HK1200820B

HK1200820B - Pyrazole aminopyrimidine derivatives as lrrk2 modulators

Info

Publication number: HK1200820B
Application number: HK15101249.0A
Authority: HK
Inventors: 查尔斯．贝克-格伦; 丹尼尔．乔恩．伯狄克; 马克．钱伯斯; 布赖恩．K．尚; 陈慧芬; 安东尼．埃斯特拉德; 扎卡里．凯文．斯威尼
Original assignee: 健泰科生物技术公司
Priority date: 2012-05-03
Filing date: 2013-04-30
Publication date: 2017-07-14

Abstract

Pyrazole compounds that are modulators of LRRK2, methods of making the compounds, and methods for using the compounds for treatment of diseases associated with LRRK2 receptor, such as Parkinson's disease.

Description

Pyrazolylaminopyrimidine derivatives as LRRK2 modulators

Technical Field

The present invention relates to compounds that modulate the function of LRRK2 and are useful for the treatment of LRRK2 mediated diseases and disorders, such as parkinson's disease.

Background

Neurodegenerative diseases such as Parkinson's disease, Lewy body dementia and Huntington's disease affect millions of individuals. Parkinson's disease is a chronic, progressive motor system disorder, with about one suffering per 1000 persons, with hereditary parkinson's disease accounting for 5-10% of all patients. Parkinson's disease is caused by progressive loss of mesencephalic dopamine neurons, giving patients impaired ability to direct and control their actions. The primary parkinson's disease symptoms are trembling, stiffness, bradykinesia, and impaired balance. Many parkinson's disease patients also experience other symptoms such as mood changes, memory loss, speech problems and sleep disturbances.

The gene encoding the leucine-rich repeat kinase 2 protein (LRRK2) has been identified as being associated with hereditary Parkinson's disease (Paisan-Ruiz et al, Neuron, Vol.44(4), 2004, pp 595-607; Zimphrich et al, Neuron, Vol.44(4), 2004, 601-607). In vitro studies have shown that parkinson-disease-associated mutations lead to increased LRRK2 kinase activity and a reduced rate of GTP hydrolysis compared to wild-type (Guo et al, Experimental Cell Research, vol.313(16), 2007, pp.3658-3670). anti-LRRK 2 antibodies have been used to label brainstem Lewy bodies associated with Parkinson's disease and cortical antibodies associated with Lewis body dementia, suggesting that LRRK2 may play an important role in Lewy body formation and pathogenesis associated with these diseases (Zhou et al, Molecular Degeneration, 2006, 1: 17 doi: 10.1186/1750-. LRRK2 has also been identified as a gene potentially associated with increased susceptibility to crohn's disease and leprosy (Zhang et al, New England j.med.vol.361(2009) pp.2609-2618.

LRRK2 is also related to: the shift from mild cognitive impairment to alzheimer's disease (WO 2007/149789); l-dopa-induced dyskinesia (Hurley et al, Eur. J. Neurosci., Vol.26, 2007, pp.171-177; CNS disorders associated with neuronal precursor differentiation (Milosevic et al, Neurodeen., Vol.4, 2009, p.25); cancers such as renal, breast, prostate, blood and lung cancer and acute myeloid leukemia (WO 2011/038572); papillary renal and thyroid cancers (Looyenga et al,www.pnas.org/cgi/doi/10.1073/pnas.1012500108) (ii) a Multiple myeloma (Chapman et al, Nature Vol.471, 2011, pp.467-472); amyotrophic Lateral Sclerosis (Shtilbans et al, Amyotropic Laterial Sclerosis "Early Online 2011, pp.1-7); rheumatoid arthritis (Nakamura et al, DNA Res.Vol.13(4), 2006, pp.169-183); and spinal rigidity (alkylosingspindylitis) (Danoy et al, PLoS Genetics, Vol.6(12), 2010, e1001195, pp.1-5).

Thus, compounds and compositions effective to modulate LRRK2 activity may provide treatment for the following diseases: neurodegenerative diseases such as parkinson's disease and lewy body dementia, CNS disorders such as alzheimer's disease and L-dopa induced dyskinesia, cancer such as renal, breast, prostate, blood, papillary and lung cancer, acute myeloid leukemia and multiple myeloma, and inflammatory diseases such as leprosy, crohn's disease, amyotrophic lateral sclerosis, rheumatoid arthritis and spinal stiffness. In particular, there is a need for compounds with LRRK2 affinity that are selective for LRRK2 over other kinases such as JAK2, which can provide effective drugs for the treatment of neurodegenerative diseases such as PD.

Disclosure of Invention

The present invention provides a compound selected from the group consisting of:

N²- (1 ', 5-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N²- (1 ', 3-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

4- (cyclopropylamino) -2- ((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-5-carbonitrile;

4- (cyclopropylamino) -2- ((3-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-5-carbonitrile;

2- ((1- (1-cyanopropyl) -5-methyl-1H-pyrazol-4-yl) amino) -4- (cyclopropylamino) pyrimidine-5-carbonitrile;

n2- (1- (1-fluoro-2-methylpropan-2-yl) -3-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

2- ((5-chloro-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -4- (ethylamino) pyrimidine-5-carbonitrile;

5-bromo-N²- (1, 5-dimethyl-1H-pyrazol-4-yl) -N⁴-methylpyrimidine-2, 4-diamine;

2-methyl-2- (3-methyl-4- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) propan-1-ol;

1- ((5-chloro-4- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) methyl) cyclopropanol;

n2- (3-chloro-1- (2- (4-ethyl-4H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (3-chloro-1- (2- (1-ethyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (3-methyl-1- (3-methyloxetan-3-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- (2- (1, 4-dimethyl-1H-imidazol-2-yl) propan-2-yl) -3-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (3-methyl-1- (2- (5-methyl)Oxazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (3-methyl-1- (3-methyloxetan-3-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (1- ((2-methoxyethyl) sulfonyl) -3-methyl-1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- ((2-methoxyethyl) sulfonyl) -3-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- ((2-methoxyethyl) sulfonyl) -5-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (5-chloro-1- (2- (4-methyl-4H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-ethyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (5-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) propyl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (3-methyl-1- (2- (pyrimidin-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (3-chloro-1- (2- (5-methyl-1, 3, 4-)Oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-ethyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N⁴-ethyl-N²- (3-methyl-1- (2- (5-methyl-1, 3, 4-)Oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-Ethyl-N2- (1- (2- (5-ethyl-1, 3, 4-)Oxadiazol-2-yl) propan-2-yl) -3-methyl-1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (1- (2- (1-isopropyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -3-methyl-1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (3-chloro-1- (2- (4-methyl-4H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-ethyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (3-chloro-1- (2- (1-methyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-ethyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (5-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (3-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (3-methyl-1- (2- (1-methyl-1H-pyrazol-5-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-methyl-N2- (3-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- (3-fluorotetrahydro-2H-pyran-4-yl) -3-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- (3-fluorotetrahydro-2H-pyran-4-yl) -5-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- ((3R) -3-Fluorotetrahydro-2H-pyran-4-yl) -5-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine

3-methyl-3- (3-methyl-4- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) butanenitrile;

2- ((3-methyl-4- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) methyl) butanenitrile;

3-methyl-3- (3-methyl-4- (4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-ylamino) -1H-pyrazol-1-yl) butanenitrile;

2- (5- ((4- (ethylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1-methyl-1H-pyrazol-3-yl) -2-methylpropanenitrile;

2-methyl-2- (1-methyl-5- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-3-yl) propionitrile;

2- ((1- (2-cyanoprop-2-yl) -3-methyl-1H-pyrazol-4-yl) amino) -4- (cyclopropylamino) pyrimidine-5-carbonitrile;

2- ((1- (2-cyanoprop-2-yl) -3-methyl-1H-pyrazol-4-yl) amino) -4- (ethylamino) pyrimidine-5-carbonitrile;

2-methyl-2- (3-methyl-4- ((4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) butanenitrile; and

5- ((4- (ethylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1-methyl-1H-pyrazole-3-carbonitrile,

or a pharmaceutically acceptable salt thereof.

The invention also provides pharmaceutical compositions comprising the compounds, methods of using the compounds, and methods of making the compounds.

Detailed Description

Definition of

Unless otherwise indicated, the following terms used in the present application, including the specification and claims, have the definitions given below. It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

"alkyl group"refers to a monovalent straight or branched chain saturated hydrocarbon moiety consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms. "lower alkyl" refers to alkyl of one to six carbon atoms, i.e., C₁-C₆An alkyl group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl, n-hexyl, octyl, dodecyl and the like.

"alkenyl" means a straight chain monovalent hydrocarbon group of two to six carbon atoms or a branched monovalent hydrocarbon group of three to six carbon atoms containing at least one double bond, such as ethenyl, propenyl, and the like.

"alkynyl" means a straight-chain monovalent hydrocarbon group of two to six carbon atoms or a branched-chain monovalent hydrocarbon group of three to six carbon atoms containing at least one triple bond, such as ethynyl, propynyl, and the like.

"alkylene" means a straight chain saturated divalent hydrocarbon group of one to six carbon atoms or a branched saturated divalent hydrocarbon group of three to six carbon atoms, such as methylene, ethylene, 2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene, and the like.

"alkoxy" and "alkyloxy" are used interchangeably to refer to a moiety of the formula-OR, wherein R is an alkyl moiety as defined herein. Examples of alkoxy moieties include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

"alkoxyalkyl" means a group of the formula R^a-O-R^bA moiety of (a) wherein R^aAnd R^bRespectively, alkyl and alkylene as defined herein. Exemplary alkoxyalkyl groups include, for example, 2-methoxyethyl, 3-methoxypropyl, 1-methyl-2-methoxyethyl, 1- (2-methoxyethyl) -3-methoxypropyl, and 1- (2-methoxyethyl) -3-methoxypropyl.

"Alkoxyalkyloxy" refers to a group of the formula-O-R-R ', where R and R' are, respectively, alkylene and alkoxy as defined herein.

"alkylcarbonyl" refers to a moiety of the formula-c (o) -R, wherein R is alkyl as defined herein.

"alkoxycarbonyl" refers to a group of formula-C (O) -R, wherein R is alkoxy as defined herein.

"Alkylcarbonylalkyl" refers to a group of the formula-R-C (O) -R, wherein R and R' are, respectively, alkylene and alkyl as defined herein.

"Alkoxycarbonylalkyl" refers to a group of the formula-R-C (O) -R, where R and R' are, respectively, alkylene and alkoxy as defined herein.

"Alkoxycarbonylalkoxy" refers to a group of the formula-O-R-C (O) -R ', wherein R and R' are, respectively, alkylene and alkoxy as defined herein.

"Hydroxycarbonylalkoxy" refers to a group of the formula-O-R-C (O) -OH, wherein R is alkylene as defined herein.

"Alkylaminocarbonylalkoxy" refers to a group of the formula-O-R-C (O) -NHR ', wherein R and R' are, respectively, alkylene and alkyl as defined herein.

"Dialkylaminocarbonylalkoxy" refers to a group of the formula-O-R-C (O) -NR 'R ", wherein R is alkylene as defined herein and R' and R" are alkyl as defined herein.

"alkylaminoalkoxy" refers to a group of the formula-O-R-NHR ', wherein R and R' are, respectively, alkylene and alkyl as defined herein.

"Dialkylaminoalkoxy" refers to a group of the formula-O-R-NR ' R ', wherein R is an alkylene group as defined herein and R ' and R "are alkyl groups as defined herein.

"alkylsulfonyl" means a compound of the formula-SO₂-a moiety of R, wherein R is alkyl as defined herein.

"Alkylsulfonylalkyl" refers to a compound of the formula-R' -SO₂A moiety of-R ", wherein R' and R" are alkylene and alkyl, respectively, as defined herein.

"Alkylsulfonylalkoxy" refers to a compound of the formula-O-R-SO₂-R 'wherein R and R' are alkylene and alkyl, respectively, as defined herein.

"amino" refers to a moiety of the formula-NRR ', where R and R' are each independently hydrogen or alkyl as defined herein. "amino" thus includes "alkylamino (where one of R and R 'is alkyl and the other is hydrogen) and" dialkylamino (where R and R' are both alkyl).

"aminocarbonyl" refers to a group of formula-C (O) -R, wherein R is amino as defined herein.

"Alkoxyamino" refers to a moiety of the formula-NR-OR 'where R is hydrogen OR alkyl and R' is alkyl, where alkyl is as defined herein.

"alkylthio" refers to a moiety of formula-SR, wherein R is alkyl as defined herein.

"aminoalkyl" refers to the group-R-R ', wherein R' and R are, respectively, amino and alkylene as defined herein. "aminoalkyl" includes aminomethyl, aminoethyl, 1-aminopropyl, 2-aminopropyl and the like. The amino moiety of "aminoalkyl" may be substituted once or twice with alkyl to provide "alkylaminoalkyl" and "dialkylaminoalkyl", respectively. "alkylaminoalkyl" includes methylaminomethyl, methylaminoethyl, methylaminopropyl, ethylaminoethyl, and the like. "dialkylaminoalkyl" includes dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl, N-methyl-N-ethylaminoethyl, and the like.

"Aminoalkoxy" refers to the group-OR-R ', where R' and R are, respectively, amino and alkylene as defined herein.

"Alkylsulfonamido" refers to a compound of the formula-NR' SO₂A moiety of-R, wherein R is alkyl and R' is hydrogen or alkyl.

"Aminocarbonyloxyalkyl" or "carbamoylalkyl" refers to a group of the formula-R-O-C (O) -NR 'R ", wherein R is alkylene and R', R" are each independently hydrogen or alkyl as defined herein.

"Alkynylalkoxy" refers to a group of the formula-O-R-R ', wherein R and R' are alkylene and alkynyl groups, respectively, as defined herein.

"aryl" refers to a monovalent cyclic aromatic hydrocarbon moiety composed of a monocyclic, bicyclic, or tricyclic aromatic ring. Aryl groups may be optionally substituted as defined herein. Examples of aryl moieties include, but are not limited to, phenyl, naphthyl, phenanthryl, fluorenyl, indenyl, pentalenyl, azulenyl, oxydiphenyl, biphenyl, methylenediphenyl, aminodiphenyl, diphenylsulfide, diphenylsulfonyl, diphenylisopropylidene, benzodiazepineAlkyl, benzofuranyl, benzodioxolyl, benzopyranyl, benzoAzinyl radical, benzoOxazinonyl (benzoxazinonyl), benzopiperidinyl (benzopiperadinyl), benzopiperazinyl, benzopyrrolidinyl, benzomorpholinyl, methylenedioxyphenyl, ethylenedioxyphenyl, and the like, including partially hydrogenated derivatives thereof, each optionally substituted.

"arylalkyl" and "aralkyl" are used interchangeably to mean the group-R^aR^bWherein R is^aAnd R^bAlkylene and aryl, respectively, as defined herein; for example, phenylalkyl such as benzyl, phenethyl, 3- (3-chlorophenyl) -2-methylpentyl and the like are examples of arylalkyl groups.

"arylsulfonyl" means a compound of the formula-SO₂-R, wherein R is aryl as defined herein.

"aryloxy" refers to a group of the formula-O-R, wherein R is aryl as defined herein.

"aralkyloxy" refers to a group of the formula-O-R-R ", wherein R and R' are, respectively, alkylene and aryl as defined herein.

"carboxy" or "hydroxycarbonyl" are used interchangeably and refer to a group of the formula-C (O) -OH.

"cyanoalkyl" refers to a moiety of the formula-R '-R', wherein R 'is alkylene as defined herein and R' is cyano or nitrile.

"cycloalkyl" refers to a monovalent saturated carbocyclic moiety consisting of a single ring or a double ring. Particular cycloalkyl groups are unsubstituted or substituted with alkyl groups. Cycloalkyl groups may be optionally substituted with one or more substituents, wherein each substituent is independently hydroxy, alkyl, alkoxy, halo, haloalkyl, amino, monoalkylamino, or dialkylamino, unless otherwise specified. Examples of cycloalkyl moieties include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, including partially unsaturated (cycloalkenyl) derivatives thereof.

"cycloalkylalkyl" refers to a moiety of the formula-R '-R', where R 'and R' are alkylene and cycloalkyl, respectively, as defined herein.

"cycloalkylalkoxy" refers to a group of the formula-O-R-R ', wherein R and R' are, respectively, alkylene and cycloalkyl as defined herein.

"Heteroalkyl" means an alkyl group as defined herein, wherein 1, 2 OR 3 hydrogen atoms have been independently selected from the group consisting of-OR^a，-NR^bR^cand-S (O)_nR^d(wherein n is an integer from 0 to 2) with the understanding that the point of attachment of the heteroalkyl group is through a carbon atom, wherein R is^aIs hydrogen, acyl, alkyl, cycloalkyl, or cycloalkylalkyl; r^bAnd R^cIndependently of one another, hydrogen, acyl, alkyl, cycloalkyl, or cycloalkylalkyl; and when n is 0, R^dIs hydrogen, alkyl, cycloalkyl, or cycloalkylalkylAnd when n is 1 or 2, R^dIs alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, monoalkylamino, or dialkylamino. Representative examples include, but are not limited to, 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2, 3-dihydroxypropyl, 1-hydroxymethylethyl, 3-hydroxybutyl, 2, 3-dihydroxybutyl, 2-hydroxy-1-methylpropyl, 2-aminoethyl, 3-aminopropyl, 2-methylsulfonylethyl, aminosulfonylmethyl, aminosulfonylethyl, aminosulfonylpropyl, methylaminosulfonylmethyl, methylaminosulfonylethyl, methylaminosulfonylpropyl, and the like.

"heteroaryl" means a monocyclic or bicyclic group of 5 to 12 ring atoms having at least one aromatic ring containing one, two or three ring heteroatoms selected from N, O or S, the remaining ring atoms being C, wherein it is understood that the point of attachment of the heteroaryl group is on the aromatic ring. The heteroaryl ring may be optionally substituted as defined herein. Examples of heteroaryl moieties include, but are not limited to, optionally substituted imidazolyl,azolyl radical, isoAn azole group, a thiazole group, an isothiazole group,oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, benzothienyl, thienyl, furyl, pyranyl, pyridyl, pyrrolyl, pyrazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl, benzothienyl, benzothiopyranyl, benzimidazolyl, benzofuranyl, benzothiophenyl, thienyl, and the likeAzolyl radical, benzoOxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranAn indolyl group, an isoindolyl group, a triazolyl group, a triazinyl group, a quinoxalinyl group, a purinyl group, a quinazolinyl group, a quinolizinyl group, a naphthyridinyl group, a pteridinyl group, a carbazolyl group, an azaazanyl groupRadical, diazaThe radicals, acridinyl and the like, including partially hydrogenated derivatives thereof, are each optionally substituted.

"heteroarylalkyl" or "heteroaralkyl" refers to a group of the formula-R-R ', wherein R and R' are alkylene and heteroaryl, respectively, as defined herein.

"Heteroarylsulfonyl" means a compound of the formula-SO₂-R, wherein R is heteroaryl as defined herein.

"heteroaryloxy" refers to a group of the formula-O-R, wherein R is heteroaryl as defined herein.

"Heteroaralkyloxy" refers to a group of the formula-O-R-R ", wherein R and R' are alkylene and heteroaryl, respectively, as defined herein.

The terms "halo", "halogen" and "halide" are used interchangeably to refer to the substituents fluorine, chlorine, bromine or iodine.

"haloalkyl" refers to an alkyl group, as defined herein, wherein one or more hydrogens have been replaced with the same or different halogen. Exemplary haloalkyl groups include-CH₂Cl，-CH₂CF₃，-CH₂CCl₃Perfluoroalkyl (e.g. -CF)₃) And the like.

"haloalkoxy" refers to a moiety of the formula-OR, wherein R is a haloalkyl moiety as defined herein. An exemplary haloalkoxy group is difluoromethoxy.

"heterocyclic amino" refers to a saturated ring wherein at least one ring atom is N, NH or N-alkyl and the remaining ring atoms form an alkylene group.

"Heterocyclyl" refers to a monovalent saturated moiety consisting of one to three rings, incorporating one, two or three or four heteroatoms (selected from nitrogen, oxygen or sulfur). The heterocyclyl ring may be optionally substituted as defined herein. Examples of heterocyclyl moieties include, but are not limited to, optionally substituted piperidinyl, piperazinyl, homopiperazinyl, azaA group, pyrrolidinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyridyl, pyridazinyl, pyrimidinyl,oxazolidinyl, isoOxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinuclidinyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazolidinyl, benzothiazolinylOxazolidinyl (benzolysidinyl), dihydrofuranyl, tetrahydrofuranyl, dihydropyranyl, tetrahydropyranyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

"Heterocyclylalkyl" refers to a moiety of the formula-R-R ', where R and R' are alkylene and heterocyclyl, respectively, as defined herein.

"Heterocyclyloxy" refers to a moiety of formula-OR, wherein R is heterocyclyl as defined herein.

"Heterocyclylalkoxy" refers to a moiety of the formula-OR-R ', where R and R' are alkylene and heterocyclyl, respectively, as defined herein.

"Hydroxyalkoxy" refers to a moiety of the formula-OR, where R is hydroxyalkyl as defined herein.

"Hydroxyalkylamino" refers to a moiety of the formula-NR-R ', wherein R is hydrogen or alkyl and R' is hydroxyalkyl as defined herein.

"Hydroxyalkylaminoalkyl" refers to a moiety of the formula-R-NR '-R ", wherein R is alkylene, R' is hydrogen or alkyl, and R" is hydroxyalkyl as defined herein.

"Hydroxycarbonylalkyl" or "carboxyalkyl" refers to a group of the formula-R- (CO) -OH, wherein R is alkylene as defined herein.

"Hydroxyalkyloxycarbonylalkyl" or "hydroxyalkoxycarbonylalkyl" refers to a group of the formula-R-C (O) -O-R-OH, wherein each R is alkylene and may be the same or different.

"hydroxyalkyl" refers to an alkyl moiety as defined herein, which is substituted with one or more, e.g., one, two, or three, hydroxyl groups, provided that the same carbon atom does not carry more than one hydroxyl group. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1- (hydroxymethyl) -2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2, 3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2, 3-dihydroxybutyl, 3, 4-dihydroxybutyl and 2- (hydroxymethyl) -3-hydroxypropyl.

"Hydroxycycloalkyl" refers to a cycloalkyl moiety as defined herein wherein one, two or three hydrogen atoms in the cycloalkyl group have been replaced with a hydroxyl substituent. Representative examples include, but are not limited to, 2-, 3-, or 4-hydroxycyclohexyl and the like.

"Alkoxyhydroxyalkyl" and "hydroxyalkoxyalkyl" are used interchangeably and refer to an alkyl group, as defined herein, substituted at least once with a hydroxy group and at least once with an alkoxy group. Thus "alkoxyhydroxyalkyl" and "hydroxyalkoxyalkyl" encompass, for example, 2-hydroxy-3-methoxy-propan-1-yl and the like.

"Urea" or "ureido" refers to a group of the formula-NR '-C (O) -NR "R'", wherein R ', R "and R'" are each independently hydrogen or alkyl.

"carbamate" refers to a group of the formula-O-C (O) -NR 'R "wherein R' and R" are each independently hydrogen or alkyl.

"carboxy" refers to a group of the formula-O-C (O) -OH.

"Sulfonamido (sulfonamide)" means a compound of the formula-SO₂A group of-NR ' R ', wherein R ', R ' and R ' are each independently hydrogen or alkyl.

"optionally substituted", when used with "aryl", "phenyl", "heteroaryl", "cycloalkyl" or "heterocyclyl", means that the aryl, phenyl, heteroaryl, cycloalkyl or heterocyclyl is optionally independently substituted with 1 to 4 substituents, for example 1 to 2 substituents, selected from alkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, hydroxyalkyl, halo, nitro, cyano, hydroxy, alkoxy, amino, acylamino, mono-alkylamino, di-alkylamino, haloalkyl, haloalkoxy, heteroalkyl, -COR, -SO₂R (wherein R is hydrogen, alkyl, phenyl or phenylalkyl), - (CR 'R')_n-COOR (wherein n is an integer from 0 to 5, R ' and R "are independently hydrogen or alkyl, and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl), or- (CR ' R ')_n-CONR^aR^b(wherein n is an integer from 0 to 5, R 'and R' are independently hydrogen or alkyl, and R^aAnd R^bIndependently of one another, hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl). Some particular optional substituents for "aryl", "phenyl", "heteroaryl", "cycloalkyl" or "heterocyclyl" include alkyl, halo, haloalkyl, alkoxy, cyano, amino and alkylsulfonyl. In one embodiment the substituents are methyl, fluoro, chloroTrifluoromethyl, methoxy, amino and methylsulfonyl.

"leaving group" means a group having the meaning conventionally associated therewith in synthetic organic chemistry, i.e., an atom or group displaceable under substitution reaction conditions. Examples of leaving groups include, but are not limited to, halogen, alkyl-or arylenesulfonyloxy, such as methanesulfonyloxy, ethanesulfonyloxy, thiomethyl, benzenesulfonyloxy, toluenesulfonyloxy, and thienyloxy, dihalophosphonooxy, optionally substituted benzyloxy, isopropyloxy, acyloxy, and the like.

"modulator" refers to a molecule that interacts with a target. Such interactions include, but are not limited to, agonism, antagonism, and the like, as defined herein.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

"disease" and "disease state" refer to any disease, condition, symptom, disorder, or sign.

By "inert organic solvent" or "inert solvent" is meant that the solvent is inert under the conditions of the reaction described in connection therewith and includes, for example, benzene, toluene, acetonitrile, tetrahydrofuran, N-dimethylformamide, chloroform, methylene chloride or dichloromethane, dichloroethane, diethyl ether, ethyl acetate, acetone, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol, tert-butanol, di-N-butanol, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol, N-dimethylformamide, NAlkanes, pyridines, and the like. Unless specified to the contrary, the solvent used in the reaction of the present invention is an inert solvent.

By "pharmaceutically acceptable" it is meant that it can be used to prepare pharmaceutical compositions that are generally safe, non-toxic, and not biologically or otherwise undesirable, and include that they are acceptable for veterinary as well as human pharmaceutical use.

By "pharmaceutically acceptable salt" of a compound is meant a pharmaceutically acceptable salt as defined herein and which possesses the desired pharmacological activity of the parent compound.

It is to be understood that all references to pharmaceutically acceptable salts include the solvent addition forms (solvates) or crystalline forms (polymorphs) of the same acid addition salt as defined herein.

"protecting group" or "protecting group" refers to a group that selectively blocks one reactive site in a polyfunctional compound so that a chemical reaction can be selectively carried out at another unprotected reactive site, in the sense conventionally associated therewith in synthetic chemistry. Certain processes of the present invention rely on protecting groups to block reactive nitrogen and/or oxygen atoms present in the reactants. For example, the terms "amino-protecting group" and "nitrogen-protecting group" are used interchangeably herein and refer to those organic groups intended to protect a nitrogen atom from unwanted reactions during synthetic procedures. Exemplary nitrogen protecting groups include, but are not limited to, trifluoroacetyl, acetamido, benzyl (Bn), benzyloxycarbonyl (carbonylbenzyloxy, CBZ), p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, tert-Butoxycarbonyl (BOC), and the like. The person skilled in the art knows how to select groups that are easy to remove and that have the ability to withstand subsequent reactions.

"solvate" refers to a solvent addition form containing a stoichiometric or non-stoichiometric amount of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thereby forming solvates. If the solvent is water, the solvate formed is a hydrate, and when the solvent is an alcohol, the solvate formed is an alcoholate. The hydrate maintains its molecular state as H with water therein via one or more water molecules₂O, such combination being capable of forming one or more hydrates.

"Parkinson's disease" refers to a degenerative disorder of the central nervous system that impairs motor skills, speech and/or cognitive function. Symptoms of parkinson's disease may include, for example, muscle stiffness, tremor, slowing of body movement (bradykinesia), and loss of body movement (akinesia).

"Lewy (Lewy) microsomia" also known as "Lewy body dementia," diffuse Lewy body disease, "cortical Lewy body disease," refers to a neurogenesis disorder that is anatomically characterized by the presence of Lewy bodies in the brain.

By "subject" is meant both mammals and non-mammals. Mammal refers to any member of the mammalian class, including but not limited to humans; non-human primates such as chimpanzees and other apes and monkey species; livestock such as cattle, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, and cats; the experimental animals include rodents such as rats, mice, guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds and the like. The term "subject" does not denote a particular age or gender.

By "therapeutically effective amount" is meant an amount of a compound that, when administered to a subject to treat a disease state, is sufficient to effect such treatment of the disease state. The "therapeutically effective amount" will vary depending on the compound, the disease state being treated, the severity or disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending medical or veterinary practitioner, and other factors.

When referring to a variable, the terms "those defined above" and "those defined herein" are incorporated by reference to the broad definition of the variable as well as the specific definitions, if any.

"treating" or "treatment" of a disease state includes, inter alia, inhibiting the disease state, i.e., arresting the development of the disease state or its clinical symptoms, and/or alleviating the disease state, i.e., temporary or permanent diminution of the disease state or its clinical symptoms.

The terms "treating", "causing.. to contact", and "causing.. to react" when referring to a chemical reaction, refer to adding or mixing two or more reagents under appropriate conditions to produce the specified and/or desired product. It will be appreciated that the reaction that produces the specified and/or desired product may not necessarily result directly from the combination of the two reagents initially added, i.e., there may be one or more intermediates produced in the mixture that ultimately result in the formation of the specified and/or desired product.

Naming and Structure

In general, the nomenclature used in this application is based on AUTONOM^TMv.4.0, used to generate the Beilstein Institute computerized system named by the IUPAC system. The chemical structures shown herein are usesVersion 2.2. Unless otherwise indicated, any open valency appearing on a carbon, oxygen, sulfur or nitrogen atom in the structures herein indicates the presence of a hydrogen atom. The heteroaryl ring containing nitrogen exhibits an open valency at the nitrogen atom, and the variables are as R^a，R^bOr R^cWhere shown on the heteroaryl ring, such variables may be bound or joined to the open-valence nitrogen. In the case where one or more chiral centers are present in the structure but no specific stereochemistry is shown for that chiral center, both enantiomers associated with each such chiral center are encompassed by the structure. Where a structure shown herein can exist in multiple tautomeric forms, all such tautomers are encompassed by the structure. The atoms represented in the structures herein are intended to encompass all natural isotopes of such atoms. Thus, for example, a hydrogen atom as represented herein is meant to include deuterium and tritium, and a carbon atom is meant to include C¹³And C¹⁴An isotope.

All patents and publications identified herein are hereby incorporated by reference in their entirety.

Compounds of the invention

One embodiment of the present invention relates to compounds of formula I:

or a pharmaceutically acceptable salt thereof,

wherein:

x is: -NH;

R¹the method comprises the following steps: c_1-6Alkyl or C_3-6A cycloalkyl group;

R²the method comprises the following steps: halogenating; cyano or halo-C_1-6An alkyl group;

R³the method comprises the following steps: hydrogen; c_1-6An alkyl group; halo-C_1-6An alkyl group; hydroxy-C_1-6An alkyl group; cyano-C_1-6An alkyl group; c_1-6An alkoxysulfonyl group; optionally substituted with R⁶Substituted one or more times by C_3-6A cycloalkyl group; c_3-6cycloalkyl-C_1-6Alkyl radical, wherein C_3-6Cycloalkyl moieties optionally substituted by R⁶One or more substitutions; optionally substituted with R⁷Heterocyclyl substituted one or more times; heterocyclyl-C_1-6Alkyl, wherein the heterocyclyl moiety is optionally substituted by R⁷One or more substitutions; optionally substituted with R⁸Heteroaryl substituted one or more times; or heteroaryl-C_1-6Alkyl, wherein the heteroaryl moiety is optionally substituted by R⁸One or more substitutions;

R⁴the method comprises the following steps: hydrogen; c_1-6An alkyl group; or halo; (ii) a

R⁵The method comprises the following steps: hydrogen; or C_1-6An alkyl group;

each R⁶Independently are: c_1-6Alkyl or hydroxy;

each R⁷Independently are: c_1-6An alkyl group; or halo; and is

Each R⁸Independently are: c_1-6Alkyl or halo-C_1-6An alkyl group;

wherein the compound is selected from:

5- ((4- (ethylamino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) -1-methyl-1H-pyrazole-3-carbonitrile.

In one aspect of the invention, there is provided a compound selected from the group consisting of:

N⁴-ethyl-N²- (3-methyl-1- (2- (5-methyl-1, 3, 4-)Oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diAn amine;

or a pharmaceutically acceptable salt thereof.

The present invention also provides a method for the treatment of a disease or condition mediated by or otherwise associated with the LRRK2 receptor, which method comprises administering to a subject in need thereof an effective amount of a compound of the present invention.

The disease may be a neurodegenerative disease such as parkinson's disease, huntington's disease or lewy body dementia.

The disease may be a CNS disorder such as Alzheimer's disease or L-dopa-induced dyskinesia.

The disease may be cancer or a proliferative disorder such as renal, breast, prostate, blood, papillary or lung cancer, acute myeloid leukemia or multiple myeloma.

The disease may be an inflammatory disease such as leprosy, crohn's disease, amyotrophic lateral sclerosis, rheumatoid arthritis, or ankylosing spondylitis.

The present invention also provides a method for enhancing cognitive memory, comprising administering to a subject in need thereof an effective amount of a compound of the present invention.

The invention also provides a compound as described herein for use as therapeutically active substance.

The invention also provides a compound as described herein for use as therapeutically active substance for the therapeutic and/or prophylactic treatment of parkinson's disease

The invention also provides the use of a compound as described herein in the therapeutic and/or prophylactic treatment of parkinson's disease.

Representative compounds of the process according to the invention are shown in the following experimental examples.

Synthesis of

The compounds of the present invention may be prepared by various methods depicted in the exemplary synthetic reaction schemes shown and described below.

The starting materials and Reagents used in preparing these compounds are generally available from commercial suppliers such as aldrich chemical co, or prepared by methods known to those skilled in the art according to procedures set forth in the references, such as Fieser and Fieser's Reagents for Organic Synthesis; wiley & Sons: new York, 1991, volume 1-15; rodd's Chemistry of Carbon Compounds, Elsevier Science Publishers, 1989, volumes 1-5 and desiccation; and Organic Reactions, Wiley & Sons: new York, 1991, volume 1-40. The following synthetic reaction schemes are merely illustrative of the methods by which the compounds of the present invention may be synthesized and various modifications may be made to these synthetic reaction schemes and will be suggested to one skilled in the art having referred to the disclosure contained in this application.

If desired, the starting materials and intermediates of the synthetic reaction schemes can be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like. Such materials may be characterized using conventional means, including physical constants and spectral data.

Unless otherwise specified, the reactions described herein may be carried out under an inert atmosphere at atmospheric pressure at a temperature in the range of from about-78 ℃ to about 150 ℃, for example from about 0 ℃ to about 125 ℃, or conveniently at about room (ambient) temperature, for example about 20 ℃.

Scheme A below illustrates one synthetic procedure that may be used to prepare certain compounds of formula I, wherein X, R¹，R²，R³，R⁴And R⁵As defined herein.

In step 1 of scheme A, dichloropyrimidine compoundaWith reagentsbReacted to give a pyrimidine compoundc. The reaction of step 1 may occur under polar solvent conditions. In which X is-O- (reagent)bIs an alcohol), the reaction of step 1 may be carried out in the presence of a base.

After step 1, performing one of steps 2a, 2b and 2cOne, the first step. In step 2a, a pyrimidine compoundcWith 4-amino-pyrazole compoundsd1The reaction is carried out to provide an aminopyrimidine compound of formula III. In step 2b, the pyrimidine compoundcWith 5-amino-pyrazole compoundsd2To provide an aminopyrimidine compound of formula IV. In step 2c, the pyrimidine compoundcWith 3-amino-pyrazole compoundsd3To obtain the aminopyrimidine compounds according to the invention. The reaction of steps 2a-2c may take place in a polar protic solvent and in the presence of an acid such as HCl.

Many variations to the procedure of scheme a are possible and will suggest themselves to those skilled in the art. Specific details for the preparation of the compounds of the present invention are described in the following examples.

Administration and pharmaceutical compositions

The present invention includes pharmaceutical compositions comprising at least one compound of the present invention, or an individual isomer, or a racemic or non-racemic mixture of isomers, or a pharmaceutically acceptable salt or solvate thereof, together with at least one pharmaceutically acceptable carrier, and optionally other therapeutic and/or prophylactic ingredients.

Typically, the compounds of the invention are administered in a therapeutically effective amount by any of the accepted modes of administration for agents used for similar purposes. Suitable dosage ranges are generally from 1 to 500 mg/day, for example from 1 to 100 mg/day, and in some embodiments from 1 to 30 mg/day, depending on a number of factors such as the severity of the disease being treated, the age and relative health of the subject, the potency of the compound used, the route and form of administration, the indication involved in the administration, and the preference and experience of the medical practitioner involved. One of ordinary skill in the art of treating such diseases, without undue experimentation and relying on personal knowledge and the disclosure of this application, will be able to determine a therapeutically effective amount of a compound of the present invention for a given disease.

The compounds of the invention may be administered as pharmaceutical formulations including those suitable for oral (including buccal and sublingual), rectal, nasal, topical, pulmonary, vaginal or parenteral (including intramuscular, intraarterial, intrathoracic, subcutaneous and intravenous) administration or in a form suitable for administration by inhalation or insufflation. One particular mode of administration is usually oral, using a convenient daily dosage regimen which may be adjusted according to the degree of affliction.

A compound or compounds of the invention, together with one or more conventional adjuvants, carriers or diluents, may be placed in the form of pharmaceutical compositions and unit dosages. The pharmaceutical compositions and unit dosage forms can be comprised of conventional ingredients in conventional proportions (with or without additional active compounds or ingredients), and the unit dosage forms can contain any suitable effective amount of the active ingredient commensurate with the desired daily dosage range to be employed. The pharmaceutical compositions may be employed as solids such as tablets or filled capsules, semisolids, powders, sustained release formulations, or liquids such as solutions, suspensions, emulsions, elixirs, or filled capsules for oral use; or in the form of suppositories for rectal or vaginal administration; or in the form of sterile injectable solutions for parenteral use. Formulations containing about one (1) milligram of active ingredient per tablet, or more broadly from about 0.01 to about one hundred (100) milligrams, are accordingly suitable representative unit dosage forms.

The compounds of the present invention may be formulated into a wide variety of oral dosage forms. Pharmaceutical compositions and dosage forms may comprise a compound or compounds of the invention or a pharmaceutically acceptable salt thereof as the active ingredient. The pharmaceutically acceptable carrier may be a solid or a liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances that may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is usually a finely divided solid, which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is usually mixed with a carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. Powders and tablets may contain from about one (1) to about seventy (70) percent of the active compound. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low melting wax, cocoa butter, and the like. The term "formulation" is intended to include the formulation of the active compound with encapsulating material as a carrier, providing a capsule in which the active ingredient (with or without a carrier) is surrounded by a carrier with which it is associated. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be as solid forms suitable for oral administration.

Other forms suitable for oral administration include liquid form preparations, including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, or solid form preparations which are intended to be converted into liquid form preparations immediately prior to use. Emulsions may be prepared in solution, for example in aqueous propylene glycol, or may contain emulsifying agents, for example lecithin, sorbitan monooleate or acacia. Aqueous solutions can be prepared by dissolving the active ingredient in water and adding suitable colorants, fragrances, stabilizers and thickeners. Aqueous suspensions may be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents. Solid form preparations include solutions, suspensions, and emulsions, and may contain, in addition to the active ingredient, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

The compounds of the invention may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion) and may be presented in unit dosage form in ampoules, pre-filled syringes, small volume infusions or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, for example, solutions in aqueous polyethylene glycol solutions. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and may contain formulatory agents such as preservatives, wetting, emulsifying or suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by sterile isolation of a sterile solid, or by lyophilization from a solution thereof which is reconstituted with a suitable vehicle, e.g., sterile pyrogen-free water, prior to use.

The compounds of the invention may be formulated for topical administration to the epidermis as an ointment, cream or lotion, or as a transdermal patch. Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. Formulations suitable for topical administration in the mouth include lozenges comprising the active agent in a flavoured base, usually sucrose and arabinose or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated for administration as suppositories. A low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active ingredient is dispersed homogeneously, for example by stirring. The molten homogeneous mixture is then poured into a conveniently sized mold, allowed to cool and solidify.

The compounds of the invention may be formulated for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing such carriers in addition to the active ingredient are known in the art to be suitable.

The subject compounds may be formulated for nasal administration. The solution or suspension is applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or nebulizer. The formulations may be provided in single or multiple dose forms. In the latter case of a dropper or pipette, this may be achieved by the patient administering an appropriate, predetermined amount of solution or suspension. In the case of a nebulizer, this can be achieved, for example, by means of a metered nebulization spray pump.

The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract, and include intranasal administration. The compounds typically have a small particle size, for example on the order of five (5) microns or less. Such particle sizes may be obtained by means known in the art, for example by micronisation. The active ingredient is provided in pressurized packs with a suitable propellant such as a chlorofluorocarbon (CFC), e.g., dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas. The aerosol may conveniently also contain a surfactant such as lecithin. The dosage of the drug may be controlled by a metering valve. Alternatively, the active ingredient may be provided in the form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP). The powder carrier forms a gel in the nasal cavity. Powder compositions may be presented in unit dosage form, for example in capsules or cartridges, for example in gelatin or film packs, from which the powder may be administered by the inhaler.

When desired, the formulations may be prepared with an enteric coating suitable for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention may be formulated in a transdermal or subcutaneous drug delivery device. These delivery systems are advantageous when sustained release of the compound is desired and patient compliance with a treatment regimen is critical. The compounds in transdermal delivery systems are often attached to a skin-adherent solid carrier. The compound of interest may also be combined with a penetration enhancer, such as azone (1-dodecylazacycloheptan-2-one). The sustained release delivery system is inserted subcutaneously into the subcutaneous layer by surgery or injection. The subcutaneous implants encapsulate the compound in a lipid-soluble membrane, such as silicone rubber, or a biodegradable polymer, such as polylactic acid.

The pharmaceutical preparation may be in unit dosage form. In such form, the preparation can be subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, a package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Moreover, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

Other suitable pharmaceutical carriers and their formulations are described in Remington: the Science and practice of Pharmacy 1995, edited by E.W Martin, Mack Publishing Company, 19th edition, Easton, Pennsylvania. Representative pharmaceutical formulations containing the compounds of the present invention are described below.

Utility of

The compounds of the invention are useful for treating LRRK2 mediated diseases or conditions, including neurodegenerative diseases such as parkinson's disease, lewy body dementia, and huntington's disease, and for enhancing cognitive memory in a subject in need thereof.

Examples

The following preparations and examples are given to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being exemplary and representative thereof.

All temperatures, including melting points (i.e., MP) are in degrees celsius (° c), unless otherwise indicated. It will be appreciated that the reaction which produces the specified and/or desired product may not necessarily result directly from the combination of the two reagents initially added, i.e. there may be one or more intermediates produced in the mixture which ultimately results in the formation of the specified and/or desired product. The following abbreviations may be used in the preparations and examples.

Abbreviations

AcOH acetic acid

AIBN 2, 2' -azobis (2-methylpropanenitrile)

Atm. atmospheric pressure

(BOC)₂Di-tert-butyl O dicarbonate

dba tris (dibenzylidene acetone)

DCM dichloromethane/methylene chloride

DIAD diisopropyl azodicarboxylate

DIPEA diisopropylethylamine

DMAP 4-dimethylaminopyridine

DME 1, 2-dimethoxyethane

DMF N, N-dimethylformamide

DMSO dimethyl sulfoxide

DPPF 1, 1' -bis (diphenylphosphino) ferrocene

Et₂O diethyl ether

EtOH ethanol/Ethyl alcohol

EtOAc ethyl acetate

HATU 2- (1H-7-azabenzotriazol-1-yl) -1, 1, 3, 3-tetramethylureaAmmonium methyl hexafluorophosphate

HBTU O-benzotriazole-1-yl-N, N, N ', N' -tetramethylureaHexafluorophosphates

HOBT 1-hydroxybenzotriazole

HPLC high pressure liquid chromatography

RP HPLC reversed-phase high-pressure liquid chromatography

i-PrOH Isopropanol/Isopropanol

LCMS liquid chromatography/Mass Spectrometry

MeOH methanol/methyl alcohol

MW microwave

NBS N-bromosuccinimide

NMP 1-methyl-2-pyrrolidone

PSI PSI

RT Room temperature

SFC supercritical fluid chromatography

TBDMS tert-butyldimethylsilyl group

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

Xphos 2-dicyclohexylphosphino-2 ', 4 ', 6 ' -triisopropylbiphenyl

Liquid chromatography-mass spectrometry method A

LC-MS was performed as follows: agilent SD-C18 columns (1.8 μm, 2.1X30mm) were used on an Agilent 1200 Series LC coupled to an Agilent 6140 quadrupole mass spectrometer with a linear gradient of 3-95% acetonitrile/water (0.05% trifluoroacetic acid in each mobile phase) over 8.5 minutes and held at 95% for 2.5 minutes.

Liquid chromatography-mass spectrometry method B

LC-MS was performed as follows: a Phenomenex Luna C18(2) column (5um, 100x4.6mm plus guard cartridge (guard cartridge)) was used on a Waters 2795Alliance HT HPLC with a Waters 2996 diode array detector combined with a Micromass ZQ, single quadrupole mass spectrometer with a linear gradient of 5-95% acetonitrile/water (0.1% formic acid in each mobile phase) over 3.5 minutes and held at 95% for 2.0 minutes.

Liquid chromatography-mass spectrometry method C

LC-MS was performed as follows: a Waters Xterra MS C18 column (5um, 100x4.6mM plus guard cylinder) was used on a Waters 2795Alliance HT HPLC with a Waters 2996 diode array detector combined with a Micromass ZQ, single quadrupole mass spectrometer, initially held at 5% acetonitrile/water (10mM ammonium bicarbonate in aqueous mobile phase) for 0.5 min, followed by a linear gradient of 5-95% over 3.5 min, then at 95% for 1.5 min.

Analytical method

¹H Nuclear Magnetic Resonance (NMR) spectroscopy was performed as follows: the solvent was used at about room temperature using a Bruker instrument, running at 400 or 500MHz, unless otherwise described. In all cases, the NMR data were consistent with the proposed structure. Characteristic chemical shifts () are given in parts per million, using the conventional abbreviations for the assignment of the main peaks: e.g., s, singlet; d, double peak; t, three peaks; q, four peaks; dd, double doublet; dt, double trimodal; br, broad peak. In the case of Thin Layer Chromatography (TLC), this means that silica gel MK6F is usedSilica gel TLC, R of the plate_fIs the distance the compound travels divided by the distance the solvent travels on the TLC plate. Flash chromatography refers to silica gel chromatography and is performed as follows: using SP4 or the Isolara 4 MPLC system (manufactured by Biotage); pre-loaded silica cartridges (supplied by Biotage); or using conventional glass column chromatography.

Preparation of compounds

Where the preparation of the starting materials is not described, they are commercially available, known in the literature, or readily available to those skilled in the art using standard procedures. Where the indicated compounds are prepared similarly to the previous examples or intermediates, it will be understood by those skilled in the art that reaction times, number of equivalents of reagents and temperatures may be varied for each particular reaction, and that it may be necessary or desirable to employ different workup or purification techniques. In the case of reactions carried out using microwave irradiation, the microwave used is Initiator 60 supplied by Biotage. The actual power applied was varied during the reaction to maintain a constant temperature.

The compounds prepared in the following examples are summarized in the following table, which shows affinity values (Ki, micromolar concentration) for LRRK2 for representative compounds along with LCMS method (M), LC retention time in minutes (RT), and mass spectrometry M/z values (molecular weight).

Intermediate 12, 5-dichloro-N-methylpyrimidin-4-amine

To a cooled (0 ℃) solution of 2, 4, 5-trichloropyrimidine (2.0g, 11mmol) in methanol (30mL) was added dropwise a 2M solution of methylamine in methanol (6.3 mL). The reaction was allowed to warm to room temperature and stirred overnight. The reaction was then concentrated and redissolved in DCM. The solution was saturated with NaHCO₃Washed with brine and Na₂SO₄Dried, filtered and concentrated. The crude product was purified by column chromatography (0-40% EtOAc in heptane) to give 2, 5-dichloro-N-methylpyrimidin-4-amine (0.9g, 50%).¹H-NMR(DMSO)：8.13(s，1H)，7.89(s，1H)，2.86(d，J＝4.5，3H)。

Intermediate 2: 5-bromo-2-chloro-N-methylpyrimidin-4-amine

To a cooled (0 ℃) solution of 5-bromo-2, 4-dichloropyrimidine (5.0g, 22mmol) in methanol (42mL) was added dropwise a 33 wt% solution of methylamine in ethanol (3.3 mL). The reaction was allowed to warm to room temperature. The reaction was then concentrated. The crude product was purified by column chromatography (0-10% methanol in DCM) to give 5-bromo-2-chloro-N-methylpyrimidin-4-amine (1.8g, 39%).¹H-NMR(DMSO)：8.22(s，1H)，7.75(s，1H)，2.85(d，J＝3.9，3H)。

Intermediate 32-chloro-N-methyl-5- (trifluoromethyl) pyrimidin-4-amine

To a cooled (-10 ℃) solution of 2, 4-dichloro-5-trifluoromethylpyrimidine (20g, 0.089mol) in methanol (100mL) was added triethylamine (12.5mL, 0.089mol) and a 2M solution of methylamine in methanol (45 mL). The reaction was allowed to warm to room temperature and stirred overnight. The reaction was then concentrated and redissolved in ethyl acetate. The solution was saturated with NaHCO₃Washed with brine and MgSO₄Dried, filtered and concentrated. The crude product was purified by column chromatography (5-25% EtOAc in heptane) to give 2-chloro-N-methyl-5- (trifluoromethyl) pyrimidin-4-amine (8.6g, 45%).¹H-NMR(DMSO)：8.37(s，1H)，7.90(s，1H)，2.90(s，3H)。

Additional intermediates prepared using a similar procedure to that described above are listed in table 1 below:

TABLE 1

Intermediates 8 and 91 ', 5-dimethyl-1' H-1, 4 '-bipyrazol-4-amine and 1', 3-dimethyl-1 'H-1, 4' -bipyramid Pyrazole-4-amines

Step 1-1 ', 5-dimethyl-4-nitro-1 ' H-1, 4 ' -bipyrazole and 1 ', 3-dimethyl-4-nitro-1 ' H-1, 4' -bipyrazoles

To a suspension of 3-methyl-4-nitro-1H-pyrazole (350mg, 2.76mmol), 1-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (859mg, 4.13mmol), and copper (II) acetate (150mg, 0.825mmol) in DMF (8mL) was added pyridine (87mg, 1.1 mmol). The mixture was stirred at 95 ℃ for 12h under oxygen. The reaction mixture was diluted with water and extracted with EtOAc (30mLx 3). The combined extracts were washed with brine, washed with Na₂SO₄Dried, filtered, and concentrated. The residue was purified by column chromatography on silica gel eluting with ethyl acetate/petroleum ether (1/5) to give a mixture of the two regioisomers (71mg, 13%) as an off-white solid.

Step 2-1 ', 5-dimethyl-1' H-1, 4 '-bipyrazole-4-amine and 1', 3-dimethyl-1 'H-1, 4' -bipyrazole- 4-amines

A mixture of 1 ', 5-dimethyl-4-nitro-1' H-1, 4 '-bipyrazole and 1', 3-dimethyl-4-nitro-1 'H-1, 4' -bipyrazole (71mg, 0.34mmol) and a suspension of 10% Pd/C (50mg) in methanol (10mL) at 55 deg.C in H₂Stirred for 3 h. Insoluble material was filtered off and the filtrate was concentrated under reduced pressure to give the two title compounds as a mixture (50mg, 83%).

Intermediate 101- (3-fluoro-tetrahydro-2H-pyran-4-yl) -3-methyl-1H-pyrazol-4-amine

Step 1-3-fluoro-tetrahydro-2H-pyran-4-ol

Reacting NaBH₄(190mg, 5.00mmol) was added to 3-fluoro-tetrahydropyran-4-one (300mg, 2.50mmol) in CH₃CN (3 mL). After stirring at 20 ℃ for 10H, H was added₂O (10 mL.) the resulting mixture was extracted with ethyl acetate (20mL × 3), dried over sodium sulfate, and concentrated under reduced pressure to give the title compound (200mg, 62%) as an oil.

Step 2-methanesulfonic acid 3-fluoro-tetrahydro-2H-pyran-4-yl ester

MsCl (189mg, 1.65mmol) was added to a mixture of 3-fluoro-tetrahydro-2H-pyran-4-ol (180mg, 1.5mmol) and triethylamine (0.5mL) at 0 ℃. The solution was stirred at room temperature for 4H, H was added₂O (10mL) to quench the reaction the resulting mixture was extracted with ethyl acetate (20mL × 3), dried over sodium sulfate, and concentrated under reduced pressure to give the title compound (300mg, 91%) as an oil.

Step 3-1- (3-fluoro-tetrahydro-2H-pyran-4-yl) -3-methyl-4-nitro-1H-pyrazole

Methanesulfonic acid 3-fluoro-tetrahydro-2H-pyran-4-yl ester (60mg, 0.50mmol), 3-methyl-4-nitro-1H-pyrazole (98mg, 0.50mmol), and Cs₂CO₃(243mg, 0.750mmol) of a mixture in DMF (2.0mL) was stirred at 100 ℃ for 2H after cooling, the resulting mixture was extracted with ethyl acetate (20mL × 3) and washed with H₂O (20mL) wash. The organic layers were combined, washed with brine (20mL), dried over sodium sulfate, and concentrated under reduced pressure to provide the title compound (60mg, 56%) as an oil. LC-MS (ESI): m/z 230(M + H)⁺。

Step 4-1- (3-fluoro-tetrahydro-2H-pyran-4-yl) -3-methyl-1H-pyrazol-4-amine

A mixture of 1- (3-fluoro-tetrahydro-2H-pyran-4-yl) -3-methyl-4-nitro-1H-pyrazole (60mg, 0.26mmol) and 10% Pd/C (30mg) in methanol (5mL) was dissolved in H₂The mixture was stirred at room temperature for 5 hours. Insoluble material was filtered off and the filtrate was concentrated under reduced pressure to give the title compound (40mg, 77%) as an oil. LC-MS (ESI): m/z 200(M + H)⁺。

Intermediates 11 and 123-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-amine and 5- Methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-amine

Step 1-2- (1H-pyrazol-3-yl) acethydrazide

A solution of 5-nitropyridin-2-ol (5.0g, 36mmol) in hydrazine hydrate (20mL) was stirred at 100 ℃ for 3 h. After cooling, the mixture was concentrated to provide the title compound (5.0g, 100%) as a red oil. LC-MS (ESI): 141(M + H) M/z⁺。

Step 2-2- (1H-pyrazol-3-yl) acetic acid

A solution of 2- (1H-pyrazol-3-yl) acetohydrazide (5.0g, 36mmol) in concentrated HCl (200mL) was stirred at 100 ℃ for 3H. Insoluble materials were filtered off, and the filtrate was concentrated to give the title compound (4.0g, 89%) as a yellow solid. LC-MS (ESI): m/z 127(M + H)⁺。

Step 3-2- (1H-pyrazol-3-yl) acetic acid ethyl ester

2- (1H-pyrazol-3-yl) acetic acid (4.0g, 32mmol) and concentrated H₂SO₄A solution (1.0mL) in ethanol (100mL) was refluxed overnight. After concentration, the residue is taken up in NaHCO₃Treating with water, extracting with ethyl acetate, and extracting with Na₂SO₄And (5) drying. The solvent was removed to give the title compound (4.6g, 94%) as a brown oil. LC-MS (ESI): 155(M + H) M/z⁺。

Step 4-ethyl 2- (1-methyl-1H-pyrazol-3-yl) acetate

To ethyl 2- (1H-pyrazol-3-yl) acetate (1.0g, 6.5mmol) and Cs₂CO₃(4.2g, 13mmol) to a mixture in DMF (5mL) was added methyl iodide (1.84g, 13.0 mmol). The mixture was stirred at 0 ℃ for 6 h. Then H is introduced₂O (20ml) was added and the resulting mixture was extracted with EtOAc (20mlx 3). The organic layers were combined, washed with brine (20mL), dried over sodium sulfate, and concentrated under reduced pressure to provide the title compound (600mg, 55%) as a yellow oil. LC-MS (ESI): 169(M + H) M/z⁺。

Step (ii) of5-2- (1-methyl-1H-pyrazol-3-yl) ethanol

To a solution of ethyl 2- (1-methyl-1H-pyrazol-3-yl) acetate (600mg, 3.57mmol) in THF (10mL) was added borane-tetrahydrofuran complex (10mL, 1.0M). The mixture was stirred at room temperature for 12 h. Adding CH₃OH (10ml) and H₂O (10ml) and the resulting mixture was extracted with EtOAc (20ml x 3). The organic layers were combined, washed with brine (20mL), dried over sodium sulfate, and concentrated under reduced pressure to provide the title compound (400mg, 89%) as a yellow oil. LC-MS (ESI): m/z 127(M + H)⁺。

Step 6-methanesulfonic acid 2- (1-methyl-1H-pyrazol-3-yl) ethyl ester

To 2- (1-methyl-1H-pyrazol-3-yl) ethanol (400mg, 3.18mmol), Et at 0 deg.C₃N (641mg, 6.35mmol) in CH₂Cl₂To the mixture in (20ml) was added methanesulfonyl chloride (579mg, 5.08mmol) dropwise. After stirring overnight at room temperature, the mixture was taken up in CH₂Cl₂Diluting with NaHCO₃Washing with aqueous solution, and adding Na₂SO₄And (5) drying. Removal of the solvent afforded the title compound (600mg, 93%) as a yellow oil. LC-MS (ESI): m/z 205(M + H)⁺。

Step 7-3-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -4-nitro-1H-pyrazole and 5-methyl- 1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -4-nitro-1H-pyrazole

2- (1-methyl-1H-pyrazol-3-yl) ethyl methanesulfonate (600mg, 2.94mmol), 3-methyl-4-nitro-1H-pyrazole (373mg, 2.94mmol), and Cs₂CO₃A mixture of (1.92g, 5.882mmol) in DMF (20mL) was stirred at 100 ℃ for 2 h. Addition of H₂O (20ml) and the resulting mixture was extracted with EtOAc (20ml x 3). The organic layers were combined, washed with brine (20mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC eluting with petroleum ether/ethyl acetate (1/1) to give the title compound (600mg, 87%) as a white solid. LC-MS (ESI): 236(M + H) M/z⁺。

Step 8-3-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-amine and 5-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -1H-pyrazol-4-amine

To a mixture of 3-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -4-nitro-1H-pyrazole and 5-methyl-1- (2- (1-methyl-1H-pyrazol-3-yl) ethyl) -4-nitro-1H-pyrazole (300mg, 1.28mmol) in methanol (25ml) was added 10% Pd/C (30 mg). The reaction mixture is reacted in H₂The mixture was stirred at room temperature for 1 hour. Insoluble material was filtered off and the filtrate was concentrated under reduced pressure to give the title compound (260mg, 99%) as a white solid. LC-MS (ESI): m/z 206(M + H)⁺。

Intermediate 133-methyl-1- (3-methyloxetan-3-yl) -1H-pyrazol-4-amine

Step 1-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) malonic acid diethyl ester

To a solution of 3-methyl-4-nitro-1H-pyrazole (3.81g, 30.0mmol) in DMF (20mL) was added K₂CO₃(8.28g, 60.0mmol) and diethyl 2-bromo-2-methylmalonate (9.10g, 36.0 mmol). The mixture was stirred at 100 ℃ for 20 h. The reaction mixture is then washed with H₂O (500mL) and extracted with ethyl acetate (20 mL. times.3). The organic layer was evaporated and the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (10/1) to give the title compound (3.6g, 41%) as an oil. LC-MS (ESI): 300(M + H) M/z⁺。

Step 2-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propane-1, 3-diol

To a solution of diethyl 2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) malonate (3588mg, 12.00mmol) in methanol (50mL) was added NaBH₄(890mg, 24.0 mmol). At 20 deg.CAfter stirring for 2H, the reaction was quenched H₂O (500mL) and extracted with ethyl acetate (20mL x 3). The organic phase was evaporated and the residue was purified by column chromatography on silica gel eluting with petroleum ether/ethyl acetate (5/1) to give the title compound (900mg, 35%) as a yellow solid. LC-MS (ESI): 216(M + H)⁺。

Step 3-4-Methylbenzenesulfonic acid 3-hydroxy-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propyl ester

To a solution of 2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propane-1, 3-diol (860mg, 4.00mmol) in pyridine (10mL) was added 4-methylbenzene-1-sulfonyl chloride (760mg, 4.00 mmol). The mixture was then refluxed for 20 h. After cooling, the mixture was concentrated. The residue was purified by column chromatography on silica gel eluting with petroleum ether/ethyl acetate (5/1) to give the title compound (670mg, 45%) as a white solid. LC-MS (ESI): m/z 370(M + H)⁺。

Step 4-3-methyl-1- (3-methyloxetan-3-yl) -4-nitro-1H-pyrazole

To a solution of 4-methylbenzenesulfonic acid 3-hydroxy-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propyl ester (660mg, 1.80mmol) in THF (5mL) at 0 ℃ was added NaH (90mg, 60%, 2.2 mmol). The mixture was then heated to reflux for 2 h. After cooling, the mixture is washed with H₂O (1 mL). After concentration, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (10/1) to give the title compound (280mg, 80%) as a white solid. LC-MS (ESI): 198(M + H) M/z⁺。

Step 5-3-methyl-1- (3-methyloxetan-3-yl) -1H-pyrazol-4-amine

To a solution of 3-methyl-1- (3-methyloxetan-3-yl) -4-nitro-1H-pyrazole (275mg, 1.40mmol) in methanol (5mL) were added Raney Ni (20mg) and hydrazine hydrate (80%, 2.0 mL). The mixture was stirred at room temperature for 2 h. Insoluble material was filtered off and the filtrate was concentrated to provide the title compound (220mg, 94%) as a yellow solid. LC-MS (ESI): m/z 168(M + H)⁺。

Intermediate 14 and 153-chloro-1- (2- (4-methyl-4H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4- Amines and 3-chloro-1- (2- (1-methyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-amine

Step 1-2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropanoic acid ethyl ester

To a solution of 3-chloro-4-nitro-1H-pyrazole (1.0g, 6.8mmol) in DMF (30mL) was added ethyl 2-bromo-2-methylpropionate (2.00g, 10.2mmol) and Cs₂CO₃(4.40g, 13.6 mmol). The mixture was stirred at 100 ℃ for 2 h. After filtration, the solution was poured into water and extracted 3 times with ethyl acetate. Combining the organic layers with anhydrous Na₂SO₄And (5) drying. The solvent was evaporated in vacuo and the residue was purified by silica gel column chromatography to give the title compound (1.2g, 68%) as a yellowish oil. LC-MS (ESI): m/z 262.0(M + H)⁺。

Step 2-2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropanoic acid

To a solution of ethyl 2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropionate (1.0g, 3.8mmol) in THF (5mL) was added LiOH (500mg, 22.8mL), iPrOH (5mL), and H₂O (5 mL). The mixture was refluxed for 2 h. The solvent was evaporated in vacuo. The resulting mixture was extracted 3 times with ethyl acetate. Combining the organic layers with anhydrous Na₂SO₄And (5) drying. The solvent was removed to give the title compound (850mg, 95% yield) as a white solid. LC-MS (ESI): 234.1(M + H) M/z⁺。

Step 3-2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropanamide

To a solution of 2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropanoic acid (800mg, 3.40mmol) in DCM (10mL) was added SOCl₂(500mg, 22.8mL) and DMF (one drop). The mixture was refluxed at 50 ℃ for 2 h. The solvent was evaporated in vacuo. To the resulting residue in DCM (10mL) was added NH₄OH (2 mL). The solution was stirred at rt for 0.5 h. After evaporation of the solvent in vacuo, the resulting residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (10: 1) to give the title compound (650mg, 81%) as a white solid. LC-MS (ESI): 233.2(M + H) with M/z⁺。

Step 4- (Z) -2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -N- ((dimethylamino) methylene) -2-methyl- Propionamide

A mixture of 2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -2-methylpropanamide (250mg, 1.10mmol) in DMF-DMA (1.5mL) was stirred at 95 ℃ for 2H. Removal of the solvent provided the title compound (250mg, 79%). LC-MS (ESI): m/z 288.1(M + H)⁺。

Step 5-3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4H-1, 2, 4-triazole

To a solution of (Z) -2- (3-chloro-4-nitro-1H-pyrazol-1-yl) -N- ((dimethylamino) methylene) -2-methyl-propionamide (250mg, 0.867mmol) in AcOH (1.5mL) was added hydrazine (aq, 85%, 1.0 mL). The mixture was stirred at 95 ℃ for 1.5 h. Then saturated NaHCO was added₃The mixture was extracted 3 times with ethyl acetate. Combining the organic layers with anhydrous Na₂SO₄And (5) drying. Removal of the solvent provided the title compound (250mg, 97%) as a yellow oil. LC-MS (ESI): m/z 257.2(M + H)⁺。

Step 6-3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4-methyl-4H-1, 2, 4-triazole and 3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1-methyl-1H-1, 2, 4-triazole

To a solution of 3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4H-1, 2, 4-triazole (250mg, 0.972mmol) in CH₃Adding Cs to CN (10mL)₂CO₃(650mg, 2.00mmol) and MeI (280mg, 2.00 mmol). Mixing the mixture inStirred at room temperature for 2 h. After concentration, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (5/1) to give the title compound (235mg, 87%) as a colorless oil. LC-MS (ESI): m/z 271.2(M + H)⁺。

Step 7-3-chloro-1- (2- (4-methyl-4H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-amine and 3- Chloro-1- (2- (1-methyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -1H-pyrazol-4-amine

To a solution of a mixture of 3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4-methyl-4H-1, 2, 4-triazole and 3- (2- (3-chloro-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1-methyl-1H-1, 2, 4-triazole (100mg, 0.370mmol) in EtOH (2mL) was added saturated NH₄Zn (1.0g) in Cl (2.0 mL). The mixture was stirred at room temperature for 1 h. After filtration, the solvent was evaporated in vacuo. The residue was redissolved in DCM and the mixture was filtered again. The filtrate was concentrated to give the title compound (80mg, 90%). LC-MS (ESI): m/z 241.2(M + H)⁺。

Intermediate 163-methyl-1- (2- (5-methyl-1, 3, 4-) Oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-amine

Step 1-N' -acetyl-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propionylhydrazine

To a mixture of 2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propionic acid (500mg, 2.38mmol) in DCM (20mL) was added acethydrazide (211mg, 2.86mmol), HATU (1.8g, 4.76mmol), and DIPEA (618mg, 4.76 mmol). The mixture was stirred at room temperature for 1 h. After concentration, the residue was purified by column chromatography on silica gel eluting with petroleum ether/ethyl acetate (1/3) to give the title product (600mg, 95%) as a pale yellow oilAnd (4) forming a substance. LC-MS (ESI): m/z 270.1(M + H)⁺。

Step 2-2-methyl-5- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1, 3, 4- Diazoles

A mixture of N' -acetyl-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propionohydrazide (500mg, 1.73mmol) in trichlorophosphoryl (2ml) was stirred at 100 ℃ for 1H. The reaction was quenched with ice-water. The mixture was extracted with ethyl acetate. Combining the organic layers with anhydrous Na₂SO₄And (5) drying. After evaporation of the solvent in vacuo, the resulting residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (3/1) to give the title compound (320mg, 68%) as a white solid. LC-MS (ESI): 252.3(M + H) M/z⁺。

Step 3-3-methyl-1- (2- (5-methyl-1, 3, 4-) Oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-amine

To 2-methyl-5- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1, 3, 4-To a mixture of diazole (100mg, 0.400mmol) in methanol (10ml) was added 10% Pd/C (50 mg). The reaction mixture is reacted in H₂Stir at rt for 2 h. Insoluble materials were filtered off and the filtrate was concentrated under reduced pressure to give the title compound (85mg, 92%) as a yellowish oil. LC-MS (ESI): 222.3(M + H)⁺。

Intermediate 171- (2- (1-isopropyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -3-methyl-1H-pyrazole-4- Amines as pesticides

Step 1- (Z) -N- ((dimethylamino) methylene) -2-methyl-2- (3-methyl-4-nitro-1H-pyrazole-1-) Mesityl-propionamide

A solution of 2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propanamide (1.60g, 7.55mmol) in DMF-DMA (8.98g, 75.5mmol) was stirred at 95 ℃ for 2H. After cooling, the mixture was concentrated to provide the title compound (2.10g, over 100%). LC-MS (ESI): m/z 268(M + H)⁺。

Step 2-3- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4H-1, 2, 4-triazole

A solution of (Z) -N- ((dimethylamino) methylene) -2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) -propionamide (2.10g, 7.55mmol) and hydrazine hydrate (6.0mL) in AcOH (15mL) was stirred at 95 ℃ for 1.5H. After cooling, the mixture was washed with NaHCO₃The aqueous solution was diluted and extracted with ethyl acetate. The combined extracts were washed with brine, over MgSO₄Drying, filtration, and concentration afforded the title compound (1.3g, 73% for 2 steps). LC-MS (ESI): 237(M + H)⁺。

Step 3-1-isopropyl-3- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1H-1, 2, 4-tris Azole

To a solution of 3- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -4H-1, 2, 4-triazole (1.3g, 5.5mmol) in CH₃To a solution in CN (30mL) was added 2-bromopropane (813mg, 6.60mmol) and Cs₂CO₃(1.79g, 5.50 mmol). The mixture was stirred at 50 ℃ overnight. After cooling, the mixture was diluted with water and extracted with ethyl acetate. The organic layer was concentrated to give the title compound (1.06g, 70%). LC-MS (ESI): m/z 279(M + H)⁺。

Step 4-1- (2- (1-isopropyl-1H-1, 2, 4-triazol-3-yl) propan-2-yl) -3-methyl-1H-pyrazol-4-amine

1-isopropyl-3- (2- (3-methyl)A mixture of yl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) -1H-1, 2, 4-triazole (600mg, 2.16mmol) and 10% Pd/C (200mg) in EtOH (25mL) was stirred under an atmosphere of hydrogen at 50 ℃ for 2H. Insoluble material was filtered off and the filtrate was evaporated to give the title compound (420mg, 80%). LC-MS (ESI): 249(M + H)⁺。

Intermediate 183-methyl-1- (2- (5-methyl) Azol-2-yl) propan-2-yl) -1H-pyrazol-4-amine

Step 1-2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) -N- (prop-2-ynyl) propionamide

To a solution of 2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propionic acid (1.00g, 4.69mmol) and prop-2-yn-1-amine (387mg, 7.04mmol) in DCM (30mL) were added DIPEA (908mg, 7.04mmol) and HTAU (2.38g, 7.04 mmol). After stirring at room temperature for 3h, the mixture was extracted with DCM and washed with water. The organic layer was concentrated to give the title compound (770mg, 66%). LC-MS (ESI): 251(M + H) M/z⁺。

Step 2-5-methyl-2- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) Azole

2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) -N- (prop-2-ynyl) propionamide (500mg, 2.00mmol) and AuCl₃(61mg, 0.20mmol) in CH₃The mixture in CN (10mL) was stirred at room temperature for 21 h. Insoluble material was filtered off and the filtrate was concentrated to provide the title compound (200mg, 40%). LC-MS (ESI): 251(M + H) M/z⁺。

Step 3-3-methyl-1- (2- (5-methyl) Azol-2-yl) propan-2-yl) -1H-pyrazol-4-amine

Mixing 5-methyl-2- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propyl-2-yl)A mixture of oxazole (200mg, 0.800mmol) and 10% Pd/C (80mg) in EtOH (10mL) was stirred at room temperature under a hydrogen atmosphere for 2 h. Insoluble material was filtered off and the filtrate was evaporated to give the title compound (175mg, 100%). LC-MS (ESI): m/z 221(M + H)⁺。

Intermediate 19 3-methyl-1- (2- (pyrimidin-2-yl) propan-2-yl) -1H-pyrazol-4-amine

Step 1-2- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) pyrimidine

2-methyl-2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propionamidine (500mg, 2.37mmol) and 1, 1, 3, 3-tetramethoxypropane (1.94mg, 11.1mmol) were added to 1, 4-bisThe solution in alkane (20mL) was stirred at 130 ℃ overnight. Insoluble material was filtered off and the filtrate was concentrated. The residue was purified by column chromatography on silica gel eluting with petroleum ether/ethyl acetate (3/1) to give the title compound (220mg, 34%). LC-MS (ESI): m/z 248(M + H)⁺。

Step 2-3-methyl-1- (2- (pyrimidin-2-yl) propan-2-yl) -1H-pyrazol-4-amine

A mixture of 2- (2- (3-methyl-4-nitro-1H-pyrazol-1-yl) propan-2-yl) pyrimidine (220mg, 0.890mmol) and 10% Pd/C (80mg) in EtOH (10mL) was stirred at room temperature under a hydrogen atmosphere for 2H. Insoluble matter is filtered off andthe filtrate was evaporated to give the title compound (160mg, 83%). LC-MS (ESI): m/z 218(M + H)⁺。

Intermediate 20 1- ((4-amino-5-chloro-1H-pyrazol-1-yl) methyl) cyclopropanol

Step 1-1- (tetrahydro-2H-pyran-2-yloxy) cyclopropanecarboxylic acid ethyl ester

To a solution of ethyl 1-hydroxycyclopropanecarboxylate (1040mg, 8.000mmol) and PTSA (137mg, 0.800mmol) in DCM (20mL) was added dropwise a solution of DHP (1344mg, 16.00mmol) in DCM (10mL) at 0 ℃. The mixture was stirred at room temperature for 20 h. After concentration, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (10/1) to give the title compound (1500mg, 85%) as an oil. LC-MS (ESI): 215(M + H) M/z⁺。

Step 2- (1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methanol

To a solution of ethyl 1- (tetrahydro-2H-pyran-2-yloxy) cyclopropanecarboxylate (1500mg, 7.000mmol) in THF (20mL) at 0 deg.C was added LiAlH₄(532mg, 14.0 mmol). After stirring at 0 ℃ for 30min, the reaction was washed with H₂O (1.0mL) was quenched. After concentration, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (3/1) to give the title compound (920mg, 77%) as a colorless oil. LC-MS (ESI): m/z 173(M + H)⁺。

Step 3-4-Nitro-1- ((1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methyl) -1H-pyrazole

To (1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methanol (515mg, 3.00mmol), 4-nitro-1H-pyrazole (407mg, 3.60mmol), and PPh at 0 deg.C₃(1180mg, 4.500mmol) to a solution in THF (10mL) was added DIAD (606mg, 3.00 mmol). The mixture was stirred at 20 ℃ for 20 h. ConcentrationAfter condensation, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (4/1) to give the title compound (186mg, 24%) as a white solid. LC-MS (ESI): m/z 268(M + H)⁺。

Step 4-5-chloro-4-nitro-1- ((1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methyl) -1H-pyrazole

To a solution of 4-nitro-1- ((1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methyl) -1H-pyrazole (186mg, 0.700mmol) in THF (5mL) at-70 deg.C in N₂LHMDS (1M in THF, 1.0mL) was added next. The mixture was warmed to-30 ℃ and stirred for 30 min. At-70 ℃ in N₂Adding C₂Cl₆(344mg, 1.40mmol) in THF (2 mL). The resulting mixture was warmed to room temperature and stirred for 1 h. The reaction was then quenched with water. After removal of volatiles, the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (5/1) to provide the title compound (102mg, 50%) as a white solid. LC-MS (ESI): 302(M + H) M/z⁺。

Step 5-1- ((5-chloro-4-nitro-1H-pyrazol-1-yl) methyl) cyclopropanol

To a solution of 5-chloro-4-nitro-1- ((1- (tetrahydro-2H-pyran-2-yloxy) cyclopropyl) methyl) -1H-pyrazole (100mg, 0.330mmol) in methanol (5mL) was added 2M HCl/bisAlkane (1 mL). The mixture was stirred for 2 h. The mixture was concentrated and the residue was purified by reverse phase Combiflash to provide the title compound (66mg, 90%) as a white solid. LC-MS (ESI): m/z 218(M + H)⁺。

Step 6-1- ((4-amino-5-chloro-1H-pyrazol-1-yl) methyl) cyclopropanol

To a solution of 1- ((5-chloro-4-nitro-1H-pyrazol-1-yl) methyl) cyclopropanol (66mg, 0.30mmol) in ethanol (5mL) was added zinc powder (39mg, 0.60mmol) and 1M NH₄Cl (1 mL). The mixture was stirred at room temperature for 2 h. Evaporating the mixtureAnd the residue was purified by reverse phase Combiflash to provide the title compound (45mg, 80%). LC-MS (ESI): 188(M + H) M/z⁺。

Examples 1 and 2 N ² - (1 ', 5-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) Pyrimidine-2, 4-diamines and N ² - (1 ', 3-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine Pyridine-2, 4-diamines

A microwave vial equipped with a magnetic stirrer was charged with a mixture of 1 ', 5-dimethyl-1' H-1, 4 '-bipyrazol-4-amine and 1', 3-dimethyl-1 'H-1, 4' -bipyrazol-4-amine (50mg, 0.28mmol), 2-chloro-N-methyl-5- (trifluoromethyl) pyrimidin-4-amine (60mg, 0.28mmol), and t-BuOH (3 mL). The mixture was heated at 100 ℃ under microwave irradiation for 1 h. After removal of volatiles, the residue was purified by preparative HPLC to afford N²- (1 ', 5-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine (17mg, 17%) as a white solid, and N²- (1 ', 3-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine (26mg, 26%) as a white solid.

N ² - (1 ', 5-dimethyl-1 ' H-1, 4 ' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-di Amines as pesticides

¹H NMR(500MHz，DMSO-d₆)9.06(s, 1H), 8.07(s, 2H), 7.90(s, 1H), 7.68(s, 1H), 6.99(br s, 1H), 3.88(s, 3H), 2.86(s, 3H), 2.24(s, 3H). LC-MS (method B): m/z 353.3(M + H)⁺4.42min, purity > 99.0%.

N ² - (1', 3-dimethyl)1 'H-1, 4' -bipyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-bis Amines as pesticides

¹H NMR(500MHz，DMSO-d₆)9.14(s, 1H), 8.28(s, 1H), 8.11(s, 1H), 8.03(s, 1H), 7.69(s, 1H), 7.07(s, 1H), 3.84(s, 3H), 2.91(d, J ═ 4.5Hz, 3H), 2.21(s, 3H). LC-MS (method B): m/z 353.3(M + H)⁺4.65min, > 99.0% purity.

The compounds prepared using the above procedure, along with low resolution mass spectrometry (M + H), proton NMR, and LRRK2K as determined by the test methods described below for selected compounds_iThe (micromolar concentration) data are shown in table 2 below.

TABLE 2

Example 8 5-bromo-N ²- (1, 5-dimethyl-1H-pyrazol-4-yl) -N⁴ -methylpyrimidine-2, 4-diamine

To a mixture of 5-bromo-2-chloro-N-methylpyrimidin-4-amine (0.201g, 0.903mmol) and 1, 5-dimethyl-1H-pyrazol-4-amine (0.12g, 1.08mmol) in 2-methoxyethanol (2mL) was added TFA (0.070mL, 0.9 mmol). The reaction was stirred in a sealed tube at 100 ℃ for 90 minutes. The resulting precipitate was collected by filtration. The isolated solid was further purified by reverse phase HPLC to give 5-bromo-N²- (1, 5-dimethyl-1H-pyrazol-4-yl) -N⁴-a methyl-pyrimidine-2,4-diamine (46mg, 17%). LCMS (method a): [ MH⁺]297.0, at 2.57 min.¹H-NMR(DMSO)：8.28(s，1H)，7.84(s，1H)，7.49(s，1H)，6.79(d，J＝3.4，1H)，3.67(s，3H)，2.82(d，J＝3.6，3H)，2.14(s，3H)。Ki＝0.017μM。

The compounds prepared using the above procedure, along with low resolution mass spectrometry (M + H), proton NMR, and LRRK2K as determined by the test methods described below for selected compounds_iThe (micromolar concentration) data are shown in table 3 below.

TABLE 3

Example 43: 3-methyl-3- (3-methyl-4- (4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-ylamino) - 1H-pyrazol-1-yl) butanenitrile

Step 1-3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butanoic acid methyl ester

A solution of 3-methyl-4-nitro-1H-pyrazole (1.00g, 7.87mmol), methyl 3-methylbut-2-enoate (2.20g, 18.9mmol), and DBU (3.10g, 20.5mmol) in DMF (5.0mL) was stirred overnight. The mixture was purified by reverse phase Combiflash to afford the title compound (130mg, 7%) as a yellow oil. LC-MS (ESI): m/z 242(M + H)⁺。

Step 2-3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butanoic acid

To methyl 3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butanoate (508mg, 2.10mmol) in EtOH (10mL) and H₂To a solution in O (10mL) was added LiOH (265mg, 6.30 mmol). The reaction mixture was stirred for 12 h. The mixture was acidified to a pH of about 5. The resulting mixture was extracted with EtOAc, washed with brine, and Na₂SO₄And (5) drying. Removal of the solvent provided the title compound (450mg, 94%). LC-MS (ESI): m/z 228.1(M + H)⁺。

Step 3-3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butanamide

To 3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butyric acid (450mg, 1.98mmol) in SOCl₂To the solution (5.0mL) was added a catalytic amount of DMF. After stirring for 3h, NH was added₃H₂O (173mg, 4.95 mmol). The mixture was stirred for a further 3 h. After removal of volatiles, the residue was extracted with EtOAc, washed with brine, and Na₂SO₄Dried and concentrated. The residue was purified by preparative TLC eluting with ethyl acetate/petroleum ether (1/3 to 1/1) to provide the title compound (372mg, 83%). LC-MS (ESI): m/z 227.1(M + H)⁺。

Step 4-3- (4-amino-3-methyl-1H-pyrazol-1-yl) -3-methylbutanamide

A mixture of 3-methyl-3- (3-methyl-4-nitro-1H-pyrazol-1-yl) butanamide (372mg, 1.64mmol) and 10% Pd/C (20mg) in MeOH (10mL) in H₂Stir under atmosphere for 4 h. After insoluble matter was filtered off, the filtrate was concentrated to give the title compound (306mg, 95%). LC-MS (ESI): m/z 197.1(M + H)⁺。

Step 5-3-methyl-3- (3-methyl-4- (4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-ylamino) -1H- Pyrazol-1-yl) butanamides

To 3- (4-amino-3-methyl-1H-pyrazol-1-yl) -3-methylbutanamide (306)mg, 1.56mmol) in t-BuOH (2mL) was added 2-chloro-N-methyl-5- (trifluoromethyl) pyrimidin-4-amine (329mg, 1.56 mmol). The reaction mixture was heated at 100 ℃ under microwave irradiation for 1 h. After concentration, the residue was purified by preparative TLC eluting with ethyl acetate/petroleum ether (1/3 to 1/1) to provide the title compound (347mg, 60%). LC-MS (ESI): 372.2(M + H) with M/z⁺。

Step 6-3-methyl-3- (3-methyl-4- (4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-ylamino) -1H- Pyrazol-1-yl) butanenitrile

To a solution of 3-methyl-3- (3-methyl-4- (4- (methylamino) -5- (trifluoromethyl) pyrimidin-2-ylamino) -1H-pyrazol-1-yl) butanamide (100mg, 0.270mmol) in DCM (10mL) was added TFA (2mL) dropwise at 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 h. After quenching with ice water, the mixture was extracted with EtOAc, washed with brine, and Na₂SO₄Dried and concentrated. The residue was purified by preparative HPLC to give the title compound (45mg, 47%).¹H NMR(500MHz，CDCl₃)8.12-8.03(m, 2H), 6.69(s, 1H), 5.21(s, 1H), 3.11(s, 3H), 2.98(s, 2H), 2.25(s, 3H), 1.76(s, 6H). LC-MS (method B): m/z 354.3(M + H)⁺5.19min, purity > 99.0%.

The compounds prepared using the above procedure, along with low resolution mass spectrometry (M + H), proton NMR, and LRRK2K determined by the test methods described below for selected compounds_iThe (micromolar concentration) data are shown in table 10 below.

TABLE 4

Example 51: in vitro LRRK2Lanthascreen binding assay

The assay is used to determine Ki_app、IC₅₀Or percent inhibition value to determine the efficacy of the compound in inhibiting the activity of LRRK 2. In 384-well microwell plates F (propiplates F) in black light well plates LRRK2, Eu-anti-GST-antibody, AlexaThe kinase tracer 236 is incubated with the test compound.

Detection of Alexa by addition of Eu-labeled anti-GST antibodyBinding of a "tracer" to a kinase. Binding of the tracer and antibody to the kinase brings about a high degree of FRET, whereas displacement of the tracer by the kinase inhibitor results in loss of FRET.

The assay conditions and materials used were as follows:

final assay conditions：

Material：

Reaction buffer solution: h₂O/50mM Tris，pH 7.4/10mM MgCl₂/1mM EGTA/0.01％Brij35。

Compound plate preparation:

test compounds (10mM starting material) were serially diluted 1: 3.16(20ul +43.2ul) in 100% DMSO. 12pt curve. Each concentration was 1: 33.3(3ul +97ul) diluted in reaction buffer. 5ul was printed onto the assay plate. The final highest concentration tested was 100 uM.

Bulk and blank preparation：

In the reaction buffer, 5ul of DMSO (3%) was added to the bulk and blank wells and 5ul of Eu-labeled anti-GST antibody (6nM) was added to the blank wells. 5ul of LRRK2(30 nM)/Eu-labeled anti-GST antibody (6nM) mix was added to the compounds and all wells.

Measurement procedure：

To all wells 5ul of kinase tracer (25.5nM) was added. The plates were incubated at room temperature for 1 hour on a plate shaker (gently shaking). Read according to the Perkin Elmer EnVision reader HTRF protocol.

Data processing:

calculating the proportion: (665/620)*10000. The average background value was subtracted from all data points. The% of control was calculated for each test value. The% of control is plotted against compound concentration. Ki values were calculated (xlfit curve fitting-Morrison equation). The results are expressed as Ki in. mu.M. The formula for Ki is:

Y＝V0*(1-((x+Ki*(1+S/Km)+Et)/(2*Et)-(((x+Ki*(1+S/Km)+Et)^2-(4*Et*x))^0.5)/(2*Et)))

wherein Et is 4nM

kd (tracer) ═ 8.5nM

Tracer concentration (S) 8.5 nM.

Example 52 in vitro LRRK2 assay

The assay is used to determine Ki_app、IC₅₀Or percent inhibition value to determine the amount of the compoundPotency in inhibiting the activity of LRRK 2. In polypropylene plates, LRRK2, fluorescently labeled peptide substrate, ATP, and test compound were incubated together. Using LabChip 3000(Caliper Life Sciences), after the reaction, the substrate was separated into two populations by capillary electrophoresis: phosphorylated and unphosphorylated. The relative amounts of each are quantified by fluorescence intensity. LRRK2Ki is determined according to the following equation:

Y＝V0*(1-((x+Ki*(1+S/Km)+Et)/(2*Et)-(((x+Ki*(1+S/Km)+Et)^2-(4*Et*x))^0.5)/(2*Et)))。

ki values in table 4 and elsewhere herein are shown in μ M.

The assay conditions and materials used were as follows:

final assay conditions：

LRRK2G2019S at 5mM MgCl₂The method comprises the following steps: 5.2nM (Invitrogen lot #567054A)

LRRK2G2019S in 1mM MnCl₂The method comprises the following steps: 11nM (Invitrogen lot #567054A)

LRRK2 wild type at 5mM MgCl₂The method comprises the following steps: 15nM (Invitrogen lot #500607F)

LRRK2I2020T at 5mM MgCl₂The method comprises the following steps: 25nM (Invitrogen lot #43594)

ATP ^app Kms：

Material：

Solid carrier: black 50 μ L volume polypropylene 384 well plate (Matrical cat # MP101-1-PP)

Kinase enzymes: LRRK2G2019S (Invitrogen cat # PV 4882). LRRK2 wild type (Invitrogen cat # PV 4874).

Substrate: 5FAM-GAGRLGRDKYKTLRQIRQ-CONH₂

A non-bonded plate: a384 well clear V-bottom polypropylene plate (Greiner cat # 781280).

ATP：10mM ATP(Cell Signaling cat#9804)。

Triton X-100：Triton X-100。

Brij-35：Brij-35(Pierce cat#20150)。

Coating reagent # 3: coating reagent #3 (Caliper).

DMSO：DMSO(Sigma cat#34869-100ML)。

Complete reaction buffer: h₂O/25mM Tris，pH 8.0/5mM MgCl₂/2mM DTT/0.01％Triton X-100。

Stopping liquid: h₂O/100mM HEPES, pH 7.2/0.015% Brij-35/0.2% coating reagent #3/20mM EDTA.

Separating buffer solution: h₂O/100mM HEPES, pH 7.2/0.015% Brij-35/0.1% coating reagent # 3/1: 200 coating reagent #8/10mM EDTA/5% DMSO.

Compound plate preparation:

for serial dilutions, 34.6. mu.l DMSO was added to columns 3-24. For assay controls, 37.5. mu.l DMSO was added to columns 1 and 2 of rows A and P, and 50. mu.l 25. mu. M G-028831(Staurosporine) was added to columns 1 and 2 of row B. For the samples: starting at 100. mu.M, 37.5. mu.l DMSO was added to columns 1 and 2, followed by 12.5. mu.l 10mM compound; starting at 10. mu.M, 78. mu.l DMSO was added to columns 1 and 2, followed by 2. mu.l 10mM compound; and starting at 1. mu.M, 25. mu.M compound (2. mu.l of 10mM compound + 798. mu.l DMSO) was added to empty columns 1 and 2. A1: 3.16 dilution series ("PLK _ BM _ serial _ halflog") was performed using precision instrumentation.

ATP preparation：

ATP was diluted to 282.1. mu.M (final concentration of 130. mu.M) in complete kinase buffer.

Bulk and blank preparation：

In complete reaction buffer, the substrate was diluted to 4 μ M. Equal volumes of complete reaction buffer and 4 μ M substrate were combined to obtain a blank. Equal volumes of complete reaction buffer and 4 μ M substrate were combined and 2X final LRRK2 concentration was added to the combined solution.

Measurement procedure：

To a 50. mu.l polypropylene plate, 5. mu.l/well buffer/substrate was added by hand to the blank well. The kinase reaction ("PLK SAR 23 ATP") was initiated using Biomek FX. The following were added to the appropriate wells:

2 μ l compound +23 μ l ATP;

5 μ l/well compound/ATP in assay plate;

5 μ l/well kinase/substrate in assay plate;

the plates were incubated for 2 hours in the dark. The kinase reaction ("PLK Stop") was stopped using Biomek FX and 10. mu.l/well Stop solution was added to the assay plate. The results were read on a LabChip 3000.

Lab Chip 3000 protocol：

The LabChip 3000 was run using the work "LRRK 2IC 50" with the following work settings:

example 53 mouse model of Parkinson's disease

Parkinson's disease can replicate in mice and primates by administering 1-methyl-4-phenyltetrahydropyridine (MPTP), a selective nigrostriatal dopaminergic neurotoxin that produces a loss of striatal Dopamine (DA) nerve terminal markers. The efficacy of the compounds of the invention in the treatment of Parkinson's disease can be evaluated using MPTP-induced neurodegeneration, generally in accordance with the protocol described by Saponito et al, J.Pharmacology (1999) Vol.288, pp.421-427.

Briefly, MPTP was dissolved in PBS at a concentration of 2-4mg/ml and mice (male C57, 20-25g in weight) were injected subcutaneously with 20-40 mg/kg. The compounds of the invention were solubilized with polyethylene glycol hydroxystearate and dissolved in PBS. Mice were given a 10ml/kg solution of the compound by subcutaneous injection 4 to 6h prior to MPTP administration, and then daily for 7 days. On the day of the last injection, mice were sacrificed and midbrain blocked and postfixed (postfix) in paraformaldehyde. Striatum was cut, weighed and stored at-70 ℃.

The striatum thus collected was evaluated for its content of dopamine and its metabolites dihydroxyphenylacetic acid and homovanillic acid by HPLC with electrochemical detection as described by Sonsalla et al, J.Pharmacol.Exp.Ther. (1987) Vol.242, pp.850-857. The striatum may also be assayed using the tyrosine hydroxylase of Okunu et al, Anal Biochem (1987) Vol.129, pp.405-411, by measuring the tyrosine hydroxylase-mediated conversion of labeled tyrosine to L-dopa¹⁴CO₂The deposition was evaluated. The striatum may also be assessed using monoamine oxidase-B as described by White et al, Life Sci, (1984), Vol.35, pp.827-833, and by detecting dopamine uptake as described by Saponito et al, (1992) Vol.260, pp.1400-1409.

While the invention has been described with reference to specific embodiments thereof, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process step or steps, to the objective spirit and scope of the present invention. All such modifications are intended to be within the scope of the appended claims.

Claims

1. A compound selected from the group consisting of:

N4-methyl-N2- (3-methyl-1- (2- (5-methyloxazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (3-chloro-1- (2- (5-methyl-1, 3, 4-oxadiazol-2-yl) propan-2-yl) -1H-pyrazol-4-yl) -N4-ethyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

N4-ethyl-N2- (1- (2- (5-ethyl-1, 3, 4-oxadiazol-2-yl) propan-2-yl) -3-methyl-1H-pyrazol-4-yl) -5- (trifluoromethyl) pyrimidine-2, 4-diamine;

n2- (1- ((3R) -3-fluorotetrahydro-2H-pyran-4-yl) -5-methyl-1H-pyrazol-4-yl) -N4-methyl-5- (trifluoromethyl) pyrimidine-2, 4-diamine;

or a pharmaceutically acceptable salt thereof.

2. A composition, comprising:

(a) a pharmaceutically acceptable carrier; and

(b) the compound of claim 1.

3. A compound according to claim 1 for use as therapeutically active substance.

4. A compound according to claim 1 for use as therapeutically active substance for the therapeutic and/or prophylactic treatment of parkinson's disease.