TWI480276B - Compounds and compositions as protein kinase inhibitors - Google Patents

Description

Compounds and compositions as protein kinase inhibitors

The present invention provides a novel class of compounds, pharmaceutical compositions comprising the same, and the use of such compounds to treat or prevent a disease or condition associated with abnormal or dysregulated kinase activity, particularly a disease or condition associated with abnormal activation of B-Raf method.

The present application claims the priority of US Provisional Patent Application No. 61/238,073, filed on Aug. 28, 2009, and U.S. Provisional Patent Application No. 61/313,039, filed on March 11, 2010. The entire disclosure of these applications is hereby incorporated by reference in its entirety in its entirety in its entirety herein in its entirety herein

Protein kinases represent a large family of proteins that play a central role in regulating a variety of cellular processes and maintaining control over cellular function. A partial, non-limiting list of such kinases includes: receptor tyrosine kinases such as platelet-derived growth factor receptor kinase (PDGF-R), nerve growth factor receptor trkB, Met, and fibroblast growth factor receptor FGFR3 Non-receptor tyrosine kinases such as Abl and fusion kinases BCR-Abl, Lck, Csk, Fes, Bmx and c-src; and serine/threonine kinases such as B-Raf, sgk, MAP kinase ( For example, MKK4, MKK6, etc.) and SAPK2α, SAPK2β and SAPK3. Abnormal kinase activity has been observed in a number of disease conditions, including benign and malignant proliferative disorders as well as diseases caused by improper activation of the immune system and nervous system.

The novel compounds of the invention inhibit the activity of B-Raf or a mutant form thereof (e.g., V600E) and are therefore contemplated for use in the treatment of B-Raf related diseases.

In one aspect, the invention provides a compound of formula I:

Wherein: Y is selected from N and CR ₆ ; R ₁ is selected from the group consisting of hydrogen, -X ₁ R _8a , -X ₁ OX ₂ R _8a , -X ₁ C(O)NR _8a R _8b , -X ₁ NR _8a X ₂ R _8b , -X ₁ NR _8a C(O)X ₂ OR _8b , -X ₁ NR _8a C(O)X ₂ NR _8a R _8b , -X ₁ NR _8a S(O) _0-2 R _8b ; each X ₁ is independently a C _1-4 alkylene group; X ₁ and 1-3 of the hydrogen is optionally substituted selected from hydroxy, halo, cyano, C _1-4 alkyl, halo-substituted C ₁ by the _a group substituted with a _-4 alkyl group, a C _1-4 alkoxy group, and a halogen-substituted C _1-4 alkoxy group; the X ₂ group is selected from the group consisting of a bond and a C _1-4 alkylene group; wherein R _8a and R _8b is independently selected from the group consisting of hydrogen, C _1-6 alkyl, halo-substituted C _1-6 alkyl, C _3-8 cycloalkyl, heteroaryl and C _3-8 heterocycloalkyl; the cycloalkyl group of R _8a or R _8b, heterocycloalkyl or heteroaryl optionally substituted with 1 to 3 substituents independently selected from amino, cyano, C _1-4 alkyl, C _1-4 alkoxy Substituting a halogen-substituted C _1-4 alkyl group and a halogen-substituted C _1-4 alkoxy group; the restriction is that when R ₁ is selected from -X ₁ NHC(O)OR _8b and -X ₁ NR _8a S(O) _0-2 R _8b , R _{8b is} not hydrogen; R ₂ , R ₃ , R ₅ and R ₆ are independently selected from hydrogen, halo, cyano, C _{1- a 4-} alkyl group, a halogen-substituted C _1-4 alkyl group, a C _1-4 alkoxy group, and a halogen-substituted C _1-4 alkoxy group; the limitation is that when R ₅ is fluorine and the R ₁ system is selected from _{hydrogen, -X 1 R 8a, -X 1} OX 2 R 8a, -X 1 C (O) NR 8a R 8b, -X 1 NR 8a X 2 R 8b, -X 1 NR 8a C (O) X ₂ OR _8b and -X ₁ NR _8a S(O) _0-2 R _8b , when R _{3 is} different from R ₆ as hydrogen; R ₄ is selected from -R ₉ and -NR ₁₀ R ₁₁ ; wherein R ₉ is selected From C _1-6 alkyl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, aryl and heteroaryl; wherein alkyl, cycloalkyl, heterocycloalkyl, aryl of R ₉ The aryl or heteroaryl group is optionally selected from 1 to 3 independently selected from halo, cyano, C _1-4 alkyl, halo substituted C _1-4 alkyl, C _1-4 alkoxy and Substituted by a halogen-substituted C _1-4 alkoxy group; and R ₁₀ and R ₁₁ are independently selected from hydrogen and R ₉ ; R ₇ is selected from hydrogen, C _1-4 alkyl, C _3-5 a cycloalkyl group and a C _3-5 heterocycloalkyl group; wherein the alkyl group, cycloalkyl group or heterocycloalkyl group of R ₇ is optionally independently selected from the group consisting of halo, cyano, hydroxy, C _{1 -4} alkyl, halo-substituted group of C _1-4 alkyl, the C _1-4 alkoxy substituted by halo and C _1-4 alkoxy groups substituted with And N-oxide derivatives, prodrug derivatives, protected derivatives, tautomers, individual isomers and mixtures of isomers; and pharmaceutically acceptable salts and solvates of such compounds (eg hydrate).

In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or an N-oxide derivative thereof, an individual isomer and a mixture of isomers or a pharmaceutically acceptable salt thereof, and one or A wide variety of suitable excipients.

In a third aspect, the invention provides a method of treating a disease in an animal, wherein inhibition of kinase activity, particularly B-Raf activity, prevents, inhibits or ameliorates the pathology and/or symptoms of the disease, the method comprising The animal is administered a therapeutically effective amount of a compound of formula I or an N-oxide derivative thereof, a mixture of individual isomers and isomers, or a pharmaceutically acceptable salt thereof.

In a fourth aspect, the invention provides the use of a compound of formula I for the manufacture of a medicament for the treatment of a disease in an animal, wherein the kinase activity, in particular the B-Raf activity, in particular the mutant B-Raf (eg V600E), contributes to the pathology of the disease and / or symptoms.

In a fifth aspect, the invention provides a compound of formula I, an N-oxide derivative thereof, a prodrug derivative, a protected derivative, an individual isomer and a mixture of isomers, and a pharmaceutically acceptable compound thereof The method of salt.

definition

The "alkyl group" which is a group and a structural component of other groups (for example, a halogen-substituted alkyl group and an alkoxy group) may be a straight chain or a branched chain. The C _1-4 alkoxy group includes a methoxy group, an ethoxy group, and the like. The C _1-4 alkyl group substituted by a halogen means any alkyl group (branched chain or unbranched chain) in which a hydrogen may be substituted by halogen. For example, a halo-substituted C _1-4 alkyl group can be a trifluoromethyl group, a difluoroethyl group, a pentafluoroethyl group, and the like. Similarly, a _C1-6 alkyl group substituted with a hydroxy group means an alkyl group (branched chain or unbranched chain) in which any hydrogen may be substituted with a hydroxy group. For example, a _C1-6 alkyl group substituted with a hydroxy group includes a 2-hydroxyethyl group and the like. Similarly, a C _1-6 alkyl group substituted by a cyano group means an alkyl group (branched chain or unbranched chain) in which any hydrogen may be substituted by a cyano group.

"Aryl" means a monocyclic or fused bicyclic aromatic ring assembly containing from 6 to 10 ring carbon atoms. For example, the aryl group can be phenyl or naphthyl, preferably phenyl. "Stretching aryl" means a divalent group derived from an aryl group.

"Cycloalkyl" means a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring containing the specified number of ring atoms. For example, a C _3-10 cycloalkyl group includes a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, and the like.

"Heteroaryl" is as defined above for aryl, wherein one or more ring members are heteroatoms. For example, heteroaryl includes pyridyl, indenyl, oxazolyl, quinoxalinyl, quinolinyl, benzofuranyl, benzopipetanyl, benzothiopiperidyl, benzo[ 1,3]dioxolyl, imidazolyl, benzimidazolyl, pyrimidinyl, furyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, thienyl, etc. .

"Heterocycloalkyl" means a cycloalkyl group as defined in the present application, with the proviso that one or more of the specified ring carbons are selected from the group consisting of -O-, -N=, -NR-, -C(O). Partial substitution of -, -S-, -S(O)- or -S(O) ₂ - (wherein R is hydrogen, C _1-4 alkyl or a nitrogen protecting group). For example, the C _3-8 heterocycloalkyl group used in the present application to describe the compound of the present invention includes 2H-piperidyl, 4H-piperidyl, piperidinyl, 1,4-dioxyl, Lolinyl, 1,4-dithiaalkyl, N-thiomorpholinyl, imidazolidin-2-one, tetrahydrofuranyl, piperazinyl, 1,3,5-trithiaalkyl, pyrrolidinyl, Pyrrolidin-2-one, piperidinyl, piperidinone, 1,4-dioxa-8-aza-spiro[4.5]dec-8-yl and the like.

"Halogen" (or halo) means chlorine, fluorine, bromine or iodine.

"pMEK" means phosphorylating Mek.

"pERK" means phosphorylated ERK.

"Treatment" means a method of alleviating or alleviating a disease and/or its accompanying symptoms.

The compounds of the invention are named using Chemdraw Ultra (version 10.0) and/or ChemAxon name generator (JChem version 5.3.1.0).

Description of the preferred embodiment

The present invention provides compounds, compositions and methods for treating kinase-related diseases, particularly B-Raf kinase-associated diseases; for example, metastatic melanoma, solid tumors, brain tumors (such as glioblastoma multiforme (GBM)), acute bone marrow Acute leukemia (AML), prostate cancer, gastric cancer, papillary thyroid cancer, low ovarian cancer, and colorectal cancer.

In one embodiment, the compounds of formula I, R ₁ is selected from -X ₁ R _8a and _{-X 1 NHC (O) OR 8b} ; wherein each X ₁ is independently a C _1-4 alkylene group; and X ₁ 1 to 3 hydrogens are optionally substituted with a group selected from the group consisting of a hydroxyl group, a halogen group, a cyano group, a C _1-4 alkyl group, and a halogen-substituted C _1-4 alkyl group; wherein R _8a and R _8b are independently It is selected from hydrogen and C _1-6 alkyl; the restriction is that when R ₁ is -X ₁ NHC(O)OR _8b , R _{8b is} not hydrogen.

In another embodiment, a compound of Formula Ia is provided:

Wherein: Y is selected from N and _{CR 6; R 2, R 3} , R 5 and R ₆ are independently selected from hydrogen, halo, cyano, C _1-4 alkyl, halo-substituted by the group C _{1- a 4-} alkyl group, a C _1-4 alkoxy group, and a halogen-substituted C _1-4 alkoxy group; the limitation is that when R ₅ is fluorine and R ₁ is selected from hydrogen, -X ₁ R _8a , -X ₁ OX ₂ R _8a , -X ₁ C(O)NR _8a R _8b , -X ₁ NR _8a X ₂ R _8b , -X ₁ NR _8a C(O)X ₂ OR _8b and -X ₁ NR _8a S(O When _0-2 R _8b , R ₃ and R _{6 are} not hydrogen; R ₄ is selected from -R ₉ and -NR ₁₀ R ₁₁ ; wherein R ₉ is selected from C _1-6 alkyl, C _3-8 a cycloalkyl group, a C _3-8 heterocycloalkyl group, an aryl group and a heteroaryl group; wherein the alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group of R ₉ is optionally 1 to 3 Independently selected from halo, cyano, C _1-4 alkyl, halo substituted C _1-4 alkyl, C _1-4 alkoxy, and halo substituted C _1-4 alkoxy a group substituted; and R ₁₀ and R ₁₁ are independently selected from hydrogen and R ₉ ; and R ₇ is selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-5 cycloalkyl, and C _3-5 heterocycloalkane Wherein the alkyl, cycloalkyl or heterocycloalkyl group of R ₇ is optionally selected from 1 to 3 independently from halo, cyano, hydroxy, C _1-4 alkane Substituents substituted with a halo-substituted C _1-4 alkyl group, a C _1-4 alkoxy group, and a halogen-substituted C _1-4 alkoxy group.

In another embodiment, R ₄ is ₉ -R < ₉ wherein R is selected from C _1-3 alkyl and C _3-8 cycloalkyl; wherein the R ₉ alkyl group or the cycloalkyl optionally substituted with 1 Substituting to three groups independently selected from a halo group and a halo-substituted C _1-4 alkyl group.

In another embodiment, R ₂ is selected from the group consisting of hydrogen and fluorine; R ₃ is selected from the group consisting of chlorine, fluorine, and methyl; R ₅ is selected from the group consisting of hydrogen, chlorine, and fluorine; and Y is selected from the group consisting of N and CR ₆ ; ₆ is selected from the group consisting of hydrogen and fluorine.

In another embodiment, a compound selected from the group consisting of N-[(2S)-1-({4-[3-(3-chloro-5-methanesulfonylamino)phenyl)-1-(propyl) is provided Methyl-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbazate; N-[(2S)-1-[(4-{ 3-[2-Fluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino Methyl propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(2-fluoro-3-methanesulfonylamino)phenyl)-1-(propyl- Methyl 2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-[(4-{3 -[3-chloro-5-(propane-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino] Methyl propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(2,6-difluoro-3-methanesulfonylamino)phenyl)-1-( Methyl-2-phenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-[(4- {3-[2,6-Difluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2- Methyl)amino]propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-{[4-(3-{2-fluoro-3-[(3,3,3-trifluoro) Propane)sulfonylamino]phenyl}-1-(propan-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl]amino} Methyl-2-yl]carbamate; N-[(2S)-1-({4-[3-(3-chloro-2-methanesulfonylaminopyridin-4-yl)-1-(propyl) Methyl-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbazate; N-[(2S)-1-({4-[ 3-(3-Fluoro-2-methanesulfonylaminopyridin-4-yl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl Methyl-2-yl]carbamate; N-[(2S)-1-({4-[3-(2-chloro-3-ethanesulfonylamino-4,5-difluorophenyl)) Methyl-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1- ({4-[3-(2,4-Difluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl }amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4-[1-(propan-2-yl)-3-(2,4,5-tri) Fluoro-3-methanesulfonylaminophenyl-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)- 1-({4-[3-(3-Methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl Methyl-2-yl]carbamate; N-[(2S)-1-({4-[3-(3-ethanesulfonylamino-2,4-difluorophenyl)-1-() Methyl-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S )-2-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl] Methyl pyrimidin-2-yl}amino)propyl]carbamic acid; N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino) Phenyl)-1-(oxacyclo-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N- [(2S)-1-[(4-{3-[2,4-Difluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H- Methyl pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(3-cyclopropane) Amidino-2,5-difluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]amino Methyl formate; N-[(2S)-1-({4-[3-(5-chloro-3-cyclopropanesulfonylamino-2-fluorophenyl)-1-(propan-2-yl)) -1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; and N-[(2S)-1-[(4-{3-[5 -Chloro-2-fluoro-3-(propane-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino Methyl propan-2-yl]carbamate.

In another embodiment, a compound of Formula Ib is provided:

Wherein R ₃ is selected from the group consisting of chlorine, fluorine and methyl; R ₅ is selected from the group consisting of fluorine and chlorine; and R ₇ is selected from the group consisting of ethyl and isopropyl.

In another embodiment, a compound selected from the group consisting of N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-)- Methyl 1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-( {4-[3-(2,5-Difluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl} Methylamino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)phenyl) Methyl 1-ethyl-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-({4-[ 3-(2-Fluoro-3-methanesulfonylamino-5-methylphenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino Methyl propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(2-chloro-3-methanesulfonylamino-5-methylphenyl)-) Methyl 1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-( {4-[3-(2-Chloro-5-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl Methylamino)propan-2-yl]carbamic acid; N-[(2R)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)phenyl )-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino) Methyl-2-yl]carbamate; N-[(2S)-1-({4-[3-(2,5-dichloro-3-methanesulfonylamino)-1-(propyl) Methyl-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbazate; and N-[(2S)-1-({4- [3-(5-Chloro-2-fluoro-3-methanesulfonylaminophenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]aminocarboxylic acid Methyl ester.

In another embodiment, a compound selected from the group consisting of N-[5-chloro-3-(4-{2-[(2-cyanoethyl)amino]pyrimidin-4-yl}-1- (propan-2-yl)-1H-pyrazol-3-yl)-2-fluorophenyl]methanesulfonamide; N-{5-chloro-3-[4-(2-{[2-(two) Methylamino)ethyl]amino}pyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-2-fluorophenyl}methanesulfonamide; N -(5-chloro-2-fluoro-3-{4-[2-(methylamino)pyrimidin-4-yl]-1-(propan-2-yl)-1H-pyrazol-3-yl} Phenyl)methanesulfonamide; and N-{3-[4-(2-aminopyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-5 -Chloro-2-fluorophenyl}methanesulfonamide.

In another embodiment, a compound selected from the examples and tables below is provided.

In another embodiment, an intermediate compound selected from the group consisting of 3-bromo-5-chloro-2-fluoroaniline; cyano-(2-methylthio-pyrimidin-4-yl)-acetic acid tert-butyl ester is provided 1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine; 2-((2-benzylidene-1-ethylfluorenyl) Methylene)-malononitrile; 1-(3-amino-1-isopropyl-1H-pyrazol-4-yl)ethanone; 1-(3-iodo-1-isopropyl-1H -pyrazol-4-yl)ethanone; 1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)ethanone; 1-(3-iodo-1-methyl-1H-pyridyl Zyzol-4-yl)ethanone; 3-(dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)prop-2-en-1-one 3-(dimethylamino)-1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)prop-2-en-1-one; 3-(dimethylamino) )-1-(3-iodo-1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one; 4-(3-iodo-1-isopropyl-1H-pyrazole) 4-yl)pyrimidin-2-amine; 4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-amine; 4-(3-iodo-1-methyl- 1H-pyrazol-4-yl)pyrimidine-2-amine; 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol; 2-chloro-4-( 3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine; (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl) Pyrimidin-2-ylamino)propan-2-ylaminocarboxylic acid Ester; (R)-methyl-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate; (S)-1-butyl 4-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamic acid; 3-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propanenitrile; 4-(3-iodo-1-isopropyl-1H -pyrazol-4-yl)-N-methylpyrimidin-2-amine; N ¹ -(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-yl -N ² ,N ² -dimethylethane-1,2-diamine; N-(3-bromo-2,4-difluorophenyl)propane-1-sulfonamide; 3-fluoro-4 -iodopyridin-2-amine; 3-chloro-4-iodopyridin-2-amine; 3-bromo-2,5,6-trifluoroaniline; 2,4-dibromo-3,6-dichloroaniline; 3-bromo-2-chloro-5-methylaniline; 3-bromo-2,5-difluoroaniline; 3-bromo-5-chloro-2-fluorobenzoic acid; 3-bromo-5-chloro-2- Tert-butyl fluorophenylaminocarbamate; tert-butyl 3-bromo-2-fluoro-5-methylphenylcarbamic acid; 5-chloro-2-fluoro-3-(4,4,5, 5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester; 2,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester; N-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) 2-yl)phenyl)propane-1-sulfonamide; 2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-di Oxyboron ; 2,5-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2,5-dichloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-chloro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-fluoro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester; 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)pyridin-2-amine; 2,3,6-trifluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)pyridin-2-amine; 3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; and 3-methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) aniline.

The invention also includes all suitable isotopic variations of the compounds of the invention or their pharmaceutically acceptable salts. An isotopic variation of a compound of the invention or a pharmaceutically acceptable salt thereof is defined as the substitution of at least one atom by an atom having the same atomic number but differing in atomic mass from the atomic mass normally found in nature. Examples of isotopes which may be incorporated into the compounds of the invention and their pharmaceutically acceptable salts include, but are not limited to, isotopes of hydrogen, carbon, nitrogen and oxygen, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ³⁵ S, ¹⁸ F, ³⁶ Cl and ¹²³ I. Certain isotopic variations of the compounds of the invention and their pharmaceutically acceptable salts, for example, with radioisotopes (such as ³ H or ¹⁴ C), are useful in drug and/or matrix tissue distribution studies. In a particular example, the ³ H and ¹⁴ C isotopes can be used for ease of preparation and detectability. In other instances, substitution with an isotope (such as ² H) may provide certain therapeutic advantages due to higher metabolic stability, such as prolonged in vivo half-life or reduced dosage requirements. Isotopic variations of a compound of the invention or a pharmaceutically acceptable salt thereof can generally be prepared by conventional procedures using suitable isotopic variations of the appropriate reagents.

In addition to enhancing MEK and ERK signaling in wild-type B-Raf cells, some Raf inhibitors also induce cell growth in cancer cell lines and promote fibroblast transformation and growth. Induction of downstream signaling has previously been attributed to the disclosed Raf path feedback loop. However, induction of pMEK and pERK can be performed within a few minutes of treatment with Raf inhibitors, even before the reported feedback phosphorylation events were observed on B-Raf and C-Raf. Both induction signaling and cell growth were performed in a two-phase mode in which low compound concentrations (0.01-0.1 μM) produced maximum induction and higher compound concentrations (1-10 μM) produced less intense induction. Such two-phase models are also observed in biochemical assays of purified wild-type B-Raf or C-Raf and indicate mechanisms involved in the interaction of two signaling subunits. In addition, Raf dimerization upregulates pMEK, which is not achieved by transphosphorylation of Raf molecules, but may be achieved by conformational activation of the kinase. Consistent with the model, Raf inhibitor treatment induces the formation of B-Raf/C-Raf dimers in cells. In addition, knockdown of A-Raf or B-Raf with siRNA did not abolish the induction of pMEK and pERK by Raf inhibitors, and knocking out C-Raf only slightly reduced the induction. Notably, knocking out K-Ras in K-Ras mutant cells also only slightly reduced this induction, suggesting that this effect is not primarily mediated by Ras. In summary, the data proposes a model in which an inhibitor that binds to a Raf molecule induces the conformational activation of the Raf molecule in the dimerization and dimer. This may explain the insensitivity of wild-type Raf and mutant Ras tumors to selective Raf kinase inhibitors, and may also be importantly associated with toxicity, as induction of strong mitogenic signaling leads to excessive proliferation of normal tissues. Understanding the Raf inhibitor induction mechanism can be used to design improved inhibitors.

The addition of a combination of a MEK inhibitor and a Raf inhibitor significantly inhibits ERK signaling and thus reduces cell proliferation and transformation. Because treatment with MEK inhibitor alone has produced dose-limiting toxicity in the clinic, the combination of a Raf inhibitor and a MEK inhibitor may represent an excellent therapeutic strategy.

The invention also includes combinations of the BRaf inhibitors disclosed herein with other agents. In particular, the invention provides a combination with a MEK1/2 inhibitor. Figure 1 illustrates that the addition of a MEK small molecule inhibitor reverses ERK signaling, cell growth and transformation induced by Raf small molecule inhibitors. For example, a compound of formula I (compound 9 of the invention, ie: (S)-1-(4-(3-(5-chloro-2-fluoro-3-(methylsulfonylamino)phenyl)) 1-Isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamic acid methyl ester) induces cell proliferation, as shown in Figure 1 using SW620 cells for Cell Titer The negative inhibition observed by the Glo assay. The Y axis shows negative inhibition and positive inhibition. Each experiment was presented in 9 serial dilution series between 10 μM and 0.002 μM. Compound Al(N-(4-methyl-3-(1-(6-(4-methylpiperazin-1-ylamino)pyrimidin-4-yl)-1H-imidazol-2-ylamino) Phenyl)-3-(trifluoromethyl)benzamide was used as a control and A3 was a MEK inhibitor (PD0325901). Compound 9 was tested in the absence and presence of 1 μM, 0.1 μM and 0.01 μM MEK inhibitor A3.

Pharmacology and utility

The compounds of the invention modulate kinase activity and are therefore useful in the treatment of diseases or conditions in which the kinase contributes to the pathology and/or symptoms of the disease. Examples of kinases that are inhibited by the compounds and compositions described herein and which are useful in the methods described herein include, but are not limited to, B-Raf, including mutant forms of B-Raf.

The mitogen-activated protein kinase (MAPK) pathway mediates the activity of a number of effector molecules that synergistically control cell proliferation, survival, differentiation, and migration. The Raf is recruited by, for example, a growth factor, a cytokine or a hormone to cause the plasma membrane-associated Ras to bind to GTP, thereby being activated to recruit Raf. This interaction induces the kinase activity of Raf, allowing direct phosphorylation of MAPK/ERK (MEK), which in turn phosphorylates extracellular signal-associated kinase (ERK). The activated ERK then phosphorylates a number of effector molecules such as kinases, phosphatases, transcription factors and cytoskeletal proteins. Thus, the Ras-Raf-MEK-ERK signaling pathway transports cells from cell surface receptors to the nucleus and is required for example for cell proliferation and survival. The regulation of this signaling cascade consists of multiple isoforms of Ras (including K-Ras, N-Ras and H-Ras), and various isoforms of Raf (A-Raf, B-Raf, C-). Raf/Raf-1), MEK's multiple isoforms (MEK-1 and MEK-2) and ERK's multiple isoforms (ERK-1 and ERK-2) were further enhanced. Because 10-20% of human cancers have oncogenic Ras mutations, and many human cancers have activated growth factor receptors, this pathway is an ideal target for intervention.

The fundamental role and location of Raf in many signaling pathways has been demonstrated by studies using dysregulated and dominant inhibitory Raf mutants in mammalian cells and by studies using biochemical and genetic techniques in model organisms. In the past, the focus of anti-tumor drug target Raf has focused on its function as a downstream effector of Ras. However, recent findings suggest that Raf may have a prominent role in certain tumor formation without the need for a carcinogenic Ras dual gene. In particular, the activation duality of B-Raf and N-Ras has been identified in approximately 70% melanoma, 40% papillary thyroid cancer, 30% ovarian low cancer, and 10% colorectal cancer. K-Ras mutations are present in approximately 90% of pancreatic cancers. Most B-Raf mutations were found to be within the kinase domain, with a single substitution (V600E) accounting for at least 80%. The mutated B-Raf protein activates the Raf-MEK-ERK pathway via higher kinase activity on MEK or via activation of C-Raf.

Therefore, the development of B-Raf kinase inhibitors is the treatment of many types of human cancers, especially metastatic melanoma, solid tumors, brain tumors (such as glioblastoma multiforme (GBM)), acute myeloid leukemia (AML). , lung cancer, papillary thyroid cancer, low ovarian cancer and colorectal cancer provide new treatment opportunities. Several Raf kinase inhibitors have been described to exhibit the efficacy of inhibiting tumor cell proliferation in vitro and/or in vivo assays (see, for example, U.S. Patent Nos. 6,391,636, 6,358,932, 6,037,136, 5,717,100, 6,458,813, Nos. 6,204,467 and 6,268,391). Other patents and patent applications teach the use of Raf kinase inhibitors to treat leukemias (see, for example, U.S. Patent Nos. 6,268,391, 6,204,467, 6,756,410, and 6, 281, 193; and U.S. Patent Application Nos. And, for example, U.S. Patent Nos. 6,358,932, 5,717,100, 6,458,813, 6,268,391, 6,204,467 and 6,911,446. The data demonstrate that Raf kinase inhibitors significantly inhibit signaling via the MAPK pathway, resulting in a significant reduction in B-Raf (V600E) tumors.

The compounds of the invention inhibit cellular processes involving B-Raf kinase by blocking the signaling cascade in such cancer cells and ultimately inducing cell arrest and/or death.

According to the foregoing, the present invention further provides a method for preventing or treating diseases of lung cancer, prostate cancer, gastric cancer, pancreatic cancer, bladder cancer, colon cancer, bone marrow disease, prostate cancer, thyroid cancer, melanoma, adenoma, and Carcinoma of the ovary, eyes, liver, biliary tract and nervous system. Further, the present invention further provides a method of preventing or treating any of the above diseases or conditions in an individual in need of such treatment, the method comprising administering to the individual a therapeutically effective amount (see "Administration and Pharmaceutical Composition" below) a compound of formula I or Its pharmaceutically acceptable salt. For any of the above uses, the dosage required will vary depending on the mode of administration, the particular condition being treated, and the desired effect.

Administration and pharmaceutical composition

In general, the compounds of the invention will be administered in a therapeutically effective amount, either alone or in combination with one or more therapeutic agents, by any of the usual and accepted modes known in the art. The therapeutically effective amount varies widely depending on the severity of the disease, the age and relative health of the individual, the potency of the compound employed, and other factors. In general, satisfactory results are obtained by systemic administration at a daily dose of from about 0.03 mg to 30 mg per kg of body weight. In larger mammals (e.g., humans), the specified daily dose is in the range of from about 0.5 mg to about 2000 mg, preferably administered, for example, in divided doses of up to four times a day or in a delayed form. Unit dosage forms suitable for oral administration comprise from about 1 mg to 500 mg of the active ingredient.

The compounds of the invention may be administered in the form of a pharmaceutical composition via any conventional route, especially by enteral administration, for example, orally, for example, in the form of a lozenge or capsule; or parenterally, for example, injectable solutions or suspensions Form; topical administration, for example, in the form of a lotion, gel, ointment or cream or in the form of a nasal or suppository. Pharmaceutical compositions comprising a compound of the invention in free form or in a pharmaceutically acceptable salt form in association with at least one pharmaceutically acceptable carrier or diluent may be prepared by mixing, granulating or coating methods in a conventional manner Manufacturing. For example, the oral composition can be a troche or gelatin capsule containing the active ingredient and a) a diluent such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine ; b) a lubricant, such as cerium oxide, talc, stearic acid, its magnesium or calcium salt and / or polyethylene glycol; for tablets, also contains c) binders, such as magnesium aluminum silicate, starch paste , gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; if necessary, d) a disintegrant such as starch, agar, alginic acid or its sodium salt, or a foaming mixture; and/or e) an adsorbent, a colorant, a flavoring agent, and a sweetener. The injectable compositions can be aqueous isotonic solutions or suspensions, and the suppository can be prepared from a fat emulsion or suspension. The compositions may be sterilized and/or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, salts for regulating osmotic pressure, and/or buffers. In addition, it may also contain other therapeutically valuable substances. For example, the compounds of the invention may be formulated as microemulsion pre concentrate (MEPC). The compound of formula I can be made up to 40 mg/ml in a mixture of 56% PEG 400, 29% cetyl alcohol EL (cremophor EL) and 15% oleic acid. A mixture containing no compound of formula I is first prepared by vortexing/shocking. The compound of the invention is added and the powder is dispersed into the vehicle using sonication. The mixture was heated to 80 ° C with stirring in a water bath for about 1 hour, and sonicated every 15 minutes. This mixture was physically and chemically stable for about 1 week at room temperature.

Formulations suitable for transdermal administration comprise an effective amount of a compound of the invention and a carrier. The carrier can include a pharmacologically acceptable absorbable solvent to aid passage through the skin of the subject. By way of example, the transdermal device is in the form of a bandage comprising a substrate element, a reservoir containing the compound and, optionally, a carrier, optionally delivered to the subject over a longer period of time at a controlled and predetermined rate. The rate of the skin controls the barrier and the means for fastening the device to the skin. A matrix transdermal formulation can also be used. Formulations suitable for topical administration (e.g., application to the skin and eyes) are preferably aqueous solutions, ointments, creams or gels well known in the art. These formulations may contain solubilizers, stabilizers, tonicity enhancing agents, buffers, and preservatives.

The compounds of the invention may be administered in a therapeutically effective amount in combination (pharmaceutical combination) with one or more therapeutic agents. For example, it may have a synergistic effect with other anti-tumor agents or anti-proliferative agents such as mitotic inhibitors, alkylating agents, antimetabolites, embedded antibiotics, growth factor inhibitors, Cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones, antiandrogens, anti-angiogenic agents, kinase inhibitors, pan kinase inhibitors or Growth factor inhibitor.

The compounds of the invention may be administered in a therapeutically effective amount in combination with one or more suitable excipients selected from the group consisting of corn starch, potato starch, tapioca starch, starch paste, pregelatinized starch, sugar, gelatin, natural gum, synthetic Gum, sodium alginate, alginic acid, tragacanth, guar gum, cellulose, ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl Methylcellulose, microcrystalline cellulose, magnesium aluminum silicate, polyvinylpyrrolidone, talc, calcium carbonate, powdered cellulose, dextrate, kaolin, mannitol, citric acid, sorbose Alcohol, agar, sodium carbonate, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, clay, sodium stearate, calcium stearate , magnesium stearate, stearic acid, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, sodium lauryl sulfate, hydrogenated vegetable oil, peanut oil, cottonseed oil, Sunflower oil, sesame oil, olives Oil, corn oil, soybean oil, zinc stearate, sodium oleate, ethyl oleate, ethyl laurate, cerium oxide, and combinations thereof.

An embodiment of the invention is the method of claim 12 or 13, which additionally comprises administering to the individual another therapeutic agent. The additional therapeutic agent comprises an anti-cancer compound, an analgesic, an antiemetic, an antidepressant or an anti-inflammatory agent. In addition, the additional therapeutic agent is a different Raf kinase inhibitor, or an inhibitor of MEK, mTOR, HSP90, AKT, PI3K, CDK9, PAK, protein kinase C, MAP kinase, MAPK kinase or ERK, and a compound of the invention Invest in individuals in parallel.

For example, the addition of a combination of a MEK inhibitor and a Raf inhibitor significantly inhibits ERK signaling and thus reduces cell proliferation and transformation. Because treatment with MEK inhibitor alone has produced dose-limiting toxicity in the clinic, the combination of a Raf inhibitor and a MEK inhibitor may represent an excellent therapeutic strategy. Examples of MEK inhibitors are AS703026 (EMD Serono), MSC1936369B (EMD Serono), GSK1120212 (GlaxoSmithKline), AZD6244 (Memorial Sloan-Kettering Cancer Center), PD-0325901 (Pfizer), ARRY-438162 (Array BioPharma), RDEA119 ( Ardea Biosciences, Inc.), GDC0941 (Genentech), GDC0973 (Genentech), TAK-733 (Millennium Pharmaceuticals, Inc.), RO5126766 (Hoffmann-La Roche), and XL-518 (Exelixis).

In another embodiment of the present invention, there is provided a combination and method for treating cancer comprising a therapeutically effective amount of a compound of the [invention] (Raf inhibitor) and at least one MEK protein kinase inhibitor.

When the compound of the present invention is administered in combination with other therapies, the dose of the compound to be administered will of course vary depending on the type of co-drug used, the particular drug used, the condition to be treated, and the like.

The invention also provides a pharmaceutical combination (e.g., kit) comprising: a) a first agent, which is a compound of the invention as disclosed herein, in free form or in a pharmaceutically acceptable salt form, and b) at least A co-agent. The kit can contain instructions for its administration.

The terms "co-administered" or "combined administration" or the like as used herein are intended to encompass the administration of a selected therapeutic agent to a single patient, and are intended to include treatment regimens that are not necessarily administered or administered simultaneously via the same administration route. .

The term "pharmaceutical combination" as used herein means a product obtained by mixing or combining more than one active ingredient, and includes both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that the active ingredient (eg, a compound of formula I) and the co-agent are administered to the patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredient (eg, a compound of formula I) and the co-agent are administered to the patient simultaneously, in parallel, or sequentially (without a specific time limit) in the form of separate entities, wherein the administration provides therapeutically effective levels in the patient. Two compounds. The latter is also suitable for cocktai1therapy, for example by administering three or more active ingredients.

Method of preparing a compound of the invention

The invention also includes methods of preparing the compounds of the invention. In the reaction, it may be necessary to protect reactive functional groups (e.g., hydroxyl, amine, imido, thio or carboxy groups, which are required in the final product) to avoid undesired participation in the reaction. Conventional protecting groups can be used according to standard specifications, for example, see T. W. Greene and P. G. M. Wuts, "Protective Groups in Organic Chemistry", John Wiley and Sons, 1991.

The compound of formula I can be prepared by carrying out the following reaction scheme I:

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₇ and Y are as defined in the Summary of the Invention, and P is a suitable protecting group (for example, MEM, MOM, SEM, R ₄ SO ₂ and the like) And M is a leaving group (e.g., chlorine, bromine, iodine, methanesulfonyloxy, p-toluenesulfonyloxy, and the like). The protecting group P can be removed by a compound of formula 2 (for example, when P is MEM, MOM or SEM, treated with a strong acid such as hydrogen chloride in the presence of a protic solvent such as methanol or water; or when P is second In the case of sulfonyl R ₄ SO ₂ , the compound of formula I is synthesized by treatment with an aqueous solution of sodium carbonate or potassium carbonate or a solution of methanol in the presence of a cosolvent such as toluene.

It can be prepared by reacting a compound of formula 3 with an alkylating agent of formula 4 in the presence of a suitable solvent such as DMF, DMSO, and the like, and a suitable base such as potassium carbonate, sodium hydride, and the like. Compound of formula 2. The reaction is carried out at a temperature in the range of from about 0 ° C to about 150 ° C and can be completed in as long as about 24 hours. The reaction mixture is further reacted as appropriate to remove any protecting groups.

Compounds of formula I can also be prepared by carrying out the following reaction scheme II:

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and Y are as defined in [Summary of the Invention]. By suitable bases (such as pyridine, triethylamine, 4-(N,N-dimethylamino)pyridine and the like) and suitable solvents (such as pyridine, dichloromethane, 2-methyl) The compound of formula I is synthesized by reacting a compound of formula 5 with a sulfonating reagent of formula 6 in the presence of tetrahydrofuran and its analogs. The reaction is carried out at a temperature in the range of from about 0 ° C to about 100 ° C and can be completed in as long as about 24 hours. The reaction mixture is further reacted as appropriate to remove any protecting groups. In some cases, the sulfonation reagent can be reacted twice to produce a bis-sulfonyl derivative. In this case, a suitable base such as sodium hydroxide or potassium hydroxide can be used in the presence of a co-solvent such as toluene or 2-methyltetrahydrofuran in the presence of a protic solvent such as methanol or water. , or sodium carbonate or potassium carbonate) to convert the bissulfonyl compound to a compound of formula a. The reaction is carried out at a temperature in the range of from about 20 ° C to about 100 ° C and can be completed in as long as about 24 hours.

Compounds of formula I can also be prepared by carrying out the following reaction scheme III:

Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and Y are as defined in the Summary of the Invention, and R ₅₀ is a leaving group (e.g., iodine, bromine, chlorine, trifluoromethanesulfonyloxy and the like) a group), each R' may be, for example, hydrogen, a methyl group, and the like, or two R' groups may be joined together to form a ring Acid ester. The two R ₉₀ groups may each be hydrogen, or the two R ₉₀ groups together may represent a suitable nitrogen protecting group (eg, one R ₉₀ may be hydrogen and the other R ₉₀ may be BOC). By suitable transition metal catalysts (such as ruthenium (triphenylphosphine) palladium (0) or PdCl ₂ (dppf)), suitable solvents (such as DME, dioxane, toluene, ethanol and the like) and The compound of formula Ia is synthesized by reacting a compound of formula 7 with a compound of formula 8 in the presence of a suitable base such as anhydrous potassium carbonate or aqueous sodium carbonate and the like. The reaction is carried out at a temperature in the range of from about 20 ° C to about 120 ° C and can be completed in as long as about 24 hours. The reaction mixture is further reacted as appropriate to remove any protecting groups.

The compound of formula 1 can also be prepared by a Suzuki reaction-like scheme in which a compound of formula 7 is reacted with a compound of formula 8a to provide a compound of formula 9. Removal of the protecting group R ₉₀ of the group (sulfonylureas reaction, as described for the reaction Scheme II), the compound of formula Ia.

Those skilled in the art will appreciate that other organometallic coupling reactions, such as the use of tin reagents (Stille coupling) or zinc reagents (Negishi coupling), may be used instead of those described in Reaction Scheme III. Suzuki coupling reaction of boron reagent.

The compound of formula 7 can also be prepared by carrying out the following reaction scheme IV:

In this case, M is a leaving group (e.g., chlorine, bromine, iodine, methanesulfonyl and the like), and R ₅₀ is a leaving group (e.g., iodine, bromine, chlorine, trifluoromethanesulfonyloxy) And a similar group thereof, and R _{7 is} as defined in [Summary of the Invention]. The compound of formula 7 can be prepared by reacting an amine compound of formula 10 with a compound of formula 11. The reaction is carried out in the presence of a suitable base such as triethylamine, potassium carbonate and the like in a solvent such as isopropanol, DMSO, NMP or dioxane at a temperature of from about 25 ° C to about 120 ° C. Go on. In some cases, the newly introduced R ₁ group can then be further converted to obtain the final desired R ₁ group.

A compound of formula 11a as a subgroup of compounds of formula 11 wherein M is methanesulfonyl can be prepared by carrying out the following reaction scheme V:

Wherein R ₇ and R _{50 are} as defined in [Summary of the Invention]. The compound of formula 12 can be reacted with a suitable oxidizing system (e.g., a solution of m-chloroperbenzoic acid in dichloromethane solvent, or a solution of OxoneTM in aqueous methanol ⁾ at a temperature of from about -78 ° C to about 50 ° C and Its analog) is reacted to prepare a compound of formula 11a. The reaction took up to about 24 hours to complete.

Further, by combining the amine compound of the formula 13 with a suitable diazonium reagent system (for example, nitrous acid combined with copper (I) halide; isoamyl nitrite in combination with copper (I) / diiodomethane iodide; nitrous acid The compound of formula 12 (wherein R ₅₀ is chlorine, bromine or iodine) is prepared by reacting isoamyl ester in combination with boron trifluoride, iodine and potassium iodide in acetonitrile; and analogs thereof. The reaction is carried out at a temperature of from about 0 ° C to about 80 ° C and takes about 1 hour to about 6 hours to complete.

The compound of formula 13 can be prepared by reacting a compound of formula 14 with a compound of formula 3, as described for Reaction Scheme I.

The compound of formula 14 can be prepared by cyclizing an enamine nitrile compound of formula 15 with a hydrazine or hydrazine salt in a suitable solvent such as ethanol and the like. The reaction is carried out at a temperature of from about 25 ° C to about 100 ° C and can take from about 1 hour to about 24 hours to complete.

Alternatively, by making the formula 15 compound with a substituted hydrazine mono-R ₇ NH-NH _2-step reaction procedure to the preparation of compounds of formula 13 formula 15 compounds. The reaction is carried out at a temperature of from about 25 ° C to about 100 ° C and can take from about 1 hour to about 24 hours to complete.

Further, Formula 15 can be prepared by reacting a compound of Formula 16 with DMF DMA or Bredereck's reagent in the presence of a cosolvent such as DMF, at a temperature of from about 50 ° C to about 150 ° C. Compound. The reaction takes about 1 hour to about 24 hours to complete.

The compound of formula 16 can be prepared by treating a compound of formula 17 with a suitable acid such as p-toluenesulfonic acid and the like in a suitable inert solvent such as toluene and the like. The reaction is carried out at a temperature of from about 50 ° C to about 120 ° C and takes about 1 hour to about 24 hours to complete.

Finally, 4-chloro- can be obtained by the presence of a suitable base (such as sodium hydride and the like) and a suitable solvent (such as DMSO and the like) at a temperature of from about 25 ° C to about 80 ° C. The compound of formula 17 is prepared by reacting 2-(methylthio)pyrimidine with t-butyl cyanoacetate. The reaction takes about 1 hour to about 24 hours to complete.

A compound of formula 11b as a subgroup of compounds of formula 11 wherein M is a halogen can be prepared by carrying out the following reaction scheme VI:

Wherein R ₇ and R _{50 are} as defined in [Summary of the Invention]. The compound of formula 11b (wherein M is a halogen) can be prepared by reacting a compound of formula 20 with a suitable diazonium reagent system such as nitrous acid in combination with a copper (I) halide; sodium nitrite in combination with p-toluenesulfonic acid and potassium iodide. . The reaction is carried out at a temperature of from about 0 ° C to about 80 ° C and takes about 1 hour to about 6 hours to complete. Alternatively, the compound of formula 20 is treated with a diazotization reagent (e.g., sodium nitrite and the like) in the presence of a carboxylic acid (e.g., trifluoroacetic acid and the like) at a temperature of from 0 ° C to 40 ° C for about 0.5 hours to about Treatment with an aqueous alkaline solution such as aqueous potassium carbonate and the like affords the compound of formula 21 (such compounds may exist in tautomeric forms). Subsequent to the presence of a base such as N,N-dimethylaniline or DIPEA, and optionally in an inert solvent such as acetonitrile or toluene and an additive such as DMF, at a temperature of from about 50 ° C to about 110 ° C. The compound of formula 21 is treated with a chlorinating agent such as phosphorus oxychloride and the like for about 1 hour to about 72 hours to provide a compound of formula 11b (wherein M is chlorine).

Further, it may be in a suitable solvent (for example, second butanol, NMP and the like) in the presence of a base such as lithium hydroxide and the like at a temperature of from about 50 ° C to about 180 ° C. The compound of formula 20 is prepared by reacting a compound of formula 22 with a hydrazine or hydrazine salt (e.g., guanidine hydrochloride or guanidinium carbonate) for from about 2 hours to about 48 hours.

The compound of formula 22 can be prepared by reacting a compound of formula 23 with DMF DMA or a Bradney's reagent at a temperature of from about 50 ° C to about 150 ° C, optionally in the presence of a cosolvent such as DMF. The reaction takes about 1 hour to about 24 hours to complete.

By combining the amine compound of formula 24 with a suitable diazonium reagent system (eg, nitrous acid in combination with a copper (I) halide; sodium nitrite in combination with p-toluenesulfonic acid and potassium iodide; or isoamyl nitrite in combination with trifluorocarbon The compound of formula 23 (wherein R ₅₀ is chlorine, bromine or iodine) is prepared by reacting boron-THF, iodine, potassium iodide and acetonitrile. The reaction is carried out at a temperature of from about 0 ° C to about 80 ° C and takes about 1 hour to about 6 hours to complete.

Further, the compound of the formula 25 can be misaligned with, for example, methyllithium, methyllithium-lithium bromide by an inert solvent such as THF, diethyl ether or cyclopropylmethyl ether at a temperature of from about 0 ° C to about 100 ° C. The organometallic reagent of the compound or the halogenated methylmagnesium reagent is reacted; then treated with a quenching aqueous solution to prepare a compound of formula 24. The reaction takes about 2 hours to about 48 hours to complete.

Compounds of formula 25 can be prepared by reacting a compound of formula 26 with a compound of formula 3, as described for Reaction Scheme I. Alternatively, it can be prepared from a compound of formula 27 wherein two R' groups together form an acid labile protecting group (eg, an imine such as benzylidene; or a carbamate such as a third butyl carbamate) A compound of formula 25. The reaction is carried out by treating a compound of formula 27 with an acidic aqueous reagent (e.g., concentrated hydrochloric acid and the like) in a solvent such as ethanol at a temperature of from about 25 ° C to about 100 ° C. "The groups preferably together form an imine such as benzylidene.

Further, an acid labile protecting group (e.g., an imine such as a benzylidene group; or a urethane such as an aminocarboxylic acid) can be formed by reacting a compound of the formula 28 with a compound of the formula 29 in which two R' groups are formed together. The third butyl ester)) is reacted to prepare a compound of formula 27. The reaction is carried out at a temperature of from about 25 ° C to about 120 ° C, optionally in the presence of a catalyst such as DMAP, in a solvent such as toluene, methanol or ethanol. And taking about 1 hour to about 24 hours to complete. The reaction is optionally in the presence of a base (such as n-butyllithium and the like) in an inert solvent at a temperature of from about -80 ° C to about 25 ° C. (for example, THF and the like) is carried out for about 0.5 hours to about 12 hours.

The compound of formula 29 can be prepared by methods known to those skilled in the art. For example, the benzaldehyde can be prepared by reacting benzaldehyde with a monosubstituted ruthenium R ₇ NHNH ₂ in a solvent such as ethanol or toluene at a temperature of from about 25 ° C to about 120 ° C for from about 1 hour to about 24 hours. a compound of formula 29 wherein two R' groups together form a benzylidene group. Alternatively, the reaction may use a monosubstituted phosphonium salt (eg, a hydrochloride or oxalate salt and the like) in combination with a base (eg, acetic acid). Sodium or triethylamine).

The compound of formula 8 or the compound of formula 8a can be prepared as described in Reaction Scheme VII below:

Wherein R ₂ , R ₃ , R ₄ , R ₅ and Y are as defined in the Summary of the Invention, and M is a leaving group (for example, iodine, bromine, chlorine, trifluoromethanesulfonyloxy and the like), and Each R' may be, for example, hydrogen, methyl, and the like, or two R' groups may be joined together to form a ring Acid ester. The two R ₉₀ groups may each be hydrogen, or the two R ₉₀ groups together may represent a suitable nitrogen protecting group (eg, one R ₉₀ may be hydrogen and the other R ₉₀ may be BOC). It can be in a suitable solvent (for example, toluene, dioxane and the like) in the presence of a suitable transition metal catalyst (for example PdCl ₂ (dppf)) and a suitable base such as potassium acetate and the like. Compounds of formula 8 or compounds of formula 8a are prepared by reacting a compound of formula 50 or a compound of formula 50a with a diboron compound, such as bis(pinacolyl)diboron and the like, respectively. The reaction is carried out at a temperature in the range of from about 20 ° C to about 120 ° C and can be completed in as long as about 24 hours. Again, the compound of formula 50 can be prepared by sulfonating a compound of formula 51 as described for Reaction Scheme II. Those skilled in the art will appreciate that a compound of formula 51 or a compound of formula 50a (e.g., 3-bromoaniline or N-BOC protected 3-bromoaniline) can be prepared by a variety of methods including, but not limited to, as further described in the Examples below. method.

The compound of formula 70 can be prepared as described in Reaction Scheme VIII below:

Wherein R _8b as [Summary] defined above and R ₅₅ is selected from the C _1-4 alkyl substituted by halo or C _1-4 alkyl. The compound of formula 71 can be treated by a suitable reduction system (e.g., hydrogenation over a palladium catalyst and the like) in a suitable solvent (e.g., ethanol or ethyl acetate) at a temperature of from about 25 ° C to about 75 ° C. The compound of formula 70 is prepared from 0.5 hours to about 12 hours.

Alternatively, the compound of formula 71 can be prepared by a two-step process consisting of converting the hydroxyl group of the compound of formula 73 to a suitable leaving group, followed by replacement with an azide anion. For the first step, suitable reagents include a combination of phosphorus tribromide or methanesulfonium chloride with a suitable base such as triethylamine. These reactions are carried out in a suitable solvent such as dichloromethane and the like at a temperature of from about 0 ° C to about 50 ° C. For the second step, the displacement is carried out in a suitable solvent (e.g., DMF or DMSO) with an azide reagent (e.g., sodium azide and the like) at a temperature of from about 25 °C to about 150 °C. The reaction takes about 1 hour to about 24 hours to complete. Alternatively, the conversion can be carried out by using a phosphine reagent (such as triphenylphosphine and the like) and azodicarboxylic acid in the presence of azide acid (formed in situ from an azide salt such as sodium azide). Treatment of the compound of formula 73 with an ester reagent (e.g., diethyl azodicarboxylate and the like) is carried out in one step. The reaction is carried out at a temperature of from about -80 ° C to about 75 ° C and takes about 1 hour to about 24 hours to complete.

The compound of formula 73 can be prepared by treating a compound of formula 74 with a chloroformate such as methyl chloroformate and the like or an alternative alkoxycarbonylating reagent such as di-t-butyl dicarboxylate. The reaction is carried out in an inert solvent such as dichloromethane and the like at a temperature of from about -80 ° C to about 25 ° C and takes about 1 hour to about 12 hours to complete. A base such as triethylamine can be used as appropriate. The reaction can also be carried out in a two-phase system consisting of a solvent such as THF or dioxane and an aqueous alkaline solution such as aqueous sodium hydrogencarbonate at a temperature of about 25 ° C for about 2 hours to about 16 hours.

Compounds of formula 70 can also be prepared as described in Reaction Scheme IX below:

Wherein R _{8b is} as defined in [Section], R ₅₅ is selected from C _1-4 alkyl or halo substituted C _1-4 alkyl, and R ₅₆ is a suitable protecting group (eg, benzyloxycarbonyl) (CBz)). The compound of formula 70 can be prepared by removal of a protecting group from a compound of formula 75. For example, it can be suitably used in a suitable solvent (such as ethanol, methanol, MTBE or ethyl acetate) at a temperature of from about 25 ° C to about 75 ° C, optionally in the presence of an acid such as hydrogen chloride. The compound of formula 70 is prepared by treating a compound of formula 75 (wherein R ₅₆ is Cbz) for about 0.5 hours to about 12 hours in a reduction system (e.g., by hydrogenation of a palladium catalyst and the like). Alternatively, the deprotection group can be carried out using a suitable hydrogen donor such as formic acid, ammonium formate or 1,4-cyclohexadiene under transfer hydrogenation conditions. Alternatively, a compound of formula 75 can be prepared by reacting a compound of formula 76 with a chloroformate such as methyl chloroformate and the like or an alternative alkoxycarbonylating reagent such as di-t-butyl dicarboxylate. . The reaction is carried out in an inert solvent such as dichloromethane and the like at a temperature of from about -80 ° C to about 25 ° C and takes about 1 hour to about 24 hours to complete. A base such as triethylamine can be used as appropriate. The reaction can also be carried out in a two-phase system consisting of a solvent such as THF or dioxane and an aqueous alkaline solution such as aqueous sodium hydrogencarbonate at a temperature of about 25 ° C for about 2 hours to about 16 hours. The compound of formula 76 can be prepared by removal of the protecting group of the compound of formula 77. The protective conditions can be selected to give, preferentially or substantially, a single isomer of formula 76. Alternatively, the following conditions can be selected: the single isomer of formula 77 is shown to be less preferred or not preferentially formed, although the compound of formula 76 can be isolated in sufficient purity for further use. The manner in which the purity is obtained can include crystallizing the free base, or salt. The compound of formula 76 (wherein R ₅₆ is Cbz) can be prepared by treating a compound of formula 77 with benzyl chloroformate. The reaction is carried out in an inert solvent such as dichloromethane and the like at a temperature of from about -80 ° C to about 25 ° C and takes about 1 hour to about 24 hours to complete. A base such as triethylamine can be used as appropriate. The reaction can also be carried out in a two-phase system consisting of a solvent such as THF or dioxane and an aqueous alkaline solution such as aqueous sodium hydrogencarbonate at a temperature of about 25 ° C for about 2 hours to about 24 hours. A salt of a compound of formula 77 can be used in combination with a base such as triethylamine or sodium carbonate as a feed to the free base.

It will be understood by those skilled in the art that the compound of formula 70 can be a single enantiomer or a mixture of enantiomers, and a single enantiomer of formula 70 can be obtained by a suitable single pair of formula 74 or formula 77. The anamorphic compound is obtained as the starting material.

Detailed examples of the synthesis of compounds of formula Ia can be found in the examples below.

Other methods of preparing the compounds of the invention

The compound can be prepared into a pharmaceutically acceptable acid addition salt by reacting the free base form of the compound of the invention with a pharmaceutically acceptable inorganic or organic acid. Alternatively, a pharmaceutically acceptable base addition salt of the compound can be prepared by reacting the free acid form of the compound of the invention with a pharmaceutically acceptable inorganic or organic base.

Alternatively, the starting materials or salts of the intermediates can be used to prepare the salt forms of the compounds of the invention.

The free acid or free base form of the compound of the invention may be prepared from the corresponding base addition salt or acid addition salt form, respectively. For example, a compound of the invention in an acid addition salt form can be converted to the corresponding free base by treatment with a suitable base such as aqueous ammonium hydroxide, sodium hydroxide and the like. The compound of the invention in the form of a base addition salt can be converted to the corresponding free acid by treatment with a suitable acid such as hydrochloric acid or the like.

A reducing agent (for example, sulfur, sulfur dioxide, triphenylphosphine, lithium borohydride, or the like) may be used in a suitable inert organic solvent (for example, acetonitrile, ethanol, aqueous dioxane or the like) at 0 to 80 ° C. The non-oxidized form of the compound of the invention is prepared from the N-oxide of the compound of the invention by treatment with sodium borohydride, phosphorus trichloride, tribromide or the like.

Prodrug derivatives of the compounds of the invention can be prepared by methods known to those of ordinary skill in the art (for example, see Saulnier et al. (1994), Bioorganic and Medicinal Chemistry Letters, Vol. 4, page 1985 for more details). For example, by derivatizing a compound of the invention with a suitable aminomethionating reagent (eg, 1,1-decyloxyalkyl chloroformate, p-nitrophenyl carbonate or the like) The reaction is carried out to prepare a suitable prodrug.

Protected derivatives of the compounds of the invention can be prepared in a manner known to those of ordinary skill in the art. A detailed description of techniques suitable for the generation and removal of protecting groups can be found in T. W. Greene, "Protecting Groups in Organic Chemistry", 3rd edition, John Wiley and Sons, Inc., 1999.

It is preferred in the process of the invention to prepare or form a compound of the invention in the form of a solvate (e.g., hydrate). The hydrate of the compound of the present invention is preferably prepared by recrystallization from an aqueous/organic solvent mixture using an organic solvent such as 1,4-dioxane, tetrahydrofuran or methanol.

The stereoisomers of the compounds of the invention can be reacted with optically active resolving agents to form a pair of diastereomeric compounds, the diastereomers are separated and the optically pure enantiomers are recovered to prepare individual stereoisomers. A compound of the invention in isomeric form. While enantiomeric resolution can be carried out using covalent diastereomeric derivatives of the compounds of the invention, the separable complexes are preferred (e.g., crystalline diastereomeric salts). Diastereomers have unique physical properties (e.g., melting point, boiling point, solubility, reactivity, etc.) and can be readily separated by utilizing such differences. The diastereomers can be separated on the basis of solubility differences by chromatography or preferably by separation/analysis techniques. The optically pure enantiomer and the resolving agent are then recovered by any practical means that does not produce racemization. A more detailed description of techniques suitable for the resolution of stereoisomers of compounds from their racemic mixtures can be found in Jean Jacques, Andre Collet, Samuel H. Wilen, "Enantiomers, Racemates and Resolutions", John Wiley And Sons, Inc. ., 1981.

In summary, the compounds of formula I can be prepared by methods involving the following:

(a) methods of Reaction Schemes I through IX; and

(b) converting a compound of the invention into a pharmaceutically acceptable salt, as appropriate;

(c) converting the salt form of the compound of the invention to a non-salt form, as appropriate;

(d) converting a non-oxidized form of a compound of the invention to a pharmaceutically acceptable N-oxide, as appropriate;

(e) converting the N-oxide form of the compound of the invention to its non-oxidized form, as appropriate;

(f) analysing individual isomers (eg, stereoisomers) of the compounds of the invention from a mixture of isomers, as appropriate;

(g) converting a non-derived compound of the invention into a pharmaceutically acceptable prodrug derivative, as appropriate;

(h) Converting a prodrug derivative of a compound of the invention to its non-derivatized form, as appropriate.

Such compounds are known in the absence of a particular description of the preparation of the starting materials, or may be prepared analogously to methods known in the art or as disclosed in the Examples below.

Those skilled in the art will appreciate that the above transformations are merely representative methods of preparing the compounds of the present invention, and that other well known methods can be similarly employed.

Instance

The present invention is further exemplified by the following intermediates and examples illustrating the preparation of the compounds of formula I of the present invention, but is not limited to such intermediates and examples.

The abbreviations used are as follows: benzyloxycarbonyl (Cbz); third butoxycarbonyl (BOC); cell proliferation (CP); dichloromethane (DCM); N,N-diisopropylethylamine (DIPEA); [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (PdCl ₂ (dppf)); 1,2-dimethoxyethane (DME); N,N- Dimethylacetamide (DMA); N,N-dimethylaminopyridine (DMAP); N,N-dimethylformamide (DMF); N,N-dimethylformamide Acetal acetal (DMF DMA); dimethyl hydrazine (DMSO); ethyl acetate (EtOAc); high pressure liquid chromatography (HPLC); isopropyl acetate (iPrOAc); methanesulfonyl (Ms); Methyltetrahydrofuran (2-methylTHF); N-methylpyrrolidone (NMP); tetrahydrofuran (THF); thin layer chromatography (TLC); and p-toluenesulfonic acid (pTsOH).

Intermediate: cyano-(2-methylthio-pyrimidin-4-yl)-acetic acid tert-butyl ester

To a suspension of sodium hydride (7.15 g, 179 mmol, 60% in oil) in DMSO (100 mL) was added <RTI ID=0.0>0> After the evolution of hydrogen ceased, 4-chloro-2-methylthiopyrimidine (13.7 g, 85 mmol) was added. The reaction was heated at 80 ° C for 16 hours. The reaction mixture was then cooled to room rt and quenched with EtOAc EtOAc. The solid was filtered and washed with water (2×200 mL). 300 mL of hexane was added to the solid, and the suspension was heated at 60 ° C for 1 hour, followed by cooling to room temperature. The solid was filtered and washed with hexane to give the title ^{compound; 1 H NMR 400 MHz (CDCl} 3) δ7.82 (d, 1H), 6.74 (d, 1H), 2.63 (s, 3H), 2.61 (s, 1H) , 1.52 (s, 9H); MS m / z: 266.0 (m + 1).

Intermediate: (2-methylthio-pyrimidin-4-yl)-acetonitrile

Add p-toluenesulfonic acid (800) to a solution of the intermediate cyano-(2-methylthio-pyrimidin-4-yl)-acetic acid tert-butyl ester (5.3 g, 20 mmol) in anhydrous toluene (100 mL) Mg). The mixture was heated to reflux for 8 h, cooled to rt and extracted with EtOAc. The organic layer was washed with aq. By silica gel chromatography (3: 1 hexanes / ethyl acetate as eluent) of the crude title ^{compound: 1 H NMR 400 MHz (CDCl} 3) δ8.56 (d, 1H), 7.12 (d, 1H ), 3.84 ( s , 2H), 2.57 ( s , 3H); MS m / z : 166.0 (M + 1).

Intermediate: 4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine

Step 1: 3-(Dimethylamino)-2-(2-(methylthio)pyrimidin-4-yl)acrylonitrile. Add N,N -dimethylformamide dimethyl acetal (30 mL) to the intermediate (2-methylthio-pyrimidin-4-yl)-acetonitrile (2.62 g, 15.7 mmol). The mixture was heated at ° C for 16 hours. The cooled reaction mixture was concentrated and the residue was used without further purification.

Step 2: 4-(2-(Methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine. Crude 3-(dimethylamino)-2-(2-(methylthio)pyrimidin-4-yl)acrylonitrile (total amount) and hydrazine monohydrate (2.36 mL) in the previous step at 80 °C A mixture of 47 mmol) in absolute ethanol (75 ml) was heated for 16 h. The reaction mixture was cooled to room temperature and concentrated. The reaction mixture was partitioned between ethyl acetate and brine. The organic layer was separated and washed with brine, dried over sodium sulfate The crude product was purified by silica gel chromatography (2% to 5% methanol in dichloromethane) to afford 4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazole-3- ^{amine: 1 H NMR 400 MHz (DMSO} -d 6) δ11.9 (s, 1H), 8.3 (s, 1H), 7.87 (s, 1H), 7.23 (s, br 1H), 6.43 (s, 1H) , 5.74 ( s , 1H), 2.53 ( s , 3H); MS m/z : 208.0 (M+1).

Intermediate: 1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine

The intermediate 4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine (10.0 g, 40 mmol) was dissolved in THF (200 mL). (6.3 mL, 63 mmol) and sodium methoxide (25% by weight solution in methanol, 14.3 ml, 63 mmol). The mixture was heated at 50 ° C under stirring for 3 days under a nitrogen atmosphere, followed by concentration under vacuum. The residue was dissolved in EtOAc (EtOAc)EtOAcEtOAcEtOAc The residue was subjected to silica gel chromatography (hexane/ethyl acetate as solvent) to give 1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyridine as a solid. Oxazol-3-amine; ¹ H NMR 400 MHz (CDCl ₃ ) δ 8.23 (d, J = 5.6 Hz, 1H), 7.60 (s, 1H), 6.83 (d, J = 5.6 Hz, 1H), 5.23 ( d, J = 2.8 Hz, 2H), 4.21-4.25 (m, 1H), 2.50 (s, 3H), 1.37 (d, J = 6.8 Hz, 6H); MS m/z : 250.1 (M+1).

Preparation of 1-ethyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine and 1-methyl-4-(2-(methylthio)pyrimidine analogously 4-yl)-1H-pyrazol-3-amine.

Intermediate: 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine

The intermediate 1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine (4.0 g, 16.0 mmol), nitrite at 100 ° C A mixture of amyl ester (13.2 g, 112 mmol) and diiodomethane (30 mL) was heated for 3 hours. The volatiles were removed in vacuo to give a dark residue by silica gel chromatography (2: 1 hexanes / ethyl acetate as eluent) to give the title compound as a solid; MS m / z: 361.1 ( M+1).

Similarly prepared 4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine; 4-(3-iodo-1-methyl-1H-pyrazole 4-yl)-2-(methylthio)pyrimidine; and 4-(3-iodo-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine.

Intermediate: 4-(3-iodo-1-(tetrahydro-2H-piperidin-2-yl)-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine

4-(3-Iodo-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine (270 mg, 0.85 mmol) and p-toluenesulfonic acid monohydrate (32 mg, 0.17) at 60 °C A solution of mmol in 3,4-dihydro-2H-pyran (1 ml) was heated for 5 h. The cooled mixture was diluted with EtOAc and EtOAc (EtOAc)EtOAc. The residue was subjected to EtOAc (EtOAc (EtOAc) MS ( m / z ): 402.7 (M + 1).

Intermediate: 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine

To the intermediate 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine (4.51 g, 12.5 mmol) in dichloromethane m-Chloroperbenzoic acid (3.65 g, 77% purity, 16.3 mmol) was added to the solution in (60 mL). The mixture was stirred at 0 ° C under N2 for 3 h then diluted with ethyl acetate and washed with brine and brine. The organic layer was dried <RTI ID=0.0> MS m / z : 393.0 (M + 1).

Similarly prepared 4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine; and 4-(3-iodo-1-methyl-1H -pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine.

Intermediate: 1-benzylidene-2-isopropyl hydrazine

Add isopropyl hydrazine hydrochloride (11.1 g, 0.1 mol) and benzaldehyde (10.6 g, 0.1 mol) to a round bottom flask containing anhydrous sodium acetate (8.2 g, 0.1 mol) in 125 ml of 50% ethanol. ). The mixture was stirred at room temperature for 20 hours. The reaction was extracted with diethyl ether (3 x 250 mL). The organic layers were combined, washed with aqueous sodium bi hydrogen sulfate and brine and dried over sodium sulfate. Filtration, concentrating and co-evaporation with toluene (3x) MS m/z 163.3 (M + 1).

Starting from ethyl oxalate as the starting material, 1-benzylidene-2-ethylguanidine was prepared by using methanol and triethylamine instead of ethanol and sodium acetate respectively; and methylhydrazine was used as the starting material, and methanol was used. As the solvent, 1-benzylidene-2-methylindole was similarly prepared without adding a base.

Intermediate: 2-((2-benzylidene-1-isopropylindenyl)methylene)-malononitrile

A solution of the intermediate 1-benzylidene-2-isopropylindole (12.9 g, 0.079 mol) in 200 mL anhydrous THF was cooled in a dry ice/acetone bath under argon. To this solution was added n-butyllithium (1.6 M in hexanes, 66 ml, 0.106 mol) via syringe. After the addition was completed, the mixture was further stirred at a dry ice temperature for 5 minutes. A solution of 2-(ethoxymethylene)malononitrile (13.6 g, 0.11 mol) in THF (30 mL). The mixture was stirred at dry ice temperature for 0.5 h then quenched with saturated aqueous sodium bicarbonate. The quenched reaction mixture was warmed to room rt and extracted with EtOAc. The organic layers were combined, dried over sodium sulfate, filtered and evaporated. The residue was suspended in ethanol by sonication. The resulting precipitate was collected by EtOAc (EtOAc) elute The mother liquor was concentrated and the residue was purified by silica gel column chromatography (1:1 hexane/ethyl acetate as solvent) to give a further portion of 2-((2-benzylidene-1- isopropyl) Methylene)malononitrile. MS m/z 239.2 (M+1).

Intermediate: 2-((2-benzylidene-1-ethylindenyl)methylene)-malononitrile

To the solution of 2-(ethoxymethylene)malononitrile (15.2 g, 0.124 mol) in 100 ml of toluene was added the intermediate 1-benzylidene-2-ethylhydrazine (18.4 g, 0.124 mol) ). The mixture was allowed to stand at room temperature and a precipitate formed. The reaction mixture was stirred for a further 16 hours. The precipitate was collected on a filter and washed with a small amount of cold ethanol to give 2-((2-benzylidene-1-ethylindolyl)methylene)malononitrile as a solid.

2-((2-Benzylidene-1-methylindenyl)-methylene)malononitrile was prepared similarly.

Intermediate: 3-amino-1-isopropyl-1H-pyrazole-4-carbonitrile

The intermediate 2-((2-benzylidene-1-isopropylindenyl)-methylene)malononitrile (9.42 g, 40 mmol), concentrated hydrochloric acid (5 ml) and ethanol The mixture (50 ml) was heated for 20 minutes. The reaction mixture was concentrated and diethyl ether (50 ml) was evaporated. The mixture was sonicated and the upper ether layer was discarded. 20 ml of 5 N aqueous sodium hydroxide solution was added to the residue, and the mixture was extracted with dichloromethane (3×). The organic layers were combined, dried over sodium sulfate, filtered and evaporated. The residue was purified by EtOAc EtOAc EtOAc elut elut elut elut elut Nitrile. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.09 (s, 1H), 5.51 (s, 2H), 4.22 (m, 1H), 1.31 (d, J = 7 Hz, 6H); MS m/ z 151.2 (M+1).

3-Amino-1-ethyl-1H-pyrazole-4-carbonitrile; and 3-amino-1-methyl-1H-pyrazole-4-carbonitrile were prepared analogously.

Intermediate: 1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)ethanone

Add methylmagnesium bromide to a solution of the intermediate 3-amino-1-isopropyl-1H-pyrazole-4-carbonitrile (5.29 g, 36.5 mmol) in 200 mL anhydrous THF at 0 °C Solution (3 M in diethyl ether, 56.5 ml, 0.17 mol). The reaction mixture was heated under reflux for 4 hours. The mixture was cooled to 0 ° C and then quenched with 10% aqueous hydrochloric acid to a neutral pH. The reaction was extracted with a large amount of 9:1 dichloromethane/isopropanol. The organic layers were combined and dried over sodium sulfate, filtered and evaporated. The crude product was purified by EtOAc EtOAc elut elut elut elut elut ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.18 (s, 1H), 5.62 (s, 2H), 4.23 (m, 1H), 2.20 (s, 3H), 1.37 (d, J = 7 Hz , 6H); MS m/z 168.2 (M + 1).

Similarly prepared 1-(3-amino-1-ethyl-1H-pyrazol-4-yl)ethanone; and 1-(3-amino-1-methyl-1H-pyrazol-4-yl Ethyl ketone.

Intermediate: 1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)ethanone

To the intermediate 1-(3-amino-1-isopropyl-1H-pyrazol-4-yl)ethanone (3.97 g, 24 mmol) and p-TsOH‧H ₂ O (9.07) g, 48 mmol, 2 eq.) A solution of sodium nitrite (2.97 g, 43 mmol, 1.8 eq.) and potassium iodide (8.0 g, 48 mmol, 2.0 eq.) in 20 ml of water was added dropwise in 150 ml of acetonitrile. . The mixture was stirred at this temperature for 10 minutes, then allowed to warm to room temperature and stirred for 3 hours. The mixture was concentrated, then diluted with water and neutralized to pH 9-10 with aqueous sodium carbonate. The mixture was extracted with ethyl acetate (3x). The combined organic layers were washed with aq. The residue was purified by EtOAc EtOAc EtOAc elut elut ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.51 (s, 1H), 4.53 (m, 1H), 2.37 (s, 3H), 1.41 (d, J = 7 Hz, 6H); MS m/z 279.1 (M+1).

Similarly prepared 1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)ethanone; and 1-(3-iodo-1-methyl-1H-pyrazol-4-yl)B ketone.

Intermediate: 3-(Dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)prop-2-en-1-one

The intermediate. 1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)ethanone (5.0 g, 18.0 mmol) and N,N-dimethylformamidine at 155 °C. A mixture of amine dimethyl acetal (50 mL) was heated for 20 hours. The mixture was concentrated under vacuum to give the crude title compound. MS m / z 334.0 (M + 1).

Similarly prepared 3-(dimethylamino)-1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)prop-2-en-1-one; and 3-(dimethyl Amino)-1-(3-iodo-1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one.

Intermediate: 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine

The crude intermediate 3-(dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl) was stirred at 110 ° C in a pyridine sealed reaction vessel. a mixture of prop-2-en-1-one (4.0 g, 12.0 mmol), guanidine hydrochloride (2.63 g, 27.6 mmol), lithium hydroxide (635 mg, 27.6 mmol) and second butanol (50 ml) 20 hours. The cooled reaction mixture was concentrated, then water was added and the mixture was extracted with ethyl acetate. The combined extracts were dried over sodium sulfate, filtered and concentrated. Ethyl acetate was added to the solid residue, and the mixture was subjected to sonication. The solid product was collected on EtOAc (EtOAc m. MS m/z 330.0 (M + 1).

Similarly prepared 4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-amine; and 4-(3-iodo-1-methyl-1H-pyrazole-4- Pyrimidine-2-amine.

Intermediate: 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol

To the intermediate 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine (500 mg, 1.52 mmol) and trifluoroacetic acid (15 ml) Sodium nitrite (314 mg, 4.55 mmol) was added portionwise to the stirred mixture. The mixture was allowed to warm to room temperature and stirred for 1 hour then the solvent was removed in vacuo. The crude mixture was diluted with EtOAc. MS m/z 331.0 (M + 1).

Similarly prepared 4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-ol; and 4-(3-iodo-1-methyl-1H-pyrazole-4- Pyrimidine-2-ol.

Intermediate: 2-chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine

The intermediate 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol (438 mg, 1.33 mmol) in phosphorus oxychloride (10 ml) at 110 °C The solution was heated for 16 hours. The mixture was concentrated under vacuum, then aqueous sodium bicarbonate was carefully added and mixture was extracted with ethyl acetate. The combined organic extracts were dried with sodium s MS m/z 349.0 (M + 1).

Similarly prepared 2-chloro-4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidine; and 2-chloro-4-(3-iodo-1-methyl-1H-pyridyl Zol-4-yl)pyrimidine.

Intermediate: (S)-1-hydroxypropan-2-ylaminocarbamate

To a solution of (S)-propanol (10 g, 130 mmol) and sodium bicarbonate (32.8 g, 390 mmol) in THF-H ₂ O (1:1, 650 mL) at 0 ° C Methyl chloroformate (11.4 mL, 143 mmol) was added. The mixture was stirred and allowed to warm to room temperature over 4 hr then extracted with ethyl acetate. The organic layer was washed with EtOAc EtOAc m. MS m/z 134.1 (M + 1).

Preparation of methyl ( R )-1-hydroxyprop-2-ylcarbamate analogously using ( R )-anilinol instead of ( S )-alanamine; and using di-tert-butyl dicarbonate instead of methyl chloroformate (S) 1,1-Dimethylethyl 1-hydroxyprop-2-ylcarbamate was prepared.

Intermediate: (S)-1-Azidopropan-2-ylaminocarbamate

A solution of the intermediate (S)-methyl 1-hydroxypropan-2-ylcarbamate (2.65 g, 20 mmol) and triethylamine (7.0 ml, 50 mmol) in anhydrous dichloromethane (100 mL) Methane sulfonium chloride (1.91 mL, 23.9 mmol) was added. The mixture was stirred at room temperature for 3 hours and then extracted with ethyl acetate. The organic layer was washed with aq. The crude mesylate product was then dissolved in dry DMF (70 mL) and sodium azide (5.2 g, 80 mmol) was then applied. The mixture was heated at 80 ° C for 2 hours with stirring. The cooled reaction mixture was concentrated and the residue was partitioned betweenEtOAc and brine. The organic layer was separated and washed with brine, dried over sodium sulfate The residue was purified by EtOAc (EtOAc:EtOAc:EtOAc) MS m/z 159.1 (M + 1).

Preparation of ( R )-1-azidopropan-2-ylcarbamic acid methyl ester; and (S)-1-azidopropan-2-ylaminocarbamate 1,1-dimethylethyl ester .

Intermediate: (S)-1-aminopropan-2-ylaminocarbamate

Add 10% palladium on carbon (wet, to a solution of the intermediate (S)-1-Azidopropan-2-ylaminocarbamate (2.86 g, 18.2 mmol) in ethyl acetate (200 ml) 286 mg). The flask was degassed, refilled with hydrogen (balloon, 1 atm), and the mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through a pad of celite and washed with ethyl acetate. The filtrate was concentrated to give crude (S)-l-aminopropan-2-ylcarbamate, which was used without further purification. ¹ H NMR 400 MHz (CDCl ₃ ) δ 4.79 ( s , 1H), 3.71-3.65 (m, 1H), 3.66 ( s , 3H), 2.75 (dd, 1H), 2.65 (dd, 1H), 1.14 ( d, 3H); MS m/z 133.1 (M + 1).

Methyl ( R )-1-aminopropan-2-ylcarbamate; and 1,1-dimethylethyl ( S )-1-aminopropan-2-ylcarbamate were prepared analogously.

Intermediate: (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylaminocarbamate

The intermediate 2-chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine (1.4 g, 4.01 mmol), (S) was placed in a pyridine sealed vessel at 125 °C. Methyl 1-aminopropan-2-ylcarbamate (0.8 g, 6 mmol) and triethylamine (2.8 ml, 20 mmol) in isopropanol (30 ml) and dioxane (20 mL) The solution was heated for 48 hours. The cooled mixture was concentrated under vacuum, and aqueous sodium bicarbonate was added to the residue. The mixture was extracted with EtOAc and EtOAc (EtOAc)EtOAc. The crude residue was purified by EtOAcqqqqqq MS m/z 445.0 (M + 1).

Preparation of (R)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylaminocarbamate (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylaminocarboxylic acid tert-butyl ester 3-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propanenitrile; 4-(3-iodo-1-isopropyl- 1H-pyrazol-4-yl)-N-methylpyrimidin-2-amine; and N ¹ -(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine-2 -yl)-N ² ,N ² -dimethylethane-1,2-diamine.

Intermediate: N-(3-bromo-2,4-difluorophenyl)propane-1-sulfonamide

3-Bromo-2,4-difluoroaniline (4.16 g, 20 mmol, EP 184384), n-propane sulfonium chloride (4.6 ml, 40 mmol), pyridine (8 ml), DMAP (97 mg) at room temperature And a solution of DCM (100 ml) was stirred for 16 hours. Aqueous sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic layer was washed with aqueous sodium bicarbonate and brine. The crude product was purified by EtOAc (EtOAc:EtOAc) MS m/z 313.9 (M + 1).

Intermediate: 3-Fluoro-4-iodopyridin-2-amine

Treatment of 2-amino-3-fluoropyridine (1.0 g, 8.9 mmol) in dry THF (40 mL) with n-butyllithium (1.6 M in hexane, 13.9 ml, 22.3 mmol). Solution in ml). After the addition, the mixture was stirred at -78 °C for 1 hour, followed by a solution of iodine (10.2 g, 40.1 mmol) in THF (20 ml). The mixture was extracted with ethyl acetate, and the organic extract was washed with sodium thio sulfate, sodium hydrogen carbonate and brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by EtOAc (EtOAc:EtOAc) MS m/z 238.9 (M + 1).

3-Chloro-4-iodopyridin-2-amine was prepared analogously.

Intermediate: 3-bromo-2,5,6-trifluoroaniline

To the microwave tube was added 1-bromo-2,3,4,5-tetrafluorobenzene (1.0 g) and a 28% aqueous ammonium hydroxide solution (5 mL). The mixture was heated at 150 ° C under microwave irradiation for 2 hours, then the mixture was poured into water and extracted with hexane. The organic layer was separated, dried over MgSO ₄ and evaporated. The residue was purified by EtOAc EtOAc (EtOAc:EtOAc ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.75 (m, 1H), 3.95 (br s, 2H) ppm; MS m / z : 226, 228 (M+H) ⁺ .

Intermediate: 2,4-dibromo-3,6-dichloroaniline

A mixture of 2,5-dichloroaniline (0.2 g), N -bromosuccinimide (0.48 g) and THF (20 mL) was stirred at room temperature for 2 hr. The solvent was removed and the residue was purified EtOAcjjjjjjj MS m / z : 318 (M+H) ⁺ .

Intermediate: 1,3-dibromo-2,5-dichlorobenzene

Heat a stirred mixture of 2,4-dibromo-3,6-dichloroaniline (5.0 g), butyl nitrite (3.3 g) and EtOH (50 mL) in a sealed tube at 50 °C. hour. The mixture was concentrated and the residue was purified EtOAcjjjjjjj MS m/z : 303 (M+H) ⁺ .

Intermediate: 3-bromo-2-chloro-5-fluoroaniline

Step 1: 3-Bromo-2-chloro-N-(diphenylmethylene)-5-fluoroaniline. 2,6-Dibromo-4-fluoro-1-chlorobenzene (865 mg, 3 mmol), benzophenone imine (0.61 ml, 3.6 mmol), Pd ₂ (dba) ₃ at 80 ° C ( A mixture of 137 mg, 0.15 mmol), sodium butoxide (432 mg, 4.5 mmol), (S)-BINAP (280 mg, 0.45 mmol) and toluene (30 ml) was heated for 16 hours. The mixture was extracted with ethyl acetate and the combined organic phases were washed with brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by EtOAc (EtOAc) elute MS m/z 388.9 (M + 1).

Step 2: 3-Bromo-2-chloro-5-fluoroaniline. A solution of 3-bromo-2-chloro-N-(diphenylmethylene)-5-fluoroaniline (1.16 g) in THF (20 mL). The mixture was stirred at room temperature for 2 hours. The mixture was extracted with ethyl acetate and the combined organic phases were washed with brine. The organic phase was dried over sodium sulfate and concentrated. The crude residue was subjected to silica gel chromatography (40:1 to 15:1hexane/ethyl acetate as solvent) to afford the title compound. MS m/z 223.9 (M + 1).

3-Bromo-2,5-dichloroaniline; 3-bromo-2-chloro-5-methylaniline; and 3-bromo-2,5-difluoroaniline were similarly prepared.

Intermediate: 3-bromo-5-chloro-2-fluorobenzoic acid

To a solution of LDA in -78 ° C in THF (prepared from diisopropylamine (3.38 ml, 24 mmol) and n-BuLi (1.6 M, 13.1 ml, 21 mmol)) was added dropwise 2-bromo-4-chloro- A solution of 1-fluorobenzene (4.31 g, 20 mmol). The mixture was stirred at -78 °C for 1 hour, then was slowly transferred (about 30-60 minutes) via cannula to dry ice and THF (40 ml) at -78 °C to stir the mixture. The mixture was stirred at -78 ° C for 1 hour, and then allowed to warm to room temperature (gas evolution). The mixture was concentrated and then treated with 50 mL of 1 N sodium hydroxide solution. The mixture was extracted with ethyl acetate (discarded). The aqueous layer was acidified with 1N aqueous hydrochloric acid and then extracted with chloroform (3×400 ml). The chloroform extract was dried over sodium sulfate, filtered and evaporated elut The product was contaminated with a small amount of the isomeric product 2-bromo-6-chloro-3-fluorobenzoic acid. ¹ H NMR of the title compound 3-bromo-5-chloro-2-fluorobenzoic acid: (400 MHz, CDCl ₃ ) δ 7.93 (dd, 1H, J = 2.8, 5.6 Hz), 7.79 (dd, 1H, J =2.8, 5.6 Hz) ppm.

3-Bromo-2,6-difluorobenzoic acid; and 3-bromo-2-fluoro-5-methylbenzoic acid were similarly prepared.

Intermediate: 3-bromo-5-chloro-2-fluorophenylcarbamic acid tert-butyl ester

The crude intermediate 3-bromo-5-chloro-2-fluorobenzoic acid (2.03 g, 8 mmol), diphenylphosphonium azide (2.07 mL, 9.6 mmol, 1.2 eq.) A solution of DIPEA (1.67 mL, 9.6 mmol, 1.2 eq.) in 1:1 EtOAc/EtOAc (EtOAc) The mixture was concentrated and then partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by EtOAc (EtOAc:EtOAc) MS m / z : 267.8 (M+H) ⁺ (M- ^t Bu).

Similarly, 3-bromo-2,6-difluorophenylcarbamic acid tert-butyl ester; and 3-bromo-2-fluoro-5-methylphenylaminocarboxylic acid tert-butyl ester were prepared.

Intermediate: 3-bromo-5-chloro-2-fluoroaniline

Stir a solution of 3-bromo-5-chloro-2-fluorophenylcarbamic acid tert-butyl ester (900 mg, 2.78 mmol) in DCM/TFA (1:1, 20 mL) at room temperature 1 hour. The reaction mixture was concentrated, then the residue was crystalljjjjjjjj The organic layer was dried with sodium MS m / z 223.9 (M + 1).

Intermediate: 5-bromo-3-methoxy-2-methylaniline

Stir a heterogeneous reaction mixture of 4-bromo-2-methoxy-6-nitrotoluene (500 mg, 2.032 mmol), acetic acid (20 ml) and iron (1135 mg, 20.32 mmol) at room temperature. hour. Ethyl acetate was added, then the mixture was filtered through celite and concentrated. The residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous layer was extracted with additional ethyl acetate. The combined organics were washed with EtOAcqqqq MS m / z : 218.0 (M+H) ⁺ .

Intermediate: 5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester

The intermediate 3-bromo-5-chloro-2-fluorophenylcarbamic acid tert-butyl ester (1.45 g, 4.46 mmol), bis(pinacolyl) diboron (100 ° C) was placed in a sealed tube. A mixture of 1.7 g, 6.69 mmol), potassium acetate (1.53 g, 15.6 mmol), PdCl ₂ (dppf)-CH ₂ Cl ₂ (163 mg, 0.22 mmol) and dioxane (100 ml) was heated for 16 hours. The crude reaction mixture was taken up in ethyl acetate and washed with aqueous sodium hydrogen sulfate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude compound was diluted with hot hexane (600 mL) and heated to 65 ° C for 30 min then cooled to room temperature. The brown mixture was filtered through celite and the cake was washed with hexane. The combined filtrate was concentrated to give the crude title compound. MS m / z 233 (- t Bu M- frequency pinacolato).

Similarly prepared 5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester; N-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) 2-yl)phenyl)propane-1-sulfonamide; 2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-di Oxyboron ; 2,5-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2,5-dichloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-chloro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 2-fluoro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) phenylaminocarbamic acid tert-butyl ester; 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)pyridin-2-amine; 2,3,6-trifluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)pyridin-2-amine; 3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl)aniline; 3-methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) aniline.

Intermediate: 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) aniline

The intermediate 2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborate was hydrogen under 1 atmosphere of hydrogen. A mixture of (500 mg), 10% palladium on carbon (50 mg) and ethyl acetate (20 ml) was stirred for 16 hours. The mixture was flushed with nitrogen and filtered. The filtrate was concentrated to give the title compound. MS m/z 237.1 (M + 1).

Example 1 N-[(2S)-1-[(4-{3-[2,6-Difluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-) Methyl 1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate (Compound 7 in Table 1)

The crude intermediate N-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) at 80 °C -2-yl)phenyl)propane-1-sulfonamide (854 mg), intermediate (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl) Methyl pyrimidin-2-ylamino)propan-2-ylcarbamate (350 mg, 90% pure), hydrazine (triphenylphosphine) palladium (0) (90 mg), 2M aqueous sodium carbonate (6) A mixture of ml), toluene (50 ml) and ethanol (6 ml) was heated for 16 hours. The cooled mixture was extracted with ethyl acetate and the combined organic extracts were washed with brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by silica gel chromatography (EtOAc:EtOAc:EtOAc:

Example 2 N-[(2S)-1-({4-[3-(2-chloro-5-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazole) Methyl 4-methyl]pyrimidin-2-yl}amino)propan-2-yl]carbamate (Compound 15 in Table 1)

Step 1: (S)-1-(4-(3-(3-Amino-2-chloro-5-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidine-2 Methylamino)propan-2-ylaminocarbamate. The crude intermediate 2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborate at 85 °C Benzylamine (214 mg), intermediate (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino) Methyl propan-2-ylcarbamate (68 mg, 0.14 mmol), hydrazine (triphenylphosphine)-palladium (0) (16 mg), 2 M aqueous sodium carbonate (3 ml), toluene (18 ml) A mixture of ethanol (3 ml) was heated for 16 hours. The mixture was extracted with ethyl acetate and the combined organic extracts were washed with brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by silica gel chromatography (60:1 to 40:1 DCM/methanol as solvent) to afford (S)-1-(4-(1-isopropyl-1H-pyrazol-4-yl) The title compound (46 mg) contaminated with methyl pyrimidin-2-ylamino)propan-2-ylcarbamate. MS m/z 462.1 (M + 1).

Step 2: (S)-1-(4-(3-(2-chloro-5-fluoro-3-(methanesulfonylamino)phenyl)-1-isopropyl-1H-pyrazole-4- Methyl pyrimidin-2-ylamino)propan-2-ylaminocarbamate. Stir a mixture of the aniline product (46 mg), pyridine (2 ml), triethylamine (1 mL), DCM (4 ml) and methanesulfonium chloride (23 μl, 0.3 mmol) from Step 1 at rt. 16 hours. The crude reaction mixture was concentrated, then the residue was dissolved in a mixture of toluene (9 ml), ethanol (1 ml), sodium carbonate (2 g) and water (10 ml). The stirred mixture was heated at 85 ° C for 16 hours. The crude product was purified by EtOAc (EtOAc:EtOAc)

Example 3 N-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)) Methyl-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate (Compound 1 in Table 1)

Step 1: (S)-1-(4-(3-(5-Chloro-2-fluoro-3-(t-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazole Methyl 4-methyl)pyrimidin-2-ylamino)propan-2-ylcarbamate. The crude intermediate 5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborate at 80 °C -2-yl) phenylaminocarbamic acid tert-butyl ester (2.0 g), intermediate (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl) Methyl pyrimidin-2-ylamino)propan-2-ylcarbamate (600 mg, 1.34 mmol), hydrazine (triphenylphosphine) palladium (0) (150 mg, 0.13 mmol), 2 M aqueous sodium carbonate A mixture of (6.7 ml, 13.5 mmol), toluene (80 mL) and ethanol (6 mL) was then evaporated. The mixture was extracted with ethyl acetate and the combined organic extracts were washed with brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by silica gel chromatography (EtOAc: EtOAc: EtOAc) MS m/z 562.1 (M + 1).

Step 2: (S)-1-(4-(3-(3-Amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidine-2 Methylamino)propan-2-ylaminocarbamate. Partially purified (S)-1-(4-(3-(5-chloro-2-fluoro-3-(t-butoxycarbonylamino)phenyl)-1-isopropyl) at room temperature Stirring of methyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (1.15 g) in DCM (50 mL) hour. The solvent was removed, followed by the addition of aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate, and then the combined organic extracts were washed with sodium hydrogen carbonate and brine. The organic phase was dried over sodium sulfate and concentrated. The product was obtained by silica gel chromatography (60:1 to 40:1 DCM / methanol as solvent); MS m/z 462.1 (M+1).

Step 3: N-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propyl-2) Methyl-l-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate. Treatment of (S)-1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H- with propane sulfonium chloride (40 mg, 0.27 mmol) A mixture of methyl pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (41 mg, 0.09 mmol), triethylamine (1 ml) and DCM (4 ml). The mixture was stirred at room temperature for 16 hours. The crude mixture was concentrated, and the residue was dissolved in a mixture of toluene (9 ml), ethanol (1 ml), sodium carbonate (2 g) and water (10 ml). The stirred mixture was heated at 85 ° C for 16 hours. The crude product was purified by EtOAc (EtOAc:EtOAc)

Example 4 N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylaminophenyl)-1-(oxacyclo-2-yl)-) Methyl 1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate (Compound 33 in Table 1) and N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1H-pyridyl) Methyl oxazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate (Compound 31 in Table 1)

Step 1: 5-Chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyridyl Zyrid-3-yl) aniline. According to the procedure of Example 1, Step 1, the intermediate 4-(3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-2-(methylthio) Pyrimidine and intermediate 5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron 2-yl) aniline was prepared as the starting material. MS m/z 420.0 (M + 1).

Step 2: N-(5-Chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)- 1H-pyrazol-3-yl)phenyl)methanesulfonamide) and N-(5-chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1 -(Tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)phenyl)-N-(methylsulfonyl)methanesulfonamide. To 5-chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-piperidin-2-yl)-1H-pyrazole- Methanesulfonium chloride (0.13 mL, 1.66 mmol) was added to a solution of EtOAc (EtOAc (EtOAc) (EtOAc) The mixture was stirred at room temperature for 16 hours to give a mixture of the title compound (LCMS). Ethyl acetate was added and the mixture was washed with EtOAc EtOAc. MS m/z of the monosulfonamide: 498.0 (M + 1); MS m/z :

Step 3: N-[(2S)-1-({4-[3-(5-Chloro-2-fluoro-3-methanesulfonylamino)-1-(oxacyclohexane-2-) Methyl-1)-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester. The crude product mixture of Step 2 was dissolved in THF-H ₂ O (1:1, 30 mL) and used at room temperature with Oxone (1.68 g, 2.75 mmol) treatment. The mixture was stirred at room temperature for 16 hours, followed by the addition of ethyl acetate. The organic layer was washed with aqueous sodium bicarbonate, water and brine then dried over sodium sulfate. The crude residue was dissolved in NMP (5 mL), and treated with intermediate (S) -1- amino-2-yl amine-carboxylate (146 mg, 1.1 mmol) and sodium carbonate (233 mg, 2.2 Mm) treatment. The mixture was heated at 110 ° C for 16 hours with stirring. The cooled reaction mixture was diluted with ethyl acetate then washed with water and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The residue was subjected to silica gel chromatography (2%MeOHMeOHMeOH)

Step 4: N-[(2S)-1-({4-[3-(5-Chloro-2-fluoro-3-methanesulfonylaminophenyl)-1H-pyrazol-4-yl]pyrimidine- Methyl 2-amino}amino)propan-2-yl]carbamate. To (2S)-1-(4-(3-(5-chloro-2-fluoro-3-(methylsulfonylamino)phenyl)-1-(tetrahydro-2H-pyran-2-yl) Add a solution of methyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (94 mg, 0.16 mmol) in MeOH (15 mL) 0.5 mL). The mixture was stirred at room temperature for 16 hours. An aqueous solution of sodium hydrogencarbonate was added to adjust the pH to 9, and the mixture was extracted with ethyl acetate. The organic layer was washed with aqueous sodium bicarbonate and brine. The crude product was purified by silica gel chromatography (30:1 to 15:1 m.

Example 5 N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazole Methyl 4-methyl]pyrimidin-2-yl}amino)propan-2-yl]carbamate (Compound 9 in Table 1)

To (S)-1-(4-(3-(3-Amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-yl Methane sulfonium chloride (0.277 mL, 3.57 mmol) was added to a solution of methylamino)propan-2-ylaminocarbamate (550 mg, 1.2 mmol) in DCM (30 mL) The mixture was stirred at room temperature for 16 hours. Aqueous sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate and brine. The organic phase was dried over sodium sulfate and concentrated. The crude product was purified by EtOAc (EtOAc:EtOAc) An alternative synthesis is described in Example 6 below.

Also isolated from the reaction mixture: N-[(2S)-2-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2) -yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl]carbamic acid methyl ester (Compound 32 in Table 1); N-{3-[4-(2- Aminopyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-5-chloro-2-fluorophenyl}methanesulfonamide (Compound 30 in Table 1) ).

Example 6 N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazole Methyl 4-methyl]pyrimidin-2-yl}amino)propan-2-yl]carbamate (Compound 9 in Table 1)

Step 1:1 - benzylidene-2-isopropyl hydrazine. Isopropyl hydrazine hydrochloride (712 g, 6.43 mol), sodium acetate (528 g, 6.43 mol) and 50% were fed to a reactor equipped with a mechanical stirrer, thermometer and addition funnel under a nitrogen purge. Ethanol (4500 mL). The mixture was stirred at 20 ° C for 5 minutes. Benzaldehyde (683 g, 6.43 mol) was added while maintaining the batch temperature below 23 °C. The mixture was stirred at 20 ° C for 20 hours. Toluene (6500 mL) was added and stirring was maintained for 5 minutes. The organic layer was separated. A saturated aqueous solution of sodium hydrogencarbonate (4800 mL) was slowly added to the vigorously stirred organic layer (Note: the pH of the aqueous layer was about 8.0). The organic layer was separated and washed with aq. sodium hydrogen sulfate (30 mL). The organic layer was separated and then at 40 ℃, concentrated in vacuo (50 → 20 Torr (torr)), to give the title compound as a yellow oil of (without further purification).

Step 2: 2-((2-Benzylmethylene-1-isopropylindenyl)methylene)-malononitrile. (Ethyloxyethylidene)malononitrile (755 g, 6.18 mol), DMAP (150 g, 1.23 mol) and a flask equipped with a mechanical stirrer, thermometer and addition funnel were fed under a nitrogen purge. Ethanol (6400 mL). The mixture was stirred to give a dark orange solution, and an endothermic process from 20 ° C to 12 ° C was observed. 1-Benzimylidene-2-isopropylindole (1101 g, crude) was slowly added over 15 min, exotherm to 32 &lt The orange suspension was heated to 50 ° C and held at 50 ° C for 30 minutes to give a dark brown suspension. Ethanol (3200 mL) was added to the mixture, and the mixture was cooled to 20 ° C and kept at 20 ° C for 1 hour. The slurry was filtered and the solid cake was washed with ethanol (3000 mL). The title compound was obtained as a yellow solid. HPLC purity >99%.

Step 3: 3-Amino-1-isopropyl-1H-pyrazole-4-carbonitrile. Feeding 2-((2-benzylidene-1-isopropylindenyl)methylene)-propyl to a flask equipped with a mechanical stirrer, thermometer, condenser and addition funnel under a nitrogen purge Dinitrile (632.6 g, 2.65 mol), MeOH (2.5 L) and concentrated (12 N) hydrochloric acid (329.0 mL, 3.94 mol). The mixture was heated to 63 ° C and held at 63 ° C for 30 minutes to give an orange solution. The mixture was cooled to 15 °C. Heptane (4 L) and MTBE (1 L) were added, and the mixture was stirred for 5 min. Water (7.5 L) was then added as a stream over 30 minutes at 15 °C to 25 °C. After the water addition was completed, the mixture was stirred at 25 ° C for 10 minutes. The heptane/MTBE layer was separated. The aqueous layer was washed with a (4:1 v/v) heptane/MTBE mixture (2 x 5 L) by stirring at 25 ° C for 10 minutes each time. Separate the layers. Solid sodium chloride (1 Kg) was added to the aqueous layer. Followed by slow addition of saturated aqueous potassium carbonate to the aqueous layer to control the CO ₂ precipitation and pH was adjusted to about 9.0. Followed by _{CH 2 Cl 2 (1 × 2.2} L, 1 × 800 mL) and the aqueous layer was extracted twice. The combined and dried over MgSO ₄ of CH ₂ Cl ₂ layer was filtered and at a bath temperature of 30 deg.] C, under vacuum (200 torr) until the residual filtrate was concentrated to a weight of about 1 Kg. Under stirring to a solution of CH ₂ Cl ₂ was slowly (over about 20 minutes) was added heptane (6.0 L) and form a slurry. The mixture was concentrated under vacuum (60 torr) at an internal temperature of 25 ° C until the residual volume was about 6.2 L. The slurry was cooled to 15 ° C and held at this temperature for 10 minutes. The product was filtered and the solid was washed with heptane (1 L). The solid was dried under vacuum (5 Torr). HPLC purity >99%.

Step 4: 1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)-ethanone. The 3-amino-1-isopropyl group was fed to a flask equipped with a mechanical stirrer, a thermometer, a reflux condenser, a heating/cooling capacity, an addition funnel and a nitrogen inlet/outlet at 20 ° C under nitrogen. 1H-pyrazole-4-carbonitrile (274 g, 1.82 mol) and cyclopentyl methyl ether (2600 mL). The suspension was cooled to -10 °C. A solution of 1.5 M methyllithium/lithium bromide complex in diethyl ether (6.0 L, 9.00 mol) was added dropwise at -10 °C to 0 °C over 2.5 hours. When the methyl lithium addition was completed, the reaction suspension was rapidly warmed to 5 ° C to 10 ° C and maintained at this temperature range for 1 hour. The mixture was cooled to 0 ° C and 2 N HCl (6.0 L) was added dropwise at 5-10 °C. The (upper) organic layer was separated and extracted with 2N EtOAc (500 mL). The combined aqueous layers were stirred at room temperature for 16 hours. The mixture was cooled to 15 ° C and basified with 50% NaOH (260.0 g) to give a pH of about 11.0. The mixture was extracted with CH ₂ Cl ₂ (1×2.0 L, 1×1 L). The combined and dried over MgSO ₄ of CH ₂ Cl ₂ layer was filtered and concentrated under vacuum to give a yellow solid (278 g). After heating to 65 ° C, the solid was dissolved in EtOAc (750 mL). The solution was cooled to room temperature and a slurry was obtained. Heptane (1500 mL) was slowly added over 40 minutes at room temperature. The slurry was cooled to -10 ° C and held at -10 ° C for 30 minutes. The slurry was filtered and the filter cake was rinsed with heptane (300 mL). The solid was dried under vacuum (5 Torr). HPLC purity >99%. Melting point: 136-139 ° C.

Step 5: 1-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-ethanone. Feeding 1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)-ethanone (250.0) to a flask equipped with a mechanical stirrer, thermometer and addition funnel under a nitrogen purge g, 1.49 mol) and acetonitrile (3725 mL). The mixture was cooled to -20 °C. BF ₃ ‧THF (313.1 g, 2.23 mol) was added dropwise while maintaining the internal temperature <-10 °C. Isoamyl nitrite (227.5 g, 1.94 mol) was added dropwise while maintaining the internal temperature <-10 °C. The mixture was warmed to 10 ° C and stirred at 10 ° C for 30 minutes. The mixture was added in a fine stream to a flask containing a mixture of I ₂ (28.5 g, 0.112 mol), KI (371.9 g, 2.24 mol) and acetonitrile (1160 mL) at 10-15 ° C under vigorous stirring. The addition causes nitrogen to evolve and is slightly exothermic. The mixture was stirred at room temperature for 30 minutes. HPLC showed no residual diazonium intermediate. A solution of sodium bisulfite (157.1 g, 1.51 mol) in 8% sodium chloride solution (4360 mL) was then added at 15-20 °C. The mixture was basified with saturated potassium carbonate to a pH of about 8.5. The upper acetonitrile layer was separated and concentrated in vacuo to give an oil residue. The oil was dissolved in iPrOAc (2770 mL) and washed with saturated aqueous sodium carbonate (1100 mL). The iPrOAc layer was separated and concentrated to a residual volume of about 1.5 L. A suspension is obtained. Heptane (5.5 L) was added to the suspension over 30 minutes at 20 °C. After the addition of heptane was completed, the suspension was stirred at 20 ° C for 10 minutes. The syrup was filtered and the solid was washed with heptane (1 L) then dried <RTI ID=0.0> Melting point: 90-92 ° C.

Step 6: 3-(Dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-prop-2-en-1-one. Feeding 1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-ethanone (640 g) to a flask equipped with a mechanical stirrer, thermometer and addition funnel under a nitrogen purge , 2.30 mol) and DMF (6.4 L). The resulting orange solution was heated to 120 °C. A Bradnec reagent (598.6 g, 3.43 mol) was added in one portion. The addition caused the batch temperature to drop to 114 ° C and the solution color turned darker orange. The mixture was stirred at 120 ° C for 20 minutes. The mixture was cooled to room temperature and then concentrated at 5 mm Hg, 60 ° C to give an oily residue. The residue was dissolved in iPrOAc (2400 mL) by warming to 74 °C. The mixture was cooled to 35 ° C and stirred to give a slurry. Heptane (6000 mL) was added over 1 hour at 35 ° C to room temperature. The mixture was cooled to -15 ° C and filtered, and the title compound was crystall HPLC purity > 98%. Melting point: 106-109 ° C.

Step 7: 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidine-2-amine. (E)-3-dimethylamino group was fed to a flask equipped with a mechanical stirrer, a thermometer, a Dean-Stark trap and a condenser under a nitrogen purge. 1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-prop-2-en-1-one (735 g, 2.2 mol), cesium carbonate (596 g, 3.3 mol) and NMP (5200 mL). The mixture was heated to 130 ° C and held at 130 ° C for 5 hours. (Note: Any low boiling fraction is collected by the Dean-Stoker trap). The mixture was cooled to 80 ° C, and a 15% aqueous sodium chloride solution (7500 mL) was added over a period of about 1 hour at 80 ° C to 35 ° C. When about half of the aqueous sodium chloride solution was added, the product began to precipitate. The mixture was further cooled to room temperature and held for 30 minutes. The title compound was obtained as a solid. HPLC purity >99%. Melting point: 167-169 ° C.

Step 8: 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-ol. TFA (748.8 mL) was fed to a flask equipped with a mechanical stirrer and a thermometer under a nitrogen purge. Add 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-amine (300 g, 0.91 mol) as a solid while maintaining the internal temperature using a cold water bath Below 30 ° C. The mixture was stirred at room temperature for 10 minutes to give a solution. The mixture was cooled to 20 ° C and sodium nitrite (79.7 g, 1.27 mol) was added portionwise over 5 hours with rapid stirring at 22-28 °C. (Note: Some gas evolution was observed with a slight exotherm, which is easy to control with a cold water bath). DCM (12 L) was added and the mixture was allowed to warm to 27 °C. Add water (4400 mL) (note: gas evolution at the beginning). A saturated solution of potassium carbonate (about 1500 mL) was slowly added to the mixture to alkalinize to a pH of about 9.0 (note: a large amount of gas was precipitated). A solution of sodium hydrogen sulfite (32 g, 0.30 mol) in water (100 mL) was added to the mixture. The mixture was stirred at 27 ° C for 15 minutes and the pH was adjusted again to about 9.0. The DCM layer was separated and concentrated under vacuum (200-100 mm Hg) at a bath temperature of 40 ° C to a residual weight of about 2300 g (about 1750 mL). MTBE (1500 mL) was added to the residue at 20 °C. The mixture was stirred at 20 ° C for 10 minutes and then filtered. The solid was dried under vacuum (5 Torr). HPLC purity >99%. Melting point: 216-218 ° C.

Step 9: 2-Chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidine. Feeding 4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidine-2- to a flask equipped with a stirrer, thermometer, condenser, addition funnel and nitrogen inlet/outlet Alcohol (311 g, 942 mmol). Acetonitrile (2500 mL) was added to the solid at 20 °C. The mixture was stirred to give a suspension. DIPEA (246.2 mL, 1.41 mol) and DMF (218.8 mL, 2.83 mol) were added sequentially to the suspension. The resulting suspension was stirred at 20 ° C for 5 minutes. POCl ₃ (217 g, 1.41 mol) was added to the suspension at 20-40 ° C to give an orange solution. The mixture was warmed to 80 ° C and maintained at 80 ° C for 3 hours. The mixture was cooled to 10 ° C and a solution of ammonium hydroxide (622 mL, 28%) in deionized water (5550 mL) was slowly added over 1.5 hr while maintaining the temperature below 20 °C. After the ammonium hydroxide addition was completed, the resulting suspension was stirred at 10-20 ° C for 40 minutes. The title compound was obtained as a brown solid. HPLC purity >99%.

Step 10: (S)-2-(Methoxycarbonylamino)propylaminocarbamic acid benzyl ester. Potassium carbonate (1190 mmol) was added to a suspension of (S)-1,2-diaminopropane dihydrochloride (50 g, 340 mmol) in dichloromethane (500 mL). The suspension was stirred and the filtrate was collected by filtration. The filtrate was cooled to 0-5 ° C and stirred while benzyl chloroformate (51 ml, 357 mmol). After the addition was completed, the reaction mixture was stirred at 0 to 5 ° C for 3 hours, then allowed to warm to room temperature and stirred overnight at room temperature. Triethylamine (71 ml, 510 mmol) was added dropwise to this mixture, and the mixture was cooled to 0-5. Methyl chloroformate (28 ml, 357 mmol) was slowly added at 0-5 ° C, and the mixture was warmed to room temperature and stirred overnight. The mixture was poured into water. The organic volatiles were removed under vacuum. The resulting aqueous slurry was filtered to give a solid, which was then washed with ethyl acetate to afford white solid (65 g, 92-94% HPLC purity). The title compound was obtained as a white solid (yield: 99.5%).

Step 11: (S)-1-Aminopropan-2-ylaminocarbamic acid methyl ester hydrochloride. A solution of (S)-2-(methoxycarbonylamino)propyl carbamic acid benzyl ester in methanol was hydrogenated over a 5% palladium on C catalyst at 50-60 psi. The reaction mixture was filtered and the~~~~~ 60 g of a colorless oil was dissolved in 200 mL of anhydrous dichloromethane and the solution was cooled to 0-5 ° C in an ice-water bath. A solution of HCl in methanol (about 75 mL) was added dropwise until the pH of the solution was <1. The resulting suspension was stirred at 0-5 ° C for 30 minutes, then the solid was collected via filtration. The title compound was obtained as a white solid.

Step 12: 3-Bromo-5-chloro-2-fluorobenzaldehyde. Cool a solution of 2,2,6,6-tetramethylpiperidine (327 g, 98%, 2.274 mol) and THF (1.9 L, HPLC grade) to -75 ° C under argon (dry ice-methanol) bath). A 1.6 M n-BuLi/hexane solution (1.47 L, 2.35 mol) was slowly added to the mixture at -72 ° C to -67 ° C over 1 hour. The mixture was stirred at -72 ° C to -67 ° C for 30 minutes to give a pale yellow suspension. 2-Bromo-4-chloro-1-fluorobenzene (435 g, 97%, 2.02 mol) was slowly added to the mixture over 30 minutes at -72 ° C to -67 ° C, then at -72 ° C to -67 ° C The mixture was stirred for another 30 minutes. To the mixture was slowly added dimethylformamide (230 g, 99.5%, 3.14 mol) at -70 ° C to -65 ° C for 30 minutes, followed by stirring the mixture at -70 ° C to -65 ° C for 30 minutes. Light brown solution. The cooling bath was removed, and then a saturated solution of ammonium chloride (720 mL) was added to the batch at -60 ° C to -30 ° C for 15 minutes to obtain a cloudy mixture. 6 N hydrochloric acid was quickly added to the mixture at -30 ° C to 10 ° C for 15 minutes to pH 1, and then ethyl acetate (2.0 L) was added at 10 ° C to 20 ° C. The layers were separated and aqueous layer was extracted with ethyl acetate (1×300 mL). The combined organic extracts were washed with EtOAc (EtOAc m. The filtrate was concentrated under vacuum (60-65 ° C) to give the title compound as a tan, vis. ¹ H NMR (CDCl ₃ ): δ 7.76-8.30 (m, 2H), 10.0-10.8 (br s, 1H); MS m/z 238.0 (M+1).

Step 13: 3-Bromo-5-chloro-2-fluorobenzoic acid. The stirred mixture of 3-bromo-5-chloro-2-fluorobenzaldehyde (415 g), tert-butanol (1.2 L) and water (1.2 L) was warmed to 30 ° C, then at 40-45 ° C. Potassium permanganate (335 g, 2.12 mol) (5 parts) was added to the batch hour. The dark purple inclusions were heated in the following stepwise manner: 45-50 ° C for 30 minutes, 50-55 ° C for 30 minutes and 55-60 ° C for 30 minutes to give a purple-brown suspension. The reaction mixture was cooled to 20 ° C, then a saturated solution of sodium sulfite was added at 22-27 ° C until a peroxide test gave a negative value. Warm water (2.5 L, about 50 ° C) and a saturated solution of sodium carbonate (100 mL) were added sequentially to the mixture over 15 minutes. The dark suspension was filtered through a 1 cm celite bed and the filter cake was washed with warm water (4 x 1 L, about 50 ° C). The combined filtrate was acidified to pH 1 with 6N aqueous hydrochloric acid to give a yellow oil. Ethyl acetate (3 L) was added to the mixture, and the mixture was stirred for 10 min. The organic layer was washed with EtOAc (EtOAc)EtOAc. Hexane (700 mL) was added to the residue and the suspension was cooled to 5-10 °C. The title compound was obtained as a yellow solid. Melting point: 150-152 ° C; HPLC purity (225 nm): 97.5%; ¹ H NMR (d ₆ -DMSO): δ 7.82 (s, 1H), 8.10 (s, 1H), 13.82 (br s, 1H) ; MS m/z 254 (M+H).

Step 14: 3-Butyl 5-bromo-5-chloro-2-fluorophenylcarbamate. Heating a mixture of 3-bromo-5-chloro-2-fluorobenzoic acid (243 g, 97.5%, 0.935 mol), triethylamine (105 g, 99.5%, 1.02 mol) and third butanol (1.4 L) To 74 ° C. A solution of diphenylphosphonium azide (260 g, 97%, 0.916 mol) in toluene (960 mL) was slowly added to the mixture over 1 hour (slow reflux) at 75-79 °C. The mixture was slowly heated to 83 ° C over 30 minutes and maintained at 83-84 ° C (slow reflux) for 1 hour. The contents were concentrated to a viscous oil under vacuum (65-70 ° C). Toluene (2 L) and water (1.5 L) were sequentially added to the batch, followed by stirring the mixture at 35 ° C for 15 minutes. Discard the water layer. The organic layer was washed with a saturated solution of sodium bicarbonate (400 mL) and water (400 mL). The organic layer was concentrated under vacuum (60-65 °C) to approximately 350 mL residue (approximately 94% purity). 10% ethyl acetate/hexane (about 1.2 L, v/v) was added to the batch, followed by heating the mixture at 50 ° C for 15 minutes to give a pale yellow homogeneous solution. Transfer the ethyl acetate/hexane solution to a 4 L Pyros B-funnel (Pyrex B Chner funnel) (prepared tannin (1.8 kg, 70-200 mesh) / hexane bed on a coarse sintered disc, 40-60 μm, 16 cm diameter, 18 cm height). The title compound was obtained as an off-white solid as the title compound was obtained eluted from EtOAc (EtOAc: EtOAc) Melting point: 87-88 ° C; HPLC purity (225 nm): 97-98%; ¹ H NMR (d ₆ -DMSO): δ 1.48 (s, 9H), 7.48-7.49 (m, 1H), 7.80-7.82 (m, 1H), 9.42 (s, 1H); MS m/z 325 (M+H).

Step 15: (S)-1-(4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylaminocarbamate . Feeding 2-chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl) into a four-necked flask equipped with a mechanical stirrer, thermometer and condenser under a nitrogen purge Pyrimidine (300.0 g), (S)-1-aminopropan-2-ylcarbamic acid methyl ester hydrochloride (174.3 g), sodium carbonate (365.7 g) and DMSO (2400 mL). The mixture was heated under stirring at an internal temperature of 90 ° C for 18 hours. The mixture was cooled to 40 °C. Toluene (3870 mL) was added with stirring at 37-43 °C. Water (7200 mL) was added at 37-43 °C. The toluene layer was separated at 37-43 °C. A 15% aqueous solution of sodium chloride (3870 mL) was added to the toluene layer, and the pH of the aqueous layer was adjusted to a pH of about 5.0 by adding a 10% aqueous citric acid solution at 37-43 °C. A pH adjustment requires about 20 mL of a 10% aqueous citric acid solution. The toluene layer was subsequently washed with a saturated aqueous solution of sodium hydrogencarbonate (2880 mL) at 37-43 °C. The toluene layer containing the title compound was used as the feed in step 17.

Step 16: 5-Chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron -2-yl) tert-butyl phenylaminocarbamate. A 3-bromo-5-chloro-2-fluorophenylcarbamic acid tert-butyl ester (33.0 g) was fed into a flask equipped with a mechanical stirrer, a thermometer, a condenser and a heating mantle under a nitrogen purge. Bis (pinacoldyl) diboron (447.0 g), potassium acetate (405.6 g) and toluene (2700 mL). The mixture was stirred at room temperature for 15 minutes and PdCl ₂ (dppf) (50.4 g) was added. The mixture was then heated to 108 ± 2 ° C (note: the mixture turned dark at 50-60 ° C). A solution of 3-bromo-5-chloro-2-fluorophenylcarbamic acid tert-butyl ester (414 g) in toluene (1770 mL) was added over a period of 70 min. The mixture was kept at 108 ± 2 ° C for 15 hours. The mixture was cooled to room temperature under a stream of nitrogen and then filtered through Celite. The filtrate containing the title compound was used as the feed in step 17.

Step 17: (S)-1-(4-(3-(5-Chloro-2-fluoro-3-(t-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazole Methyl 4-methyl)pyrimidin-2-ylamino)propan-2-ylcarbamate. Feeding (S)-1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylaminocarboxylic acid to the flask Methyl ester (3870 ml toluene solution, approximately 382 g, 0.861 mol) and 5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboron 2-Benzyl) phenylaminocarbamic acid tert-butyl ester (4470 ml of toluene solution, about 467.0 g, 1.26 mol). A solution of sodium carbonate (349.8 g, 3.30 mol) in water (1400 mL) was added to the obtained brown solution. To the mixture was added PdCl ₂ (dppf) (34.5 g, 0.047 mol). The mixture was warmed to 80 ° C with stirring and maintained at this temperature for 2 hours. The mixture was cooled to 40 ° C and filtered through celite. The layers in the filtrate were separated. The toluene layer containing the title compound was used directly as the feed in step 18.

Step 18: (S)-1-(4-(3-(3-Amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidine-2 Methylamino)propan-2-ylaminocarbamate. (S)-1-(4-(3-(5-chloro-2-fluoro-3-(t-butoxycarbonylamino)phenyl) was fed into the flask at 20 ° C under a nitrogen atmosphere. Methyl-1--1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (about 7.3 L in toluene, ca. 483.3 g, 0.86 mol). 12 N HCl (574.3 mL, 6.95 mol) was added to the solution over about 20 minutes while maintaining the temperature below 25 °C. Hydrochloric acid addition causes an exothermic process from 19 ° C to 24 ° C. The mixture was stirred at 20-23 ° C for 1 hour. Water (3100 mL) was added to the reaction mixture. The mixture was stirred at 20 ° C for 10 minutes. The aqueous layer was separated and washed with 2-methyltetrahydrofuran (3100 mL). A saturated aqueous solution of potassium carbonate (about 778 mL) was slowly added to the aqueous layer. The pH of the aqueous layer is about 8.5. The aqueous layer was extracted with 2-methyltetrahydrofuran (3825 mL). The 2-methyltetrahydrofuran layer was concentrated under vacuum (60 mm Hg, 40 ° C). The residue was diluted with 2-methyltetrahydrofuran (~3800 mL) to give the title compound. HPLC purity: 95%.

Step 19: (S)-1-(4-(3-(5-Chloro-2-fluoro-3-(N-(methylsulfonyl)methylsulfonyl)phenyl)-1-iso Methyl propyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate. (S)-1-(4-(3-(3-Amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazole-) was fed into the flask at 20 °C. Methyl 4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (a solution of about 3.8 L of methyltetrahydrofuran, about 396.5 g, 0.86 mol). Triethylamine (435.0 g, 4.3 mol) was added to the solution. Cool the solution to 0 to -5 °C. Methanesulfonium chloride (246.0 g, 2.15 mol) was added dropwise to the solution over 20 minutes with stirring at 0 to -5 °C. The mixture was allowed to warm to 18-20 ° C and held at this temperature for 20 minutes. Water (2115 mL) was added to the reaction mixture over 30 minutes at 18-20 °C. After the addition was completed, the mixture was stirred at 20 ° C for 10 minutes. The pH was then adjusted to between 6.0 and 6.5 with 2 N HCl (about 1230 mL). The pH was then adjusted to 7-7.5 using a saturated aqueous solution of sodium bicarbonate. The mixture was stirred at 20 ° C for 10 minutes. Separate the layers. The 2-methyltetrahydrofuran layer containing the title compound was used directly in step 20.

Step 20: N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H Methyl pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate. (S)-1-(4-(3-(5-chloro-2-fluoro-3-(N-(methylsulfonyl)methylsulfonylamino)phenyl)-1 was fed into the flask Methyl isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (approximately 3.8 L solution of methyltetrahydrofuran, approximately 531.5 g, 0.86 mol). A 3 N aqueous sodium hydroxide solution (1782.8 mL, 5.34 mol) was added to the solution under stirring at 15-20 °C. The mixture was vigorously stirred at 20-23 ° C for 30 minutes and stirring was stopped. Discard the water layer. 2 N HCl (about 410 mL) was added to the organic layer to adjust the pH to 6.0-6.5, followed by a saturated aqueous solution of sodium hydrogencarbonate (about 300 mL) to adjust the pH to about 8.5. Discard the water layer. The organic layer was washed with a 15% aqueous sodium chloride solution (2000 mL). The organic layer was concentrated under vacuum (40 Torr) at a bath temperature of 45 ° C to give a brown solution (780 g). The solution was diluted with 2-methyltetrahydrofuran (3500 mL) followed by a suspension of PICA HP 120N activated carbon (90 g, CDH 858) in 2-methyltetrahydrofuran (1 L). The resulting black suspension was allowed to warm to 60 ° C and maintained at 60 ° C for 16 hours. After 16 hours, the Pd content was 309 ppm. The mixture was cooled to 20 ° C and filtered through a pad of Celite (pre-wet with 2-methyltetrahydrofuran). The reaction flask was rinsed with 2-methyltetrahydrofuran (500 mL). This rinse was then poured through the PICA/diatomaceous earth filter cake. PL-TMT resin (90 g) was added to the filtrate. The resulting suspension was heated to 60 ° C with stirring and maintained at this temperature for 4 hours. After 4 hours at 60 ° C, the Pd content was 2.3 ppm. The mixture was cooled to 20 ° C and stirred at 20 ° C overnight. The mixture was filtered through a pad of celite (pre-wet with 2-methyltetrahydrofuran). The filtrate was concentrated under vacuum (100-80 tolu.). This residual oil was dissolved in 3 L of 200 proof ethanol by raising the temperature to 78 °C. The resulting clear orange solution was then cooled to 20 ° C over 3 hours and a precipitate formed. The mixture was then cooled to 0 ° C and maintained at 0 ° C for 1 hour. The mixture was filtered and the filter cake was washed with ethanol (300 mL). The solid was dried at 40 ° C for 14 hours to give the title compound: mp.

Similar to the above examples, the compounds of Table 1 below were prepared using the appropriate starting materials:

Example 122 B-Raf V600E/Mek Enhanced Luminous Proximity Verification (B-Raf V600E Biochemical Verification)

B-Raf (V600E; 4 pM) and biotinylated Mek (lethal kinase; 10 nM) were combined in assay buffer (50 mM Tris (pH 7.5), 15 mM MgCl ₂ , 0.01% at 2× final concentration) BSA and 1 mM DTT) were dispensed in assay culture plates (Greiner 384-well white assay plate, No. 781207) containing 0.5 μl of the compound of the invention diluted in 100% DMSO, 10 μl per well. The plates were incubated for 60 minutes at room temperature.

B-Raf kinase activity was initiated by the addition of 10 μl of 2×ATP (10 μM) diluted in assay buffer per well. After 3 hours, 10 μl of stop reagent (60 mM EDTA, 0.01% Tween 20) was added to terminate the reaction. Rabbit anti-p-MEK (Cell Signaling, No. 9121) antibody and Alpha Screen IgG (Protein A) detection kit (PerkinElmer, No. 6760617R) were used, by adding 30 μL of antibody-containing antibody to the well (1:2000 dilution) Phosphorylated products were measured in a bead buffer (50 mM Tris (pH 7.5), 0.01% Tween 20) which was a mixture of beads (both beads diluted 1:1000). The addition is carried out under dark conditions to protect the beads from light. The lid was placed on top of the plate and the plate was incubated for 1 hour at room temperature. Luminescence was read on a PerkinElmer Envision instrument. By using XL Fit data analysis software for non-linear regression to calculate the 50% inhibitory concentration of each compound (IC _50).

The compounds of the invention in free form or in a pharmaceutically acceptable salt form exhibit valuable pharmacological properties, for example, as indicated by the in vitro test described in the present application. For example, the compound of the invention exhibits an IC ₅₀ for V600E B-Raf in the range of from 1 x 10 ^-10 to 1 x 10 ^-5 M, preferably less than 500 nM, 250 nM, 100 nM and 50. nM.

For example, homogeneous luminescent proximity assay some of the compounds of the invention IC ₅₀ data are shown in the table above.

Example 123 A375 Cell Proliferation Assay (A375 CP)

A375 is a melanoma cell line with a B-Raf V600E mutation. A375-luc cells engineered to express luciferase were plated at 1,500 cells/50 μl/well in DMEM containing 10% FBS in a 384-well white clear bottom plate. The compound of the present invention dissolved in 100% DMSO at an appropriate concentration was transferred to the cells by a robotic pin tool (100 nl). The cells were incubated for 2 days at 25 ° C, then 25 μl of BrightGlo ^TM was added to each well and the plate data was read via luminescence. By using XL Fit data analysis software for non-linear regression to calculate the 50% inhibitory concentration of each compound (IC _50).

The compounds of the invention in free form or in a pharmaceutically acceptable salt form exhibit valuable pharmacological properties, for example, as indicated by the in vitro test described in the present application. For example, the compounds of the invention and wild-type B-Raf the V600E B-Raf IC ₅₀ of the illustrated preferred less than 500 nM, 250 nM, 100 nM, and the range of 50 nM.

For example, A375 cell proliferation assay of some compounds of the invention IC ₅₀ data are shown in the table below.

Example 124 Immunoassay

Cells were seeded at a density of 30 x 10 ³ cells per well (96 wells) in tissue culture treated plates in 1640 RPMI + 10% FBS, followed by 37 ° C and 5% CO ₂ prior to compound addition. Cultivate for 24 hours. Test compounds were serially diluted with DMSO, then added to the cells (final DMSO concentration was 0.1%) and incubated for 3 hours at 37 ° C and 5% CO ₂ . The pMEK and pERK levels were measured using a Meso Scale Discovery kit. The culture supernatant was removed and the cells were lysed by adding cold lysis buffer (provided in the kit) over 30 minutes with gentle shaking. To detect pMEK1/2 (Ser217/221) and pERK1/2 (Thr/Tyr202/204, Thr/Tyr185/187), add lysate to the plate coated with the blocked antibody and have a set, and Cultivate overnight at 4 ° C under shaking. Plates were washed and phosphoprotein was detected using the provided labeled antibodies and data was read on a Sector 6000 instrument.

Example 125 SW620 cell viability assay

SW620 cells were seeded at a density of 1500 cells per well (96 wells) in tissue culture treated black wall clear bottom plates in 1640 RPMI + 10% FBS. Test compounds were serially diluted with DMSO, then added to the cells (final DMSO concentration 0.1%) and incubated for 4 days at 37 ° C and 5% CO ₂ . To measure cell viability, the cell culture plates were allowed to reach room temperature, the medium was removed, and 200 μl of Cell Titer-Glo reagent (Promega) was added to each well, and the kit components were mixed according to the manufacturer's protocol, followed by growth medium. Dilute 1:2). The plates were shaken for 5 minutes, then incubated for 5 minutes at room temperature, and luminescence (Trilux, Perkin Elmer) was measured.

Example 126 Rat1 soft agarose test

Rat1 cells were suspended in 1% agarose (Lonza) at a density of 1000 cells per well (96 wells). The agarose/cell mixture is allowed to set. Test compounds were serially diluted with DMSO, then added to the agarose cell mixture (final DMSO concentration 0.2%) and incubated at 37 ° C and 5% CO ₂ . After 17 days, community growth was determined by incubating cells with alamarBlue (TREK Diagnostics) and measuring the metabolic activity using a Spectramax plate reader (Molecular Devices, Inc; measuring absorbance at 562 nm). Happening.

Example 127 Liver microsomal clearance test

In vitro microsomal clearance assays were designed to assess the potential risks associated with hepatic metabolic stability of the compounds. Test compound (1 μM) with liver microsomes (0.5 mg/mL) of different species (mouse, rat, monkey, dog and human) and 100 mM potassium phosphate containing NADPH (1 mM) at 37 °C The buffer is incubated together. At specific reaction time points (0, 5, 10 and 30 minutes), the reaction aliquot was removed and the reaction was stopped by the addition of ice-cold acetonitrile containing the internal standard for mass spectrometry. The sample was centrifuged and the supernatant was analyzed by LC-MS/MS. The in vitro metabolic half-life (t _1/2 , min) and intrinsic clearance (CLint, μL/min/mg) are based on the metabolic rate and extent of the test compound as determined by the disappearance of the parent compound from the reaction mixture. These values can be scaled to predict hepatic metabolic clearance (CLh, mL/min/kg) and extraction rate (ER, expressed as the ratio of predicted hepatic metabolic clearance to hepatic blood flow in the species). It is generally believed that compounds with higher CLint or ER predicted in vitro are at higher risk of exposing exposure-limiting metabolism in vivo.

The following table gives the measured extraction rates for some of the compounds of the invention.

Example 128 A549 p38α MAP kinase Bright-Glo reporter gene assay

A549 cells were stably transfected with the IL-8 promoter driven reporter pGL3-IL8-Luc. Cells were plated at 4 x 10 ⁵ /ml in 384-well solid white plates (40 μl per well, 5% CD-FBS, 1 x P/S, DMEM) and incubated overnight at 37 °C (18- 20 hours). Test compounds were serially diluted with DMSO, and then 50 nl of test solution (final DMSO concentration of 0.1%) was added to the culture. After incubation with the test compound for 30 minutes, the cells were stimulated with 1 ng/ml IL-1β (10 μl 5 ng/ml solution per well). After 7-8 hours of stimulation, Bright-Glo (25 μl per well) was added to measure luciferase performance. The table below gives some of the compounds of the invention IC ₅₀ data.

Example 129 In vivo pharmacokinetic assay

Complete pharmacokinetic study: male Balb/c mice (n=3, body weight 22-25 g) or male Wistar rats (n=3, body weight 250-300 g) via the lateral tail vein The test compound is administered orally or by gavage. The formulation is typically a solution of 2.5 mg/mL compound in 75% PEG 300 and 25% D5W. After the administration, six blood samples (50 μL each) were continuously collected over 24 hours. The blood sample is centrifuged to separate the plasma. Plasma samples were analyzed and quantified by LC-MS/MS.

Rapid pharmacokinetic studies: male Balb/c mice (n=3, body weight 22-25 g) or male Wistar rats (n=3, body weight 250-300 g) via the lateral tail vein intravenously (IV The test compound is administered orally or via a gavage method (PO). The formulation is typically a solution of 2.5 mg/mL compound in 75% PEG 300 and 25% D5W. After the administration, six blood samples (50 μL each) were continuously collected over 24 hours. The blood sample was centrifuged, and plasma was separated, and plasma at each time point of each of the three administration routes was pooled. Plasma samples were analyzed and quantified by LC-MS/MS.

For complete pharmacokinetic studies and rapid pharmacokinetic studies, the following parameters were calculated by non-compartmental regression analysis using Winnonlin 5.0 software (Pharsight, Mountain View, CA, USA): plasma clearance (Cl), plasma maximum concentration (Cmax), plasma concentration-time curve area (AUC _0-inf ) and oral bioavailability percentage (F%).

The table below gives some pharmacokinetic parameters of the compounds of the invention in mice.

The compound of formula I in free form or in a pharmaceutically acceptable salt form exhibits valuable pharmacological properties, for example, as indicated by in vitro and in vivo tests as described herein. For example, the compounds of formula I IC cell proliferation assay in A375 CP shown in the preferred ₅₀ within the range of 250 nM or better, preferably less than 200 nM, 150 nM, 100 nM and 50 nM.

The 2-(methoxycarbonylamino)-1-propyl group at R ₁ provides a better level of activity and selectivity over other kinases, including p38. For example, compound 9 with an increased activity of more than 30 times between 29 wherein A375 IC ₅₀ were 2 nM and 76 nM.

Substituted phenyl-type compounds of formula Ib for the metabolic stability (in the mouse and human ER) in terms of the best, wherein the fluoro or chloro at the R ₅ position, and fluoro, chloro or methyl group at the R ₃ position. In contrast, for example, the ER (human) of compounds 9 and 6 were <0.21 and <0.69, respectively.

2- of the R ₁ (methoxycarbonyl amino) -1-methyl group at the ₄ combinations propyl, R ₃ / R ₅ substituent and R has the type of impact on the very surprising total drug exposure (AUC). See, for example, Compound 9 (compared to Compounds 1, 3, 4, 6, and 7) with an AUC of 30 ± 4 μM‧h at an oral dose of 10 mg/kg.

Example 130 In vivo efficacy - 14 days A375 mouse xenograft model

A375 cells were grown for 2 to 4 weeks under aseptic conditions in a 37 ° C incubator containing 5% CO ₂ . The cells were cultured in RPMI-1640 medium supplemented with 10% FBS. The cells were subcultured twice a week with 0.05% trypsin/EDTA. On the day of implantation, cells were harvested in HBSS (Hank's Balanced Salt Solution). On day 1, 0.2 mL of matrigel subcutaneously (SQ) containing A375 cells was implanted subcutaneously (SQ) into the right abdomen of female Nu/Nu mice (Charles River, 10-11 weeks at the start of the study), each Mice 5 x 10 ⁶ A375 cells. Nine days after implantation, mice were randomized into 6 groups (9 mice per group) with an average tumor volume of 215 mm ³ and an average body weight of 24 g. From day 19, the test compound was administered twice daily for a dose of 0.2 mL per dose for 14 days in which the compound was formulated at the appropriate concentration to achieve the desired dose. Clinical observations are performed daily. Tumor volume and body weight were measured twice a week. End point: Any individual animal or group exhibits a weight loss that exceeds 25% of the initial weight and/or a tumor volume of more than 3000 mm ³ .

According to the above protocol, Compound 9 (mixed in 20% PEG300/3% ETPGS/77% water) was administered using the following dosing regimen:

Group 1: Media, once a day × 14

Group 2: 1 mg/kg of compound 9, twice daily × 14

Group 3: 3 mg/kg of compound 9, twice daily × 14

Group 4: 10 mg/kg of compound 9, twice daily × 14

Group 5: 20 mg/kg of compound 9, twice daily × 14

Tumor volume results were assessed 14 days after the first dose. Tumor growth was determined as a partial response to 20-50% of the control tumor growth. The final tumor volume is defined as a stable disease within +/- 20% of the initial tumor size. The final tumor volume was less than 80% of the initial tumor volume and was determined to be partially resolved.

Group 2: partial reaction

Group 3: Stable disease

Group 4: Partial regression

Group 5: Partial regression

It is understood that the examples and embodiments described herein are intended to be illustrative only, and that various modifications and changes may be made by those skilled in the art, and such modifications and variations are included in the spirit and scope of the present application. Included in the scope of the patent application. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety herein

Figure 1 illustrates the inhibition produced by the addition of a MEK inhibitor to a compound of formula I of the present invention.

(no component symbol description)

Claims (22)

a compound of formula Ia, Wherein: Y is selected from N and CR ₆ ; R ₂ and R ₆ are independently selected from hydrogen, halo and C _1-4 alkyl; R ₃ is selected from halo and C _1-4 alkyl; R ₅ Is selected from the group consisting of hydrogen, halo and C _1-4 alkyl; R ₄ is selected from -R ₉ and -NR ₁₀ R ₁₁ ; wherein R ₉ is selected from C _1-6 alkyl, C _3-8 cycloalkyl , C _3-8 heterocycloalkyl, aryl and heteroaryl; wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group of R ₉ is optionally selected from 1 to 3 Substituted by a group of a halo group, a cyano group, a C _1-4 alkyl group, a halogen-substituted C _1-4 alkyl group, a C _1-4 alkoxy group, and a halogen-substituted C _1-4 alkoxy group And R ₁₀ and R ₁₁ are independently selected from hydrogen and R ₉ ; R ₇ is selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-5 cycloalkyl, and C _3-5 heterocycloalkyl; the alkyl group of _7, cycloalkyl or heterocycloalkyl optionally substituted with 1 to 3 substituents independently selected from halo, cyano, hydroxy, C _1-4 alkyl, substituted C _1-4 alkoxy group of halo Substituted by a group of a C _1-4 alkoxy group and a halogen-substituted C _1-4 alkoxy group; or a stereoisomer or a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein R ₄ is -R ₉ ; wherein R ₉ is selected from the group consisting of C _1-3 alkyl and C _3-8 cycloalkyl; wherein the alkyl or cycloalkyl group of R ₉ is optionally One to three groups independently selected from a halogen group and a halogen-substituted C _1-4 alkyl group are substituted. The compound of claim 2, wherein: R ₂ is selected from the group consisting of hydrogen and fluorine; R ₃ is selected from the group consisting of chlorine, fluorine, and methyl; R ₅ is selected from the group consisting of hydrogen, chlorine, and fluorine; and Y is selected from the group consisting of N and CR ₆ ; And R ₆ is selected from the group consisting of hydrogen and fluorine. The compound of claim 1, which is selected from the group consisting of: N-[(2S)-1-({4-[3-(3-chloro-5-methanesulfonylamino)phenyl)-1-(propan-2) -yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-[(4-{3- [3-Chloro-5-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propyl Methyl-2-yl]carbamate; N-[(2S)-1-({4-[3-(2-chloro-3-ethanesulfonylamino-4,5-difluorophenyl)) Methyl-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1- ({4-[1-(propan-2-yl)-3-(2,4,5-trifluoro-3-methanesulfonylaminophenyl)-1H-pyrazol-4-yl]pyrimidine-2 Methyl-amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-2-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino) Phenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl]aminocarbamic acid methyl ester; N-[(2S)-1- ({4-[3-(5-Chloro-2-fluoro-3-methanesulfonylaminophenyl)-1-(oxacyclo-2-yl)-1H-pyrazol-4-yl] Methyl pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(3-cyclopropanesulfonylamino-2,5) -difluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propane-2 -yl]methyl carbamate; N-[(2S)-1-({4-[3-(5-chloro-3-cyclopropanesulfonylamino-2-fluorophenyl)-1-(propyl) Methyl-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)- 1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propane-2 -yl]methyl carbamate; and N-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propan-1-sulfonylamino)phenyl]] Methyl-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate. A compound of claim 1 which has the formula Ib: Wherein R ₃ is selected from the group consisting of chlorine, fluorine and methyl; R ₅ is selected from the group consisting of fluorine and chlorine; and R ₇ is selected from the group consisting of ethyl and isopropyl. The compound of claim 5, which is selected from the group consisting of: N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)-1-) (propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4 -[3-(2,5-Difluoro-3-methanesulfonylaminophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino Methyl propan-2-yl]carbamate; N-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonylamino)phenyl)-1 -ethyl-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4-[3- (2-fluoro-3-methanesulfonylamino-5-methylphenyl)-1-(propan-2-yl)- N-[(2S)-1-({4-[3-(2-chloro)] -3-Methanesulfonylamino-5-methylphenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl Methyl carbazide; N-[(2S)-1-({4-[3-(2-chloro-5-fluoro-3-methanesulfonylamino)phenyl)-1-(prop-2- Methyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2R)-1-({4-[3-( 5-Chloro-2-fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propane-2 -yl]methyl carbamate; and N-[(2S)-1-({4-[3-(2,5-dichloro-3-methanesulfonylamino)-1-(propyl)- Methyl 2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate. A compound selected from the group consisting of: N-[(2S)-1-({4-[3-(3-chloro-2-methanesulfonylaminopyridin-4-yl)-1-(propan-2) -yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4-[3- (3-Fluoro-2-methanesulfonylaminopyridin-4-yl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propane-2 -yl]methyl carbamate; N-[(2S)-1-({4-[3-(2,4-difluoro-3-methanesulfonylamino)phenyl)-1-(propan-2) -yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)-1-({4-[3- (3-methanesulfonylaminophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid Ester; N-[(2S)-1-({4-[3-(3-ethanesulfonylamino-2,4-difluorophenyl)-1-(propan-2-yl)-1H-) Methyl pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; N-[(2S)-1-[(4-{3-[2,4-di) Fluoro-3-(propane-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2- Methyl carbazate; N-[(2S)-1-[(4-{3-[2-fluoro-3-(propane-1-sulfonamide) Methyl)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propyl-2-yl]carbamic acid methyl ester; N-[( 2S)-1-({4-[3-(2-Fluoro-3-methanesulfonylamino)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidine-2 Methyl-amino)propan-2-yl]aminocarbamate; N-[(2S)-1-({4-[3-(2,6-difluoro-3-methanesulfonylamino)benzene Methyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamic acid methyl ester; N-[(2S)- 1-[(4-{3-[2,6-difluoro-3-(propan-1-sulfonylamino)phenyl]-1-(propan-2-yl)-1H-pyrazole-4- Methyl}pyrimidin-2-yl)amino]propan-2-yl]carbamic acid methyl ester; and N-[(2S)-1-{[4-(3-{2-fluoro-3-[(3) ,3,3-trifluoropropane)sulfonylamino]phenyl}-1-(propan-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}propan-2- Methyl carbazate. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 in admixture with at least one pharmaceutically acceptable excipient. The pharmaceutical composition according to claim 8, wherein the excipient is selected from the group consisting of corn starch, potato starch, tapioca starch, starch paste, pregelatinized starch, sugar, gelatin, natural rubber, synthetic rubber, Sodium alginate, alginic acid, tragacanth, guar gum, cellulose, ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl group Cellulose, microcrystalline cellulose, magnesium aluminum silicate, polyvinylpyrrolidone, talc, calcium carbonate, powdered cellulose, dextrate, kaolin, mannitol, citric acid, sorbitol, Agar, sodium carbonate, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, clay, sodium stearate, calcium stearate, hard Fatty acid Magnesium, stearic acid, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, sodium lauryl sulfate, hydrogenated vegetable oil, peanut oil, cottonseed oil, sunflower oil, sesame oil , olive oil, corn oil, soybean oil, zinc stearate, sodium oleate, ethyl oleate, ethyl laurate, cerium oxide and combinations thereof. A pharmaceutical composition according to claim 8 which additionally comprises another therapeutic agent. The pharmaceutical composition according to claim 10, wherein the other therapeutic agent is selected from the group consisting of an anticancer compound, an analgesic, an antiemetic, an antidepressant, and an anti-inflammatory agent. A compound according to any one of claims 1 to 7 for use in the treatment of cancer. The compound of claim 12, wherein the cancer to be treated is selected from the group consisting of lung cancer, pancreatic cancer, bladder cancer, colon cancer, bone marrow disease, prostate cancer, thyroid cancer, melanoma, adenoma, and ovary Cancer of the eyes, liver, biliary tract and nervous system. A use of a compound according to any one of claims 1 to 7 for the manufacture of a medicament for the treatment of cancer. The use of claim 14, wherein the cancer is selected from the group consisting of lung cancer, pancreatic cancer, bladder cancer, colon cancer, bone marrow disease, prostate cancer, thyroid cancer, melanoma, adenoma, and ovary, eyes, Cancer of the liver, biliary tract and nervous system. The use of claim 14 or 15, wherein the medicament additionally comprises another therapeutic agent. The use of claim 16, wherein the additional therapeutic agent comprises an anti-cancer compound, an analgesic, an antiemetic, an antidepressant or an anti-inflammatory agent. The use of claim 16, wherein the additional therapeutic agent is a different Raf kinase Inhibitor, or inhibitor of MEK, mTOR, HSP90, AKT, PI3K, CDK9, PAK, protein kinase C, MAP kinase, MAPK kinase or ERK. The use of claim 18, wherein the MEK inhibitor is selected from the group consisting of: AS703026, MSC1936369B, GSK1120212, AZD6244, PD-0325901, ARRY-438162, RDEA119, GDC0941, GDC0973, TAK-733, RO5126766, and XL-518. A use of a compound according to any one of claims 1 to 7 for the manufacture of a medicament for the treatment of a condition mediated by Raf kinase. The use of claim 20, wherein the Raf kinase is a mutant B-Raf kinase. The use of claim 21, wherein the mutant B-Raf kinase is B-Raf (V600E).