TW201605857A - Therapeutic inhibitors of CDK8 and uses thereof - Google Patents

Abstract

The present invention relates to compounds formula (I): and to salts thereof, wherein R1-R4 have any of he values defined in the specification, and compositions and uses thereof. The compounds are useful as inhibitors of CDK8. Also included are pharmaceutically acceptable compositions comprising the compounds of the present invention and methods of using said compositions in the treatment of various disorders.

Description

Medical inhibitor of CDK8 and its use

This invention relates to compounds useful as CDK8 inhibitors.

CDK8 is component ² of the mediator complex. The mediator complex plays an important role in transcription by coupling sequence-specific transcription factors with transcriptional machinery. CDK8, together with cyclin C, MED12 and MED13, forms a CDK8 module of the complex. It phosphorylates the C-terminal domain of RNA polymerase II, the serine 10 of histone H3, and the serine acid 206 of Smadl ³ as well as many other receptors. It regulates the transcriptional pathway, including beta-soda, Notch, p53, serum response factors, and SMAD. It also acts indirectly on β- ^{sodamer 4,5} by phosphorylating E2F1 and thereby reversing its inhibition of β-sodacin activity.

CDK8 is an oncogene ^6,7 . It is amplified in colon cancer and is overexpressed. In addition, it is capable of transforming NIH3T3 cells as evidenced by loss of contact inhibition, fixation-independent growth, and tumor growth in vivo. It is worth noting that the transformation does not occur in the case of kinase-death CDK8 and, therefore, it appears to be dependent on kinase activity. The CDK8 blocking gene expression of a cell line with an increased number of copies will inhibit cell proliferation. Further, inhibition of tumor growth was observed in a murine xenograft model ⁸ having inducible expression of genes in block CDK8. One proposed mechanism for CDK8 mediated inhibition in colon cancer is to down-regulate the beta-sucrose pathway in cancers with higher CDK8 levels. However, blocking gene expression results in the ability to discern the backbone of CDK8 and the importance of kinase function.

CDK8 and its agonist cyclin C are dysregulated in human cancers ⁹ . Retrospective analysis of human colon tumor samples showed that CDK8 performance was associated with beta-sodium hydroxyl activation and poor prognosis ^10,11 . CDK8 also found to induce undesirable enrichment of the differentiated colon tumors of embryonic stem cells and is associated with poor prognosis tag ^8. Other cell lines, murine xenografts, and human tumor tissue studies have also linked CDK8 to melanoma ¹³ . The therapeutic hypothesis based on this study is that small molecule inhibitors of CDK8 will inhibit tumor subset growth that exhibits CDK8 expansion and/or overexpression. Possible indications include colon cancer and melanoma.

There is a current need for compounds that inhibit CDK8 in the treatment of hyperproliferative diseases.

One aspect includes a compound of formula (I): Or a salt thereof, wherein: R ¹ is selected from the group consisting of: And wherein R ¹ is independently substituted by 1, 2 or 3 R ^x groups; R ² and R ^{3 are} independently H, C _1-12 alkyl, C _2-12 alkenyl, C _2-12 Alkynyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, halo, -OR ^a , -SR ^a , -N(R ^a ) ₂ , -CN, -NO ₂ , -C(O)R ^a , -CO ₂ R ^a , -C(O)N(R ^a ) ₂ , -C(O)SR ^a , -C(O)C(O)R ^a , -C(O)CH ₂ C(O R ^a , -C(S)N(R ^a ) ₂ , -C(S)OR ^a , -S(O)R ^a , -SO ₂ R ^a , -SO ₂ N(R ^a ) ₂ , -N (R ^a )C(O)R ^a , -N(R ^a )C(O)N(R ^a ) ₂ , -N(R ^a )SO ₂ R ^a , -N(R ^a )SO ₂ N(R ^a ) ₂ , -N(R ^a )N(R ^a ) ₂ , -N(R ^a )C(=N(R ^a ))N(R ^a ) ₂ , -C(=N)N(R ^a ) ₂ , -C=NOR ^a , -C(=N(R ^a ))N(R ^a ) ₂ , -OC(O)R ^a or - OC(O)N(R ^a ) ₂ , wherein R ² and R ³ of each of C _1-12 alkyl, C _2-12 alkenyl group, C _2-12 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl groups are each independently system optionally substituted with one or more groups a group R ^x substituted; or R ¹ and R ² together with the atom to which they are attached form a 4, 5, 6, 7 or 8 membered carbocyclyl, heterocyclyl, heteroaryl or aryl group, which is a heterocyclic or heterocyclic ring. group, aryl group and heteroaryl group optionally substituted with one or more radicals R ^x taken ; Each R ^a and R ^b are independently H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl, wherein each C _1-6 alkyl, C _3-6 alkenyl, C _3-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclic are independently substituted by one or more groups R ^x as appropriate ; or R ^a and R ^b form a 3-6 membered heterocyclic ring together with the nitrogen to which they are attached, which heteroaryl ring optionally substituted with oxo or halo by oxo, halo or C _1-6 alkyl optionally of Substituted; each R ^{v is} independently hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl, wherein each C _{1 -6} alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl group optionally substituted with one or more groups, one or more of the group The group is independently selected from the group consisting of a pendant oxy group, a halogen group, an amine group, a hydroxyl group, and optionally a C ₁ -C ₆ alkyl group substituted with one or more groups independently selected from the pendant oxy group and a halogen group; or two forming a R ^v together with the nitrogen to which they are attached 3-6 membered heterocyclic group, the heterocyclic group optionally substituted by one or more groups, the one or more groups independently selected from side-based oxygen , Halo and optionally substituted with one or more of the substituents independently selected from halo and oxo group of the C _1-3 alkyl group; and each R ^x is independently oxo, C _1-6 alkyl , C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 haloalkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, -F, -Cl, -Br, -I, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N( R ^v ) ₂ , -OR ^v , -SR ^v , -OC(O)-R ^v , -OC(O)-OR ^v , -C(O)-R ^v , -C(O)-OR ^v ,- S(O)-R ^v , -S(O) ₂ -R ^v , -OC(O)-N(R ^v ) ₂ , -N(R ^v )-C(O)-OR ^v , -N(R ^v )-C(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v , -N(R ^v ) -S(O)-R ^v , -N(R ^v )-S(O) ₂ -R ^v , -N(R ^v )-S(O)-N(R ^v ) ₂ or -N(R ^v ) -S(O) ₂ -N(R ^v ) ₂ , wherein any C _1-6 alkyl group, C _2-6 alkenyl group, C _2-6 alkynyl group, C _1-6 haloalkyl group, carbocyclic group, aromatic The base, heteroaryl and heterocyclic groups are, independently, optionally substituted by one or more groups selected from the group consisting of pendant oxy, halo, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -OR ^v ,- SR ^v , -OC(O)-R ^v , -C(O)-R ^v , -C(O)-OR ^v , -S(O)-R ^v , -S(O) ₂ -R ^v , -C(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v , -N(R ^v )-S(O)-R ^v , -N(R ^v And -S(O) ₂ -R ^v and, optionally, a C _1-6 alkyl group substituted with one or more groups independently selected from the group consisting of pendant and halogen; and wherein when R ² is F and R ³ In the case of hydrogen, R ^{x is} not methyl, ethyl, trifluoromethyl or 2-fluorophenyl.

Another aspect includes a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant, carrier or vehicle.

Another aspect includes a method of treating a condition associated with CDK8 activity comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in therapy. Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the treatment of a hyperproliferative disorder (in one example, cancer).

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a disease associated with CDK8 activity.

Another aspect includes compounds for studying kinases such as CDK8, studying intracellular signal transduction pathways mediated by the CDK8, and comparatively evaluating CDK8 modulators.

Another aspect includes a method of preparing a compound of Formula I or a salt thereof.

Compounds and definitions

Definitions and terms are described in more detail below. The chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics , 75th edition.

Unless otherwise stated, a compound of formula (I) includes the enantiomeric, diastereomeric, and geometric (or conformational) isomeric forms of the specified structure. For example, including R and S configurations for each asymmetric center, Z and E double bond isomers, Z and E configuration isomers, single stereochemical isomers, and enantiomeric, diastereomeric Heterogeneous and geometric (or conformational) mixtures. All tautomeric forms of the structures depicted herein are included unless otherwise stated. Additionally, unless otherwise stated, structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. By way of example, include a compound of formula I wherein one or more of the following independent substitutions or enrichment are carried out: replacement or enrichment of hydrogen with hydrazine or hydrazine, replacement or enrichment of carbon with ¹³ C or ¹⁴ C carbon, replacement with ¹⁵ N nitrogen Or enrichment of nitrogen, displacement or enrichment of sulfur with ³³ S, ³⁴ S or ³⁶ S sulfur, or replacement or enrichment of oxygen with ¹⁷ O or ¹⁸ O oxygen. Such compounds are useful, for example, as analytical tools, probes in biological assays, or therapeutic agents.

Where a particular enantiomer is described, in certain embodiments, an enantiomer that is substantially free of the corresponding enantiomer may be provided and may also be referred to as "optically enriched." As used herein, "optically enriched" means a mixture of enantiomers consisting of a significantly larger proportion of one enantiomer and which may be described by enantiomeric excess (ee%). In certain embodiments, the enantiomeric mixture consists of at least about 90% by weight of the designated enantiomer (about 90% ee). In other embodiments, the enantiomeric mixture consists of at least about 95%, 98%, or 99% by weight of the designated enantiomer (about 95%, 98%, or 99% ee). The enantiomers and diastereomers can be separated from the racemic mixture by any method known to those skilled in the art, including recrystallization from a solvent (wherein one stereoisomer has a solubility greater than the other a method of high pressure liquid chromatography (HPLC), supercritical fluid chromatography (SFC), formation of a palmitic salt and crystallization thereof, followed by separation by or by any of the above methods Asymmetric synthesis and further enrichment as appropriate. See, for example, Jacques et al, Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al, Tetrahedron 33: 2725 (1977); Eliel, EL Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SH Tables of Resolving Agents and Optical Resolutions, p. 268 (edited by ELEliel, Univ. of Notre Dame Press, Notre Dame, IN 1972).

The term "heteroatom" means any atom independently selected from an atom other than carbon or hydrogen, such as one or more of oxygen, sulfur, nitrogen, phosphorus or antimony (including any oxidation of nitrogen, sulfur, phosphorus or antimony). Form; and any quaternary ammonium form of nitrogen).

The terms "halo" and "halogen" as used herein mean selected from fluoro (fluoro, -F), chloro (chloro, -Cl), bromo (bromo, -Br) and iodine (iodo). , the atom of -I). Specific examples of the halogen group are a fluorine group and a chlorine group.

The term "unsaturated" as used herein means that the moiety has one or more units of unsaturation.

The term "carbocyclyl" used alone or as part of a larger moiety refers to a saturated, partially unsaturated or aromatic ring system having from 3 to 20 carbon atoms. In one embodiment, a carbocyclic group comprises from 3 to 12 carbon atoms (C ₃ -C _12). In another embodiment, the carbocyclic group comprises C ₃ -C ₈ , C ₃ -C ₁₀ or C ₅ -C ₁₀ . In other embodiments, as a form of monocyclic carbocyclic groups include _{C 3 -C 8, C 3 -C} 6 or C ₅ -C _6. In another embodiment, the carbocyclic group in the form of a bicyclic ring comprises C ₇ -C ₁₂ . In another embodiment, the form of the spiro ring system carbocyclic groups include C ₅ -C _12. Examples of the monocyclic carbocyclic group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a 1-cyclopent-1-enyl group, a 1-cyclopent-2-enyl group, a 1-cyclopent-3-enyl group, Cyclohexyl, deuterated cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl , cyclodecyl, cyclodecyl, cyclodecyl, phenyl and cyclododecyl; bicyclic carbocyclyl having 7 to 12 ring atoms including [4,3], [4,4], [4,5], [5,5], [5,6] or [6,6] ring systems, such as bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane, naphthalene and bicyclo [ 3.2.2] decane; and the spiro carbocyclic group includes spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane and spiro[4.5]decane, each carbon The ring group is independently substituted with one or more of the groups described herein, as appropriate. The term carbocyclyl includes an aryl ring system as defined herein.

The term "alkyl" as used herein refers to a saturated straight or branched chain monovalent hydrocarbon group. In one embodiment, an alkyl group of 1 to 18 carbon atoms (C ₁ -C _18). In other embodiments, the alkyl group is C ₀ -C ₆ , C ₀ -C ₅ , C ₀ -C ₃ , C ₁ -C ₁₂ , C ₁ -C _{10 ,} C ₁ -C ₈ , C ₁ -C ₆ , C ₁ -C ₅ , C ₁ -C ₄ or C ₁ -C ₃ . C ₀ alkyl means a bond. Examples of the alkyl group include a methyl group (Me, -CH ₃ ), an ethyl group (Et, -CH ₂ CH ₃ ), a 1-propyl group (n-Pr, n-propyl group, -CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, isopropyl, -CH(CH ₃ ) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl 1-propyl (i-Bu, isobutyl, -CH ₂ CH(CH ₃ ) ₂ ), 2-butyl (s-Bu, s-butyl, -CH(CH ₃ )CH ₂ CH ₃ ) , 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH ₃ ) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ) , 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C(CH) ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH ( CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2 -hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2- Pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH (CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), heptyl, octyl, decyl, decyl, undecyl And dodecyl. A specific example of a C _1-12 alkyl group is a C _1-6 alkyl group. Specific examples of the C _1-6 alkyl group are a methyl group and an ethyl group.

The term "alkenyl" as used herein denotes a straight or branched chain monovalent hydrocarbon radical having at least one double bond. Alkenyl groups include groups having "cis" and "trans" orientations or "E" and "Z" orientations. In one example, the alkenyl group is 2 to 18 carbon atoms (C ₂ -C ₁₈ ). In other examples, the alkenyl group is C ₂ -C ₁₂ , C ₂ -C ₁₀ , C ₂ -C ₈ , C ₂ -C ₆ or C ₂ -C ₃ . Examples include, but are not limited to, ethenyl or vinyl (-CH=CH ₂ ), prop-1-enyl (-CH=CHCH ₃ ), prop-2-enyl (-CH ₂ CH=CH ₂ ), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, butane-1,3-dienyl, 2-methylbutyl- 1,3-diene, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl and hexa-1,3-dienyl.

The term "alkynyl" as used herein refers to a straight or branched chain monovalent hydrocarbon radical having at least one carbon-carbon triple bond. In one example, the alkynyl group is 2 to 18 carbon atoms (C ₂ -C ₁₈ ). In other examples, the alkynyl group is C ₂ -C ₁₂ , C ₂ -C ₁₀ , C ₂ -C ₈ , C ₂ -C ₆ or C ₂ -C ₃ . Examples include, but are not limited to, ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH ₃ ), prop-2-ynyl (propargyl, -CH ₂ C≡CH), 1-ynyl, but-2-ynyl and but-3-ynyl.

The term "alkoxy" refers to a straight or branched chain monovalent group represented by the formula -OR wherein R is alkyl, alkenyl, alkynyl or carbocyclyl. Alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, monofluoromethoxy, difluoromethoxy, and trifluoromethoxy and cyclopropoxy. A specific example of an alkoxy group is a methoxy group.

The term "haloalkyl" as used herein, refers to an alkyl group, as defined herein, substituted by one or more (eg 1, 2, 3 or 4) halo groups. Specific examples of haloalkyl groups are trifluoromethyl and difluoromethyl.

The term "partially used alone or as part of a larger moiety (such as "arylalkyl", "arylalkoxy" or "aryloxyalkyl") refers to a monocyclic, bicyclic or tricyclic carbocyclic ring system, It includes a fused ring wherein at least one of the rings in the system is aromatic. The term "aryl" is used interchangeably with the term "aryl ring". In one embodiment, the aryl group includes a group having 6 to 18 carbon atoms. In another embodiment, the aryl group includes a group having 6-10 carbon atoms. Examples of the aryl group include phenyl, naphthyl, anthracenyl, biphenyl, phenanthryl, fused tetraphenyl, 1,2,3,4-tetrahydronaphthyl, 1H-fluorenyl, 2,3-dihydro- 1H-indenyl and the like, which may be substituted, independently or independently, with one or more substituents as described herein. One particular aryl group is a phenyl group. In another embodiment, the aryl group includes an aryl ring fused to one or more carbocyclic rings, such as indanyl, phthalimido, naphthoquinone, phenanthryl or tetrahydrogen. Naphthalene And a group thereof, wherein the linking group or point is on the aromatic ring.

The term "heteroaryl" used alone or as part of a larger moiety, such as "heteroarylalkyl" or "heteroarylalkoxy", refers to a monocyclic, bicyclic, or 5- to 14 ring atom. A tricyclic system wherein at least one ring is aromatic and contains at least one hetero atom. In one embodiment, the heteroaryl group comprises a 4-6 membered monocyclic aromatic group wherein one or more ring atoms are nitrogen, sulfur or oxygen and are independently substituted as appropriate. In another embodiment, the heteroaryl group comprises a 5-6 membered monocyclic aromatic group wherein one or more ring atoms are independently substituted nitrogen, sulfur or oxygen. Examples of heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetra Azyl, thiatriazole, oxatriazole, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolo[1,5-b]pyridazinyl, imidazole [1,2-a]pyrimidinyl, fluorenyl, benzoxazolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, fluorenyl , 1,3-thiazol-2-yl, 1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl, 1 , 2,4-oxadiazol-5-yl, 1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl, 1,2,3-triazol-5-yl and pyridine -2-yl N-oxide. The term "heteroaryl" also includes groups in which a heteroaryl group is fused to one or more aryl, carbocyclyl or heterocyclyl rings wherein the linking group or point is on the heteroaryl ring. Non-limiting examples include mercapto, isodecyl, benzothienyl, benzofuranyl, dibenzofuranyl, oxazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquine Olinyl group, Lolinyl, pyridazinyl, quinazolinyl, quinoxalinyl, 4 H -quinazinyl, oxazolyl, acridinyl, phenazinyl, phenothiazine, phenoxazinyl, tetrahydroquinoline Base, tetrahydroisoquinolyl and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. The heteroaryl group can be monocyclic, bicyclic or tricyclic.

As used herein, the term "heterocyclyl" refers to a "carbocyclyl" group as defined herein, wherein one or more (eg 1, 2, 3 or 4) carbon atoms have been heteroatoms (eg O, N or S) replacement. The heterocyclic group may be optionally substituted with one or more substituents independently selected from the substituents defined herein.

In one example, a heterocyclyl includes 3-12 ring atoms and includes monocyclic, bicyclic, tricyclic, and spiro ring systems wherein the ring atom is carbon and 1 to 5 ring atoms are selected from nitrogen, sulfur, or oxygen. A hetero atom, which is independently substituted with one or more groups, as appropriate. In one example, a heterocyclic group includes from 1 to 4 heteroatoms. In another example, a heterocyclic group includes a 3 to 7 membered monocyclic ring having one or more heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, a heterocyclic group includes a 4 to 6 membered monocyclic ring having one or more heteroatoms selected from nitrogen, sulfur, or oxygen. In another example, a heterocyclic group includes a 3-membered single ring. In another example, a heterocyclic group includes a 4-membered single ring. In another example, a heterocyclic group includes a 5-6 membered monocyclic ring. In one example, a heterocyclic group includes 0 to 3 double bonds. Any nitrogen or sulfur heteroatom may be oxidized (eg, NO, SO, SO ₂ ) as appropriate, and any nitrogen heteroatom may be ammonium quaternized as appropriate (eg, [NR ₄ ] ⁺ Cl ^- , [NR ₄ ] ⁺ OH ^- ). Exemplary heterocyclic groups include oxiranyl, aziridine, thietyl, azetidinyl, oxetanyl, thietane, 1,2-disulfide Heterocyclic butane, 1,3-dithiacycloalkyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothienyl, tetrahydrothiophenyl, imidazolidinyl , piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-di-oxy-thiomorpholinyl, dihydropiperidyl, tetrahydropyranyl, hexahydrosulfur Piperacinyl, hexahydropyrimidinyl, oxazepine, thiazepine, thioxanthyl, homopiperazinyl, homopiperidinyl, azepanyl, oxygen Heterocyclic heptyl, thiaheptanyl, oxazinyl, oxazepine, diazepine, 1,4-diazepanyl, diazenium , thiazolidine, thiazepine, tetrahydrothiopiperidyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-di- oxyisothiazolidinone, Oxazolidinone, imidazolidinone, 4,5,6,7-tetrahydro[2H]carbazolyl, tetrahydrobenzimidazolyl, 4,5,6, 7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl,oxazinyl, thiodiazepine Heterocyclohexane, oxadiazepine, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, Dihydropyrimidinyl, tetrahydropyrimidinyl, 1-pyrroline, 2-pyrolinyl, 3-pyrroyl, porphyrin, thiopyranyl, 2H-pyranyl, 4H-pyranyl, Dioxoalkyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithiaalkyl, dithiolanyl, pyrimidinone, pyrimidindione, pyrimidine-2 , 4-dione, piperazinone, piperazinedione, pyrazolyl imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3,6-diazabicyclo[3.1.1 Heptyl, 6-azabicyclo[3.1.1]heptyl, 3-azabicyclo[3.1.1]heptyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2 Hexyl, 2-azabicyclo[3.2.1]octyl, 8-azabicyclo[3.2.1]octyl, 2-azabicyclo[2.2.2]octyl, 8-azabicyclo[2.2. 2] Octyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]fluorenyl, aza Snail [2.5] octyl, azaspiro[4.5]decyl, 1-azaspiro[4.5]non-2-one, azaspiro[5.5]undecyl, tetrahydroindenyl, octahydro Indenyl, tetrahydroisodecyl, tetrahydrocarbazolyl, 1,1-di-oxy hexahydrothiopiperidyl. Examples of a 5-membered heterocyclic group containing 1 sulfur or oxygen atom and 1 to 3 nitrogen atoms are thiazolyl (including thiazol-2-yl and thiazol-2-yl N-oxide), thiadiazolyl (including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl), oxazolyl (such as oxazol-2-yl) and oxadiazolyl (such as 1, 3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl). Exemplary 5-membered ring heterocyclyl containing 2 to 4 nitrogen atoms includes imidazolyl, such as imidazol-2-yl; triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3 a triazol-5-yl, 1,2,4-triazol-5-yl; and a tetrazolyl group such as a 1H-tetrazol-5-yl group. Exemplary benzo-fused 5-membered heterocyclic groups are benzoxazol-2-yl, benzothiazol-2-yl and benzimidazol-2-yl. Exemplary 6 membered heterocyclic groups contain 1 to 3 nitrogen atoms and optionally sulfur or oxygen atoms, such as pyridyl, such as pyridin-2-yl, pyridin-3-yl and pyridin-4-yl; pyrimidinyl, Such as pyrimidin-2-yl and pyrimidin-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, in detail Pyridazin-3-yl; and pyrazinyl. Pyridine N-oxides and pyridazine N-oxides and pyridyl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazinyl and 1,3,4-triazin-2-yl are other exemplary heterocyclic rings base. Specific examples of heterocyclic groups are azetidinyl, pyrrolidinyl, piperazinyl and morpholinyl, more specifically azetidin-1-yl, pyrrolidin-1-yl, piperazine -1-yl and morpholin-4-yl.

As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond between ring atoms and the ring moiety is not aromatic.

As used herein, the term "inhibitor" refers to a compound that binds to and inhibits the CDK8 enzyme with a measurable biochemical affinity and activity. In certain embodiments, an inhibitor of biochemical IC ₅₀ and / or the binding constant of less than about 50 M, less than about [mu] M, less than about 500 nM, less than about 100nM, or less than about 10nM.

"Pharmaceutically acceptable salts" include acid and base addition salts. "Pharmaceutically acceptable acid addition salt" means a salt which retains the biological effectiveness and properties of the free base and which is not biologically or otherwise undesirable, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid. The inorganic acid of carbonic acid, phosphoric acid and the like is formed, and the organic acid may be selected from the group consisting of aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic acid and sulfonic acid organic acids, such as formic acid, acetic acid, and C. Acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamine Acid, o-aminobenzoic acid, benzoic acid, cinnamic acid, mandelic acid, en-poic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.

"Pharmaceutically acceptable base addition salts" include salts derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese salts. , aluminum salts and their analogues. Specifically, the base addition salts are ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of the following: primary amines, secondary amines and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines and basic ions Exchange resin such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethylolaminomethane, dicyclohexylamine, lysine, fine Aminic acid, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, cocoa beans Base, hydrazine, piperazine, piperidine, N-ethylpiperidine, polyamine resin and the like. Specifically, the organic non-toxic base is isopropylamine, diethylamine, ethanolamine, trimethylolaminomethane, dicyclohexylamine, choline and caffeine.

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as proton transfer tautomers) include interconversion via proton transfer, such as keto-enol and imine-enamine isomerization. Valence bond tautomers include interconversions by some bond electron recombination.

"Solvate" means an association or complex of one or more solvent molecules with a compound of the invention. Examples of the solvent include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

"Therapeutically effective amount" means (i) treating a particular disease, condition or condition, (ii) reducing, ameliorating or eliminating one or more symptoms of a particular disease, condition or condition, or (iii) preventing or delaying the treatment described herein. The amount of a compound of the invention that is one or more of the symptoms of a particular disease, condition or condition. In the case of cancer, a therapeutically effective amount of the drug reduces the number of cancer cells; reduces the size of the tumor; inhibits (ie, to some extent, and preferably terminates) the infiltration of the cancer cells into the surrounding organs; To some extent, and preferably terminates) tumor metastasis; inhibits tumor growth to some extent; and/or relieves one or more symptoms associated with cancer to some extent. For cancer therapy, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and/or determining the response rate (RR). In the case of an immune disorder, the therapeutically effective amount is an amount sufficient to reduce or alleviate the symptoms of an allergic condition, an autoimmune and/or inflammatory disease, or a symptom of an acute inflammatory response, such as asthma. In some embodiments, the therapeutically effective amount is an amount of a chemical entity described herein sufficient to significantly reduce the activity or number of drug resistant cancer cells or drug resistant refractory cancer cells.

"Treatment" (and variants such as "treat" or "treating") refers to the natural process of attempting to alter the individual or cell being treated and may be for preventive purposes or in clinical pathology. Clinical intervention performed during the process. The ideal effect of treatment includes one or more of the following: preventing the occurrence or recurrence of the disease, alleviating the symptoms, reducing any direct or indirect pathological consequences of the disease, stabilizing (ie, not worsening) the disease state, pre- Preventing metastasis, reducing the rate of disease progression, improving or mitigating the disease state, prolonging survival time and ameliorating or improving prognosis compared to the expected survival time when no treatment is received. In certain embodiments, a compound of Formula I is used to delay or slow the progression of a disease or condition. The individual in need of treatment includes an individual who has a condition or disorder as well as an individual who is prone to have a condition or disorder (e.g., by mutation or abnormal expression of a gene or protein) or an individual to be prevented from developing a condition or disorder.

Exemplary value

One embodiment includes a compound of formula (I): Or a salt thereof, wherein: R ¹ is selected from the group consisting of: And wherein R ¹ is independently substituted by 1, 2 or 3 R ^x groups; R ² and R ^{3 are} independently H, C _1-12 alkyl, C _2-12 alkenyl, C _2-12 Alkynyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, halo, -OR ^a , -SR ^a , -N(R ^a ) ₂ , -CN, -NO ₂ , -C(O)R ^a , -CO ₂ R ^a , -C(O)N(R ^a ) ₂ , -C(O)SR ^a , -C(O)C(O)R ^a , -C(O)CH ₂ C(O R ^a , -C(S)N(R ^a ) ₂ , -C(S)OR ^a , -S(O)R ^a , -SO ₂ R ^a , -SO ₂ N(R ^a ) ₂ , -N (R ^a )C(O)R ^a , -N(R ^a )C(O)N(R ^a ) ₂ , -N(R ^a )SO ₂ R ^a , -N(R ^a )SO ₂ N(R ^a ) ₂ , -N(R ^a )N(R ^a ) ₂ , -N(R ^a )C(=N(R ^a ))N(R ^a ) ₂ , -C(=N)N(R ^a ) ₂ , -C=NOR ^a , -C(=N(R ^a ))N(R ^a ) ₂ , -OC(O)R ^a or -OC(O)N(R ^a ) ₂ , wherein R ² and R ³ of each of C _1-12 alkyl, C _2-12 alkenyl group, C _2-12 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl groups are each independently system optionally substituted with one or more groups a group R ^x substituted; or R ¹ and R ² together with the atom to which they are attached form a 4, 5, 6, 7 or 8 membered carbocyclyl, heterocyclyl, heteroaryl or aryl group, which is a heterocyclic or heterocyclic ring. The base, heteroaryl and aryl are optionally substituted by one or more groups R ^x Each of R ^a and R ^{b is} independently H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclic, each of which C _1-6 alkyl, C _3-6 alkenyl, C _3-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclic are independently substituted by one or more groups R ^x as appropriate ; or R ^a and R ^b form a 3-6 membered heterocyclic ring together with the nitrogen to which they are attached, which heteroaryl ring optionally substituted with oxo or halo by oxo, halo or C _1-6 alkyl optionally of Substituted; each R ^{v is} independently hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl, wherein each C _{1 -6} alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl group optionally substituted with one or more groups, one or more of the group The group is independently selected from the group consisting of a pendant oxy group, a halogen group, an amine group, a hydroxyl group, and optionally a C ₁ -C ₆ alkyl group substituted with one or more groups independently selected from the pendant oxy group and a halogen group; or two forming a R ^v together with the nitrogen to which they are attached 3-6 membered heterocyclic group, the heterocyclic group optionally substituted by one or more groups, the one or more groups independently selected from side-based oxygen , Halo and optionally substituted with one or more of the substituents independently selected from halo and oxo group of the C _1-3 alkyl group; and each R ^x is independently oxo, C _1-6 alkyl , C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 haloalkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, -F, -Cl, -Br, -I, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N( R ^v ) ₂ , -OR ^v , -SR ^v , -OC(O)-R ^v , -OC(O)-OR ^v , -C(O)-R ^v , -C(O)-OR ^v ,- S(O)-R ^v , -S(O) ₂ -R ^v , -OC(O)-N(R ^v ) ₂ , -N(R ^v )-C(O)-OR ^v , -N(R ^v )-C(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v , -N(R ^v ) -S(O)-R ^v , -N(R ^v )-S(O) ₂ -R ^v , -N(R ^v )-S(O)-N(R ^v ) ₂ or -N(R ^v ) -S(O) ₂ -N(R ^v ) ₂ , wherein any C _1-6 alkyl group, C _2-6 alkenyl group, C _2-6 alkynyl group, C _1-6 haloalkyl group, carbocyclic group, aromatic The base, heteroaryl and heterocyclic groups are, independently, optionally substituted by one or more groups selected from the group consisting of pendant oxy, halo, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -OR ^v ,- ^{SR v, -OC (O) -R} v ^{-C (O) -R v, -C} (O) -OR v, -S (O) -R v, -S (O) 2 -R v, -C (O) -N (R v) 2, -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v , -N(R ^v )-S(O)-R ^v , -N(R ^v ) -S(O) ₂ -R ^v and, optionally, a C _1-6 alkyl group substituted with one or more groups independently selected from the group consisting of pendant and halogen; and wherein when R ² is F and R ³ is In the case of hydrogen, R ^{x is} not a methyl group, an ethyl group, a trifluoromethyl group or a 2-fluorophenyl group.

A particular embodiment relates to a compound of formula (I) or a salt thereof, wherein: R ¹ is selected from the group consisting of: R ² and R ^{3 are} independently H, C _1-12 alkyl, halo or -OR ^a , wherein each C _1-12 alkyl of R ² and R ³ is independently independently one or more groups, as appropriate R ^{x is} substituted; each R ^a and R ^{b is} independently H, C _1-6 alkyl, wherein each C _1-6 alkyl group is independently substituted with one or more groups R ^x as appropriate; each R ^v independently Is a C _1-6 alkyl group; and each R ^{x is} independently a heterocyclic group, -F or -N(R ^v ) ₂ , wherein any heterocyclic group is independently selected from one or more selected from C _{1- a} group of ₆ alkyl groups; and wherein when R ² is F and R ³ is hydrogen, then R ^{x is} not methyl, ethyl, trifluoromethyl or 2-fluorophenyl.

In certain embodiments, R ² and R ^{3 are} independently halo, trifluoromethyl, C _1-6 alkyl or C _1-6 alkoxy, wherein the alkyl and alkoxy are independently considered The case is substituted by -N(R ^v ) ₂ or a heterocyclic group, wherein the heterocyclic group is optionally substituted by a pendant oxy group, a halogen group or a C _1-6 alkyl group optionally substituted by a halogen group or a pendant oxy group.

In certain embodiments, R ² is hydrogen, chloro, methyl, (4-methylpiperazin-1-yl)methyl, trifluoromethoxy, difluoromethoxy, 4-nitroheterocycle Butan-1-ylmethyl, morpholin-4-ylmethyl, dimethylaminomethyl, pyrrolidin-1-ylmethyl or 4-ethylpiperazin-1-ylmethyl.

In certain embodiments, R ³ is hydrogen, trifluoromethyl, fluoro or chloro.

In certain embodiments, each of R ^a and R ^{b is} independently hydrogen or C _1-6 alkyl.

In certain embodiments, R ^a is hydrogen.

In certain embodiments, R ^b is methyl.

In certain embodiments, R ^v is hydrogen or C _1-6 alkyl.

In certain embodiments, R ^v is methyl.

In certain embodiments, R ² is hydrogen, chloro, methyl, (4-methylpiperazin-1-yl)methyl, trifluoromethoxy, difluoromethoxy, 4-nitroheterocycle Butan-1-ylmethyl, morpholin-4-ylmethyl, dimethylaminomethyl, pyrrolidin-1-ylmethyl or 4-ethylpiperazin-1-ylmethyl; R ³ Is hydrogen, trifluoromethyl or chloro; R ^a and R ^{b are} independently hydrogen or methyl.

Another embodiment includes a compound having the formula:

Or a salt thereof, wherein R ¹ , R ² and R ^{3 are} as defined for any embodiment of a compound of formula I.

Another embodiment comprises a compound selected from the group consisting of 1-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidine 3-yl]-3-[3-(trifluoromethyl)phenyl]urea; 1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3S)-1- ([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; 1-(3-chlorophenyl)-3-[(3S) 1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; 1-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]-3-[3-(trifluoro Methyl)phenyl]urea; 1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3R)-1-([1,2,4]triazolo[4, 3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; 1-(3-chlorophenyl)-3-[(3R)-1-([1,2,4]triazolo [4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; (S)-1-(1-([1,2,4]triazolo[4,3-b] Pyridazin-6-yl)pyrrolidin-3-yl)-3-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea; 1-[(3S)-1-(2-Aminopyrimidin-4-yl)pyrrolidin-3-yl]-3-(3-chloro-4-methyl-phenyl)urea; 1-[(S --1-(2-amino-pyrimidin-4-yl)-pyrrolidin-3-yl]-3-(4-chloro-3-trifluoromethyl-phenyl)-urea; 1-[4- Chloro-3-(trifluoromethyl)phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]urea; 1-( 3-chloro-4-methyl-phenyl)-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]urea; 1-[( 3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]-3-[4-(trifluoromethoxy)phenyl]urea; 1-[4- (difluoromethoxy)phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]urea; (S)-1- (4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2- (methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-ylmethyl)-3-fluorophenyl) -3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-((dimethylamino) Methyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-ylmethyl)-3-chlorophenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) Pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-(1-(2- (Methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl) Phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-(N-? Polinylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-((2) Methylamino)methyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-ylmethyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidine) -4-yl)pyrrolidin-3-yl)urea; (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4 -(pyrrolidin-1-ylmethyl)-3-(trifluoromethyl)phenyl)urea; (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrole Pyridin-3-yl)-3-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea; (S)-1-(4) -((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) (r)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-(N-? Lolinylmethyl)-3-(trifluoromethyl)phenyl)urea; 1-(4-azetidin-1-ylmethyl-3-trifluoromethyl-phenyl)-3-[ (S)-1-(2-methylamino-pyrimidin-4-yl)-pyrrolidin-3-yl]-urea; (S)-1-(1-([1,2,4]triazole And [4,3-b]pyridazin-6-yl)pyrrolidin-3-yl)-3-(3-chloro-4-methylphenyl)urea; and a pharmaceutically acceptable salt thereof.

Another embodiment includes a compound selected from the group consisting of:

And its salt.

Use, formulation and administration Pharmaceutically acceptable composition

Another aspect includes a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the composition further comprises a pharmaceutically acceptable carrier, adjuvant or vehicle. In another embodiment, the composition further comprises an amount of a compound that is effective to moderately inhibit CDK8. In certain embodiments, the composition is formulated for administration to a patient in need thereof.

The term "patient" or "individual" as used herein refers to an animal, such as a mammal, such as a human. In one embodiment, the patient or system refers to a human.

The term "pharmaceutically acceptable carrier, adjuvant or vehicle" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles which can be used in the compositions of the invention include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin; serum proteins, such as human serum white eggs White; buffer substances such as phosphate; glycine; sorbic acid; potassium sorbate; a mixture of partial glycerides of saturated plant fatty acids; water; salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, Sodium chloride, zinc salt; colloidal cerium oxide; magnesium tricaprate; polyvinylpyrrolidone; cellulose-based material; polyethylene glycol; sodium carboxymethyl cellulose; polyacrylate; wax; Polyoxypropylene-block polymer; polyethylene glycol and lanolin.

A composition comprising a compound of formula I or a salt thereof, can be administered orally, parenterally, by inhalation spray, topically, transdermally, rectally, nasally, buccally, sublingually, transvaginally, intraperitoneally, via Intrapulmonary, intradermal, epidural or via an implantable reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, topical, and intracranial injection or infusion techniques.

In one embodiment, a composition comprising a compound of Formula I or a salt thereof is formulated as a solid dosage form for oral administration. Solid dosage forms for oral administration include capsules, troches, pills, powders, and granules. In certain embodiments, a solid oral dosage form comprising a compound of Formula (I) or a salt thereof further comprises one or more of the following: (i) a pharmaceutically acceptable inert excipient or carrier, Such as sodium citrate or dicalcium phosphate; and (ii) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol or citric acid; (iii) binders such as carboxymethylcellulose, alginic acid Salt, gelatin, polyvinylpyrrolidone, sucrose or gum arabic; (iv) humectants such as glycerin; (v) disintegrants such as agar, calcium carbonate, potato or tapioca, alginic acid, certain citrate Or sodium carbonate; (vi) solution blocker, such as paraffin; (vii) absorption enhancer, such as a quaternary ammonium salt; (viii) a wetting agent, such as cetyl or glyceryl monostearate; (ix) An adsorbent such as kaolin or bentonite; and (x) a lubricant such as talc, calcium stearate, magnesium stearate, polyethylene glycol or sodium lauryl sulfate. In certain embodiments, a solid oral dosage form is formulated as a capsule, lozenge or pill. In certain embodiments, the solid oral dosage form further comprises a buffer. In certain embodiments, such compositions for solid oral dosage forms can be formulated to contain one or more excipients (such as lactose or milk sugar, polyethylene glycol, and the like). Soft filled gelatin Filler in pouches and hard-filled gelatin capsules.

In certain embodiments, lozenges, dragees, capsules, pills, and granules comprising a composition of a compound of Formula I or a salt thereof, optionally comprise a coating or shell, such as an enteric coating. It may optionally comprise an opacifying agent, and may also be a composition which will release the active ingredient in a delayed manner, as appropriate or preferentially, in a certain portion of the intestinal tract. Examples of the embedding composition include polymers and waxes, which can also be used as soft-filled gelatin capsules and hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycols and the like. Filler.

In another embodiment, the composition comprises a microencapsulated compound of formula (I) or a salt thereof, and optionally further comprises one or more excipients.

In another embodiment, the composition comprises a liquid dosage formulation comprising a compound of Formula I or a salt thereof for oral administration, and optionally further comprising one or more pharmaceutically acceptable emulsions, microemulsions, solutions, Suspensions, syrups and tinctures. In certain embodiments, the liquid dosage form optionally further comprises one or more inert diluents such as water or other solvents, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Benzoyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oil (in detail, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerin , tetrahydrofurfuryl alcohol, polyethylene glycol or sorbitan fatty acid esters and mixtures thereof. In certain embodiments, the liquid oral compositions further comprise one or more adjuvants such as wetting agents, suspending agents, sweetening agents, flavoring agents, and flavoring agents, as appropriate.

Injectable preparations, for example, sterile injectable aqueous or oily suspensions, may be employed in accordance with known techniques using dispersible or wetting agents and suspending agents. The sterile injectable preparation may be in the form of a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example, in the form of a solution of 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, Use any non-irritating fixed oil, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtration through a bacterial retention filter or by incorporating a sterilizing agent which can be dissolved or dissolved in sterile water or other sterile injectable medium before use in the form of a sterile solid composition.

In order to prolong the effect of the compound of formula (I), it is generally desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of a compound depends on its rate of dissolution, which in turn depends on crystal size and crystalline form. Alternatively, absorption of the parenterally administered compound form is delayed by dissolving or suspending the compound in an oil vehicle. The injectable depot form is prepared by forming a microcapsule matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. The rate of release of the compound can be controlled depending on the ratio of compound to polymer and the nature of the particular polymer employed. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). A bulk injectable formulation is also prepared by embedding the compound in a liposome or microemulsion that is compatible with body tissues.

In certain embodiments, a composition for rectal or vaginal administration is formulated as a suppository, which may be prepared by mixing a compound of formula (I) or a salt thereof with a suitable non-irritating excipient or carrier. Such excipients or carriers are, for example, cocoa butter, polyethylene glycol or suppository wax, for example solid at ambient temperature but liquid at body temperature and thus thawed in the rectum or vaginal cavity and release the compound of formula (I) Excipient or carrier.

Exemplary dosage forms for topical or transdermal administration of a compound of formula (I) include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The compound of formula (I) or a salt thereof is admixed under sterile conditions with a pharmaceutically acceptable carrier and optionally a preservative or buffer. Examples of other formulations include ophthalmic formulations, ear drops, eye drops, transdermal patches. Transdermal dosage forms can be made by dissolving or dispersing a compound of formula (I) or a salt thereof in a medium such as ethanol or dimethylhydrazine. Can also use absorption enhancer to increase The flow of the compound across the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

Nasal aerosol or inhalation formulations of a compound of formula (I) or a salt thereof may employ benzyl alcohol or other suitable preservatives, absorption enhancers for improving bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents. It was prepared as a physiological saline solution.

In certain embodiments, the pharmaceutical composition can be administered with or without food. In certain embodiments, a pharmaceutically acceptable composition can be administered without food. In certain embodiments, the pharmaceutically acceptable compositions of the invention can be administered with food.

The specific dosage and treatment regimen for any particular patient will depend on a number of factors, including age, weight, general health, sex, diet, time of administration, rate of excretion, combination of drugs, judgment of the treating physician, and the particular condition being treated The severity. The amount of a compound of formula I or a salt thereof provided in the composition will also depend on the particular compound in the composition.

In one embodiment, the therapeutically effective amount of a compound of the invention administered parenterally in each dose will range from about 0.01 to 100 mg/kg, or from about 0.1 to 20 mg/kg of patient body weight per day, wherein A typical initial range for the compound is 0.3-15 mg/kg/day. In another embodiment, oral unit dosage forms such as tablets and capsules contain from about 5 to about 100 mg of a compound of the invention.

An exemplary tablet dosage form comprises about 2 mg, 5 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg of a compound of formula (I) or a salt thereof, and further comprises about 95-30 mg of anhydrous lactose, about 5-40 mg of crosslinked carboxymethyl. Cellulose sodium, about 5-30 mg of polyvinylpyrrolidone (PVP) K30 and about 1-10 mg of magnesium stearate. A method of formulating a tablet comprises mixing the powdered ingredients together and further mixing with the PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate, and compressed into a tablet form using conventional equipment. About 2-500 mg of a compound of formula I or a salt thereof can be dissolved in a suitable buffer solution (eg, phosphate buffer) and An example of an aerosol formulation is prepared by adding a osmotic agent, such as a salt, such as sodium chloride, as needed. The solution can be filtered, for example, using a 0.2 micron filter to remove impurities and contaminants.

Use of the compound and pharmaceutically acceptable composition

Another aspect includes the use of a compound of formula (I) or a salt thereof for inhibiting CDK8, thereby treating a disease, such as cancer.

Another aspect includes a method of treating or preventing a disease in a patient that is responsive to inhibition of CDK8 activity. The method comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I) or a salt thereof.

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in therapy. Another aspect includes the use of a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in therapy.

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the treatment of a disease associated with CDK8 activity.

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a disease associated with CDK8 activity.

Another aspect includes a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a therapeutically active substance.

Another aspect includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use as a therapeutically active substance for the treatment of a hyperproliferative disorder.

In certain embodiments, the disease or condition is hyperproliferative disease, cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, multiple sclerosis, Alzheimer's disease, cystic fiber , viral disease, autoimmune disease, atherosclerosis, restenosis, psoriasis, osteoarthritis, inflammatory bowel disease, asthma, allergic conditions, inflammation, neurological disorders, hormone-related diseases, associated with organ transplantation Symptoms, immunodeficiency disorders, destructive skeletal disorders, proliferative disorders, infectious diseases, and Pathogenesis-related conditions, thrombin-induced platelet aggregation, liver disease, pathological immunological conditions involving T cell activation, CNS disorders, or spinal proliferative disorders.

In certain embodiments, the treatment can be administered after one or more symptoms have been suffered. In other embodiments, the treatment can be administered in the absence of symptoms. For example, treatment can be administered to a susceptible individual prior to the onset of symptoms (eg, based on a history of symptoms and/or according to genetic or other susceptibility factors). Treatment can also continue after the symptoms have subsided, for example to prevent or delay their recurrence.

Another aspect includes a method for treating, ameliorating or preventing cancer, drug-resistant cancer, or another proliferative disorder by administering an effective amount to a mammal (eg, a human) in need of such treatment ( I) a compound or a salt thereof. In certain embodiments, the disease to be treated is a cancer or a drug resistant cancer.

Examples of cancers that can be treated using the compounds and methods described herein include, but are not limited to, adrenal cancer, acinar cell carcinoma, acoustic neuroma, acral freckle-like melanoma, acromegaly, acute eosinophils Leukemia, acute erythrocytic leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monocytic leukemia, acute promyelocytic leukemia, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenoma-like teeth Primary tumor, adenosquamous carcinoma, adipose tissue neoplasm, adrenocortical carcinoma, adult T cell leukemia/lymphoma, invasive NK cell leukemia, AIDS-associated lymphoma, alveolar rhabdomyosarcoma, alveolar soft sarcoma, glazed cells Fibroids, degenerative large cell lymphoma, degenerative thyroid cancer, androgen-dependent cancer, vascular immune blast T cell lymphoma, angiomyolipoma, angiosarcoma, astrocytoma, atypical teratoid rod Tumor, B cell chronic lymphocytic leukemia, B cell lymphoma, basal cell carcinoma, biliary tract cancer, bladder cancer, blastoma, bone cancer, cloth Brenner tumor, Brown tumor, Burkitt's lymphoma, breast cancer, brain cancer, carcinoma, carcinoma in situ, carcinosarcoma, cartilage tumor, cementum, spinal sarcoma, Chondroma, chordoma, choriocarcinoma, choroid plexus papilloma, renal clear cell sarcoma, craniopharyngioma, skin T-cell lymphoma, cervical cancer, colorectal cancer, Degos disease, connective tissue proliferative small round cell tumor, diffuse large B-cell lymphoma, embryonic dysplastic neuroepithelial neoplasm, no Stromal cell, embryonic carcinoma, endocrine adenoma, endoderm sinus tumor, enteropathy-associated T-cell lymphoma, esophageal cancer, fetal mid-fetal, fibroid, fibrosarcoma, follicular lymphoma, follicular thyroid cancer, Ganglion cell tumor, gastrointestinal cancer, germ cell tumor, gestational choriocarcinoma, giant cell fibroblastoma, giant cell tumor of bone, glial tumor, glioblastoma multiforme, glioma, brain neuroglioma Disease, glioma, gonadal tumor, granulosa cell tumor, amphoteric blastoma, gallbladder cancer, gastric cancer, hemangioblastoma, head and neck cancer, vascular epithelioma, hematological malignancies, hepatoblastoma, liver and spleen T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, invasive lobular carcinoma, colon cancer, kidney cancer, laryngeal cancer, malignant sputum, leukemia Leydig cell tumor, liposarcoma, lung cancer, lymphangioma, lymphangiosarcoma, lymphatic epithelial carcinoma, lymphoma, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, liver cancer, small Cell lung cancer, non-small cell lung cancer, MALT lymphoma, malignant fibrous histiocytoma, malignant peripheral nerve sheath tumor, malignant neoplasm, mantle cell lymphoma, marginal zone B cell lymphoma, mast cell leukemia, mediastinal germ cell tumor, Breast myeloid carcinoma, myeloid thyroid cancer, medulloblastoma, melanoma, meningioma, merkel cell cancer, mesothelioma, metastatic urothelial carcinoma, mixed mullerian tumor (mixed Mullerian tumor), mucinous tumor, multiple myeloma, muscle tissue tumor, mycosis fungoid, mucinous liposarcoma, myxoma, mucinous sarcoma, nasopharyngeal carcinoma, schwannomas, neuroblastoma, fibromas , neuroma, melanoma, eye cancer, oligodendroglioma, oligodendroglioma, eosinophilic adenoma, optic nerve sheath meningioma, optic nerve Tumor, oral cancer, osteosarcoma, ovarian cancer, Pancoast tumor, papillary thyroid carcinoma, paraganglioma, pineal blastoma, pineal blastoma, pituitary cell tumor, pituitary gland Tumor, pituitary tumor, plasmacytoma, multi-embryo, pre- Drive T-lymphoblastic lymphoma, primary central nervous system lymphoma, primary exudative lymphoma, primary peritoneal cancer, prostate cancer, pancreatic cancer, pharyngeal cancer, peritoneal pseudomyxoma, renal cells Cancer, renal myeloma, retinoblastoma, rhabdomyosarcoma, rhabdomyosarcoma, Richter's transformation, rectal cancer, sarcoma, Schwannomatosis, seminoma, and Settle's cells Tumor, reproductive gonad stromal cell tumor, signet ring cell carcinoma, skin cancer, small blue round cell tumor, small cell carcinoma, soft tissue sarcoma, somatostatin tumor, soot, spinal cord tumor, spleen marginal lymphoma, squamous cell Cancer, synovial sarcoma, Sezary's disease, small bowel cancer, gastric cancer, T-cell lymphoma, testicular cancer, vesicular cell tumor, thyroid cancer, transitional cell carcinoma, pharyngeal cancer, urachal cancer, genitourinary Cancer, urothelial carcinoma, uveal melanoma, uterine cancer, verrucous carcinoma, visual pathway glioma, vulvar tumor, vaginal cancer, Waldenstrom's macroglobulinemia , Warthin's tumor and Wilms' tumor.

Another embodiment includes a method for treating a benign proliferative disorder. Examples of benign proliferative disorders (but not limited to) benign soft tissue tumors, bone tumors, brain and spinal cord tumors, orbital and orbital tumors, granuloma, lipoma, meningiomas, multiple endocrine neoplasms, nasal polyps, pituitary tumors, prolactin Tumor, pseudo-brain tumor, seborrheic keratosis, gastric polyps, thyroid nodules, pancreatic bladder tumor, hemangioma, vocal cord nodules, polyps and cysts, Castleman disease, chronic Problems, cutaneous fibroids, hair cysts, purulent granuloma and adolescent polyp syndrome.

Another embodiment comprises the use of a compound of formula I or a salt thereof for the manufacture of a pharmaceutical combination useful for the treatment and/or prevention and/or amelioration of a disease, disorder, condition and/or condition as referred to herein. Things.

Another embodiment comprises the use of a compound of formula I or a salt thereof for the manufacture of a disease and/or condition useful for the treatment and/or prevention of response or inhibition of CDK8, in particular as described above. A pharmaceutical composition (such as cancer).

The compound of formula (I) or a salt thereof can be administered in any amount and in any route of administration effective to treat or alleviate the severity of the condition. Depending on the patient's species, age, and general condition, such as the severity of the condition, the particular compound, its mode of administration, and its analogs, the exact amount required will vary from patient to patient. The total daily usage of the compound of formula (I) for a given patient will be determined by the attending physician within the scope of sound medical judgment. The particular effective dosage level for any particular patient will depend on a number of factors, including the severity of the condition or condition being treated; the activity of the particular compound employed; the particular composition employed; the age, weight, and general Health status, gender and diet; time of administration, route of administration and rate of excretion of the particular compound employed; duration of treatment; drugs used or compatible with the particular compound employed; and similar factors well known in the medical arts.

Another embodiment includes a method of inhibiting CDK8 activity in a biological sample comprising contacting the biological sample with a compound of Formula I or a salt thereof.

The term "biological sample" as used herein includes, without limitation, cells, cell cultures or extracts thereof; biopsies obtained from mammals or extracts thereof; and blood, saliva, urine, feces, Semen, tears or other body fluids or their extracts.

Co-administering compounds and other pharmaceutical agents

The compound of formula (I) or a salt thereof can be used alone or in combination with other therapeutic agents. For example, a second agent of a pharmaceutical combination formulation or dosing regimen can have complementary activities with a compound of formula (I) such that it does not adversely affect each other. The compounds can be administered together or separately in the overall pharmaceutical composition. In one embodiment, the compound or pharmaceutically acceptable salt can be co-administered with a cytotoxic agent to treat a proliferative disease and cancer.

The term "co-administered" means the simultaneous administration of a compound of formula (I) or a salt thereof and other active pharmaceutical ingredients (including cytotoxic agents and radiation treatment) or any of their sequential administration. If not administered at the same time, the compounds are administered at a time close to each other. Furthermore, it is not important even if the compound is administered in the same dosage form, for example one compound may be administered topically and the other compound may be administered orally.

Typically, any agent that is active against the disease or condition being treated can be co-administered. Examples of such agents can be found in Cancer Principles and Practice of Oncology, V.T. Devita and S. Hellman (ed.), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. Those skilled in the art should be able to discern which combination of agents will be applicable based on the particular characteristics of the drug and the disease involved.

In one embodiment, the method of treatment comprises co-administering a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one cytotoxic agent. The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cell function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (eg, radioactive isotopes of At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ^{212 ,} and Lu); Therapeutic agents; growth inhibitors; enzymes and fragments thereof, such as nucleolytic enzymes; and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.

Exemplary cytotoxic agents can be selected from the group consisting of anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and Hormone analogues, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutics, pro-apoptotic agents, LDH-A inhibitors; fatty acid biosynthesis inhibitors; cell cycle signaling inhibitors HDAC inhibitors, proteasome inhibitors; and cancer metabolism inhibitors.

"Chemotherapeutic agents" include compounds that are useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA ^® , Genentech/OSI Pharm.), bortezomib (VELCADE ^® , Millennium Pharm.), disulfiram, epidermis Catechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol , lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX ^® , AstraZeneca), sunitini (Sunitent ^® , Pfizer / Sugen), letrozole (FEMARA) ^® , Novartis), imatinib mesylate (GLEEVEC ^® , Novartis), finasunate (VATALANIB ^® , Novartis), oxaliplatin (ELOXATIN ^® , Sanofi), 5-FU (5-fluorouracil), formazan tetrahydrofolate, rapamycin (Sirolimus, RAPAMUNE ^® , Wyeth), Lapatinib (TYKERB ^® , GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336), Solafenib (NEXAVAR ^® , Bayer Labs), gefitinib (IRESSA ^{® )} , AstraZeneca), AG1478; alkylating agents, such as Thiotepa (Thiotepa) and CYTOXAN ^® cyclophosphamide; alkyl sulfonates such as busulfan (busulfan), where English C Shu (improsulfan) piperazine and poise Piposulfan; aziridine, such as benzodopa, carboquone, meturedopa and uredopa; ethyl imino and hydroxymethyl Melamine, including altretamine, tri-ethyl melamine, tri-ethylphosphoniumamine, tri-ethyl thiophosphonamide and trimethylol melamine; lychee lactone (especially cloth) Bullatacin (bullatacinone); camptothecin (including topotecan and irinotecan); bryostatin; calstatin ); CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (detailed spirulina 1) And spirulina 8); adrenal steroids (including prednisone and prednisolone); cyproterone ac Etate); 5α-reductase, including finasteride and dutasteride; vorinostat, romidepsin, panobinostat, Valproic acid, mocetinostat, dolastatin, arsenic, talc, and synthetic analogues KW-2189 and CB1-TM1; eleutherobin ); pancratistatin; sarcodictyin; spongistatin; nitrogen mustard, such as chlorambucil, chlomaphazine, chlorophosphamide, female Nitrogen mustard, ifosfamide, methotrexate, nitrous oxide mustard hydrochloride, amphetamine hydrogenated mustard, neombibichin, phenesterine, prednimustine ), trofosfamide, uracil mustard; nitrosourea, such as carmustine, chlorozotocin, fotemustine, lomustine ( Lomustine), nimustine and ranimnustine; antibiotics, such as enediyne antibiotics ( The calicheamicin, especially calicheamicin γ1I and calicheamicin ω1I (Angew Chem.Intl.Ed.Engl 1994 33:. 183-186); of the neomycin (dynemicin), comprising of the ADM A; bisphosphonates, such as clodronate; esperamicin; and new carotenoid chromophores and related chromoprotein diacetylene antibiotic chromophores, aclacinomysins ), actinomycin, authramycin, azaserine, bleomycins, cactinomycin, caraceptin, magentia Caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5- oxo-L-normal leucine, ADRIAMYCIN ^® (doxorubicin), N-morpholinyl doxorubicin, cyano (N-morpholinyl) doxorubicin, 2-pyrroline-based doxorubicin and Deoxytetracycline), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins (such as Mitomycin) C), mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, three Quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, net Zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as dimethyl folate, methotrexate, pteridophyte (pteropterin), trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as cyclocytidine, Cytosine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, dexifluridine, enocitabine, fluorouridine; androgen , such as dimethyltestosterone, dromostanolone propionate, epitiostanol, metoprolol (mepitio) Stane), testosterone; anti-adrenal, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid; acenolactone; Aminoglycoside; alanine; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; Defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; Gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansine, such as maytansin and ansamitocins; mitoguazone; mitoxantrone; Mopidamnol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic Acid); 2-ethyl hydrazine; procarbazine; PSK ^® polysaccharide complex (JHS Natural P Roducts, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; Triaziquone; 2,2',2"-trichlorotriethylamine; trichothecene (especially T-2 toxin, verracurin A, roridin A) And angudin; urethan; vindesine; dacarbazine; mannitol mustard; dibromomannitol; dipothalol (mitolactol); Pipobroman; gacytosine; arabinoside ("Ara-C");cyclophosphamide;thiotepa; taxane, such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE ^® (no hydrogenated castor oil), paclitaxel albumin engineered nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Ill.) and TAXOTERE ^® (docetaxel, docetaxel ( doxetaxel); Sanofi-Aventis); chlorambucil; GEMZAR ^® (gemcitabine); 6-thioguanine; mercaptopurine; carbamoyl butterfly Platinum analogues such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); methotrexate; mitoxantrone; vincristine ); NAVELBINE ^® (vinorelbine); Novantrone; teniposide; edacoxacin; daunorubicin; aminopterin; capecitabine (XELODA ^® ); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; Any of the pharmaceutically acceptable salts, acids and derivatives.

Chemotherapeutic agents also include: (i) antihormonal agents that can be used to modulate or inhibit the hormonal effects on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX) ^® ; tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, Keoxifene, LY117018, onapristone, and FARESTON ^® (toremifine citrate); (ii) aromatase that inhibits the regulation of estrogen-producing aromatase in the adrenal gland Inhibitors such as 4(5)-imidazole, amine ubmetazole, MEGASE ^® (Megestrol), AROMASIN ^® (Ethistein; Pfizer), formestanie, fadrozole, RIVISOR ^® (vorozole), FEMARA ^® (letrozole; Novartis) and ARIMIDEX ^® (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide Nilutamide), bicalutamide, leuprolide and goserelin; Buscheri (buserelin), tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and Troxacitabine (1,3-dioxolan nucleoside cytosine analog); (iv) protein kinase inhibitor; (v) lipid kinase inhibitor; (vi) antisense oligonucleotide, In particular, antisense oligonucleotides that inhibit the expression of genes involved in the signaling pathway of abnormal cell proliferation, such as PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (eg, ANGIOZYME ^® And (egiii) vaccines, such as gene therapy vaccines such as ALLOVECTIN ^® , LEUVECTIN ^® and VAXID ^® ; PROLEUKIN ^® , rIL-2; topoisomerase 1 inhibitors such as LURTOTECAN ^® ; ABARELIX ^® rmRH; And (ix) pharmaceutically acceptable salts, acids and derivatives of any of the above.

Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); Pani Monoclonal antibody (panitumumab) (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab Trastuzumab (HERCEPTIN®, Genentech), tositumomab (Bexxar, Corixia) and antibody drug conjugate gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Other humanized monoclonal antibodies having therapeutic potential as agents in combination with the compounds of the invention include: apolizumab, aselizumab, atlizumab, bapi Betabeuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, certolizumab pegol, cefozumab (cidfusituzumab), cidtuzumab, daclizumab, eculizumab, efalizumab, etrapuzumab, erlib Monoclonal antibody (erlizumab), felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, irizhu Monoclonal antibody (ipilimumab), labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab , motolizumab, natalizumab, nimotuzumab (nimotuzum) Ab), nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, Pacozhu Monoclonal antibody (pascolizumab), paxicilizumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, Reslivizumab, reslizumab, resviviumab, rovelizumab, ruplizumab, sirolizumab ( Sibrotuzumab), sicilizumab, sotuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tifibrate Tufibazumab, tocilizumab, toralizumab, tucotuzumab celmoleukin, tucusituzumab, ummazhudan Anti-(umavizumab), utoxazumab, ustekinumab, visilizumab and Interleukin -12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), which is a recombinant exclusively human sequence, i.e., genetically modified to recognize the full-length interleukin -12 p40 protein of the IgG ₁ λ antibodies.

A chemotherapeutic agent also includes an "EGFR inhibitor", which refers to a compound that binds or directly interacts with EGFR and prevents or reduces its signaling activity, and is alternatively referred to as an "EGFR antagonist." Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al). And variants thereof, such as chimeric 225 (C225 or Cetuximab; ERBUTIX ^® ) and reconstituted human 225 (H225) (see WO 96/40210, Imclone Systems Inc.); IMC-11F8, ie Full-length human EGFR-targeting antibody (Imclone); an antibody that binds to type II mutant EGFR (U.S. Patent No. 5,212,290); humanized and chimeric antibodies that bind EGFR as described in U.S. Patent No. 5,891,996; Antibodies such as ABX-EGF or Panitumumab (see WO 98/50433, Abgenix/Amgen); EMD 55900 (Stragliotto et al, Eur . J. Cancer 32A: 636-640 (1996)); EMD 7200 (Mato Mauzumab (matuzumab), a humanized EGFR antibody (EMD/Merck) against EGFR that competes with EGF and TGF-α for EGFR binding; human EGFR antibody HuMax-EGFR (GenMab); known as E1.1, E2. 4. Full length human antibodies of E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3 and described in US 6,235,883; MDX-447 (Medarex Inc And mAb 806 or humanized mAb 806 (Johns et al, J. Biol. Chem. 279(29): 30375-30384 (2004)). The anti-EGFR antibody can bind to a cytotoxic agent, thus producing an immunoconjugate (see, for example, EP 659, 439 A2, Merck Patent GmbH). EGFR antagonists include small molecules, such as those described in U.S. Patent Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534. , Nos. 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5,866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, No. 6,002,008 and 5,747,498, and the following PCT publications: WO 98/14451, WO 98/50038, WO 99/09016, and WO 99/24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA ^® (Genentech/OSI Pharmaceuticals); PD 183805 (CI 1033, 2-propenylamine, N-[4-[(3- Chloro-4-fluorophenyl)amino]-7-[3-(4-morpholino)propoxy]-6-quinazolinyl]-dihydrochloride, Pfizer Inc.); ZD1839, Kyrgyzstan IRESSA® 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-(N-morpholinyl)propoxy)quinazoline, AstraZeneca ZM 105180 ((6-Amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)) -N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166((R)-4-[ 4-[(1-Phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4 -[(1-phenethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785(N-[4-[(3-bromophenyl)amino]-6- Quinazolinyl]-2-butynylamine); EKB-569(N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy- 6-quinolinyl]-4-(dimethylamino)-2-butenoxime) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); double EGFR/HER2 tyrosine kinase inhibition Agent Such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2-methylsulfonyl)) Ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine).

Chemotherapeutic agents also include "tyrosine kinase inhibitors" which include the EGFR-targeted drugs indicated in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors such as TAK165 available from Takeda; ErbB2 receptor tyramine Oral selective inhibitors of acid kinases CP-724, 714 (Pfizer and OSI); dual HER inhibitors, such as EKB-569 (available from Wyeth), which preferentially bind to EGFR but inhibit cells that overexpress HER2 and EGFR; Nie (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors such as carnitinib (CI-1033; Pharmacia); Raf-1 inhibition Agents, such as the antisense agent ISIS-5132 (available from ISIS Pharmaceuticals) that inhibits Raf-1 signaling; non-HER targeted TK inhibitors, such as imatinib mesylate (GLEEVEC®, available from Glaxo SmithKline); Multi-targeted tyrosine kinase inhibitors, such as sunitinib (SUTENT®, available from Pfizer); VEGF receptor tyrosine kinase inhibitors, such as vatalanib (PTK787/ZK222584, available) From Novartis/Schering AG); MAPK extracellular regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazoline, such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidine; Pyrimidopyrimidine; pyrrolopyrimidines such as CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin Wei 醯 methane, 4,5-bis(4-fluoroanilino) quinone imine); tyrosine phosphorylation inhibitor containing nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecule (eg An antisense molecule that binds to a HER-encoding nucleic acid; quinoxaline (U.S. Patent No. 5,804,396); tyrosine phosphorylation inhibitor (U.S. Patent No. 5,804,396); ZD6474 (Astr a Zeneca); PTK-787 (Novartis/Schering AG); pan-HER inhibitors such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis/Lilly); imatinib mesylate (GLEEVEC®); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis/Schering AG); </ RTI> </ RTI> <RTIgt; 1998/43960 (American Cyanamid); WO 1997/38983 (Warner Lambert); WO 1999/06378 (Warner Lambert); WO 1999/06396 (Warner Lambert); WO 1996/30347 (Pfizer, Inc); WO 1996/33978 ( Zeneca); WO 1996/3397 (Zeneca); and WO 1996/33980 (Zeneca).

Chemotherapeutic agents also include dexamethasone, interferon, colchicum Alkali, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol ), amifostine, arsenic trioxide, aspartate, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, ar Darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, elotinib, filgrastim, Histrelin acetate, ibritumomab, interferon alpha-2a, interferon alpha-2b, lenalidomide, levamisole, mesna , methoxsaren, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate (pamidronate), pegademase, pegaspargase, pegfilgrastim Pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, sargramostim , temozolomide, VM-26, 6-TG, toremifene, retinoic acid, ATRA, valrubicin, zoledronate and zoledronic acid and their pharmaceutically acceptable Salt.

Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide, and koji Triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, acetone albino Fluocinolone acetonide), betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocorticosterone-17-butyrate, hydrocortinone -17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate (clobetasone-17-butyrate), clobetasol-17-propionate, fluconazole hexanoate, fluconazole valerate and fluprednidene acetate; immune selection Anti-inflammatory peptides (ImSAIDs), such as phenylalanine-glutamic acid-gan Amino acid (FEG) and its D-isomeric form (feG) (IMULAN BioTherapeutics, LLC); antirheumatic drugs such as azathioprine, cyclosporine (cyclosporin A), D-penicillamine (D-penicillamine) ), gold salt, hydroxychloroquine, leflunomide, minocycline, sulfasalazine; tumor necrosis factor alpha (TNFα) blockers, such as etasoxi Eternercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), Goley Golimumab (Simponi); interleukin 1 (IL-1) blocker, such as anakinra (Kineret); T cell co-stimulatory blocker, such as abatacept (abatacept) Orencia); interleukin 6 (IL-6) blocker, such as tocilizumab (ACTEMERA®); interleukin 13 (IL-13) blocker, such as leflunibumab (lebrikizumab); interferon alpha (IFN a blocking agent, such as Rontalizumab; a beta7 integrin blocker, such as rhuMAb β7; an IgE pathway blocker, such as an anti-M1 initial vaccine; secretes homotrimeric LTa3 And a heterologous membrane-bound trimer LTa1 / β2 blockers, such as an anti-lymphotoxin α (LTa); radioactive isotopes (e.g. ^{At 211, I 131, I 125} , Y 90, Re 186, Re 188, Sm 153, Bi ²¹² , P ³² , Pb ²¹² and Lu radioisotopes); various research agents, such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH ₃ or farnesyltransferase inhibitors ( L-739749, L-744832); polyphenols, such as quercetin, resveratrol, piceatannol, epigallocatechin gallate, Theaflavins, flavanols, procyanidins, betulinic acid and derivatives thereof; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; acetyl-camptothecin, scopolectin and 9-aminocamptothecin ); podophyllotoxin; tegafur (UFTORAL®); bexarotene (TARGRETIN®); bisphosphonates such as clodronate (eg BONEFOS®) OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid/zoledronate (ZOMETA®), alendronate (FOSAMAX®), Pa Pamidronate (AREDIA®), tiludronate (SKELID®) or risedronate (ACTONEL®); and epidermal growth factor receptor (EGF-R); Vaccines, such as THERATOPE® vaccine; perifosine, COX-2 inhibitors (eg celecoxib or etoricoxib), proteasome inhibitors (eg PS341); CCI-779 Tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors, such as oblimersen sodium (GENASENSE®); pixantrone; farnesyltransferase inhibitors, such as Los that farnesyl (lonafarnib) (SCH 6636, SARASAR TM); and any of the foregoing the acceptable pharmaceutically salts, acids and derivatives thereof; and said two or the combination of the above, such as of CHOP, an abbreviation of cyclophosphamide, doxorubicin, vincristine, and prednisolone combination therapy of; and FOLFOX, namely the use of oxaliplatin (ELOXATIN ^TM) Acronym regimen of 5-FU and leucovorin combination of A XI.

Chemotherapeutic agents also include non-steroidal anti-inflammatory drugs with analgesic, antipyretic and anti-inflammatory effects. NSAIDs include non-selective inhibitors of enzyme cyclooxygenase. Specific examples of NSAIDs include aspirin; propionic acid derivatives such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, ol. Oxprozin and naproxen; acetic acid derivatives such as indomethacin (indomethacin), sulindac, etodolac, diclofenac; enolic acid derivatives such as piroxicam, meloxicam, tenoxicam , droxicam, lornoxicam and isoxicam; fenamic acid derivatives such as mefenamic acid, meclofenamic acid ), flufenamic acid, tolfenamic acid; and COX-2 inhibitors such as celecoxib, etoricoxib, lumiracoxib, parecoxib , rofecoxib, rofecoxib and valdecoxib. NSAID can be indicated for use in, for example, osteoarthritis, osteoarthritis, inflammatory joint disease, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headache And migraine, post-operative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction and renal colic symptoms are alleviated.

Chemotherapeutic agents also include treatments for Alzheimer's disease, such as donepezil hydrochloride and rivastigmine; for the treatment of Parkinson's Disease, such as L-DOPA/ Carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, triheximide Trihexephendyl) and amantadine; agents for the treatment of multiple sclerosis (MS), such as beta interferon (eg Avonex ^® and Rebif ^® ), glatiramer acetate and mitoxantrone; For the treatment of asthma, such as albuterol and montelukast sodium; agents for the treatment of schizophrenia, such as zyprexa, risperdal, and spleen Seroquel and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide and sulfasalazine; immunomodulators and immunosuppression Agents, such as cyclosporine, tacrolimus, Rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibition Agents, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole and anti-Parkinsonian agents; agents for the treatment of cardiovascular diseases, such as beta blockers Agents, ACE inhibitors, diuretics, nitrates, calcium channel blockers and statins; agents for the treatment of liver diseases, such as corticosteroids, cholestyramine, interferons and antiviral agents An agent for treating a blood disorder, such as a corticosteroid, an anti-leukemia agent, and a growth factor; and an agent for treating an immunodeficiency disorder, such as gamma globulin.

Additionally, the chemotherapeutic agent includes a pharmaceutically acceptable salt, acid or derivative of any of the chemotherapeutic agents described herein, and combinations of two or more thereof.

The amount of the compound of formula (I) or a salt thereof and the additional agent (in a composition comprising an additional therapeutic agent as described above) which may be combined with the carrier material in a single dosage form will depend on the subject being treated and the particular mode of administration. And change. In certain embodiments, the compositions of the present invention are formulated so as to be administered at a dose of between 0.01 and 100 mg of the compound of the invention per kg body weight per day.

The additional therapeutic agent and the compound of formula (I) can act synergistically. Thus, the amount of additional therapeutic agent in such compositions may be less than that required in a single therapy using only the therapeutic agent, or if a lower dose is used, there may be less side effects to the patient. In certain embodiments, a dose between 0.01 and 1,000 micrograms of additional therapeutic agent per kilogram of body weight per day may be administered in the compositions.

In particular, a method of treating cancer in an individual comprising administering to the individual (a) a compound of formula (I) or a pharmaceutically acceptable salt thereof and (b) a cytotoxic agent (eg, targeted therapy) is provided herein. , chemotherapy and / or radiation therapy).

Provided herein are methods of treating cancer in a subject, wherein the cancer treatment comprises administering to the individual (a) an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and (b) an effective amount of a cytotoxic agent, Wherein the cancer treatment comprises administering an effective amount of a cytotoxic agent without (absence of) the efficacy of a compound of formula (I) or a pharmaceutically acceptable salt thereof (e.g., standard care treatment).

In certain embodiments of any of the methods, the cytotoxic agent is radiation therapy.

In certain embodiments of any of the methods, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered with a cytotoxic agent (eg, targeted therapy, chemotherapy, and/or radiation therapy). In certain embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered prior to and/or concurrently with a cytotoxic agent (eg, targeted therapy, chemotherapy, and/or radiation therapy).

illustration

As depicted in the following examples, in certain exemplary embodiments, compounds are prepared according to the following general procedures, wherein R ¹ , R ² , R ³ and R ^a and R ^{b are} as defined for Formula I and its alternate embodiments. It will be appreciated that while the general methods depict the synthesis of certain compounds of the invention, the following general methods and other methods known to those skilled in the art are applicable to all of the compounds and subclasses of each of these compounds as described herein. And the type.

(a) 2-iodoacetonitrile, K ₂ CO ₃ , acetonitrile, 90 ° C. (b) 1-Tertioxy-N,N,N',N'-tetramethyl-methylenediamine, DMF, 90 °C. (c) Aniline hydrochloride, DMF, 110 ° C. (d) R ^a -胍, K ₂ CO ₃ , 1-butanol, DMF, 120 °C. (e) Fe, AcOH, 40 ° C. (f) DIEA, CH ₂ Cl ₂ .

The embodiments are illustrated by the following examples.

Instance HPLC separation method A:

Instrument: Gilson-281

Column: Gemini 150×30mm, 5μm particle size

Mobile phase: ACN 25%-55% H ₂ O (+0.05% NH ₄ OH)

Rate: 25ml/min

Monitoring wavelength: 220nm/254nm

Running time: 12min/15min

Column temperature: 30 ° C

HPLC separation method B:

Instrument: BH

Column: Phenomenex Gemini C18 200×25mm×10μm

Mobile phase: MeCN: 25%-55%; H ₂ O (+0.05% NH ₄ OH)

Rate: 25mL/min

Monitoring wavelength: 220nm/254nm

Running time: 12min

HPLC separation method C:

Instrument: Gilson GX281

Column: YMC-Actus Triart C18 150×30mm

Mobile phase: MeCN: 37%-67%; H ₂ O (+0.05% ammonia solution)

Rate: 25ml/min

Monitoring wavelength: 220nm/254nm

Running time: 10min/15min

Example 1: 1-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]-3-[3-(trifluoro Methyl)phenyl]urea

Step 1a) N-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]carbamic acid tert-butyl ester

6-Chloro-[1,2,4]triazolo[4,3-b]pyridazine (200 mg, 1.2940 mmol), N-[(3S)-pyrrolidin-3-yl]amine at 70 °C A mixture of tert-butyl carboxylic acid (265 mg, 1.4234 mmol) and N,N-diisopropylethylamine (0.29 mL, 1.6822 mmol) in ethanol (6 mL, 98.6 mmol) was stirred for 6 hr. The mixture was then cooled and concentrated. The residue was wet-milled in water and the solid obtained was collected by filtration and washed with water. This material was dried under vacuum to give 240 mg of N-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrole as a white solid. Tributyl butyl-3-yl]carbamic acid. ^{1 H NMR (400MHz, DMSO)} δ 9.18 (s, 1H), 8.03 (d, J = 10.0Hz, 1H), 7.26 (d, J = 6.6Hz, 1H), 7.03 (d, J = 10.1Hz, 1H ), 4.14 (dd, J =11.7, 6.0 Hz, 1H), 3.75-3.43 (m, 4H), 2.14 (dq, J = 13.4, 7.0 Hz, 1H), 1.91 (dt, J = 12.5, 6.5 Hz, 1H), 1.39 (s, 9H).

Step 1b) (3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidine-3-amine hydrochloride

N-[(3S)-1-([1,2,4]triazole in a solution of hydrogen chloride (4 mol/L) in 1,4-dioxane (9.8 mL, 39.42 mmol) at room temperature [4,3-b]Pyridazin-6-yl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (240 mg, 0.7884 mmol) was stirred for 30 min. Concentration and drying of the mixture in vacuo gave 190 mg of (3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidine as a yellow solid. 3-Amine hydrochloride, which was used in the next step without purification.

Step 1c) 1-[(3S)-1-([1,2,4]Triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]-3-[3- (trifluoromethyl)phenyl]urea

Addition of 1-isocyanate-3-(trifluoromethyl)benzene (28 mg, 0.150 mmol) to (3S)-1-([1,2,4]triazolo[4,3-b]pyridazine- A solution of 6-yl)pyrrolidine-3-amine hydrochloride (30 mg, 0.124 mmol) and N-ethyldiisopropylamine (0.065 mL, 0.372 mmol) in dichloromethane (1 mL). The mixture was stirred at room temperature for 1 hour. The mixture was concentrated and then be by HPLC (conditions) to give 21.6mg as a white solid of 1 - [(3S) -1 - ([1,2,4] triazolo [4,3-b] pyridazine -6-yl)pyrrolidin-3-yl]-3-[3-(trifluoromethyl)phenyl]urea. LCMS m/z = 392.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 9.24-9.17 (s, 1H), 8.83-8.69 (s, 1H), 8.14-8.02 (d, J = 10.0Hz, 1H), 8.02-7.94 (s, 1H), 7.53-7.38 (m, 2H), 7.29-7.18 (d, J = 7.0 Hz, 1H), 7.15-7.05 (d, J = 10.0 Hz, 1H), 6.80-6.67 (d, J = 6.6 Hz, 1H) , 4.42-4.26 (q, J = 5.7 Hz, 1H), 3.82-3.68 (dd, J = 10.9, 6.0 Hz, 1H), 3.66.3.51 (m, 2H), 3.48-3.37 (dd, J = 10.9, 4.1 Hz, 1H), 2.30-2.16 (dq, J = 13.6, 7.0 Hz, 1H), 2.05-1.90 (dq, J = 12.4, 6.2 Hz, 1H).

Example 2: 1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazine-6 -yl)pyrrolidin-3-yl]urea

LCMS m/z = 426.1 [M + 1] ⁺ . ¹ H NMR (400 MHz, DMSO) δ 9.26-9.13 (s, 1H), 8.93-8.79 (s, 1H), 8.16-7.97 (m, 2H), 7.65-7.45 (s, 2H), 7.16-6.98 (d , J=10.0Hz, 1H), 6.87-6.75 (d, J=6.6Hz, 1H), 4.43-4.26 (q, J=5.5Hz, 1H), 3.79-3.66 (dd, J=11.0, 6.1Hz, 1H), 3.66-3.50 (q, J = 7.1, 6.2 Hz, 2H), 3.47-3.37 (m, 1H), 2.30-2.17 (dd, J = 13.2, 6.5 Hz, 1H), 2.08-1.90 (dd, J=12.2, 6.1 Hz, 1H).

Example 3: 1-(3-Chlorophenyl)-3-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidine- 3-based urea

LCMS m/z = 358.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 9.23-9.12 (s, 1H), 8.84.8.68 (s, 1H), 8.11-7.99 (d, J = 10.0Hz, 1H), 7.71-7.62 (d, J = 2.2 Hz, 1H), 7.30-7.14 (m, 2H), 7.11-7.04 (d, J = 10.1 Hz, 1H), 6.98-6.90 (d, J = 7.6 Hz, 1H), 6.91-6.83 (d, J = 6.7 Hz, 1H), 4.41-4.25 (q, J = 5.6 Hz, 1H), 3.79-3.65 (dd, J = 10.9, 6.0 Hz, 1H), 3.65-3.50 (q, J = 6.6, 5.9 Hz, 2H) ), 3.46-3.37 (dd, J = 10.9, 4.1 Hz, 1H), 2.29-2.16 (dq, J = 13.6, 7.2 Hz, 1H), 2.07-1.88 (dq, J = 12.6, 6.2 Hz, 1H).

Example 4: 1-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]-3-[3-(trifluoro Methyl)phenyl]urea

LCMS m/z = 392.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 9.26-9.18 (s, 1H), 8.78-8.65 (s, 1H), 8.12-8.02 (d, J = 10.0Hz, 1H), 8.03-7.93 (s, 1H), 7.53-7.38 (m, 2H), 7.31-7.19 (d, J = 7.0 Hz, 1H), 7.14 - 7.01 (d, J = 10.0 Hz, 1H), 6.80-6.65 (d, J = 6.8 Hz, 1H) , 4.45-4.27 (q, J = 5.7 Hz, 1H), 3.80-3.68 (dd, J = 10.9, 6.1 Hz, 1H), 3.65-3.52 (m, 2H), 3.49-3.37 (dd, J = 10.6, 4.0 Hz, 1H), 2.30-2.16 (dd, J = 12.9, 6.5 Hz, 1H), 2.05-1.92 (dd, J = 12.6, 6.4 Hz, 1H).

Example 5: 1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazine-6 -yl)pyrrolidin-3-yl]urea

LCMS m/z = 426.1 [M + 1] ⁺ . ¹ H NMR (400 MHz, DMSO) δ 9.25-9.11 (s, 1H), 8.94-8.82 (s, 2H), 8.12-8.00 (m, 3H), 7.61-7.49 (s, 3H), 7.15-6.99 (d , J = 10.0 Hz, 2H), 6.89-6.75 (d, J = 6.8 Hz, 2H), 2.30-2.15 (m, 2H), 4.41-4.25 (q, J = 5.6 Hz, 2H), 3.78-3.68 ( m, 2H), 2.07-1.91 (m, 2H), 3.64-3.51 (q, J = 6.9, 6.1 Hz, 3H), 3.48-3.37 (d, J = 4.1 Hz, 1H).

Example 6: 1-(3-Chlorophenyl)-3-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl Urea

LCMS m/z = 358.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 9.24-9.15 (s, 1H), 8.75-8.61 (s, 1H), 8.12-8.00 (d, J = 10.0Hz, 1H), 7.70-7.61 (s, 1H), 7.29-7.14(m,2H),7.13-7.03(d,J=10.0Hz,1H), 6.97-6.89(m,1H),6.84-6.73(d,J=6.6Hz,1H),4.41-4.23( q, J = 5.5 Hz, 1H), 3.79 - 3.65 (dd, J = 10.8, 6.0 Hz, 1H), 3.65 - 3.50 (q, J = 6.4, 5.5 Hz, 2H), 3.45 - 3.36 (dd, J = 10.4, 3.8 Hz, 1H), 2.30-2.14 (dq, J = 13.7, 7.2 Hz, 1H), 2.03-1.89 (dt, J = 12.4, 6.3 Hz, 1H).

Example 7: (S)-1-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl)-3-(4-((4) -ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea

LCMS m/z = 518.3 [M + 1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 9.24-9.12 (m, 1H), 8.72 (s, 1H), 8.05 (dd, J = 10.0,0.9Hz, 1H), 7.92 (d, J = 2.2Hz, 1H) , 7.60-7.52 (m, 1H), 7.51-7.43 (m, 1H), 7.08 (d, J = 10.0 Hz, 1H), 6.72 (d, J = 6.6 Hz, 1H), 4.34 (d, J = 5.6 Hz, 1H), 3.73 (dd, J = 11.0, 6.0 Hz, 1H), 3.58 (d, J = 7.0 Hz, 2H), 3.49 (s, 2H), 3.41 (dd, J = 11.0, 4.2 Hz, 1H) ), 2.43-2.16 (m, 10H), 1.98 (t, J = 6.1 Hz, 1H), 0.97 (t, J = 7.1 Hz, 3H).

Example 8: 1-[(3S)-1-(2-Aminopyrimidin-4-yl)pyrrolidin-3-yl]-3-(3-chloro-4-methyl-phenyl)urea

Step 8a) N-[(3S)-1-(2-Aminopyrimidin-4-yl)pyrrolidin-3-yl]carbamic acid tert-butyl ester

N-[(3S)-pyrrolidin-3-yl]carbamic acid tert-butyl ester (200 mg, 1.074 mmol), 4-chloropyrimidin-2-amine (167 mg, 1.29 mmol) and N, at 90 °C. A mixture of N'-diisopropylethylamine (0.28 mL, 1.61 mmol) in isopropyl alcohol (3 mL) was stirred. The mixture was then cooled to room temperature and water was added. The solid which precipitated was collected by filtration, washed with water and dried in vacuo to give crystals of <RTIgt; </RTI> <RTIgt; </RTI> N-[(3S)-1-(2-aminopyrimidin-4-yl)pyrrolidin-3-yl] Tert-butyl carbamic acid. This product was used in the next step without purification.

Step 8b) 4-[(3S)-3-Aminopyrrolidin-1-yl]pyrimidin-2-amine

N-[(3S)-1-(2-Aminopyrimidin-4-yl) in a solution of hydrogen chloride (4 mol/l) in 1,4-dioxane (2.7 mL, 10.74 mmol) at room temperature Pyrrolidin-3-yl]carbamic acid tert-butyl ester (300 mg, 1.074 mmol) was stirred for 1 hour. The reaction was then concentrated and dried <RTI ID=0.0> This product was used in the next step without purification.

Step 8c) 1-[(3S)-1-(2-Aminopyrimidin-4-yl)pyrrolidin-3-yl]-3-(3-chloro-4-methyl-phenyl)urea

2-Chloro-4-isocyanate-1-methyl-benzene (56 mg, 0.278 mmol) was added to 4-[(3S)-3-aminopyrrolidin-1-yl]pyrimidin-2-amine hydrochloride (50 mg, 0.232 mmol) and a solution of N-ethyldiisopropylamine (0.204 mL, 1.459 mmol) in dichloromethane (3 mL). The mixture was stirred at room temperature for 1 hour. The mixture was then concentrated and be by HPLC (conditions) to give 55.4mg as a white solid of 1 - [(3S) -1- ( 2- amino-4-yl) pyrrolidin-3-yl] -3 -(3-Chloro-4-methyl-phenyl)urea. LCMS m/z = 347.2 [M + 1] ⁺ . ¹ H NMR (400 MHz, DMSO) δ 8.83 (s, 1H), 8.12 - 8.00 (m, 1H), 7.73 (d, J = 5.8 Hz, 1H), 7.54 (d, J = 1.4 Hz, 2H), 6.72 (d, J = 6.6 Hz, 1H), 5.90 (s, 2H), 5.74 (d, J = 5.8 Hz, 1H), 4.26 (s, 1H), 3.58 (t, J = 8.1 Hz, 1H), 3.42 (s, 2H), 2.15 (dt, J = 13.8, 6.9 Hz, 1H), 1.91 (dd, J = 11.2, 6.0 Hz, 1H).

Example 9: 1-[(S)-1-(2-amino-pyrimidin-4-yl)-pyrrolidin-3-yl]-3-(4-chloro-3-trifluoromethyl-phenyl)-urea

LCMS m/z = 401.1 [M + 1] ⁺ . ¹ H NMR (400 MHz, DMSO) δ 8.83 (s, 1H), 8.12 - 8.00 (m, 1H), 7.73 (d, J = 5.8 Hz, 1H), 7.54 (d, J = 1.4 Hz, 2H), 6.72 (d, J = 6.6 Hz, 1H), 5.90 (s, 2H), 5.74 (d, J = 5.8 Hz, 1H), 4.26 (s, 1H), 3.58 (t, J = 8.1 Hz, 1H), 3.42 (s, 2H), 2.15 (dt, J = 13.8, 6.9 Hz, 1H), 1.91 (dd, J = 11.2, 6.0 Hz, 1H).

Example 10: 1-[4-Chloro-3-(trifluoromethyl)phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl] Urea

LCMS m/z = 415.2 [M+1] ⁺ . ¹ H NMR (400 MHz, DMSO) δ 8.83 (s, 1H), 8.06 (s, 1H), 7.76 (d, J = 5.8 Hz, 1H), 7.54 (d, J = 1.4 Hz, 2H), 6.73 (d) , J = 6.7 Hz, 1H), 6.32 (d, J = 5.0 Hz, 1H), 5.73 (d, J = 5.9 Hz, 1H), 4.27 (s, 1H), 3.52 (d, J = 64.8 Hz, 4H ), 2.73 (d, J = 4.8 Hz, 3H), 2.16 (dd, J = 12.5, 6.5 Hz, 1H), 1.92 (t, J = 8.1 Hz, 1H).

Example 11: 1-(3-Chloro-4-methyl-phenyl)-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]urea

LCMS m/z = 361.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 8.41 (s, 1H), 7.76 (d, J = 5.8Hz, 1H), 7.63 (d, J = 2.1Hz, 1H), 7.22-7.12 (m, 1H), 7.08 (dd, J = 8.3, 2.2 Hz, 1H), 6.54 (d, J = 6.8 Hz, 1H), 6.32 (q, J = 4.8 Hz, 1H), 5.73 (d, J = 5.8 Hz, 1H), 4.25 (s, 1H), 3.59 (s, 1H), 3.43 (s, 3H), 2.73 (d, J = 4.7 Hz, 3H), 2.23 (s, 3H), 2.14 (dt, J = 13.9, 7.7 Hz, 1H), 1.89 (s, 1H).

Example 12: 1-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]-3-[4-(trifluoromethoxy)phenyl]urea

LCMS m/z = 397.2 [M + 1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 8.53 (s, 1H), 7.76 (d, J = 5.8Hz, 1H), 7.54-7.39 (m, 2H), 7.22 (d, J = 8.5Hz, 2H), 6.56 (d, J = 6.8 Hz, 1H), 6.32 (d, J = 5.3 Hz, 1H), 5.73 (d, J = 5.8 Hz, 1H), 4.27 (s, 1H), 3.59 (s, 1H), 3.44 (s, 2H), 2.73 (d, J = 4.7 Hz, 3H), 2.16 (dd, J = 13.0, 6.8 Hz, 1H), 1.89 (s, 1H).

Example 13: 1-[4-(Difluoromethoxy)phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]urea

LCMS m/z =379.2 [M+1] ⁺ . ^{1 H NMR (400MHz, DMSO)} δ 8.39 (s, 1H), 7.76 (d, J = 5.8Hz, 1H), 7.47-7.33 (m, 2H), 7.11-6.99 (m, 2H), 6.50 (d, J = 6.8 Hz, 1H), 6.32 (d, J = 5.3 Hz, 1H), 5.73 (d, J = 5.8 Hz, 1H), 4.26 (s, 1H), 3.57 (s, 4H), 2.73 (d, J = 4.7 Hz, 3H), 2.23 - 2.05 (m, 1H), 1.89 (s, 1H).

Example 14: (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidine-3 -based urea

Step 14a) 1-(4-Nitrobenzyl)azetidine

Over 30 minutes azetidine hydrochloride (1.4g, 15mmol) and DIPEA (1.9g, 15mmol) in the in CH ₃ CN (30mL) was added dropwise a solution of 1- (bromomethyl) -4- Nitrobenzene (1.07 g, 5 mmol). The reaction mixture was stirred for 3 hours under a N ₂ atmosphere at room temperature, then diluted with DCM (80mL). The organic layer was washed with EtOAc EtOAc m.

Step 14b) 4-(Azetidin-1-ylmethyl)aniline

Was added a solution of 1- (4-nitrobenzyl) azetidine (0.77g, 4mmol) in AcOH (10mL) in a solution of Fe (2.24g, 40mmol), and at room temperature in N ₂ atmosphere The reaction mixture was stirred for 3 hours. When the starting material was consumed, the reaction mixture was concentrated and basified to about pH 9 then extracted with DCM (3×30 mL). The combined organics were concentrated to give the title compound, m.

Step 14c) (S)-(1-(2-chloropyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester

A solution of (S)-(1-(2-chloropyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester (14 g, 94.6 mmol) in propan-2-ol (50 mL) DIPEA (14.64 g, 113.51 mmol) was added, and the mixture was cooled to 0 ° C, then (S)-pyrrolidin-3-ylaminocarbamic acid tert-butyl ester (17.6 g, 94.6 mmol) was added portionwise to the solution. ). The reaction solution was stirred at 25 ° C for 30 minutes. The mixture was then concentrated, and the residue was purified (jjjjjjjjjjjj 3-butyl) butyl methacrylate (22.5 g, yield 80%) and (S)-(1-(4-chloropyrimidin-2-yl)pyrrolidin-3-yl)carbamic acid Tributyl ester (3.0 g, yield 10%).

(S)-(1-(2-chloropyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester

_{^{1 H NMR (CDCl 3, 400MHz}} ): δ 7.97 (d, J = 6.0,1H), 6.15 (d, J = 6.0,1H), 4.79 (s, 1H), 4.31 (s, 1H), 3.22-3.82 (m, 4H), 2.25 (s, 1H), 1.91-2.03 (m, 1H), 1.42 (s, 9H); MS (ESI): m/z 298.9 [M+1] ⁺ .

(S)-(1-(4-chloropyrimidin-2-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester

_{^{1 H NMR (CDCl 3, 400MHz}} ): δ 8.09 (d, J = 5.2,1H), 6.45 (d, J = 5.2,1H), 4.60 (s, 1H), 4.26 (s, 1H), 3.74-3.79 (m, 1H), 3.53-3.63 (m, 2H), 3.36-3.40 (m, 1H), 2.15-2.23 (m, 1H), 1.85-1.93 (m, 1H), 1.38 (s, 9H); MS (ESI): m/z 298.9 [M+1] ⁺ .

Step 14d) (S)-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester

A solution of (S)-(1-(2-chloropyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid tert-butyl ester (22 g, 74 mmol) in methylamine / THF (2M, 100 mL) CuSO ₄ (1.18 g, 7.4 mmol) was added. The reaction solution was stirred in an autoclave at 80 ° C for 16 hours. The mixture was concentrated and the residue was dissolved in ethyl acetate (600ml), washed with NH ₄ OH solution (3 × 20ml). The organic layer was dried over sodium sulfate and concentrated. The residue was purified on a silica gel column (solvent with 45% ethyl acetate / petroleum ether) to give (S)-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl Tributyl methacrylate (21 g, yield 97%).

_{^{1 H NMR (DMSO-d 6}} , 400MHz): δ 7.73 (d, J = 5.6,1H), 7.18 (d, J = 5.6,1H), 6.33 (d, J = 4.4,1H), 5.68 (d, J = 5.6, 1H), 4.02-4.05 (m, 1H), 3.54 (m, 4H), 2.72 (d, J = 4.8, 3H), 2.07-2.08 (m, 1H), 1.78-1.84 (m, 1H) ), 1.39 (s, 9H) ; MS (ESI): m / z 294.1 [m + 1] +.

Step 14e) (S)-4-(3-Aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine

(S)-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)carbamic acid in 4N HCl/dioxane (50 mL) at rt Tributyl ester (21 g, 71.5 mmol) was stirred for 1 hour. The reaction mixture was concentrated to give (S)-4-(3-aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine (18, yield over 100%).

_{^{1 H NMR (DMSO-d 6}} , 400MHz): δ 7.74 (d, J = 5.6,1H), 7.34 (d, J = 4.8,1H), 5.68 (d, J = 5.2,1H), 3.63-3.67 ( m, 1H), 3.34 - 3.57 (m, 4H), 2.72 (d, J = 4.8, 3H), 2.07 - 2.09 (m, 1H), 1.77-1.85 (m, 1H).

Step 14f) (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrole Pyridin-3-yl)urea

To a solution of 4-(azetidin-1-ylmethyl)aniline (162 mg, 1 mmol) in EtOAc (EtOAc) The reaction was stirred at room temperature for 30 min then (S)-4-(3-aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine (332 mg, 1.0 mmol). The mixture was stirred for a further 2 hours. After the starting material was consumed, the crude product was purified by preparative HPLC under Condition A to give (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3- (1-(2-(Methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea (99.2 mg, 25.9%).

^{1 H NMR (400MHz, DMSO-} d6) δ 8.28 (s, 1 H), 7.76 (d, J = 5.6Hz, 1 H), 7.29 (d, J = 8.4Hz, 2 H), 7.09 (d, J = 8.4 Hz, 2 H), 6.49 (d, J = 6.8 Hz, 1 H), 6.35 (d, J = 4.8 Hz, 1 H), 5.74 (d, J = 6.0 Hz, 1 H), 4.24 (brs , 1 H), 3.68-3.21 (m, 6 H), 3.05 (t, J = 6.8 Hz, 4 H), 2.81-2.65 (m, 3 H), 2.12 (m, 1 H), 1.97-1.82 ( m, 3 H). LCMS (ESI): m / z 382.2 [M + H +].

Example 15: (S)-1-(4-(azetidin-1-ylmethyl)-3-fluorophenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) Pyrrolidin-3-yl)urea

Step 15a) N-(4-(azetidin-1-ylmethyl)-3-fluorophenyl)-2,2,2-trifluoroacetamide

Over 30 minutes azetidine hydrochloride (1.4g, 15mmol) and DIPEA (1.9g, 15mmol) in the in CH ₃ CN (30mL) was added dropwise a solution of N- (4- (bromomethyl) 3-fluorophenyl)-2,2,2-trifluoroacetamide (1.5 g, 5 mmol). The reaction mixture was stirred at room temperature for 3 hours under N ₂ atmosphere. The reaction mixture was diluted with EtOAc EtOAc EtOAc. %).

Step 15b) 4-(Azetidin-1-ylmethyl)-3-fluoroaniline

To N- (4- (azetidin-1-ylmethyl) -3-fluorophenyl) -2,2,2-trifluoro-acetyl amine (1.1g, 3.9mmol) in CH ₃ OH ( A solution of K ₂ CO ₃ (1.62 g, 11.7 mmol) in water (10 ml) was added to a solution in 30 ml), and the reaction mixture was stirred at 75 ° C for 3 hours under N ₂ atmosphere. The reaction mixture was diluted with EtOAc EtOAc EtOAc. ).

Step 15c) (S)-1-(4-(azetidin-1-ylmethyl)-3-fluorophenyl)-3-(1-(2-(methylamine) Pyrimidin-4-yl)pyrrolidin-3-yl)urea

Add TEA (303 mg, 3 mmol) and CDI (194 mg, 1.3 mmol) to a solution of 4-(azetidin-1-ylmethyl)-3-fluoroaniline (180 mg, 1 mmol) in DMF (4 mL) . The reaction was stirred at room temperature for 30 minutes. (S)-4-(3-Aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine (332 mg, 1.0 mmol) was added, and the mixture was further stirred for 2 hr. The crude product was purified using EtOAc (EtOAc)

^{1 H NMR (400MHz, DMSO-} d 6) δ 8.54 (s, 1 H), 7.81-7.65 (m, 1H), 7.41-6.85 (m, 3 H), 6.65-6.22 (m, 2 H), 5.74 (d, J = 7.2 Hz, 1 H), 4.26 (brs, 1 H), 3.82-3.01 (m, 9 H), 2.89-2.42 (m, 4 H), 2.32-1.71 (m, 4 H). LCMS (ESI): m / z 400.2 [M + H +].

Example 16: (S)-1-(3-chloro-4-((dimethylamino)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidine -3-yl)urea

Step 16a) N-(3-Chloro-4-methylphenyl)-2,2,2-trifluoroacetamide

Pyridine (75 ml, 0.80 mol) was added to a solution of 3-chloro-4-methylaniline (14.2 g, 100 mmol) in DCM (250 ml), and trifluoroacetic acid anhydride (16.5 ml) was added dropwise at 0 °C. 112.5 mmol). The mixture was stirred at room temperature for 1 hour. The reaction mixture was washed with 10% aq. HCl then water. The organic phase was dried and concentrated over Na ₂ SO _4, to give the title compound, which was used without further purification (22g, 92.8%).

Step 16b) N-(4-(bromomethyl)-3-chlorophenyl)-2,2,2-trifluoroacetamide

Add BPO (6.7 g, 27.7) to a solution of N-(3-chloro-4-methylphenyl)-2,2,2-trifluoroacetamide (22 g, 92.2 mmol) in CCI ₄ (740 mL) Methyl) and NBS (19.7 g, 110.6 mmol). The mixture was allowed to warm to 100 ° C and was illuminated with a high pressure lamp for 1 hour. The reaction mixture was stirred at 100 ° C for 36 hours. After cooling to 0 ° C, the reaction mixture was filtered and washed with brine, then concentrated. The crude product was washed with EtOAc (EtOAc)EtOAc.

Step 16c) N-(3-Chloro-4-((dimethylamino)methyl)phenyl)-2,2,2-trifluoroacetamide

N-(4-(bromomethyl) dissolved in MeCN (150 ml) was added dropwise to a solution of dimethylamine hydrochloride (4.6 g, 56.4 mmol) in MeCN (150 mL) at EtOAc. 3-chlorophenyl)-2,2,2-trifluoroacetamide (3.0 g, 9.4 mmol), and stirred at 0 ° C for 30 min. Subsequently, 100 mL of saturated NaHCO _{3 was} added and the mixture was partitioned between DCM and water. The organic layer was washed with water (15mL), dried over Na ₂ SO _4, and concentrated to be purified by preparative TLC (petroleum ether: EtOAc = 2: 1), to give the title compound (2.2g, 83%).

Step 16d) 3-Chloro-4-((dimethylamino)methyl)aniline

To N-(3-chloro-4-((dimethylamino)methyl)phenyl)-2,2,2-trifluoroacetamide (2.2 g, crude) in MeOH (30 mL) K ₂ CO ₃ (3.25 g, 23.6 mmol) was added to the solution in H ₂ O (30 mL). The mixture was then stirred at 80 ° C for 1 hour. The reaction mixture was concentrated to give title crystall

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.48 (d, J = 8.0 Hz, 1H), δ 6.91 (d, J = 2.4 Hz, 1H), 6.79-6.77 (m, 1H), δ 3.77 (s, 2H) ), δ 3.52 (s, 2H), δ 2.47 (d, J = 10.8 Hz, 7H), δ 2.28 (s, 3H).

Step 16e) (S)-1-(3-Chloro-4-((dimethylamino)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) Pyrrolidin-3-yl)urea

Add CDI (224 mg, 1.4 mmol) and DIPEA to a solution of 3-chloro-4-((dimethylamino)methyl)aniline (200 mg, 1.08 mmol) in anhydrous DMF (5 mL) 413.2 mg, 3.26 mmol). The mixture was stirred at 25 ° C for 2 hours. Then (S)-4-(3-Aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine (208 mg, 1.08 mmol) was added, and the mixture was stirred at 25 ° C for 16 hours. The mixture was purified by preparative EtOAc (EtOAc)

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.42 (s, 1H), δ 7.74 (d,j=6.0 Hz, 1H), 7.62 (d,j=2.0 Hz, 1H), 7.24 (d,j= 8.4 Hz, 1H), 7.13 - 7.11 (m, 1H), 6.55 (d, j = 6.8 Hz, 1H), 6.33 (d, j = 4.4 Hz, 1H), 5.71 (d, j = 5.6 Hz, 1H) , 4.23 (s, 1H), 3.55 (d, j = 2.8 Hz, 2H), 3.34 (s, 3H), 2.71 (d, j = 4.8 Hz, 3H), 2.12 (s, 8H), 1.87 (d, j = 9.6 Hz, 1H).

LCMS (ESI): m / z 404.0 [M + H +].

Example 17: (S)-1-(4-(azetidin-1-ylmethyl)-3-chlorophenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) Pyrrolidin-3-yl)urea

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.09 (brs, 1 H), 7.67 (d, J = 6.0Hz, 1 H), 7.49 (s, 1 H), 7.31-7.17 (m, 2 H), 6.67 (brs, 1 H), 5.57 (d, J = 6.0 Hz, 1 H), 4.78 (brs, 1 H), 4.50 (brs, 1 H), 3.62 (s, 1 H), 3.62-2.85 (m, 9 H), 2.35-2.11 (m, 4 H), 1.27 (s, 2 H). LCMS (ESI): m / z 416.2 [M + H +].

Example 18: (S)-1-(3-chloro-4-((4-methylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidine-4 -based pyrrolidin-3-yl)urea

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.48 (s, 1H), δ 7.74 (d, j = 5.6 Hz, 1H), 7.62 (d, j = 2.4 Hz, 1H), 7.25 (d, j = 8.4 Hz, 1H), 7.13-7.10 (m, 1H), 6.58 (d, j = 6.8 Hz, 1H), 5.71 (d, j = 6.0 Hz, 1H), 4.23 (s, 1H), 3.561 (d, j=3.6 Hz, 1H), 3.42 (s, 3H), 2.71 (d, j = 4.8 Hz, 3H), 2.50 (d, j = 2.0 Hz, 2H), 2.35-2.28 (m, 8H), 2.14 ( s, 1H), 2.11 (s, 3H), 1.89-1.84 (m, 1H).

LCMS (ESI): m / z 459.0 [M + H +].

Example 19: (S)-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidine-4) -based pyrrolidin-3-yl)urea

Step 19a) N-(3-Chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-2,2,2-trifluoroacetamide

To N-(4-(bromomethyl)-3-chlorophenyl)-2,2,2-trifluoroacetamide (2.0 g, 6.35 mmol) in MeCN (150 mL) To the solution was added dropwise 1-ethylpiperazine (4.3 g, 38.1 mmol) dissolved in MeCN (150 ml). The mixture was stirred at 0 ° C for 30 minutes. After the addition of 100mL of saturated NaHCO _3, the mixture was partitioned between DCM and water. The organic layer was washed with water (15ml), dried over Na ₂ SO _4, and concentrated to be purified by preparative TLC (PE: EA = 2: 1 ), to give the title compound, which was used without further purification in the next step ( 1.8g, 81.8%).

Step 19b) 3-Chloro-4-((4-ethylpiperazin-1-yl)methyl)aniline

Prepare 3-chloro-4-(4-4- in a manner similar to 3-chloro-4-((dimethylamino)methyl)aniline with 1-ethylpiperazine instead of dimethylamine hydrochloride Ethyl piperazin-1-yl)methyl)aniline.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.01 (d, J = 7.6Hz, 1H), 6.55 (d, J = 1.2Hz, 1H), 6.45 (m, 1H), 5.23 (s, 2H), 2.23- 2.33 (m, 10 H), 0.93 (m, 3H).

Step 19c) (phenyl 3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)carbamate

To a solution of 3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenylamine (300 mg, 1.18 mmol) in anhydrous THF (12 mL) Ester (185 mg, 1.18 mmol). The mixture was stirred at 65 ° C for 30 minutes. After cooling to 25 &lt;0&gt;C, <RTI ID=0.0>

Step 19d) (S)-1-(3-Chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino)) Pyrimidin-4-yl)pyrrolidin-3-yl)urea

Add to a solution of (3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)carbamic acid phenyl ester (290 mg) in anhydrous DMF (3 mL) DIPEA (102.7 mg, 0.79 mmol). The mixture was stirred at room temperature for 0.5 hours. (S)-4-(3-Aminopyrrolidin-1-yl)-N-methylpyrimidin-2-amine (150 mg, 0.40 mmol) was added, and the mixture was stirred at room temperature for 16 hr. The mixture was purified by preparative HPLC (Method C) to give the title compound (18.0 mg, 3.2%).

^{1 H NMR (400MHz, DMSO- d} 6) δ 8.45 (s, 1H), 7.74 (d, J = 6.0Hz, 1H), 7.62 (d, J = 2.0Hz, 1H), 7.25 (d, J = 8.8 Hz, 1H), 7.12-7.10 (m, 1H), 6.55 (d, J = 6.8 Hz, 1H), 6.33-6.30 (m, 1H), 5.71 (d, J = 6.4 Hz, 1H), 3.57-3.54 (m, 2H), 3.41 (s, 3H), 3.37-3.32 (m, 2H), 3.27 (s, 1H), 2.71 (d, J = 4.8 Hz, 3H), 2.52 (d, 1H), 2.42- 2.23 (m, 10H), 2.13-2.10 (m, 1H), 1.87 (d, J = 7.2 Hz, 1H), 0.94 (t, J = 7.2 Hz, 3H).

LCMS (ESI): m / z 473.0 [M + H +].

Example 20: (S)-1-(3-chloro-4-(N-morpholinylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidine-3 -based urea

According to the manufacture of (S)-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino) This compound was prepared by the general procedure of pyrimid-4-yl)pyrrolidin-3-yl)urea (Example 19).

^{1 H NMR (400MHz, DMSO- d} 6) δ 7.65-7.62 (m, 2 H), 7.36 (d, J = 8.4Hz, 1 H), 7.22 (dd, J = 2.4,8.4Hz, 1 H), 6.17 (s, 1 H), 4.46-4.40 (m, 1 H), 3.95-3.82 (m, 2 H), 3.71-3.68 (m, 8 H), 3.48-3.47 (m, 1 H), 2.96 ( s, 3 H), 2.61-2.59 (m, 4 H), 2.38-2.32 (m, 1 H), 2.12-2.04 (m, 1 H).

LCMS (ESI): m / z 446.0 [M + H +].

Example 21: (S)-1-(4-((Dimethylamino)methyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidine-4) -based pyrrolidin-3-yl)urea

According to the manufacture of (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) This compound was prepared by the general procedure of pyrrolidin-3-yl)urea (Example 14).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.22 (br s, 1 H), 7.72-7.51 (m, 4 H), 7.59-7.45 (m, 2 H), 6.83 (br s, 1 H), 5. 5.57 (m, 1 H), 4.99 (brs, 1 H), 4.53 (brs, 1 H), 3.82-3.25 (m, 6 H), 2.93 (d, J = 4.4 Hz, 3 H), 2.36-2.05 (m, 8 H). LCMS (ESI): m / z 438.2 [M + H +].

Example 22: (S)-1-(4-(azetidin-1-ylmethyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidine) -4-yl)pyrrolidin-3-yl)urea

According to the manufacture of (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) This compound was prepared by the general procedure of pyrrolidin-3-yl)urea (Example 14).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.22 (brs, 1 H), 7.73-7.61 (m, 2 H), 7.59-7.45 (m, 2 H), 6.82 (brs, 1 H), 5.62-5.51 ( m,1 H), 4.84 (brs, 1 H), 4.51 (brs, 1 H), 3.71 (s, 2 H), 3.72-3.05 (m, 8 H), 2.93 (d, J = 4.8 Hz, 3 H), 2.32 - 2.05 (m, 4 H). LCMS (ESI): m / z 450.2 [M + H +].

Example 23: (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-(pyrrolidin-1-ylmethyl)-3- (trifluoromethyl)phenyl)urea

According to the manufacture of (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) This compound was prepared by the general procedure of pyrrolidin-3-yl)urea (Example 14).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.12 (brs, 1 H), 7.72-7.52 (m, 4 H), 7.59-7.45 (m, 2 H), 6.89 (brs, 1 H), 5.64 - 5.62 ( m,1 H), 5.39 (brs, 1 H), 4.59 (brs, 1 H), 3.76 (s, 3 H), 3.72-3.41 (m, 3 H), 2.92 (d, J = 4.0 Hz, 3 H), 2.58 (s, 4 H), 2.36-1.85 (m, 6 H). LCMS (ESI): m / z 464.2 [M + H +].

Example 24: (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-((4-methylpiperazin-1-yl)) Methyl)-3-(trifluoromethyl)phenyl)urea

According to the manufacture of (S)-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino) This compound was prepared by the general procedure of pyrimid-4-yl)pyrrolidin-3-yl)urea (Example 19).

^{1 H NMR (400MHz, DMSO- d} 6) δ 8.83 (s, 1 H), 8.16 (s, 2 H), 7.90 (d, J = 2.0Hz, 1 H), 7.74 (d, J = 6.0Hz, 1 H), 7.52 (d, J = 8.4 Hz, 1 H), 7.46-7.44 (m, 1 H), 6.79 (d, J = 7.2 Hz, 1 H), 6.48 (brs, 1 H), 5.73 ( d, J = 6.0 Hz, 1 H), 4.24 (s, 2 H), 3.58-3.55 (m, 2 H), 3.43 (s, 3 H), 2.71 (d, J = 4.8 Hz, 3 H), 2.48-2.37 (m, 8 H), 2.20 (s, 3 H), 2.15-2.13 (m, 1 H), 1.89-1.86 (m, 1 H).

LCMS (ESI): m / z 493.1 [M + H +].

Example 25: (S)-1-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamine) Pyrimidin-4-yl)pyrrolidin-3-yl)urea

According to the manufacture of (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) This compound was prepared by the general procedure of pyrrolidin-3-yl)urea (Example 14).

¹ H NMR (400 MHz, Methanol- d ₄ )) δ 7.81 (s, 1 H), 7.74 (d, J = 6.0 Hz, 1 H), 7.65 (d, J = 8.8 Hz, 1 H), 7.53 (d) , J = 8.0 Hz, 1 H), 5.84 (d, J = 6.4 Hz, 1 H), 4.62-4.51 (m, 2 H), 4.49-4.32 (m, 1 H), 3.61-3.37 (m, 5) H), 2.88 (s, 3 H), 2.65-1.95 (m, 11 H), 1.12 (t, J = 7.2 Hz, 3 H). LCMS (ESI): m / z 507.2 [M + H +].

Example 26: (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-(N-morpholinylmethyl)-3-( Trifluoromethyl)phenyl)urea

According to the manufacture of (S)-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino) This compound was prepared by the general procedure of pyrimid-4-yl)pyrrolidin-3-yl)urea (Example 19).

^{1 H NMR (400MHz, DMSO- d} 6) δ 8.80 (s, 1 H), 7.93 (s, 1 H), 7.77 (d, J = 6.0Hz, 1 H), 7.58 (d, J = 8.4Hz, 1 H), 7.49-7.47 (m, 1 H), 6.76 (d, J = 6.4 Hz, 1 H), 5.81 (d, J = 6.0 Hz, 1 H), 4.27 (s, 2 H), 3.65- 3.62 (m, 1 H), 3.58-3.56 (m, 5 H), 3.51 (s, 3 H), 2.75 (d, J = 8.8 Hz, 3 H), 2.35 (d, J = 4.4 Hz, 4 H ), 2.18-2.17 (m, 1 H), 1.92-1.91 (m, 1 H).

LCMS (ESI): m / z 480.2 [M + H +].

Example 27: 1-(4-Azetidin-1-ylmethyl-3-trifluoromethyl-phenyl)-3-[(S)-1-(2-methylamino-pyrimidin-4-yl) )-pyrrolidin-3-yl]-urea

According to the manufacture of (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) This compound was prepared by the general procedure of pyrrolidin-3-yl)urea (Example 14).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.22 (brs, 1 H), 7.73-7.61 (m, 2 H), 7.59-7.45 (m, 2 H), 6.82 (brs, 1 H), 5.62-5.51 ( m,1 H), 4.84 (brs, 1 H), 4.51 (brs, 1 H), 3.71 (s, 2 H), 3.72-3.05 (m, 8 H), 2.93 (d, J = 4.8 Hz, 3 H), 2.32 - 2.05 (m, 4 H). LCMS (ESI): m / z 450.2 [M + H +].

Assessing the inhibitory effect of test compounds on CDK8 activity

CDK8 inhibition ¹ was assessed in a competitive ATP site binding assay. The full-length recombinant human His-tagged CDK8 is pre-mixed with the cyclin C dimer and the europium-labeled anti-His (Life Technologies, Madison, WI), and added to the test compound and the comparative compound containing the diluted to a certain concentration range. In the analysis board. Tracer 236, the exclusive kinase inhibitor labeled with Alexa 647 (Life Technologies), was added and incubated for an additional 60 minutes. The combination of the tracer and CDK8 was detected by time-lapse fluorescence resonance energy transfer (TR-FRET). The enthalpy is excited by light at 340 nm, causing the energy to transfer to Alexa 647, and then emitting 665 nm of fluorescence when the two fluorophores are in close proximity by binding interaction. Fluorescence intensity (excitation at 340 nm and emission at 615 and 665 nm) was read on a ViewLux multimode plate reader (PerkinElmer, Waltham, MA). The TR-FRET signal was calculated by dividing the fluorescence intensity at 665 nm by the fluorescence intensity at 615 nm.

Analysis was performed on a 1536-well black microplate in a volume of 4 μl/well. The final concentrations of CDK8/cyclin C dimer, anti-His and Tracer 236 were 5, 4 and 10 nM, respectively. The assay buffer was 50 mM HEPES pH 7.5, 10 mM MgCl ₂ , 0.01% Brij 35, 1 mM EGTA, 0.1% bovine gamma globulin and 2 mM dithiothreitol.

Maximum tracer binding and the resulting maximum TR-FRET signal were obtained in control wells with DMSO as the sample. The tracer displacement caused by the compound causes a decrease in signal. Very small signals were obtained in control wells lacking CDK8/cyclin C dimer. The percent inhibition of tracer binding in the presence of the compound was calculated and plotted as a function of compound concentration. The resulting data with a four parameter Hill curve (Hill curves) fitting, and determines the effectiveness (IC ₅₀ value) using Genedata Screener software (Genedata, Basel, Switzerland).

references:

1. Lebakken CS, Riddle SM, Singh U, Frazee WJ. Et al. Development and applications of a braod-coverage, TR-FRET-based kinase binding assay platform. J. Biomol Screen.

2. Casamassimi A, Napoli C. Mediator complexes and eukaryotic transcription regulation: an overview. Biochemie (200&) 89 , 1439-1446.

3. Alarcon C, Zaromytidou AI, Xi Q, Gao S, Yu J, et al. Nuclear CDKs drive Smad transcriptional activation and turnover in BMP and TGF-β pathways. Cell (2009) 139 , 757-769.

4. Morris EJ, Ji JY, Yang F, Di Stefano L, Herr A, et al. E2F1 represses β-catenin transcription and is antagonized by both pRB and CDK8. Nature (2008) 455 , 552-558.

5. Zho J, Ramos R, and Demma M. CDK8 regulates E2F1 transcriptional activity through S375 phosphorylation. Oncogene (2012), 1-11.

6. Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, et al. CDK8 is a colorectal cancer oncogene that regulates β-catenin activity. Nature (2008) 455 , 547-551.

7. Firestein R and Hahn WC. Revving the throttle on an oncogene: CDK8 takes the driver seat. Cancer Res (2009) 69 , 7899-7901.

8. Adler AS, McCleland ML, Truong T et al. CDK8 maintains tumor de-differentiation and embryonic stem cell pluripotency. Cancer Research (2012)

9. Xu W and Ji JY. Dysregulation of CDK8 and Cyclin C in tumorigenesis. J Genetics Genomics (2011) 38 , 439-452.

10. Firestein R, Shima K, Nosho K, Irahara N, et al. CDK8 expression in 470 colorectal cancers in relation to β-catenin activation, other molecular alterations and patient survival. International Journal of Cancer (2010) 126 , 2863-2873 .

11. Seo JO, Han SI, and Lim SC. Role of CDK8 and β-catenin in colorectal adenocarcinoma. Oncology Reports (2010) 24 , 285-291.

12. Adler AS, McCleland ML, Truong T et al. CDK8 maintains tumor de-differentiation and embryonic stem cell pluripotency. Research (2012).

13. Kapoor A, Goldberg MS, Cumberland LK, Ratnakumar K, et al.

Although a number of embodiments have been described, such examples can be modified to provide other embodiments utilizing the compounds and methods described herein. Therefore, the scope of the invention is defined by the scope of the appended claims rather than the specific embodiments represented by the examples.

Claims (16)

a compound of formula (I), Or a salt thereof, wherein: R ¹ is selected from the group consisting of: And wherein R ¹ is independently substituted by 1, 2 or 3 R ^x groups; R ² and R ^{3 are} independently H, C _1-12 alkyl, C _2-12 alkenyl, C _2-12 Alkynyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, halo, -OR ^a , -SR ^a , -N(R ^a ) ₂ , -CN, -NO ₂ , -C(O)R ^a , -CO ₂ R ^a , -C(O)N(R ^a ) ₂ , -C(O)SR ^a , -C(O)C(O)R ^a , -C(O)CH ₂ C(O R ^a , -C(S)N(R ^a ) ₂ , -C(S)OR ^a , -S(O)R ^a , -SO ₂ R ^a , -SO ₂ N(R ^a ) ₂ , -N (R ^a )C(O)R ^a , -N(R ^a )C(O)N(R ^a ) ₂ , -N(R ^a )SO ₂ R ^a , -N(R ^a )SO ₂ N(R ^a ) ₂ , -N(R ^a )N(R ^a ) ₂ , -N(R ^a )C(=N(R ^a ))N(R ^a ) ₂ , -C(=N)N(R ^a ) ₂ , -C=NOR ^a , -C(=N(R ^a ))N(R ^a ) ₂ , -OC(O)R ^a or -OC(O)N(R ^a ) ₂ , wherein R ² and R each of C ³ _1-12 alkyl, C _2-12 alkenyl group, C _2-12 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl each line is independently optionally substituted with one or more groups a group R ^x substituted; or R ¹ and R ² together with the atom to which they are attached form a 4, 5, 6, 7 or 8 membered carbocyclyl, heterocyclyl, heteroaryl or aryl group, which is a heterocyclic or heterocyclic ring. The base, heteroaryl and aryl are optionally substituted by one or more groups R ^x ; Each of R ^a and R ^{b is} independently H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclic, wherein each C _1-6 alkyl, C _3-6 alkenyl, C _3-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl are, independently, optionally substituted with one or more groups R ^x ; or R ^a and R ^b form a 3-6 membered heterocyclic ring together with the nitrogen to which they are attached, which heteroaryl ring optionally with oxo (oxo), halo or optionally substituted with oxo or halo C _1-6 of Alkyl substituted; each R ^{v is} independently hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl or heterocyclyl, each of which C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, aryl, heteroaryl and heterocyclyl are optionally optionally selected from the group consisting of Substituting: a pendant oxy group, a halogen group, an amine group, a hydroxyl group, and optionally a C ₁ -C ₆ alkyl group substituted with one or more groups independently selected from the pendant oxy group and a halogen group; or two R ^v The attached nitrogens together form a 3-6 membered heterocyclic group which is optionally substituted with one or more groups independently selected from the group consisting of pendant oxy groups, halo groups, and optionally Or a plurality of C _1-3 alkyl groups independently substituted with a group selected from a pendant oxy group and a halogen group; and each R ^{x is} independently a pendant oxy group, a C _1-6 alkyl group, a C _2-6 alkenyl group, C _2-6 alkynyl, C _1-6 haloalkyl, carbocyclyl, aryl, heteroaryl, heterocyclyl, -F, -Cl, -Br, -I, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -OR ^v , -SR ^v , -OC(O)-R ^v , -OC(O)-OR ^v , -C(O)-R ^v , -C(O)-OR ^v , -S(O)-R ^v , -S(O) ₂ -R ^v , -OC(O)-N(R ^v ) ₂ , -N(R ^v )-C(O)-OR ^v , -N(R ^v )-C(O)- N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v , -N(R ^v )-S(O)-R ^v , -N(R ^v )-S(O) ₂ -R ^v , -N(R ^v )-S(O)-N(R ^v ) ₂ or -N(R ^v )-S(O) ₂ -N (R ^v ) ₂ , wherein any C _1-6 alkyl group, C _2-6 alkenyl group, C _2-6 alkynyl group, C _1-6 haloalkyl group, carbocyclic group, aryl group, heteroaryl group and heterocyclic ring The substrate is independently substituted with one or more groups selected from the group consisting of pendant oxy, halo, -NO ₂ , -N(R ^v ) ₂ , -CN, -C(O)-N(R). ^v ) ₂ , -S(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -OR ^v , -SR ^v , -OC(O)-R ^v , -C (O)-R ^v , -C(O)-OR ^v , -S(O)-R ^v ,- S(O) ₂ -R ^v , -C(O)-N(R ^v ) ₂ , -S(O) ₂ -N(R ^v ) ₂ , -N(R ^v )-C(O)-R ^v And -N(R ^v )-S(O)-R ^v , -N(R ^v )-S(O) ₂ -R ^v and optionally one or more selected from the group consisting of pendant and halogen groups a group substituted with a C _1-6 alkyl group; and wherein when R ² is F and R ³ is hydrogen, then R ^{x is} not methyl, ethyl, trifluoromethyl or 2-fluorophenyl. The compound of claim 1 or a salt thereof, wherein R ¹ is selected from the group consisting of: R ² and R ^{3 are} independently H, C _1-12 alkyl, halo or -OR ^a , wherein each C _1-12 alkyl of R ² and R ³ is independently independently one or more groups, as appropriate R ^{x is} substituted; each R ^a and R ^{b is} independently H, C _1-6 alkyl, wherein each C _1-6 alkyl group is independently substituted with one or more groups R ^x as appropriate; each R ^v independently Is a C _1-6 alkyl group; and each R ^{x is} independently a heterocyclic group, -F or -N(R ^v ) ₂ , wherein any heterocyclic group is independently selected from C _1-6 by one or more a group substituted with an alkyl group; and wherein when R ² is F and R ³ is hydrogen, then R ^{x is} not methyl, ethyl, trifluoromethyl or 2-fluorophenyl. The compound of claim 1 or 2, wherein R ² and R ^{3 are} independently halo, trifluoromethyl, C _1-6 alkyl or C _1-6 alkoxy, wherein the alkyl and alkoxy are independently Substituted by -N(R ^v ) ₂ or a heterocyclic group; wherein the heterocyclic group is optionally substituted by a pendant oxy group, a halogen group or, optionally, a C _1-6 alkyl group substituted by a halo group or a pendant oxy group . The compound of claim 1 or 2, wherein R ² is hydrogen, chloro, methyl, (4-methylpiperazin-1-yl)methyl, trifluoromethoxy, difluoromethoxy, 4- Azetidin-1-ylmethyl, morpholin-4-ylmethyl, dimethylaminomethyl, pyrrolidin-1-ylmethyl or 4-ethylpiperazin-1-ylmethyl . A compound according to claim 1 or 2, wherein R ³ is hydrogen, trifluoromethyl, fluoro or chloro. The compound of claim 1 or 2, wherein R ^a and R ^{b are} independently hydrogen or C _1-6 alkyl. A compound of claim 1 or 2 wherein R ^a is hydrogen. The compound of claim 1 or 2, wherein R ^b is methyl. A compound according to claim 1 or 2 which is selected from the group consisting of 1-[(3S)-1-([1,2,4]triazolo[4,3-b]pyridazine-6- Pyrrolidin-3-yl]-3-[3-(trifluoromethyl)phenyl]urea; 1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3S -1([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; 1-(3-chlorophenyl)-3- [(3S)-1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]urea; 1-[(3R)-1- ([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidin-3-yl]-3-[3-(trifluoromethyl)phenyl]urea; -[4-chloro-3-(trifluoromethyl)phenyl]-3-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazine-6- Pyrrolidin-3-yl]urea; 1-(3-chlorophenyl)-3-[(3R)-1-([1,2,4]triazolo[4,3-b]pyridazine -6-yl)pyrrolidin-3-yl]urea; (S)-1-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrolidine 3-yl)-3-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)urea; 1-[(3S)-1- (2-aminopyrimidin-4-yl)pyrrolidin-3-yl]-3-(3-chloro-4-methyl-phenyl)urea; 1-[(S)-1-(2-amino) -pyrimidin-4-yl)-pyrrolidin-3-yl]-3-(4-chloro-3-trifluoromethyl-phenyl)-urea; 1-[4-chloro-3-(trifluoromethyl) Phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidine-4- Pyrrolidin-3-yl]urea; 1-(3-chloro-4-methyl-phenyl)-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl] Pyrrole Pyridin-3-yl]urea; 1-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidin-3-yl]-3-[4-(trifluoromethoxy) Phenyl]urea; 1-[4-(difluoromethoxy)phenyl]-3-[(3S)-1-[2-(methylamino)pyrimidin-4-yl]pyrrolidine- 3-yl]urea; (S)-1-(4-(azetidin-1-ylmethyl)phenyl)-3-(1-(2-(methylamino)pyrimidine-4- () pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-ylmethyl)-3-fluorophenyl)-3-(1-(2-( Methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-((dimethylamino)methyl)phenyl)-3- (1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-ylmethyl)- 3-chlorophenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-( (4-methylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S )-1-(3-chloro-4-((4-ethylpiperazin-1-yl)methyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl) Pyrrolidin-3-yl)urea; (S)-1-(3-chloro-4-(N-morpholinylmethyl)phenyl)-3-(1-(2-(methylamino)) (S)-1-(4-((dimethylamino)methyl)-3-(trifluoromethyl)benzene -(3-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(4-(azetidin-1-yl) Methyl)-3-(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1 -(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-(pyrrolidin-1-ylmethyl)-3-(trifluoromethyl) Phenyl)urea; (S)-1-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-((4-methylperidine) (X)-1-(4-((4-ethylpiperazin-1-yl)methyl)-3) -(trifluoromethyl)phenyl)-3-(1-(2-(methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)urea; (S)-1-(1-( 2-(Methylamino)pyrimidin-4-yl)pyrrolidin-3-yl)-3-(4-(N-morpholinomethyl)-3-(trifluoromethyl)phenyl Urea; 1-(4-azetidin-1-ylmethyl-3-trifluoromethyl-phenyl)-3-[(S)-1-(2-methylamino-pyrimidine- 4-(yl)-pyrrolidin-3-yl]-urea; (S)-1-(1-([1,2,4]triazolo[4,3-b]pyridazin-6-yl)pyrrole Pyridin-3-yl)-3-(3-chloro-4-methylphenyl)urea; and pharmaceutically acceptable salts thereof. A compound according to claim 1 or 2, which is selected from the group consisting of: And its salt. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant, carrier or vehicle. Use of a compound according to any one of claims 1 to 10 for treatment drug. A use of a compound according to any one of claims 1 to 10 for the manufacture of a medicament for the treatment of a hyperproliferative disorder. A use of a compound according to any one of claims 1 to 10 for the manufacture of a medicament for the treatment of a disease associated with CDK8 activity. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, for use as a therapeutically active substance. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, for use as a therapeutically active substance for the treatment of a hyperproliferative disorder.