25) afforded the pure title compound (86.8 mg, 63% yield). Retention time = 1.39 min (Analytical Method 2); MS (ESI) m / z: 312.1 [M−H] + , [MH] + Calculated value: 312.1. 1 H NMR (400MHz, DMSO-d6) δ 9.91(s,1H), 8.06~7.84(m,2H), 7.65~7.37(m,5H), 7.06(s,1H), 7.02~6.92(m,2H), 3.76(s,3H).

[0108] Step 2. Synthesis of tert-butyl 5-[4-(4-methoxyanilino)-6-phenyl-pyrimidin-2-yl]-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate C.2) [ka]

[0109] The title compound was obtained according to the general procedure C described above using intermediate B.2 (100 mg, 0.32 mmol) and intermediate 2 (119.0 mg, 0.38 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 80 / 20) afforded the pure title compound (64.7 mg, 44% yield). Retention time = 2.37 min (analytical method 2); MS (ESI) m / z 459.6 [M−H] + , [MH] + Calculated value: 459.2. 1H NMR(400MHz,CDCl3)δ 8.05~7.97(m,2H), 7.46~7.37(m,3H), 7.31(t,J=6.5Hz,2H), 6.98~6.90(m,2H), 6.71(s,1H), 6.68~ 6.46(m,1H), 3.84(s,3H), 3.66(d,J=9.5Hz,2H), 2.82~2.60(m,2H), 2.02~1.86(m,2H), 1.56(s,9H).

[0110] Step 3. Synthesis of tert-butyl 3-[4-(4-methoxyanilino)-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.2) [ka]

[0111] The title compound was obtained using intermediate C.2 (65.0 mg, 0.14 mmol) according to method 2 of the general procedure Reaction D described above. Final normal phase purification (hexane / TBME 100 / 0 to 80 / 20) afforded the pure title compound (40 mg, 62% yield). Retention time = 1.94 min (Analytical Method 2); MS (ESI) m / z: 461.2 [M−H] + , [MH] + Calculated value: 461.2. 1 H NMR(400MHz,CDCl3)δ 8.07~7.80(m,2H), 7.46~7.38(m,3H), 7.26(s,1H), 7.01~6.89(m,2H), 6.77(s,1H), 6.71(s,1H), 3.84(s,3H) ), 3.01~2.68(m,2H), 2.23(d,J=12.3Hz,1H), 1.80(qd,J=13.0, 3.8Hz,2H), 1.73~1.52(m,4H), 1.47(s,9H).

[0112] Step 4. Synthesis of N-(4-methoxyphenyl)-6-phenyl-2-(3-piperidyl)pyrimidin-4-amine (Compound 9) [ka]

[0113] The title compound was obtained using intermediate D.2 (40 mg, 0.09 mmol) following the general procedure reaction E described above. Final normal phase purification (DCM / DCM:NH3 1M MeOH 4:1 85 / 15 to 65 / 35) afforded the pure title compound (31 mg, 99% yield). Retention time = 0.49 min (Analytical Method 2); MS (ESI) m / z: 361.6 [M−H] + , [MH] + Calculated value: 361.2. 1 H NMR(400MHz,DMSO-d6)δ 9.60(s,1H), 8.15~7.90(m,2H), 7.59(d,J=8.4Hz,2H), 7.56~7.47(m,3H), 7.05(d,J=4.2Hz,1H), 7.00~6.90(m,2H) , 3.76(s,3H), 3.60(d,J=11.9Hz,1H), 3.33~3.11(m,3H), 2.96~2.91(m,1H), 2.26~2.19(m,1H), 1.97~1.65(m,2H).

[0114] Example 3. N3,N3-dimethyl-N1-[6-phenyl-2-(3-piperidyl)pyrimidin-4-yl]benzene-1,3-diamine (Compound 10) [ka]

[0115] Step 1. Synthesis of N1-(2-chloro-6-phenyl-pyrimidin-4-yl)-N3,N3-dimethyl-benzene-1,3-diamine (Intermediate B.3) [ka]

[0116] The title compound was obtained by following the general procedure Reaction B described above using Intermediate A (300 mg, 1.13 mmol) and N1,N1-dimethylbenzene-1,3-diamine (181.5 mg, 1.33 mmol). Final normal phase purification (cyclohexane / TBME 100 / 0 to 80 / 20) afforded the pure title compound (246 mg, 57% yield). Retention time = 1.73 min (Analytical Method 2); MS (ESI) m / z: 325.1 [M−H] + , [MH] + Calculated value: 325.1. 1 H NMR(400MHz,DMSO-d6)δ 9.93(s,1H), 8.14~7.82(m,2H), 7.60~7.47(m,3H), 7.29~7.09(m,2H), 7.04 (s,1H), 6.99~6.85(m,1H), 6.51(ddd,J=8.4, 2.5, 0.8Hz,1H), 2.92(s,6H).

[0117] Step 2. Synthesis of tert-butyl 5-[4-[3-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate C.3) [ka]

[0118] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.3 (175 mg, 0.54 mmol) and Intermediate 2 (199.9 mg, 0.65 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 85 / 15) afforded the pure title compound (109.2 mg, 43% yield). Retention time = 2.65 min (Analytical Method 2); MS (ESI) m / z 472.3 [M−H] + , [MH] + Calculated value: 472.3. 1H NMR(400MHz,DMSO-d6)δ 9.34(s,1H), 8.40(s,1H), 8.04(dd,J=7.7, 1.9Hz,2H), 7.59~7.44(m,4H), 7.11(t,J=8.0Hz,2H), 6.99(s,1H), 6.40 (dd,J=9.1, 2.5Hz,1H), 3.59(t,J=5.6Hz,2H), 2.92(s,6H), 2.65~2.59(m,2H), 1.88(p,J=6.0Hz,2H), 1.50(s,9H).

[0119] Step 3. Synthesis of tert-butyl 3-[4-[3-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.3) [ka]

[0120] The title compound was obtained using intermediate C.3 (105 mg, 0.22 mmol) according to method 2 of the general procedure reaction D described above. Final normal phase purification (hexane / AcOEt 100 / 0 to 80 / 20) afforded the pure title compound (12 mg, 12% yield). Retention time = 2.35 min (analytical method 2); MS (ESI) m / z: 474.6 [M−H] + , [MH] + Calculated value: 474.3. 1 H NMR (400MHz, CDCl3)δ 8.04~7.93(m,2H), 7.49~7.37(m,4H), 7.04(s,1H), 6.78(s,1H), 6.69(d,J=7.7Hz,1H), 6.59(d,J=8.5Hz,1H), 4.20~4.08(m,1H), 3.23 ~3.16(m,1H), 2.99(s,6H), 2.98~2.91(m,1H), 2.88~2.77(m,J=14.3Hz,1H), 2.31~2.20(m,1H), 1.82~1.58(m,2H), 1.51~1.45(m,11H).

[0121] Step 4. Synthesis of N3,N3-dimethyl-N1-[6-phenyl-2-(3-piperidyl)pyrimidin-4-yl]benzene-1,3-diamine (Compound 10) [ka]

[0122] The title compound was obtained using intermediate D.3 (34 mg, 0.08 mmol) following general procedure reaction E. Final normal phase purification (DCM / DCM:NH₃ 1M MeOH 4:1 95 / 5 to 45 / 55) afforded the pure title compound (16 mg, 61% yield). Retention time = 2.07 min (Analytical Method 1); MS (ESI) m / z: 374.5 [M−H] + , [MH] + Calculated value: 374.2. 1 H NMR(400MHz,DMSO-d6)δ 9.50(s,1H), 8.01(dd,J=7.8, 1.8Hz,2H), 7.56~7.48(m,3H), 7.46(s,1H), 7.12(t,J=8 .1Hz,1H), 7.10~7.06(m,1H), 6.90(d,J=8.0Hz,1H), 6.40(dd,J=8.2, 2.5Hz,1H), 3.28 ~3.14(m,1H), 2.94(s,6H), 2.94~2.87(m,1H), 2.84~2.75(m,2H), 2.46(dd,J=12.1, 2. 9Hz,1H), 2.16~2.03(m,1H), 1.90~1.72(m,1H), 1.71~1.58(m,1H), 1.57~1.39(m,1H).

[0123] Example 4. N-(3-Methoxyphenyl)-6-phenyl-2-(3-piperidyl)pyrimidin-4-amine (Compound 11) [ka]

[0124] Step 1. Synthesis of 2-chloro-N-(3-methoxyphenyl)-6-phenyl-pyrimidin-4-amine (Intermediate B.4) [ka]

[0125] The title compound was obtained by following the general procedure reaction B described above using intermediate A (300 mg, 1.13 mmol) and m-methoxyaniline (154 μl, 1.33 mmol). Final normal phase purification (cyclohexane / TBME 100 / 0 to 85 / 15) afforded the pure title compound (150 mg, 36% yield). Retention time = 1.52 min (analytical method 2); MS (ESI) m / z: 312.1 [M−H] + , [MH] + Calculated value: 312.1. 1 H NMR(400MHz,DMSO-d6)δ 10.07(s,1H), 8.00~7.90(m,2H), 7.59~7.50(m,3H), 7.34(t,J=2.3Hz,1H), 7.2 9(t,J=8.1Hz,1H), 7.18(s,2H), 6.69(ddd,J=8.2, 2.5, 0.9Hz,1H), 3.77(s,3H).

[0126] Step 2. Synthesis of tert-butyl 5-[4-(3-methoxyanilino)-6-phenyl-pyrimidin-2-yl]-3,4-dihydro-2H-pyridine-1-carboxylate (Intermediate C.4) [ka]

[0127] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.4 (100 mg, 0.32 mmol) and Intermediate 2 (119.0 mg, 0.38 mmol). Final normal phase purification (cyclohexane / TBME 100 / 0 to 80 / 20) afforded the pure title compound (65.0 mg, 44% yield). Retention time = 2.40 min (Method 2); MS (ESI) m / z 459.6 [M−H] + , [MH] + Calculated value: 459.2.

[0128] Step 3. Synthesis of tert-butyl 3-[4-(3-methoxyanilino)-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.4) [ka]

[0129] The title compound was obtained using intermediate C.4 (65.0 mg, 0.14 mmol) according to method 2 of the general procedure Reaction D described above. Final normal phase purification (hexane / AcOEt 100 / 0 to 80 / 20) afforded the pure title compound (32.6 mg, 50% yield). Retention time = 2.13 min (Analytical Method 2); MS (ESI) m / z: 461.6 [M−H] + , [MH] + Calculated value: 461.2. 1 H NMR(400MHz,CDCl3)δ 8.04~7.92(m,2H), 7.45(p,J=3.9, 3.2Hz,3H), 7.30(t,J=8.1Hz,1H), 7.08(t,J= 2.2Hz,1H), 6.99(s,1H), 6.95(dd,J=7.9, 2.0Hz,1H), 6.89(s,1H), 6.72(dd,J=8 .3, 2.4Hz,1H), 3.84(s,3H), 3.20(s,1H), 2.99~2.88(m,1H), 2.80(t,J=12.5Hz, 1H), 2.26(d,J=12.5Hz,1H), 1.93~1.74(m,2H), 1.75~1.51(m,3H), 1.47(s,9H).

[0130] Step 4. Synthesis of N-(3-methoxyphenyl)-6-phenyl-2-(3-piperidyl)pyrimidin-4-amine (Compound 11) [ka]

[0131] The title compound was obtained using intermediate D.4 (61 mg, 0.13 mmol) following general procedure reaction E. Final normal phase purification (DCM / DCM:NH₃M MeOH 4:1 85 / 15 to 60 / 40) afforded the pure title compound (39 mg, 82% yield). Retention time = 1.91 min (Analytical Method 1); MS (ESI) m / z: 361.6 [M−H] + , [MH] + Calculated value: 361.2. 1 H NMR(400MHz,DMSO-d6)δ 9.89(s,1H), 8.17~7.94(m,2H), 7.67~7.44(m,4H), 7.34~7.12(m,3H), 6.61(dt,J=5.4, 2.4Hz,1H), 3.79(s,3H) ), 3.63(d,J=8.1Hz,1H), 3.27~3.18(m,3H), 2.89(d,J=14.1Hz,1H), 2.26(d,J=11.7Hz,1H), 2.01~1.67(m,3H).

[0132] Example 5. N4,N4-Dimethyl-N1-[6-phenyl-2-(4-piperidyl)pyrimidin-4-yl]benzene-1,4-diamine (Compound 12) [ka]

[0133] Step 1. Synthesis of tert-butyl 4-[4-[4-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (Intermediate C.5) [ka]

[0134] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.1 (100 mg, 0.35 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (117.8 mg, 0.37 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 85 / 15) afforded the pure title compound (107.4 mg, 74% yield). Retention time = 2.58 min (Analytical Method 2); MS (ESI) m / z 472.4 [M−H] + , [MH] + Calculated value: 472.3. 1 H NMR(400MHz,DMSO-d6)δ 9.28(s,1H), 8.12~7.96(m,2H), 7.64~7.42(m,5H), 7.15(s,1H), 6.96(s,1H), 6.86~6.69(m,2H) , 4.11(d,J=4.0Hz,2H), 3.55(t,J=5.7Hz,2H), 2.87(s,6H), 2.67(d,J=7.0Hz,2H), 1.44(s,9H).

[0135] Step 2. Synthesis of N4,N4-dimethyl-N1-[6-phenyl-2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-4-yl]benzene-1,4-diamine (Intermediate D'.5) [ka]

[0136] The title compound was obtained (84 mg, 99% yield) using intermediate C.5 (106 mg, 0.22 mmol) following general procedure reaction D'. Retention time = 0.72 min (Analytical Method 2); MS (ESI) m / z: 372.5 [M−H] + , [MH] + Calculated: 372.2. The solid obtained was used in the next step without any further purification procedure.

[0137] Step 3. Synthesis of N4,N4-dimethyl-N1-[6-phenyl-2-(4-piperidyl)pyrimidin-4-yl]benzene-1,4-diamine (Compound 12) [ka]

[0138] The title compound was obtained using intermediate D'.5 (84 mg, 0.22 mmol) following method 1 of general procedure reaction E' described above. Final normal phase purification (DCM / DCM:NH3 1M MeOH 4:1 70 / 30 to 50 / 50) afforded the pure title compound (54 mg, 64% yield). Retention time = 1.98 min (Analytical Method 1); MS (ESI) m / z: 374.2 [M−H] + , [MH] + Calculated value: 374.2. 1 H NMR (400MHz, DMSO-d6). δ 9.25(s,1H), 8.17~7.82(m,2H), 7.69~7.37(m,5H), 6.93(s,1H), 6.83~6.66(m,2H), 3.08(dt,J=12.2, 3.4Hz,2H), 2.87(s,6H) , 2.77(tt,J=11.5, 3.8Hz,1H), 2.66(td,J=12.1, 2.6Hz,2H), 1.93(dd,J=13.4, 3.5Hz,2H), 1.77(qd,J=13.0, 12.5, 4.0Hz,2H).

[0139] Example 6. N-(4-Methoxyphenyl)-6-phenyl-2-(4-piperidyl)pyrimidin-4-amine (Compound 13) [ka]

[0140] Step 1. Synthesis of tert-butyl 4-[4-(4-methoxyanilino)-6-phenyl-pyrimidin-2-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (Intermediate C.6) [ka]

[0141] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.2 (80 mg, 0.26 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (98.1 mg, 0.31 mmol). Final normal phase purification (cyclohexane / AcOEt 95 / 5 to 75 / 25) afforded the pure title compound (104.7 mg, 89% yield). Retention time = 2.24 min (Analytical Method 2); MS (ESI) m / z 459.3 [M−H] + , [MH] + Calculated value: 459.2. 1 H NMR(400MHz,DMSO-d6)δ 9.44(s,1H), 8.12~8.02(m,2H), 7.68~7.60(m,2H), 7.58~7.46(m,3H), 7.17(s,1H), 7.01(s,1H), 6.99~ 6.92(m,2H), 4.12(s,2H), 3.75(s,3H), 3.56(t,J=5.7Hz,2H), 2.68(q,J=4.0, 3.4Hz,2H), 1.44(s,9H).

[0142] Step 2. Synthesis of N-(4-methoxyphenyl)-6-phenyl-2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-4-amine (Intermediate D'.6) [ka]

[0143] The title compound was obtained (80 mg, 99% yield) using intermediate C.6 (102 mg, 0.22 mmol) following general procedure reaction D'. Retention time = 0.60 min (analytical method 2); MS (ESI) m / z: 359.2 [M−H] + , [MH] + Calculated: 359.2. The obtained solid was used in the next step without any further purification procedure.

[0144] Step 3. Synthesis of N-(4-methoxyphenyl)-6-phenyl-2-(4-piperidyl)pyrimidin-4-amine (Compound 13) [ka]

[0145] The title compound was obtained using intermediate D'.6 (80 mg, 0.22 mmol) following method 1 of general procedure reaction E' described above. Final normal phase purification (DCM / DCM:NH3 1M MeOH 4:1 85 / 15 to 50 / 50) afforded the pure title compound (52.5 mg, 64% yield). Retention time = 1.87 min (Analytical Method 1); MS (ESI) m / z: 361.3 [M−H] + , [MH] + Calculated value: 361.2. 1 H NMR (400MHz, DMSO-d6). δ 9.44(s,1H), 8.14~7.89(m,2H), 7.64(d,J=8.9Hz,2H), 7.58~7.38(m,3H), 6.99(s,1H), 6.96~6.91(m,2H), 3.75(s,3H), 3.13(d,J=1 2.4Hz,2H), 2.83(tt,J=11.4, 3.8Hz,1H), 2.73(td,J=12.2, 2.7Hz,2H), 1.97(dd,J=12.8, 3.4Hz,2H), 1.81(qd,J=12.2, 4.0Hz,2H).

[0146] Example 7. N3,N3-Dimethyl-N1-[6-phenyl-2-(4-piperidyl)pyrimidin-4-yl]benzene-1,3-diamine (Compound 14) [ka]

[0147] Step 1. Synthesis of tert-butyl 4-[4-[3-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (Intermediate C.7) [ka]

[0148] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.3 (120 mg, 0.37 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (129.5 mg, 0.41 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 80 / 20) afforded the title compound (155.1 mg, 89% yield). Retention time = 2.48 min (Analytical Method 2); MS (ESI) m / z 472.3 [M−H] + , [MH] + Calculated value: 472.3. 1 H NMR(400MHz,DMSO-d6)δ 9.44(s,1H), 8.17~7.95(m,2H), 7.67~7.41(m,3H), 7.31(s,1H), 7.21(s,1H), 7.14(t,J=8.1Hz,1H), 7.11(s,1H), 7.03~6.93 (m,1H), 6.42(ddd,J=8.3, 2.7, 0.8Hz,1H), 4.11(s,2H), 3.56(t,J=5.6Hz,2H), 2.93(s,6H), 2.81~2.62(m,2H), 1.44(s,9H).

[0149] Step 2. Synthesis of tert-butyl 4-[4-[3-(dimethylamino)anilino]-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.7) [ka]

[0150] The title compound was obtained using intermediate C.7 (150 mg, 0.32 mmol) according to method 1 of the general procedure reaction D described above. Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 80 / 20) afforded the pure title compound (149 mg, 99% yield). Retention time = 2.30 min (analytical method 2); MS (ESI) m / z: 474.4 [M−H] + , [MH] + Calculated value: 474.3. 1 H NMR (400 MHz, DMSO-d6) δ 9.43(s,1H), 8.08~7.94(m,2H), 7.58~7.44(m,3H), 7.40(s,1H), 7.11( t,J=8.1Hz,1H), 7.06(s,1H), 6.88(ddd,J=7.9, 2.1, 0.8Hz,1H), 6.41(d dd,J=8.4, 2.5, 0.8Hz,1H), 4.05(d,J=13.1Hz,2H), 1.98(dd,J=13.6, 3.5Hz,2H), 2.95~2.87(m,9H), 1.73(qd,J=12.5, 4.2Hz,2H), 1.42(s,9H).

[0151] Step 3. Synthesis of N3,N3-dimethyl-N1-[6-phenyl-2-(4-piperidyl)pyrimidin-4-yl]benzene-1,3-diamine (Compound 14) [ka]

[0152] The title compound was obtained using intermediate D.7 (80 mg, 0.22 mmol) following the general procedure Reaction E described above. Final normal phase purification (DCM / DCM:NH₃ 1M MeOH 4:1 95 / 5 to 60 / 40) afforded the pure title compound (39.6 mg, 34% yield). Retention time = 1.97 min (Analytical Method 1); MS (ESI) m / z: 374.6 [M−H] + , [MH] + Calculated value: 374.2. 1H NMR (400MHz, DMSO-d6). δ 9.42(s,1H), 8.01(d,J=7.0Hz,2H), 7.51(d,J=7.0Hz,3H), 7.40(s,1H), 7.12(t,J=8.1Hz,1H), 7.07(s,1H), 6.94(d,J=8.0Hz,1H), 6.51~6. 33(m,1H), 3.03(d,J=11.9Hz,2H), 2.93(s,6H), 2.85~2.70(m,1H), 2.61(t,J=11.9Hz,2H), 2.00~1.85(m,2H), 1.75(qd,J=12.3, 4.3Hz,2H).

[0153] Example 8. N-(3-Methoxyphenyl)-6-phenyl-2-(4-piperidyl)pyrimidin-4-amine (Compound 15) [ka]

[0154] Step 1. Synthesis of tert-butyl 4-[4-(3-methoxyanilino)-6-phenyl-pyrimidin-2-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (Intermediate C.8) [ka]

[0155] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.4 (180 mg, 0.58 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (202.4 mg, 0.64 mmol). Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 85 / 15) afforded the title compound (241.2 mg, 91% yield). Retention time = 2.31 min (Analytical Method 2); MS (ESI) m / z 459.3 [M−H] + , [MH] + Calculated value: 459.2. 1H NMR(400MHz,DMSO-d6)δ 9.63(s,1H), 8.19~8.03(m,2H), 7.62~7.46(m,4H), 7.33~7.22(m,2H), 7.20(s,1H), 7.11(s,1H), 6.6 7~6.54(m,1H), 4.13(d,J=3.3Hz,2H), 3.79(s,3H), 3.57(t,J=5.7Hz,2H), 2.71(s,2H), 1.44(s,9H).

[0156] Step 2. Synthesis of tert-butyl 4-[4-(3-methoxyanilino)-6-phenyl-pyrimidin-2-yl]piperidine-1-carboxylate (Intermediate D.8) [ka]

[0157] The title compound was obtained using intermediate C.8 (240 mg, 0.52 mmol) according to method 1 of the general procedure Reaction D described above. Final normal phase purification (cyclohexane / AcOEt 100 / 0 to 80 / 20) afforded the pure title compound (120 mg, 50% yield). Retention time = 2.11 min (Analytical Method 2); MS (ESI) m / z: 461.4 [M−H] + , [MH] + Calculated value: 461.2. 1 H NMR (400 MHz, DMSO-d6) δ 9.61(s,1H), 8.07~7.94(m,2H), 7.63(t,J=2.2Hz,1H), 7.57~7.45(m,3H), 7.22(t,J=8.0Hz,1H), 7.17(dt,J=8.3, 1.4Hz,1H), 7.08(s,1H), 6.59(ddd ,J=8.0, 2.5, 1.1Hz,1H), 4.12~3.96(m,2H), 3.77(s,3H), 2.93(tt,J=11.4 , 3.7Hz, 3H), 2.06~1.95 (m, 2H), 1.72 (qd, J=12.4, 4.2Hz, 2H), 1.42 (s, 9H).

[0158] Step 3. Synthesis of N-(3-methoxyphenyl)-6-phenyl-2-(4-piperidyl)pyrimidin-4-amine (Compound 15) [ka]

[0159] The title compound was obtained using intermediate D.8 (115 mg, 0.25 mmol) following the general procedure Reaction E described above. Final normal phase purification (DCM / DCM:NH₃M MeOH 4:1 95 / 5 to 50 / 50) afforded the pure title compound (80.1 mg, 89% yield). Retention time = 1.85 min (Analytical Method 1); MS (ESI) m / z: 361.6 [M−H] + , [MH] + Calculated value: 361.2. 1 H NMR (400 MHz, DMSO-d6). δ 9.58(s,1H), 8.13~7.92(m,2H), 7.65(t,J=2.0Hz,1H), 7.59~7.39(m,3H ), 7.33~7.15(m,2H), 7.07(s,1H), 6.58(dt,J=7.2, 2.3Hz,1H), 3.78(s, 3H), 3.04(dt,J=12.2, 3.3Hz,2H), 2.80(tt,J=11.6, 3.8Hz,1H), 2.61(t d,J=12.1, 2.5Hz,2H), 2.03~1.85(m,2H), 1.74(qd,J=12.2, 4.0Hz,2H).

[0160] Example 9. N1,N1-dimethyl-N4-(6-phenyl-2-tetrahydropyran-4-yl-pyrimidin-4-yl)benzene-1,4-diamine (Compound 16) [ka]

[0161] Step 1. N1-[2-(3,6-Dihydro-2H-pyran-4-yl)-6-phenyl-pyrimidin-4-yl]-N4,N4-dimethyl-benzene-1,4-diamine (Intermediate C.9) [ka]

[0162] The title compound was obtained by following the general procedure Reaction C described above using Intermediate B.1 (115 mg, 0.35 mmol) and 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (91.1 mg, 0.42 mmol). Final normal phase purification (cyclohexane / AcOEt 90 / 10 to 70 / 30) afforded the title compound (37.1 mg, 28% yield). Retention time = 1.70 min (Analytical Method 2); MS (ESI) m / z 373.5 [M−H] + , [MH] + Calculated value: 373.2. 1 H NMR(400MHz,DMSO-d6)δ 9.29(s,1H), 8.03(dd,J=7.7, 1.9Hz,2H), 7.58~7.43(m,5H), 7.25~7.12(m,1H), 6.96(s,1H), 6 .81~6.71(m,2H), 4.32(q,J=2.7Hz,2H), 3.83(t,J=5.4Hz,2H), 2.87(s,6H), 2.71~2.58(m,2H).

[0163] Step 2. N4,N4-dimethyl-N1-(6-phenyl-2-tetrahydropyran-4-yl-pyrimidin-4-yl)benzene-1,4-diamine (Compound 16) [ka]

[0164] The title compound was obtained using intermediate C.9 (35 mg, 0.09 mmol) according to method 2 of the general procedure reaction D described above. Final normal phase purification (cyclohexane / AcOEt 95 / 5 to 75 / 25) afforded the pure title compound (27 mg, 77% yield). Retention time = 1.48 min (analytical method 2); MS (ESI) m / z: 375.5 [M−H] + , [MH] + Calculated value: 375.2. 1H NMR(400MHz,DMSO-d6)δ 9.27(s,1H), 8.05~7.91(m,2H), 7.63~7.40(m,4H), 6.93(s,1H), 6.83~6.68(m,2H), 3.96(dt,J=11.4, 2.6 Hz,2H), 3.48(td,J=11.1, 3.5Hz,2H), 2.93(dq,J=10.7, 5.8, 5.3Hz,1H), 2.88(s,5H), 1.96~1.77(m,4H).

[0165] Example 10. N-(4-Methoxyphenyl)-6-phenyl-2-tetrahydropyran-4-yl-pyrimidin-4-amine (Compound 17) [ka]

[0166] Step 1. 2-(3,6-Dihydro-2H-pyran-4-yl)-N-(4-methoxyphenyl)-6-phenyl-pyrimidin-4-amine (Intermediate C.10) [ka]

[0167] The title compound was obtained according to the general procedure C described above using intermediate B.2 (90 mg, 0.29 mmol) and 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (74.3 mg, 0.35 mmol). Final normal phase purification (cyclohexane / TBME 100 / 0 to 80 / 20) afforded the title compound (44.2 mg, 43% yield). Retention time = 1.54 min (analytical method 2); MS (ESI) m / z 360.5 [M−H] + , [MH] + Calculated value: 360.2. 1H NMR(400MHz,DMSO-d6)δ 9.45(s,1H), 8.12~8.01(m,2H), 7.65(d,J=8.9Hz,2H), 7.57~7.45(m,5H), 7.01(s,1 H), 6.98~6.92(m,1H), 4.33(d,J=2.9Hz,2H), 3.76~3.72(m,5H), 2.71~2.61(m,2H).

[0168] Step 2. N-(4-Methoxyphenyl)-6-phenyl-2-tetrahydropyran-4-yl-pyrimidin-4-amine (Intermediate 17) [ka]

[0169] The title compound was obtained using intermediate C.10 (42 mg, 0.12 mmol) according to method 2 of the general procedure reaction D described above. Final normal phase purification (cyclohexane / TBME 90 / 10 to 70 / 30) afforded the pure title compound (16 mg, 38% yield). Retention time = 1.31 min (analytical method 2); MS (ESI) m / z: 362.5 [M−H] + , [MH] + Calculated value: 362.2. 1 H NMR(400MHz,DMSO-d6)δ 9.43(s,1H), 8.11~7.88(m,2H), 7.63(d,J=8.5Hz,2H), 7.55~7.45(m,3H), 6.98(s,1H), 6.96~6.90(m,2H), 3.96 (ddd,J=11.3, 4.2, 2.3Hz,2H), 3.75(s,3H), 3.47(td,J=11.3, 3.0Hz,2H), 3.03~2.88(m,1H), 1.98~1.77(m,4H).

[0170] Cell viability assay Cells were seeded at 10,000 cells / well in 96-well plates and incubated at 37°C with 5% CO for 24 hours. After overnight incubation, cells were treated with inhibitors (1.25 μM to 50 μM) alone or with 20 μM cisplatin for 24 hours. Cell viability was determined using the Cell-Titer-Glo Luminescent Cell Viability Assay Kit (Promega, Madison, WI) according to the manufacturer's instructions. Luminescence was measured using a DTX 800 microplate reader (Coulter). Half-maximal inhibitory concentration (IC50) values ​​were calculated using GraphPad Prism software. Assays were performed in triplicate.

[0171] The results are shown in Table 1.

[0172] [Table 2] TIFF0007776068000055.tif205169TIFF0007776068000056.tif213169TIFF0007776068000057.tif204169TIFF0007776068000058.tif202169

[0173] Flow cytometry analysis of apoptosis Approximately 1 x 10 6Melanoma cells were treated with inhibitors for 24 hours, then trypsinized and washed with PBS. Single-cell suspensions were incubated with Alexa Fluor-annexin V and propidium iodide (PI) (V13245; Invitrogen) according to the manufacturer's protocol and subjected to flow cytometry analysis. In all cases, cell debris was excluded (gated out) based on forward and side scatter analysis. Data were analyzed using FlowJo (Ashland, OR). The results are shown in Figure 1, in which WM3248 melanoma cells were treated with the indicated doses of Compound 1 (ARN12405), and apoptosis was measured by flow cytometry using annexin V and PI.

[0174] Cdc42 activity assay Cdc42 activation assays were performed according to the manufacturer's protocol (Cell Biolabs, San Diego, CA). Briefly, inhibitor-treated cells were lysed and loaded with GDP or GTPγS. Agarose beads conjugated with the PAK1 PBD domain pulled down Cdc42, Rac, or RhoJ only when GTP-bound. Lysates were then immunoblotted with the indicated Abs. The results are shown in Figure 2. Specifically, the Cdc42 activation assay was used to determine whether inhibitors blocked the ability of RhoJ to interact with PAK-coupled beads. Briefly, cells were treated with compound 1 (ARN12405), cell lysates were generated, and then incubated with PAK-coupled beads. Immunoprecipitated proteins were subjected to SDS-PAGE and immunoblotted with (A) RhoJ, (B) Cdc42, and (C) Rac1 antibodies. (D) Cells were treated with one of a panel of RhoJ interaction inhibitors at 10 μM or 50 μM concentrations (see Figure 2, Panels A-D). Lysates were prepared and incubated with PAK-coupled beads, and immunoprecipitated proteins were immunoblotted with RhoJ antibodies.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, X １ and X ２ are independently CH ２ , N.R. ２ and O, with the proviso that X １ and X ２ Both are NR ２ not both O, not X 1 =NR 2 and X 2 =O, and not X 1 =O and X 2 =NR 2; Y is selected from the group consisting of CH and N; A and A′ are independently: a 6-membered aromatic heterocycle containing 1 or 2 nitrogen atoms; - at any position, "C 1-6 alkyl, C 1-6 alkenyl, C 1-6 alkynyl, halogen, halo-C １～６ C substituted with alkyl, hydroxyl, alkoxy groups １～６ Alkyl, amino, one C １～６ Amino substituted with alkyl groups and two C １～６ a 6-membered aromatic ring optionally substituted with one or more substituents selected from the group consisting of amino substituted with alkyl groups, or A is: a 6-membered aromatic heterocycle containing 1 or 2 nitrogen atoms; -C 1-6 alkyl, C 1-6 alkenyl, C 1-6 alkynyl, halogen, halo-C at any position １～６ C substituted with alkyl, hydroxyl, alkoxy groups １～６ Alkyl, amino, one C １～６ Amino substituted with alkyl groups and two C １～６ a 6-membered aromatic ring optionally substituted with one or more substituents selected from the group consisting of amino substituted with alkyl groups; A' is a methoxy group and two C groups at the para or meta positions. ２ a 6-membered aromatic ring substituted with a substituent selected from the group consisting of amino substituted with alkyl groups; R １ is hydrogen and C １～６ alkyl, R ２ is hydrogen, C １～６ C substituted with alkyl or alkoxy groups １～６ Alkyl, —CO—C １～６ Alkyl and -CO-C １～６ alkyl-alkoxy), or A compound having the formula: 【Chemistry 2】 or a pharmaceutically acceptable salt or solvate thereof; at least one pharmaceutically acceptable excipient; A pharmaceutical composition comprising:

2. A and A' are independently a 6-membered aromatic heterocycle containing one nitrogen atom; C substituted with halogen or alkoxy group at any position １～６ Alkyl, amino, one C １～６ Amino substituted with alkyl groups and two C １～６ a 6-membered aromatic ring optionally substituted with a substituent selected from the group consisting of amino substituted with an alkyl group; selected from the group consisting of R １ is hydrogen, R ２ The pharmaceutical composition of claim 1, wherein is hydrogen.

3. X １ is CH ２ , NH and O; X ２ is CH ２ and NH; Y is CH ２ and A is, a 6-membered aromatic heterocycle containing one nitrogen atom at the 2- or 3-position; C substituted with halogen and alkoxy １～３ a 6-membered aromatic ring optionally substituted with a substituent selected from the group consisting of alkyl; selected from the group consisting of A' is a C substituted with halogen or alkoxy at the meta or para position. １～６ Alkyl and two C １～６ a 6-membered aromatic ring optionally substituted with a substituent selected from the group consisting of an amino substituted with an alkyl group; R １ is hydrogen, R ２ The pharmaceutical composition according to claim 1 or 2, characterized in that is hydrogen.

4. 2. The pharmaceutical composition according to claim 1, wherein A is selected from the group consisting of a 6-membered aromatic heterocycle containing one nitrogen atom and an unsubstituted 6-membered aromatic ring.

5. The compound is Table 1 The pharmaceutical composition according to claim 1 or 2, selected from the group consisting of:

6. 6. The pharmaceutical composition of any one of claims 1 to 5, further comprising a chemotherapeutic agent selected from the group consisting of cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, dacarbazine, and temozolamide.

7. The pharmaceutical composition according to claim 6, which is a combined preparation for simultaneous, sequential or separate administration.

8. The pharmaceutical composition according to any one of claims 1 to 7, for use in treating a disease or disorder associated with an increase in expression or function relative to physiological or desired RhoJ / Cdc42 levels.

9. The pharmaceutical composition according to any one of claims 1 to 7, for use in the treatment of primary and metastatic neoplastic diseases, precancerous conditions (e.g., hyperplasia, metaplasia, or dysplasia), cancer, cancer metastasis, benign tumors, hyperproliferative disorders, cardiomyopathies, and retinal disorders.

10. The pharmaceutical composition of claim 9 , wherein the cancer is melanoma.

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