# Patent

## Title
Aminopyrimidine derivatives as LRRK2 modulators

## Abstract

Compounds of the formula I:




or pharmaceutically acceptable salts thereof,

wherein A, X, R1, R2, R3 and R4 are as defined herein. Also disclosed are methods of making the compounds and using the compounds for treatment of diseases associated with LRRK2 receptor, such as Parkinson's disease.

## Background
CROSS REFERENCE TO RELATED APPLICATION
This Application claims the benefit under 35 USC §119 of U.S. Provisional Application Ser. No. 61/564,759 filed on Nov. 29, 2011, the disclosure of which is incorporated herein by reference.
FIELD OF THE INVENTION
This invention pertains to compounds that modulate the function of LRRK2 and are useful for treatment of LRRK2-mediated diseases and conditions such as Parkinson's disease.
BACKGROUND OF THE INVENTION
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 that afflicts approximately one out of every 1000 people, with hereditary Parkinson's disease accounting for 5-10% of all of patients. Parkinson's disease is caused by progressive loss of mid-brain dopamine neurons, leaving patients with impaired ability to direct and control their movements. The primary Parkinson's disease symptoms are trembling, rigidity, slowness of movement, and impaired balance. Many Parkinson's disease patients also experience other symptoms such as emotional changes, memory loss, speech problems, and sleeping disorders.
The gene encoding the leucine-rich repeat kinase 2 protein (LRRK2) has been identified in association with hereditary Parkinson's disease (Paisan-Ruiz et al., Neuron, Vol. 44(4), 2004, pp 595-600; Zimprich et al., Neuron, Vol. 44(4), 2004, 601-607). In-vitro studies show that Parkinson's disease-associated mutation leads to increased LRRK2 kinase activity and decreased rate of GTP hydrolysis compared to wild-type (Guo et al., Experimental Cell Research, Vol. 313(16), 2007, pp. 3658-3670. Anti-LRRK2 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 Lewie body formation and pathogenesis associated with these diseases (Zhou et al., Molecular Degeneration, 2006, 1:17 doi:10.1186/1750-1326-1-17). LRRK2 has also been identified as a gene potentially associated with increased susceptibility to Crohn's disease and susceptibility to leprosy (Zhang et al., New England J. Med. Vol. 361 (2009) pp. 2609-2618.
LRRK2 has also been associated with the transition of mild cognitive impairment to Alzheimer's disease (WO2007/149789); L-Dopa induced dyskinesia (Hurley et al., Eur. J. Neurosci., Vol. 26, 2007, pp. 171-177; CNS disorders associated with neuronal progenitor differentiation (Milosevic et al., Neurodegen., Vol. 4, 2009, p. 25); cancers such as kidney, breast, prostate, blood and lung cancers and acute myelogenous leukemia (WO2011/038572); papillary renal and thyroid carcinomas (Looyenga et al., www.pnas.org/cgi/doi/10.1073/pnas.1012500108); multiple myeloma (Chapman et al., Nature Vol. 471, 2011, pp. 467-472); amyotrophic lateral sclerosis (Shtilbans et al., Amyotrophic Lateral Sclerosis “Early Online 2011, pp. 1-7); rheumatoid arthritis (Nakamura et al., DNA Res. Vol. 13(4), 2006, pp. 169-183); and ankylosing spondylytis (Danoy et al., PLoS Genetics, Vol. 6(12), 2010, e1001195, pp. 1-5).
Accordingly, compounds and compositions effective at modulating LRRK2 activity may provide a treatment for neurodegenerative diseases such as Parkinson's disease and Lewie body dementia, for CNS disorders such as Alzheimer's disease and L-Dopa induced dyskinesia, for cancers such as kidney, breast, prostate, blood, papillary and lung cancers, acute myelogenous leukemia and multiple myeloma, and for inflammatory diseases such as leprosy, Crohn's disease, amyotrophic lateral sclerosis, rheumatoid arthritis, and ankylosing spondylytis. Particularly, there is a need for compounds with LRRK2 affinity that are selective for LRRK2 over other kinases, such as JAK2, and which can provide effective drugs for treatment of neurodegenerative disorders such as Parkinson's disease.
SUMMARY OF THE INVENTION
The invention provides compounds of the formula I:




or pharmaceutically acceptable salts thereof,

wherein:

A is a five- or six-membered saturated or unsaturated ring that includes one or two heteroatoms selected from O, N and S, which is substituted once with R5, and which is optionally substituted one, two or three times with R6;
X is: —NRa—; —O—; or —S(O)r— wherein r is from 0 to 2 and Ra is hydrogen or C1-6alkyl;
R1 is: C1-6alkyl; C1-6alkenyl; C1-6alkynyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; hydroxy-C1-6alkyl; amino-C1-6alkyl; C1-6alkylsulfonyl-C1-6alkyl; C3-6cycloalkyl optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; oxetanyl; or oxetan-C1-6alkyl;
or R1 and Ra together with the atoms to which they are attached may form a three to six membered ring that may optionally include an additional heteroatom selected from O, N and S, and which is substituted with oxo, halo or C1-6alkyl;
R2 is: halo; C1-6alkoxy; cyano; C1-6alkynyl; C1-6alkenyl; halo-C1-6alkyl; halo-C1-6alkoxy; C3-6cycloalkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; acetyl; oxetanyl; or oxetan-C1-6alkyl;
one of R3 and R4 is: halo; C1-6alkyl; C1-6alkoxy; C3-6cycloalkyloxy; halo-C1-6alkyl; or halo-C1-6alkoxy, and the other is hydrogen;
R5 is; oxo; C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; or —C(O)—NRbRc wherein Rb and Rc each independently is hydrogen or —C1-6alkyl, or Rb and Rc together with the atoms to which they are attached may form a heterocyclyl group that optionally includes an additional heteroatom selected from O, N and S and which is optionally substituted one or more times with R6; and
each R6 is independently: C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; oxo; or —C(O)—NRbRc.
The invention also provides and pharmaceutical compositions comprising the compounds, methods of using the compounds, and methods of preparing the compounds.


DETAILED DESCRIPTION OF THE INVENTION
Definitions
Unless otherwise stated, the following terms used in this 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” means the monovalent linear or branched saturated hydrocarbon moiety, consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms. “Lower alkyl” refers to an alkyl group of one to six carbon atoms, i.e. C1-C6alkyl. 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 linear monovalent hydrocarbon radical of two to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms, containing at least one double bond, e.g., ethenyl, propenyl, and the like.
“Alkynyl” means a linear monovalent hydrocarbon radical of two to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbon atoms, containing at least one triple bond, e.g., ethynyl, propynyl, and the like.
“Alkylene” means a linear saturated divalent hydrocarbon radical of one to six carbon atoms or a branched saturated divalent hydrocarbon radical of three to six carbon atoms, e.g., methylene, ethylene, 2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene, and the like.
“Alkoxy” and “alkyloxy”, which may be used interchangeably, mean 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 moiety of the formula Ra—O—Rb—, where Ra is alkyl and Rb is alkylene as defined herein. Exemplary alkoxyalkyl groups include, by way of example, 2-methoxyethyl, 3-methoxypropyl, 1-methyl-2-methoxyethyl, 1-(2-methoxyethyl)-3-methoxypropyl, and 1-(2-methoxyethyl)-3-methoxypropyl.
“Alkoxyalkoxy” means a group of the formula —O—R—R′ wherein R is alkylene and R′ is alkoxy as defined herein.
“Alkylcarbonyl” means a moiety of the formula —C(O)—R, wherein R is alkyl as defined herein.
“Alkoxycarbonyl” means a group of the formula —C(O)—R wherein R is alkoxy as defined herein.
“Alkylcarbonylalkyl” means a group of the formula —R—C(O)—R wherein R is alkylene and R′ is alkyl as defined herein.
“Alkoxycarbonylalkyl” means a group of the formula —R—C(O)—R wherein R is alkylene and R′ is alkoxy as defined herein.
“Alkoxycarbonylalkoxy” means a group of the formula —O—R—C(O)—R′ wherein R is alkylene and R′ is alkoxy as defined herein.
“Hydroxycarbonylalkoxy” means a group of the formula —O—R—C(O)—OH wherein R is alkylene as defined herein.
“Alkylaminocarbonylalkoxy” means a group of the formula —O—R—C(O)—NHR′ wherein R is alkylene and R′ is alkyl as defined herein.
“Dialkylaminocarbonylalkoxy” means a group of the formula —O—R—C(O)—NR′R′′ wherein R is alkylene and R′ and R′′ are alkyl as defined herein.
“Alkylaminoalkoxy” means a group of the formula —O—R—NHR′ wherein R is alkylene and R′ is alkyl as defined herein.
“Dialkylaminoalkoxy” means a group of the formula —O—R—NR′R′ wherein R is alkylene and R′ and R′′ are alkyl as defined herein.
“Alkylsulfonyl” means a moiety of the formula —SO2—R, wherein R is alkyl as defined herein.
“Alkylsulfonylalkyl means a moiety of the formula —R′—SO2—R′′ where where R′ is alkylene and R′′ is alkyl as defined herein.
“Alkylsulfonylalkoxy” means a group of the formula —O—R—SO2—R′ wherein R is alkylene and R′ is alkyl as defined herein.
“Amino means a moiety of the formula —NRR′ wherein R and R′ each independently is 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” means a group of the formula —C(O)—R wherein R is amino as defined herein.
“Alkoxyamino” means a moiety of the formula —NR—OR′ wherein R is hydrogen or alkyl and R′ is alkyl as defined herein.
“Alkylsulfanyl” means a moiety of the formula —SR wherein R is alkyl as defined herein.
“Aminoalkyl” means a group —R—R′ wherein R′ is amino and R is 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” means a group —OR—R′ wherein R′ is amino and R is alkylene as defined herein.
“Alkylsulfonylamido” means a moiety of the formula —NR′SO2—R wherein R is alkyl and R′ is hydrogen or alkyl.
“Aminocarbonyloxyalkyl” or “carbamylalkyl” means a group of the formula —R—O—C(O)—NR′R′′ wherein R is alkylene and R′, R′′ each independently is hydrogen or alkyl as defined herein.
“Alkynylalkoxy” means a group of the formula —O—R—R′ wherein R is alkylene and R′ is alkynyl as defined herein.
“Aryl” means a monovalent cyclic aromatic hydrocarbon moiety consisting of a mono-, bi- or tricyclic aromatic ring. The aryl group can 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, diphenylsulfidyl, diphenylsulfonyl, diphenylisopropylidenyl, benzodioxanyl, benzofuranyl, benzodioxylyl, benzopyranyl, benzoxazinyl, benzoxazinonyl, benzopiperadinyl, benzopiperazinyl, benzopyrrolidinyl, benzomorpholinyl, methylenedioxyphenyl, ethylenedioxyphenyl, and the like, of which may be optionally substituted as defined herein.
“Arylalkyl” and “Aralkyl”, which may be used interchangeably, mean a radical-RaRb where Ra is an alkylene group and Rb is an aryl group as defined herein; e.g., phenylalkyls such as benzyl, phenylethyl, 3-(3-chlorophenyl)-2-methylpentyl, and the like are examples of arylalkyl.
“Arylsulfonyl means a group of the formula —SO2—R wherein R is aryl as defined herein.
“Aryloxy” means a group of the formula —O—R wherein R is aryl as defined herein.
“Aralkyloxy” means a group of the formula —O—R—R′′ wherein R is alkylene and R′ is aryl as defined herein.
“Carboxy” or “hydroxycarbonyl”, which may be used interchangeably, means a group of the formula —C(O)—OH.
“Cyanoalkyl” means a moiety of the formula —R′—R′′, where R′ is alkylene as defined herein and R′′ is cyano or nitrile.
“Cycloalkyl” means a monovalent saturated carbocyclic moiety consisting of mono- or bicyclic rings. Particular cycloalkyl are unsubstituted or substituted with alkyl. Cycloalkyl can optionally be substituted as defined herein. Unless defined otherwise, cycloalkyl may be optionally substituted with one or more substituents, wherein each substituent is independently hydroxy, alkyl, alkoxy, halo, haloalkyl, amino, monoalkylamino, or dialkylamino. 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” means a moiety of the formula —R′—R′′, where R′ is alkylene and R′′ is cycloalkyl as defined herein.
“Cycloalkylalkoxy” means a group of the formula —O—R—R′ wherein R is alkylene and R′ is cycloalkyl as defined herein.
“Heteroaryl” means a monocyclic or bicyclic radical 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, with the understanding that the attachment point of the heteroaryl radical will be on an 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, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, benzothienyl, thiophenyl, furanyl, pyranyl, pyridyl, pyrrolyl, pyrazolyl, pyrimidyl, quinolinyl, isoquinolinyl, benzofuryl, benzothiophenyl, benzothiopyranyl, benzimidazolyl, benzooxazolyl, benzooxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranyl, indolyl, isoindolyl, triazolyl, triazinyl, quinoxalinyl, purinyl, quinazolinyl, quinolizinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl and the like, each of which may be optionally substituted as defined herein.
“Heteroarylalkyl” or “heteroaralkyl” means a group of the formula —R—R′ wherein R is alkylene and R′ is heteroaryl as defined herein.
“Heteroarylsulfonyl means a group of the formula —SO2—R wherein R is heteroaryl as defined herein.
“Heteroaryloxy” means a group of the formula —O—R wherein R is heteroaryl as defined herein.
“Heteroaralkyloxy” means a group of the formula —O—R—R′′ wherein R is alkylene and R′ is heteroaryl as defined herein.
The terms “halo”, “halogen” and “halide”, which may be used interchangeably, refer to a substituent fluoro, chloro, bromo, or iodo.
“Haloalkyl” means alkyl as defined herein in which one or more hydrogen has been replaced with same or different halogen. Exemplary haloalkyls include —CH2Cl, —CH2CF3, —CH2CCl3, perfluoroalkyl (e.g., —CF3), and the like.
“Haloalkoxy” means a moiety of the formula —OR, wherein R is a haloalkyl moiety as defined herein. An exemplary haloalkoxy is difluoromethoxy.
“Heterocycloamino” means 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” means a monovalent saturated moiety, consisting of one to three rings, incorporating one, two, or three or four heteroatoms (chosen 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, morpholinyl, thiomorpholinyl, azepinyl, pyrrolidinyl, azetidinyl, tetrahydropyranyl, tetrahydrofuranyl, oxetanyl and the like. Such heterocyclyl may be optionally substituted as defined herein.
“Heterocyclylalkyl” means a moiety of the formula —R—R′ wherein R is alkylene and R′ is heterocyclyl as defined herein.
“Heterocyclyloxy” means a moiety of the formula —OR wherein R is heterocyclyl as defined herein.
“Heterocyclylalkoxy” means a moiety of the formula —OR—R′ wherein R is alkylene and R′ is heterocyclyl as defined herein.
“Heterocyclylsulfonyl” means a group of formula —SO2—R wherein R is heterocyclyl as defined herein.
“Hydroxyalkoxy” means a moiety of the formula —OR wherein R is hydroxyalkyl as defined herein.
“Hydroxyalkylamino” means a moiety of the formula —NR—R′ wherein R is hydrogen or alkyl and R′ is hydroxyalkyl as defined herein.
“Hydroxyalkylaminoalkyl” means 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” means a group of the formula —R—(CO)—OH where R is alkylene as defined herein.
“Hydroxycarbonylalkoxy” means a group of the formula —O—R—C(O)—OH wherein R is alkylene as defined herein.
“Hydroxyalkyloxycarbonylalkyl” or “hydroxyalkoxycarbonylalkyl” means a group of the formula —R—C(O)—O—R—OH wherein each R is alkylene and may be the same or different.
“Hydroxyalkyl” means an alkyl moiety as defined herein, substituted with one or more, for example, one, two or three hydroxy groups, provided that the same carbon atom does not carry more than one hydroxy 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” means a cycloalkyl moiety as defined herein wherein one, two or three hydrogen atoms in the cycloalkyl radical have been replaced with a hydroxy substituent. Representative examples include, but are not limited to, 2-, 3-, or 4-hydroxycyclohexyl, and the like.
“Alkoxy hydroxyalkyl” and “hydroxy alkoxyalkyl”, which may be used interchangeably, means an alkyl as defined herein that is substituted at least once with hydroxy and at least once with alkoxy. “Alkoxy hydroxyalkyl” and “hydroxy alkoxyalkyl” thus encompass, for example, 2-hydroxy-3-methoxy-propan-1-yl and the like.
“Urea” or “ureido” means a group of the formula —NR′—C(O)—NR′′R′′′ wherein R′, R′′ and R′′′ each independently is hydrogen or alkyl.
“Carbamate” means a group of the formula —O—C(O)—NR′R′′ wherein R′ and R′′ each independently is hydrogen or alkyl.
“Carboxy” means a group of the formula —O—C(O)—OH.
“Sulfonamido” means a group of the formula —SO2—NR′R′′ wherein R′, R′′ and R′′′ each independently is hydrogen or alkyl.
“Optionally substituted” when used in association with an “aryl”, phenyl”, “heteroaryl” “cycloalkyl” or “heterocyclyl” moiety means that such moiety may be unsubstituted (i.e., all open valencies are occupied by a hydrogen atom) or substituted with specific groups as related herein.
“Leaving group” means the group with the meaning conventionally associated with it 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, alkane- or arylenesulfonyloxy, such as methanesulfonyloxy, ethanesulfonyloxy, thiomethyl, benzenesulfonyloxy, tosyloxy, and thienyloxy, dihalophosphinoyloxy, optionally substituted benzyloxy, isopropyloxy, acyloxy, and the like.
“Modulator” means a molecule that interacts with a target. The interactions include, but are not limited to, agonist, antagonist, 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 the event or circumstance occurs and instances in which it does not.
“Disease” and “Disease state” means any disease, condition, symptom, disorder or indication.
“Inert organic solvent” or “inert solvent” means the solvent is inert under the conditions of the reaction being described in conjunction therewith, including for example, benzene, toluene, acetonitrile, tetrahydrofuran, N,N-dimethylformamide, chloroform, methylene chloride or dichloromethane, dichloroethane, diethyl ether, ethyl acetate, acetone, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol, tert-butanol, dioxane, pyridine, and the like. Unless specified to the contrary, the solvents used in the reactions of the present invention are inert solvents.
“Pharmaceutically acceptable” means that which is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable and includes that which is acceptable for veterinary as well as human pharmaceutical use.
“Pharmaceutically acceptable salts” of a compound means salts that are pharmaceutically acceptable, as defined herein, and that possess the desired pharmacological activity of the parent compound.
It should be understood that all references to pharmaceutically acceptable salts include solvent addition forms (solvates) or crystal forms (polymorphs) as defined herein, of the same acid addition salt.
“Protective group” or “protecting group” means the group which selectively blocks one reactive site in a multifunctional compound such that a chemical reaction can be carried out selectively at another unprotected reactive site in the meaning conventionally associated with it in synthetic chemistry. Certain processes of this invention rely upon the protective 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 the nitrogen atom against undesirable reactions during synthetic procedures. Exemplary nitrogen protecting groups include, but are not limited to, trifluoroacetyl, acetamido, benzyl (Bn), benzyloxycarbonyl (carbobenzyloxy, CBZ), p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, tert-butoxycarbonyl (BOC), and the like. The artisan in the art will know how to chose a group for the ease of removal and for the ability to withstand the following reactions.
“Solvates” means solvent additions forms that contain either stoichiometric or non stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one of the substances in which the water retains its molecular state as H2O, such combination being able to form one or more hydrate.
“Parkinson's disease” means 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 rigidity, tremor, slowing of physical movement (bradykinesia) and loss of physical movement (akinesia).
“Lewie body disease” also called “Lewie body dementia”, diffuse Lewy body disease”, cortical Lewie body disease”, means a neurogenerative disorder characterized anatomically by the presence of Lewie bodies in the brain.
“Subject” means mammals and non-mammals. Mammals means any member of the mammalia class including, but not limited to, humans; non-human primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, and 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 sex.
“Therapeutically effective amount” means an amount of a compound that, when administered to a subject for treating a disease state, is sufficient to effect such treatment for the disease state. The “therapeutically effective amount” will vary depending on the compound, disease state being treated, the severity or the disease 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.
The terms “those defined above” and “those defined herein” when referring to a variable incorporates by reference the broad definition of the variable as well as particular 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 relieving the disease state, i.e., causing temporary or permanent regression of the disease state or its clinical symptoms.
The terms “treating”, “contacting” and “reacting” when referring to a chemical reaction means adding or mixing two or more reagents under appropriate conditions to produce the indicated and/or the desired product. It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product.
Nomenclature and Structures
In general, the nomenclature and chemical names used in this Application are based on ChembioOfficeTM by CambridgeSoftTM. Any open valency appearing on a carbon, oxygen sulfur or nitrogen atom in the structures herein indicates the presence of a hydrogen atom unless indicated otherwise. Where a nitrogen-containing heteroaryl ring is shown with an open valency on a nitrogen atom, and variables such as Ra, Rb or Rc are shown on the heteroaryl ring, such variables may be bound or joined to the open valency nitrogen. Where a chiral center exists in a structure but no specific stereochemistry is shown for the chiral center, both enantiomers associated with the chiral center are encompassed by the structure. Where a structure shown herein may 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 naturally occurring isotopes of such atoms. Thus, for example, the hydrogen atoms represented herein are meant to include deuterium and tritium, and the carbon atoms are meant to include C13 and C14 isotopes.
Compounds of the Invention
The invention provides compounds of formula I:




or pharmaceutically acceptable salts thereof,

wherein:

A is a five- or six-membered saturated or unsaturated ring that includes one or two heteroatoms selected from O, N and S, which is substituted once with R5, and which is optionally substituted one, two or three times with R6;
X is: —NRa—; —O—; or —S(O)r— wherein r is from 0 to 2 and Ra is hydrogen or C1-6alkyl;
R1 is: C1-6alkyl; C1-6alkenyl; C1-6alkynyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; hydroxy-C1-6alkyl; amino-C1-6alkyl; C1-6alkylsulfonyl-C1-6alkyl; C3-6cycloalkyl optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; oxetanyl; or oxetan-C1-6alkyl;
or R1 and Ra together with the atoms to which they are attached may form a three to six membered ring that may optionally include an additional heteroatom selected from O, N and S, and which is substituted with oxo, halo or C1-6alkyl;
R2 is: halo; C1-6alkoxy; cyano; C1-6alkynyl; C1-6alkenyl; halo-C1-6alkyl; halo-C1-6alkoxy; C3-6cycloalkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; acetyl; oxetanyl; or oxetan-C1-6alkyl;

    
    
        one of R3 and R4 is: halo; C1-6alkyl; C1-6alkoxy; C3-6cycloalkyloxy; halo-C1-6alkyl; or halo-C1-6alkoxy, and the other is hydrogen;
    
    


R5 is; oxo; C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; or —C(O)—NRbRc wherein Rb and Rc each independently is hydrogen or —C1-6alkyl, or Rb and Rc together with the atoms to which they are attached may form a heterocyclyl group that optionally includes an additional heteroatom selected from O, N and S and which is optionally substituted one or more times with R6; and

    
    
        each R6 is independently: C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; oxo; or —C(O)—NRbRc.
    
    


In certain embodiments of formula I, R1 and Ra together with the atoms to which they are attached may form a three to six membered ring that may optionally include an additional heteroatom selected from O, N and S, and which may be optionally substituted with oxo, halo or C1-6alkyl.
In certain embodiments of formula I, R1 and Ra together with the atoms to which they are attached form a five or six membered ring.
In certain embodiments of formula I, R1 and Ra together with the atoms to which they are attached form a pyrrolidinyl, piperidinyl or oxazoladinonyl group.
In certain embodiments of formula I, R2 is acetyl.
In certain embodiments of formula I, when R1 is cyclopropyl, cyclobutyl, cyclopropyl-C1-6alkyl or cyclobutyl-C1-6alkyl, then X is —O—.
In certain embodiments of formula I, r is 0.
In certain embodiments of formula I, r is 2.
In certain embodiments of formula I, X is —NRa— or —O—.
In certain embodiments of formula I, X is —NRa.
In certain embodiments of formula I, X is —O—.
In certain embodiments of formula I, X is —S(O)n—.
In certain embodiments of formula I, X is —NH— or —O—.
In certain embodiments of formula I, Ra is hydrogen.
In certain embodiments of formula I, Ra is C1-6alkyl.
In certain embodiments of formula I, R1 is: C1-6alkyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; amino-C1-6alkyl; C1-6alkylsulfonyl-C1-6alkyl; C3-6cycloalkyl; or C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R1 is: C1-6alkyl; C3-6cycloalkyl optionally substituted with C1-6alkyl; or C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl.
In certain embodiments of formula I, R1 is: C1-6alkyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; amino-C1-6alkyl; C1-6alkylsulfonyl-C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; oxetanyl; or oxetan-C1-6alkyl.
In certain embodiments of formula I, R1 is: C1-6alkyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; amino-C1-6alkyl; or C1-6alkylsulfonyl-C1-6alkyl.
In certain embodiments of formula I, R1 is C1-6alkyl.
In certain embodiments of formula I, R1 is halo-C1-6alkyl.
In certain embodiments of formula I, R1 is C1-6alkoxy-C1-6alkyl.
In certain embodiments of formula I, R1 is amino-C1-6alkyl.
In certain embodiments of formula I, R1 is C1-6alkylsulfonyl-C1-6alkyl optionally substituted with C1-6alkyl.
In certain embodiments of formula I, R1 is C3-6cycloalkyl optionally substituted with C1-6alkyl.
In certain embodiments of formula I, R1 is C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl.
In certain embodiments of formula I, R1 is tetrahydrofuranyl.
In certain embodiments of formula I, R1 is tetrahydrofuranyl-C1-6alkyl; oxetanyl.
In certain embodiments of formula I, R1 is or oxetan-C1-6alkyl.
In certain embodiments of formula I, R1 is: methyl; ethyl; n-propyl; isopropyl; isobutyl; 3,3-dimethylpropyl; cyclopropyl; cyclobutyl; cyclopentyl; cyclohexyl; cyclopropylmethyl; cyclobutylmethyl; cyclopentylmethyl; cyclopropylethyl; methoxyethyl; oxetanyl; or tetrahydrofuranylmethyl.
In certain embodiments of formula I, R1 is: methyl; ethyl; n-propyl; isopropyl; isobutyl; 3,3-dimethylpropyl; cyclopentyl; cyclohexyl; cyclopropylmethyl; cyclobutylmethyl; cyclopentylmethyl; cyclopropylethyl; methoxyethyl; oxetanyl; or tetrahydrofuranylmethyl.
In certain embodiments of formula I, R1 is: methyl; ethyl; n-propyl; isopropyl; isobutyl; 3,3-dimethylpropyl; cyclopentyl; cyclohexyl; cyclopentylmethyl; methoxyethyl; oxetanyl; or tetrahydrofuranylmethyl.
In certain embodiments of formula I, R1 is: methyl; ethyl; n-propyl; isopropyl; or isobutyl.
In certain embodiments of formula I, R1 is methyl or ethyl.
In certain embodiments of formula I, R1 is methyl.
In certain embodiments of formula I, R1 is ethyl.
In certain embodiments of formula I, R1 is: cyclopropyl; cyclobutyl; cyclopentyl; cyclohexyl; cyclopropylmethyl; cyclobutylmethyl; cyclopentylmethyl; or cyclopropylethyl.
In certain embodiments of formula I, R1 is: cyclopentyl; cyclohexyl; or cyclopentylmethyl.
In certain embodiments of formula I, R2 is: halo; C1-6alkoxy; halo-C1-6alkyl; halo-C1-6alkoxy; C3-6cycloalkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; oxetanyl; or oxetan-C1-6alkyl.
In certain embodiments of formula I, R2 is: halo; C1-6alkoxy; halo-C1-6alkyl; cyano; C1-6alkynyl; C1-6alkenyl; C3-6cycloalkyl; or C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R2 is: halo; C1-6alkoxy; halo-C1-6alkyl; cyano; C3-6cycloalkyl; or C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R2 is: halo; C1-6alkoxy; halo-C1-6alkyl; C3-6cycloalkyl; or C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R2 is: halo; halo-C1-6alkyl; or cyano.
In certain embodiments of formula I, R2 is: halo; or halo-C1-6alkyl.
In certain embodiments of formula I, R2 is halo.
In certain embodiments of formula I, R2 is C1-6alkoxy.
In certain embodiments of formula I, R2 is halo-C1-6alkoxy.
In certain embodiments of formula I, R2 is halo-C1-6alkyl.
In certain embodiments of formula I, R2 is C3-6cycloalkyl.
In certain embodiments of formula I, R2 is C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R2 is tetrahydrofuranyl.
In certain embodiments of formula I, R2 is tetrahydrofuranyl-C1-6alkyl.
In certain embodiments of formula I, R2 is oxetanyl.
In certain embodiments of formula I, R2 is oxetan-C1-6alkyl.
In certain embodiments of formula I, R2 is halo, trifluoromethyl or cyano.
In certain embodiments of formula I, R2 is chloro, trifluoromethyl or cyano.
In certain embodiments of formula I, R2 is chloro or trifluoromethyl.
In certain embodiments of formula I, R2 is fluoro, chloro or bromo.
In certain embodiments of formula I, R2 is chloro.
In certain embodiments of formula I, R2 is fluoro.
In certain embodiments of formula I, R2 is bromo.
In certain embodiments of formula I, R2 is trifluoromethyl.
In certain embodiments of formula I, R2 is methoxy.
In certain embodiments of formula I, R2 is cyano.
In certain embodiments of formula I, R2 is C1-6alkynyl.
In certain embodiments of formula I, R2 is C1-6alkenyl.
In certain embodiments of formula I, R3 is: hydrogen.
In certain embodiments of formula I, R3 is: C1-6alkyl.
In certain embodiments of formula I, R3 is halo.
In certain embodiments of formula I, R3 is C1-6alkyl.
In certain embodiments of formula I, R3 is C1-6alkoxy.
In certain embodiments of formula I, R3 is halo or C1-6alkoxy.
In certain embodiments of formula I, R3 is C3-6cycloalkyloxy.
In certain embodiments of formula I, R3 is halo-C1-6alkyl.
In certain embodiments of formula I, R3 is halo-C1-6alkoxy.
In certain embodiments of formula I, R3 is halo or methoxy.
In certain embodiments of formula I, R3 is fluoro, chloro or methoxy.
In certain embodiments of formula I, R3 is fluoro or chloro.
In certain embodiments of formula I, R3 is methoxy.
In certain embodiments of formula I, R3 is methyl
In certain embodiments of formula I, R3 is chloro.
In certain embodiments of formula I, R3 is fluoro.
In certain embodiments of formula I, R4 is: hydrogen.
In certain embodiments of formula I, R4 is: C1-6alkyl;
In certain embodiments of formula I, R4 is halo.
In certain embodiments of formula I, R4 is C1-6alkyl.
In certain embodiments of formula I, R4 is C1-6alkoxy.
In certain embodiments of formula I, R4 is halo-C1-6alkyl.
In certain embodiments of formula I, R4 is halo-C1-6alkoxy.
In certain embodiments of formula I, R4 is halo or methoxy.
In certain embodiments of formula I, R4 is R4 is fluoro, chloro, methyl or methoxy.
In certain embodiments of formula I, R4 is fluoro, chloro or methoxy.
In certain embodiments of formula I, R4 is fluoro or chloro.
In certain embodiments of formula I, R4 is methoxy.
In certain embodiments of formula I, R4 is methyl
In certain embodiments of formula I, R4 is chloro.
In certain embodiments of formula I, R4 is fluoro.
In certain embodiments of formula I, R4 is C3-6cycloalkyloxy.
In certain embodiments of formula I, R5 is: oxo; C1-6alkyl; or —C(O)—NRbRc.
In certain embodiments of formula I, R5 is oxo.
In certain embodiments of formula I, R5 is C1-6alkyl.
In certain embodiments of formula I, R5 is C3-6cycloalkyl.
In certain embodiments of formula I, R5 is C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R5 is —C(O)—NRbRc.
In certain embodiments of formula I, each R6 is independently: C1-6alkyl, hydroxy-C1-6alkyl, C1-6alkoxy-C1-6alkyl, heterocyclyl, oxo, or —C(O)—NRbRc.
In certain embodiments of formula I, R6 is C1-6alkyl.
In certain embodiments of formula I, R6 is C3-6cycloalkyl.
In certain embodiments of formula I, R6 is C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula I, R6 is halo.
In certain embodiments of formula I, R6 is halo-C1-6alkyl.
In certain embodiments of formula I, R6 is hydroxy-C1-6alkyl.
In certain embodiments of formula I, R6 is C1-6alkoxy-C1-6alkyl.
In certain embodiments of formula I, R6 is heterocyclyl.
In certain embodiments of formula I, R6 is oxo.
In certain embodiments of formula I, R6 is —C(O)—NRbRc.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form a heterocyclyl group that optionally includes an additional heteroatom selected from O, N and S and which is optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, azetidinyl, azepinyl, oxazepinyl or diazepinyl, each optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form piperidinyl, piperazinyl, or morpholinyl, each optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form piperidinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form piperazinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form morpholinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form thiomorpholinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form pyrrolidinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form azetidinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form azepinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form oxazepinyl optionally substituted one or more times with R6.
In certain embodiments of formula I, Rb and Rc together with the atoms to which they are attached form diazepinyl, optionally substituted one or more times with R6.
In embodiments of the invention wherein R6 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be azepinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, pyrrolidinyl, tetrahydrofuranyl, azetidinyl or oxetanyl.
In embodiments of the invention wherein R6 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, azetidinyl, azepinyl, oxazepinyl, or pyrrolidinyl.
In embodiments of the invention wherein R6 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl.
In certain embodiments of formula I, R6 is oxetanyl.
In certain embodiments of the invention, the subject compounds are of formula IIa or IIb:




wherein:

m is: 0; 1 or 2;
Z is; —C(R7)2—; —NR8— or —O—; or Z is absent;
each R7 is independently: hydrogen; or C1-6alkyl; and
each R8 is independently: hydrogen; C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; or —C(O)—NRbRc; and
X, R1, R2, R3 and R4 are as defined herein for formula I.
In certain embodiments, the compounds are of formula IIa.
In certain embodiments, the compounds are of formula IIb.
In certain embodiments of formula IIa and IIb, when Z is absent, m is 1 or 2.
In certain embodiments of formula IIa and IIb, m is 0.
In certain embodiments of formula IIa and IIb, m is 1
In certain embodiments of formula IIa and IIb, m is 2.
In certain embodiments of formula IIa and IIb, m is 0 or 1
In certain embodiments of formula IIa and IIb, m is 1 or 2.
In certain embodiments of formula IIa and IIb, Z is —C(R7)2—.
In certain embodiments of formula IIa and IIb, Z is —NR8—.
In certain embodiments of formula IIa and IIb, Z is —O—.
In certain embodiments of formula IIa and IIb, Z is absent.
In certain embodiments of formula IIa and IIb, R7 is hydrogen.
In certain embodiments of formula IIa and IIb, R7 is C1-6alkyl.
In certain embodiments of formula IIa and IIb, each R8 is independently: C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; or —C(O)—NRbRc.
In certain embodiments of formula IIa and IIb, each R8 is independently: C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; oxetanyl; or —C(O)—NRbRc.
In certain embodiments of formula IIa and IIb, R8 is C1-6alkyl.
In certain embodiments of formula IIa and IIb, R8 is C3-6cycloalkyl.
In certain embodiments of formula IIa and IIb, R8 is C3-6cycloalkyl-C1-6alkyl.
In certain embodiments of formula IIa and IIb, R8 is halo.
In certain embodiments of formula IIa and IIb, R8 is halo-C1-6alkyl.
In certain embodiments of formula IIa and IIb, R8 is hydroxy-C1-6alkyl.
In certain embodiments of formula IIa and IIb, R8 is C1-6alkoxy-C1-6alkyl.
In certain embodiments of formula IIa and IIb, R8 is heterocyclyl.
In certain embodiments of formula IIa and IIb, R8 is —C(O)—NRbRc.
In embodiments of the invention wherein R8 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be azepinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, pyrrolidinyl, tetrahydrofuranyl, azetidinyl or oxetanyl.
In embodiments of the invention wherein R8 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, azetidinyl, azepinyl, oxazepinyl, or pyrrolidinyl.
In embodiments of the invention wherein R8 is heterocyclyl or contains a heterocyclyl moiety, such heterocyclyl may be piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl.
In certain embodiments of formula I, R8 is oxetanyl.
In certain embodiments of the invention, the subject compounds are of formula IIIa or IIIb:




wherein:

R9 is: hydrogen; C1-6alkyl; C3-6cycloalkyl; or C3-6cycloalkyl-C1-6alkyl; and
X, R1, R2, R3, Rb and Rc are as defined herein for formula I.
In certain embodiments of the invention, the subject compounds are of formula IIIa.
In certain embodiments of the invention, the subject compounds are of formula IIIb.
In certain embodiments of formula IIIa and IIIb, R9 is: hydrogen; or C1-6alkyl.
The invention also provides a method for treating a disease or condition mediated by or otherwise associated with the LRRK2 receptor, the method comprising administering to a subject in need thereof an effective amount of a compound of the invention.
The disease may be a neurodegenerative disease such as Parkinson's disease, Huntington's disease or Lewie body dementia.
The disease may be a CNS disorder such as Alzheimer's disease and L-Dopa induced dyskinesia.
The disease may be a cancer or proliferative disorder such as kidney, breast, prostate, blood, papillary or lung cancer, acute myelogenous leukemia, or multiple myeloma.
The disease may be an inflammatory disease such as leprosy, Crohn's disease, amyotrophic lateral sclerosis, rheumatoid arthritis, and ankylosing spondylytis.
The invention also provides a method for enhancing cognitive memory, the method comprising administering to a subject in need thereof an effective amount of a compound of the invention.
Representative compounds in accordance with the methods of the invention are shown in the experimental examples below.
Synthesis
Compounds of the present invention can be made by a variety of methods depicted in the illustrative synthetic reaction schemes shown and described below.
The starting materials and reagents used in preparing these compounds generally are either available from commercial suppliers, such as Aldrich Chemical Co., or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser's Reagents for Organic Synthesis; Wiley & Sons: New York, 1991, Volumes 1-15; Rodd's Chemistry of Carbon Compounds, Elsevier Science Publishers, 1989, Volumes 1-5 and Supplementals; and Organic Reactions, Wiley & Sons: New York, 1991, Volumes 1-40. The following synthetic reaction schemes are merely illustrative of some methods by which the compounds of the present invention can be synthesized, and various modifications to these synthetic reaction schemes can be made and will be suggested to one skilled in the art having referred to the disclosure contained in this Application.
The starting materials and the intermediates of the synthetic reaction schemes can be isolated and purified if desired using conventional techniques, including but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such materials can be characterized using conventional means, including physical constants and spectral data.
Unless specified to the contrary, the reactions described herein may be conducted under an inert atmosphere at atmospheric pressure at a reaction temperature range of from about −78° C. to about 150° C., for example, from about 0° C. to about 125° C., or conveniently at about room (or ambient) temperature, e.g., about 20° C.
Scheme A below illustrates one synthetic procedure usable to prepare specific compounds of formula I or formula II, wherein X, m, R1, R2, R3, R4 and R5 are as defined herein.




In step 1 of Scheme A, dichloropyrimidine compound a is reacted with reagent b to afford pyrimidine compound c. The reaction of step 1 may take place under polar solvent conditions. In embodiments of the invention where X is —O— (i.e., reagent b is an alcohol), the reaction of step 1 may be carried out in the presence of base.
In step 2, pyrimidine compound c undergoes reaction with aniline compound d to provide a phenylaminopyridine compound of formula I in accordance with the invention. The reaction of step 2 may take place in polar protic solvent and in the presence of acid such as HCl. Many aniline compound d are commercially available or can be easily prepared from nitrobenzenes as shown in the Examples below.
Many variations on the procedure of Scheme A are possible and will suggest themselves to those skilled in the art. Specific details for producing compounds of the invention are described in the Examples below.
Administration and Pharmaceutical Composition
The invention includes pharmaceutical compositions comprising at least one compound of the present invention, or an individual isomer, 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.
In general, the compounds of the invention will be administered in a therapeutically effective amount by any of the accepted modes of administration for agents that serve similar utilities. Suitable dosage ranges are typically 1-500 mg daily, for example 1-100 mg daily, and most preferably 1-30 mg daily, depending upon numerous factors such as the severity of the disease to be treated, the age and relative health of the subject, the potency of the compound used, the route and form of administration, the indication towards which the administration is directed, and the preferences and experience of the medical practitioner involved. One of ordinary skill in the art of treating such diseases will be able, without undue experimentation and in reliance upon personal knowledge and the disclosure of this Application, to ascertain a therapeutically effective amount of the compounds of the present invention for a given disease.
Compounds of the invention may be administered as pharmaceutical formulations including those suitable for oral (including buccal and sub-lingual), rectal, nasal, topical, pulmonary, vaginal, or parenteral (including intramuscular, intraarterial, intrathecal, subcutaneous and intravenous) administration or in a form suitable for administration by inhalation or insufflation. A particular manner of administration is generally oral using a convenient daily dosage regimen which can 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 into the form of pharmaceutical compositions and unit dosages. The pharmaceutical compositions and unit dosage forms may be comprised of conventional ingredients in conventional proportions, with or without additional active compounds or principles, and the unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended 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 or, more broadly, about 0.01 to about one hundred (100) milligrams, per tablet, are accordingly suitable representative unit dosage forms.
The compounds of the invention may be formulated in a wide variety of oral administration dosage forms. The pharmaceutical compositions and dosage forms may comprise a compound or compounds of the present invention or pharmaceutically acceptable salts thereof as the active component. The pharmaceutically acceptable carriers may be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier may be one or more substances which may also act as diluents, flavouring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier generally is a finely divided solid which is a mixture with the finely divided active component. In tablets, the active component generally is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. The 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, gelatine, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “preparation” is intended to include the formulation of the active compound with encapsulating material as carrier, providing a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges may 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 shortly before use to liquid form preparations. Emulsions may be prepared in solutions, for example, in aqueous propylene glycol solutions or may contain emulsifying agents, for example, such as lecithin, sorbitan monooleate, or acacia. Aqueous solutions can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents. Aqueous suspensions can 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 component, 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, for example bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion 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. 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 preserving, wetting, emulsifying or suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution for constitution before use with a suitable vehicle, e.g., sterile, pyrogen-free water.
The compounds of the invention may be formulated for topical administration to the epidermis as ointments, creams or lotions, 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 containing 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 active agents in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatine and glycerine or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.
The compounds of the 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 component is dispersed homogeneously, for example, by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool, and to solidify.
The compounds of the invention may be formulated for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate.
The subject compounds may be formulated for nasal administration. The solutions or suspensions are applied directly to the nasal cavity by conventional means, for example, with a dropper, pipette or spray. The formulations may be provided in a single or multidose form. In the latter case of a dropper or pipette, this may be achieved by the patient administering an appropriate, predetermined volume of the solution or suspension. In the case of a spray, this may be achieved for example by means of a metering atomizing spray pump.
The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract and including intranasal administration. The compound will generally have a small particle size for example of the order of five (5) microns or less. Such a particle size may be obtained by means known in the art, for example by micronization. The active ingredient is provided in a pressurized pack with a suitable propellant such as a chlorofluorocarbon (CFC), for example, dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas. The aerosol may conveniently also contain a surfactant such as lecithin. The dose of drug may be controlled by a metered valve. Alternatively the active ingredients may be provided in a form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidine (PVP). The powder carrier will form a gel in the nasal cavity. The powder composition may be presented in unit dose form for example in capsules or cartridges of e.g., gelatine or blister packs from which the powder may be administered by means of an inhaler.
When desired, formulations can be prepared with enteric coatings adapted for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention can be formulated in transdermal or subcutaneous drug delivery devices. These delivery systems are advantageous when sustained release of the compound is necessary and when patient compliance with a treatment regimen is crucial. Compounds in transdermal delivery systems are frequently attached to an skin-adhesive solid support. The compound of interest can also be combined with a penetration enhancer, e.g., Azone (1-dodecylazacycloheptan-2-one). Sustained release delivery systems are inserted subcutaneously into the subdermal layer by surgery or injection. The subdermal implants encapsulate the compound in a lipid soluble membrane, e.g., silicone rubber, or a biodegradable polymer, e.g., polylactic acid.
The pharmaceutical preparations may be in unit dosage forms. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, 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, Pa. Representative pharmaceutical formulations containing a compound of the present invention are described below.
Utility
The compounds of the invention are useful for treatment of LRRK2-mediated diseases or conditions, including neurodegenerative diseases such as Parkinson's disease, Lewy body dementia and Huntington's disease, and for enhancement of cognitive memory generally in subjects in need thereof.
EXAMPLES
The following preparations and examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.
Unless otherwise stated, all temperatures including melting points (i.e., MP) are in degrees celsius (° C.). It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product. The following abbreviations may be used in the Preparations and Examples.
List of Abbreviations


    AcOH Acetic acid
    AIBN 2,2′-Azobis(2-methylpropionitrile)
    Atm. Atmosphere
    (BOC)2O di-tert-Butyl dicarbonate
    DavePhos 2-Dicyclohexylphosphino-2′-(N,N-dimethylamino)biphenyl
    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-tetramethyl uronium hexafluorophosphate Methanaminium
    HBTU O-Benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate
    HOBT 1-Hydroxybenzotriazole
    HPLC High pressure liquid chromatography
    RP HPLC Reverse phase high pressure liquid chromatography
    i-PrOH Isopropanol/isopropyl alcohol
    LCMS Liquid Chromatograph/Mass Spectroscopy
    MeOH Methanol/Methyl alcohol
    MW Microwaves
    NBS N-Bromosuccinimide
    NMP 1-Methyl-2-pyrrolidinone
    PSI Pound per square inch
    RT Room temperature
    TBDMS tert-Butyldimethylsilyl
    TFA Trifluoroacetic acid
    THF Tetrahydrofuran
    TLC Thin layer chromatography


Preparation 1
2-chloro-5-fluoro-N-methylpyrimidin-4-amine




To a 250 mL round bottom flask equipped with a stir bar was added 9.0 g 5-fluoro-2,4-dichloro-pyrimidine, 40 mL methanol and 15 mL of 8M methylamine in ethanol. The reaction heated up (mild exo-therm) and was allowed to stir at room temperature for  ̃30 minutes. A check by TLC (1:1 EtOAc:heptane) and LCMS showed complete reaction. The reaction was concentrated down to give 9.77 g crude material which was purified on a silica column running a gradient of 1% to 10% MeOH in DCM over 35 minutes to give 6.77 g pure 2-chloro-5-fluoro-N-methylpyrimidin-4-amine.
The same method was used to make the compounds shown in Table 1 below, using the appropriate commercially available substituted 2,4-dichloro-pyrimidines and amines.









TABLE 1







 1
2-chloro-5-chloro-N- methylpyrimidin-4- amine






 


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






 


 3
2-chloro-5-trifluoromethyl-N- methylpyrimidin-4-amine






 


 6
2-chloro-5-methoxy-N- methylpyrimidin-4- amine






 


 8
2-chloro-5-fluoro-N,N- dimethylpyrimidin- 4-amine






 


 9
2-chloro-5-chloro-N- ethylpyrimidin-4- amine






 


10
2-chloro-5-chloro-N- propylpyrimidin-4- amine






 


11
2-chloro-5-chloro-N- isopropylpyrimidin-4- amine






 


12
2-chloro-5-chloro-N- isobutylpyrimidin-4- amine






 


13
4-(2,5-dichloropyrimidin- 4-yl)morpholine






 


14
2,5-dichloropyrimidin- 4-amine






 


15
2,5-dichloro-N,N- dimethylpyrimidin-4- amine






 


16
4-(azetidin-1-yl)-2,5- dichloropyrimidine






 


17
2,5-dichloro-4- (pyrrolidin-1-yl)pyrimidine






 


18
2,5-dichloro-4- (piperidin-1-yl)pyrimidine






 


19
2,5-dichloro-4-(2- (methoxymethyl) piperidin-1-yl)pyrimidine






 


20
2,5-dichloro-4-(4- (methoxymethyl) piperidin-1-yl)pyrimidine






 


21
2,5-dichloro-N- (cyclopropylmethyl) pyrimidin-4-amine






 


22
2,5-dichloro-N- (cyclobutylmethyl) pyrimidin-4-amine






 


23
2,5-dichloro-N- (cyclopentylmethyl) pyrimidin-4-amine






 


24
2-chloro-N- methylpyrimidin-4-amine






 


25
2,5-dichloro-N- (2-methoxyethyl)pyrimidin- 4-amine













Preparation 2
2,5-dichloro-4-methoxypyrimidine




To a 250 mL round bottom flask equipped with a stir bar was added 1 g 5-chloro-2,4-dichloro-pyrimidine, and 15 mL of diethyl ether. The mixture was cooled to 0° C. in an ice bath and then 1 equivalent of sodium methoxide in methanol (prepared from reacting 120 mg of sodium with 4 mL of methanol at room temperature) was slowly added. The reaction was stirred over night at room temperature and checked by LCMS. The white precipitate was filtered and the solid washed with cold methanol. After drying, 0.98 g of pure 2,5-dichloro-4-methoxypyrimidine was obtained and this material was used without further purification.
The same method was used to make the compounds shown in Table 2 below, using the appropriate commercially available alcohols and the appropriately substituted 2,4-dichloro-pyrimidines.









TABLE 2







1
2,5-dichloro-4-ethoxypyrimidine 






 


2
2,5-dichloro-4-propoxypyrimidine






 


3
2,5-dichloro-4-isoprpoxypyrimidine






 


6
5-bromo-2-chloro-4- methoxypyrimidine






 


7
2-chloro-5-iodo-4- methoxypyrimidine













Preparation 3
5-Amino-6-methoxy-2-methylisoindolin-1-one
Step 1: 5-Methoxy-2-methyl-4-nitrobenzonitrile




To 5-methoxy-2-methyl-4-nitroaniline (4.9 g, 26.8 mmol) in a mixture of acetone (17.5 mL) and water (19 mL) at 0° C. was added conc. HCl (5.6 mL). A solution of sodium nitrite (2.25 g, 32.6 mmol) in water (7.5 mL) was added dropwise and the mixture was allowed to stir at 0° C. for 30 min. The mixture was then added dropwise to a mixture of copper cyanide (3.75 g, 42 mmol) and sodium cyanide (5.5 g, 112 mmol) in water (25 mL) and EtOAc (12.5 mL). The mixture was allowed to stir at RT for 2 h and then water (50 mL) was added. The mixture was extracted with EtOAc (3×100 mL) and the combined organic fractions were washed with 2 M NaOH(aq) (50 mL) and brine (50 mL). The organic fraction was passed through a hydrophobic frit and the solvent was removed in vacuo. The residue was triturated with iso-hexane and dried to give 5-methoxy-2-methyl-4-nitrobenzonitrile (4.62 g, 90%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M]−=192 at 3.19 min. 1H NMR (400 MHz, CDCl3): δ 7.72 (s, 1H), 7.31 (s, 1H), 3.98 (s, 3H), 2.55 (s, 3H).
Step 2: 5-Methoxy-2-methyl-4-nitrobenzoic acid




A mixture of 5-methoxy-2-methyl-4-nitrobenzonitrile (2 g, 10.4 mmol) in AcOH (20 mL), water (20 mL) and conc. sulphuric acid (20 mL) was heated to 120° C. for 5 h. Water (100 mL) and DCM (100 mL) were added and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo to give 5-methoxy-2-methyl-4-nitrobenzoic acid (1.27 g, 58%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M−H]−=210 at 3.37 min. 1H NMR (400 MHz, CDCl3): δ 7.75 (s, 1H), 7.70 (s, 1H), 3.99 (s, 3H), 2.62 (s, 3H).
Step 3: Methyl 5-methoxy-2-methyl-4-nitrobenzoate




To 5-methoxy-2-methyl-4-nitrobenzoic acid (1.27 g, 6 mmol) in methanol (15 mL) was added thionyl chloride (0.44 mL, 6 mmol) dropwise. The mixture was heated to reflux for 3 h and then allowed to cool. The solvent was removed in vacuo and the residue was partitioned between DCM (20 mL) and saturated aqueous NaHCO3 (20 mL). The organic phase was passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via silica gel column chromatography (0-100% EtOAc in iso-hexane) gave methyl 5-methoxy-2-methyl-4-nitrobenzoate (1.11 g, 82%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=226 at 3.92 min. 1H NMR (400 MHz, CDCl3): δ 7.68 (s, 1H), 7.60 (s, 1H), 3.98 (s, 3H), 3.95 (s, 3H), 2.55 (s, 3H).
Step 4: Methyl 2-(bromomethyl)-5-methoxy-4-nitrobenzoate




A mixture of methyl 5-methoxy-2-methyl-4-nitrobenzoate (1.11 g, 5 mmol) and NBS (1.07 g, 6 mmol) in MeCN (20 mL) was degassed and then AIBN (18 mg, 0.11 mmol) was added. The mixture was heated to 70° C. for 18 h. DCM (50 mL) and water (50 mL) were added and the organic phase was passed through a hydrophobic frit. The solvent was removed in vacuo and purification of the residue via silica gel column chromatography (0-100% EtOAc in iso-hexane) in order to remove baseline impurities gave methyl 2-(bromomethyl)-5-methoxy-4-nitrobenzoate (477 mg, 31%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M−Br]−=224 at 3.92 min. 1H NMR (400 MHz, CDCl3): δ 7.92 (s, 1H), 7.66 (s, 1H), 4.88 (s, 2H), 4.02 (s, 3H), 4.00 (s, 3H).
Step 5: 6-Methoxy-2-methyl-5-nitroisoindolin-1-one




To methyl 2-(bromomethyl)-5-methoxy-4-nitrobenzoate (212 mg, 0.7 mmol) in methanol (6 mL) was added Et3N (0.12 mL, 0.84 mmol) and methylamine in EtOH (0.11 mL,  ̃8 M in EtOH, 0.84 mmol). The mixture was heated to 70° C. for 5 h. and then EtOAc (30 mL) and 2M HCl(aq) (30 mL) was added. The layers were separated and the aqueous layer was extracted with EtOAc (2×30 mL). The combined organic fractions were passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via silica gel column chromatography (0-100% EtOAc in iso-hexane then EtOAc-10% MeOH in DCM) gave 6-methoxy-2-methyl-5-nitroisoindolin-1-one (117 mg, 75%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=223 at 3.02 min. 1H NMR (400 MHz, CDCl3): δ 7.86 (s, 1H), 7.53 (s, 1H), 4.40 (s, 2H), 4.02 (s, 3H), 3.24 (s, 3H).
Step 6: 5-Amino-6-methoxy-2-methylisoindolin-1-one




To 6-methoxy-2-methyl-5-nitroisoindolin-1-one (111 mg, 0.5 mmol) in ethanol (6 mL) and water (0.6 mL) was added SnCl2.2H2O (451 mg, 2 mmol) and the mixture was heated to 65° C. for 5 h. DCM (20 mL) and 2M NaOH(aq) (10 mL) were added and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo to give 5-amino-6-methoxy-2-methylisoindolin-1-one (68 mg, 71%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=193 at 2.32 min. 1H NMR (400 MHz, DMSO): δ 7.01 (s, 1H), 6.74 (s, 1H), 5.44 (br s, 2H), 4.24 (s, 2H), 3.85 (s, 3H), 3.02 (s, 3H).
Preparation 4
6-Amino-5-methoxy-2-methylisoindolin-1-one
Step 1: 2-Methyl-4,5-dinitrobenzoic acid




To concentrated sulfuric acid (10 mL) at 0° C. was added concentrated nitric acid (10 mL) dropwise and the mixture was allowed to stir at 0° C. for 10 min. 2-Methyl-4-nitrobenzic acid (1.81 g, 10 mmol) was added and the mixture was allowed to stir at RT for 5 d. The mixture was poured into ice-cold water and the solid was removed by filtration and dried to give 2-methyl-4,5-dinitrobenzoic acid (1.93 g, 85%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M−H]−=225 at 3.10 min. 1H NMR (400 MHz, d6-DMSO): δ 8.53 (s, 1H), 8.26 (s, 1H), 2.71 (s, 3H).
Step 2: 4-Methoxy-2-methyl-5-nitrobenzoic acid




2-Methyl-4,5-dinitrobenzoic acid (1.93 g, 8.5 mmol) was added to KOH (2.38 g, 42.5 mmol) in methanol (50 mL) and the mixture was heated to 70° C. for 1.5 h. The mixture was acidified with 2 M HCl(aq) and the solid was removed by filtration and dried to give 4-methoxy-2-methyl-5-nitrobenzoic acid (1.08 g, 60%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M−H]−=210 at 3.23 min. 1H NMR (400 MHz, d6-DMSO): δ 8.40 (s, 1H), 7.36 (s, 1H), 4.02 (s, 3H), 2.67 (s, 3H).
Step 3: Methyl 4-methoxy-2-methyl-5-nitrobenzoate




To 4-methoxy-2-methyl-5-nitrobenzoic acid (1.06 g, 5 mmol) in methanol (10 mL) was added thionyl chloride (0.36 mL, 5 mmol) dropwise. The mixture was heated to reflux for 5 h and then allowed to cool. The solvent was removed in vacuo and the residue was partitioned between DCM (20 mL) and saturated aqueous NaHCO3 (20 mL). The organic phase was passed through a hydrophobic frit and the solvent was removed in vacuo to give methyl 4-methoxy-2-methyl-5-nitrobenzoate (998 mg, 89%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M+H]+=226 at 3.25 min. 1H NMR (400 MHz, CDCl3): δ 8.56 (s, 1H), 6.91 (s, 1H), 4.01 (s, 3H), 3.90 (s, 3H), 2.71 (s, 3H).
Step 4: Methyl 2-(bromomethyl)-4-methoxy-5-nitrobenzoate




A mixture of methyl 4-methoxy-2-methyl-5-nitrobenzoate (994 mg, 4.41 mmol) and NBS (943 mg, 5.3 mmol) in MeCN (20 mL) was degassed and then AIBN (15 mg, 0.09 mmol) was added. The mixture was heated to 70° C. for 18 h. Further AIBN (15 mg, 0.09 mmol) was added and the mixture was heated to 70° C. for 24 h. Further AIBN (15 mg, 0.09 mmol) and NBS (300 mg, 1.7 mmol) were added and the mixture heated to 70° C. for 24 h. Further AIBN (15 mg, 0.09 mmol) was added and the mixture was heated to 70° C. for 24 h. DCM (50 mL) and water (50 mL) were added and the organic phase was passed through a hydrophobic frit. The solvent was removed in vacuo to give crude methyl 2-(bromomethyl)-4-methoxy-5-nitrobenzoate that was used directly in the next reaction without any purification.
Step 5: 5-Methoxy-2-methyl-6-nitroisoindolin-1-one




To crude 2-(bromomethyl)-4-methoxy-5-nitrobenzoate (<4 mmol) in methanol (36 mL) was added Et3N (0.68 mL, 4.8 mmol) and methylamine in EtOH (0.6 mL,  ̃8 M in EtOH, 4.8 mmol). The mixture was heated to 70° C. for 5 h. and then EtOAc (30 mL) and 2M HCl(aq) (30 mL) was added. The layers were separated and the aqueous layer was extracted with EtOAc (2×30 mL). The combined organic fractions were passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via silica gel column chromatography (0-100% EtOAc in iso-hexane then EtOAc-10% MeOH in DCM) gave impure 5-methoxy-2-methyl-6-nitroisoindolin-1-one (487 mg) as a brown solid. The crude material was used directly in the next reaction without any further purification.
Step 6: 6-Amino-5-methoxy-2-methylisoindolin-1-one




To crude 5-methoxy-2-methyl-6-nitroisoindolin-1-one (121 mg, 0.54 mmol) in ethanol (6 mL) and water (0.6 mL) was added SnCl2.2H2O (487 mg, 2.16 mmol) and the mixture was heated to 65° C. for 24 h. DCM (10 mL) and 2M NaOH(aq) (10 mL) were added and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo to give impure 6-amino-5-methoxy-2-methylisoindolin-1-one (101 mg, 97%) as an off-white solid. The crude material was used directly in the next reaction without any further purification. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=193 at 2.00 min.
Preparation 5
5-Amino-6-chloro-2-methylisoindolin-1-one
Step 1: 5-Chloro-2-methyl-4-nitrobenzonitrile




To 5-chloro-2-methyl-4-nitroaniline (5 g, 26.8 mmol) in a mixture of acetone (17.5 mL) and water (19 mL) at 0° C. was added conc. HCl (5.6 mL). A solution of sodium nitrite (2.25 g, 32.6 mmol) in water (7.5 mL) was added dropwise and the mixture was allowed to stir at 0° C. for 30 min. The mixture was then added dropwise to a mixture of copper cyanide (3.75 g, 42 mmol) and sodium cyanide (5.5 g, 112 mmol) in water (25 mL) and EtOAc (12.5 mL). The mixture was allowed to stir at RT for 1 h and then water (50 mL) was added. The mixture was extracted with EtOAc (3×100 mL) and the combined organic fractions were washed with 2 M NaOH(aq) (50 mL) and brine (50 mL). The organic fraction was passed through a hydrophobic frit and the solvent was removed in vacuo. The residue was suspended in a 1:1 mixture of diethyl ether:iso-hexane. The solid formed was removed by filtration and washed with iso-hexane and dried. Further fractions were isolated from the filtrate and combined with the initial solid fraction to give 5-chloro-2-methyl-4-nitrobenzonitrile (3.22 g, 61%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=197 at 3.97 min. 1H NMR (400 MHz, CDCl3): δ 7.80 (s, 1H), 7.80 (s, 1H), 2.63 (s, 3H).
Step 2: 4-Amino-5-chloro-2-methylbenzonitrile




To 5-chloro-2-methyl-4-nitrobenzonitrile (192 mg, 1 mmol) in ethanol (6 mL) and water (0.6 mL) was added SnCl2.2H2O (903 mg, 4 mmol) and the mixture was heated to 65° C. for 5 h. DCM (10 mL) and 2M NaOH(aq) (10 mL) were added, and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo to give 4-amino-5-chloro-2-methylbenzonitrile (131 mg, 81%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=163 at 3.37 min. 1H NMR (400 MHz, CDCl3): δ 6.89 (s, 1H), 6.52 (s, 1H), 4.21 (br s, 2H), 3.84 (s, 3H), 2.38 (s, 3H).
Step 3: 5-Chloro-2-methyl-4-nitrobenzonitrile




A mixture of 5-chloro-2-methyl-4-nitrobenzonitrile (1 g, 5 mmol) in AcOH (10 mL), water (10 mL) and conc. sulphuric acid (10 mL) was heated to 120° C. for 5 h. Water (100 mL) was added and the solid was removed by filtration and dried to give 5-chloro-2-methyl-4-nitrobenzoic acid (905 mg, 84%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M−H]−=214 at 2.12 min. 1H NMR (400 MHz, CDCl3): δ 8.22 (s, 1H), 7.76 (s, 1H), 2.70 (s, 3H).
Step 4: Methyl 5-chloro-2-methyl-4-nitrobenzoate




To 5-chloro-2-methyl-4-nitrobenzoic acid (900 mg, 4.15 mmol) in methanol (10 mL) was added thionyl chloride (0.3 mL, 4.15 mmol) dropwise. The mixture was heated to reflux for 5 h and then allowed to cool. The solvent was removed in vacuo and the residue was partitioned between DCM (20 mL) and saturated aqueous NaHCO3 (20 mL). The organic phase was passed through a hydrophobic frit and the solvent was removed in vacuo to give methyl 5-chloro-2-methyl-4-nitrobenzoate (880 mg, 92%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=230 at 4.25 min. 1H NMR (400 MHz, CDCl3): δ 8.08 (s, 1H), 7.73 (s, 1H), 3.95 (s, 3H), 2.64 (s, 3H).
Step 5: Methyl 2-(bromomethyl)-5-chloro-4-nitrobenzoate




A mixture of methyl 5-chloro-2-methyl-4-nitrobenzoate (862 mg, 4 mmol) and NBS (854 mg, 4.8 mmol) in MeCN (16 mL) was degassed and then AIBN (14 mg, 0.08 mmol) was added. The mixture was heated to 70° C. for 18 h. The solvent was removed in vacuo and the residue was partitioned between DCM (50 mL) and water (50 mL). The organic phase was passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via silica gel column chromatography (0-100% EtOAc in iso-hexane) in order to remove baseline impurities gave methyl 2-(bromomethyl)-5-chloro-4-nitrobenzoate (1.09 g, 89%) as a brown oil. 1H NMR (400 MHz, CDCl3): δ 8.14 (s, 1H), 7.98 (s, 1H), 4.91 (s, 2H), 4.00 (s, 3H).
Step 6: 6-Chloro-2-methyl-5-nitroisoindolin-1-one




To methyl 2-(bromomethyl)-5-chloro-4-nitrobenzoate (617 mg, 2 mmol) in methanol (18 mL) was added Et3N (0.34 mL, 2.4 mmol) and methylamine in EtOH (0.3 mL,  ̃d 8 M in EtOH, 2.4 mmol). The mixture was heated to 70° C. for 3 h. and then EtOAc (30 mL) and 2M HCl(aq) (30 mL) was added. The layers were separated and the aqueous layer was extracted with EtOAc (2×30 mL). The combined organic fractions were passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via silica gel column chromatography (0-10% methanol in DCM) gave 6-chloro-2-methyl-5-nitroisoindolin-1-one (347 mg, 77%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M+H]+=227 at 2.89 min. 1H NMR (400 MHz, CDCl3): δ 8.00 (s, 1H), 7.90 (s, 1H), 4.46 (s, 2H), 3.24 (s, 3H).
Step 7: 5-Amino-6-chloro-2-methylisoindolin-1-one




To 6-chloro-2-methyl-5-nitroisoindolin-1-one (340 mg, 1.5 mmol) in ethanol (9 mL) and water (0.9 mL) was added SnCl2.2H2O (1.35 g, 6 mmol) and the mixture was heated to 65° C. for 5 h. DCM (20 mL) and 2M NaOH(aq) (10 mL) were added and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo to give 5-amino-6-chloro-2-methylisoindolin-1-one (251 mg, 85%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M+H]+=197 at 2.31 min. 1H NMR (400 MHz, d6-DMSO): δ 7.45 (s, 1H), 6.90 (s, 1H), 5.99 (br s, 2H), 4.32 (s, 2H), 3.01 (s, 3H).
Preparation 6
5-Amino-4-methoxy-2-methylisoindolin-1-one
Step 1: 3-Hydroxy-2-methyl-4-nitrobenzoic acid




To a solution of 3-hydroxy-2-methylbenzoic acid (5.00 g, 25.3 mmol) in concentrated H2SO4 (98%, 10 mL) was added concentrated HNO3 (63%, 10 mL) dropwise. The mixture was stirred at −40° C. for 10 min. The mixture was poured into ice-water and extracted by EA (3 times). The organic layers were combined, dried over anhydrous Na2SO4. Then the solvent was evaporated in vacuo to afford 3-hydroxy-2-methyl-4-nitrobenzoic acid as a yellow solid, which was used for next step without further purification.
Step 2: Methyl 3-methoxy-2-methyl-4-nitrobenzoate




To a solution of the crude 3-hydroxy-2-methyl-4-nitrobenzoic acid in acetone (300 mL) was added K2CO3 (15.8 g, 115 mmol) and Me2SO4 (8.7 g, 69 mmol). The mixture was refluxed at 60° C. for 2 h. After the solvent was evaporated in vacuo, the resulting residue was purified by silica gel-column (eluting with PE/EA=10:1) to afford methyl 3-methoxy-2-methyl-4-nitrobenzoate as a yellowish solid (2.7 g, 33% yield, two steps). m/z (ES+APCI)+: [M+H]+226.1.
Step 3: Methyl 2-(bromomethyl)-3-methoxy-4-nitrobenzoate




To a solution of methyl 3-methoxy-2-methyl-4-nitrobenzoate (800 mg, 3.6 mmol) in CH3CN (10 mL) was added NBS (770 mg, 4.3 mmol) and AIBN (12 mg, 0.072 mmol) under the protection of N2. The mixture was refluxed at 80° C. for overnight. After the solvent was evaporated in vacuo, the resulting residue was purified by silica gel-column (eluting with PE/EA=15:1) to afford methyl 2-(bromomethyl)-3-methoxy-4-nitrobenzoate as yellow oil (910 mg, yield 84%). m/z (ES+APCI)+: [M−Br]+224.1.
Step 4: 4-Methoxy-2-methyl-5-nitroisoindolin-1-one




To a solution of methyl 2-(bromomethyl)-3-methoxy-4-nitrobenzoate (300 mg, 1.0 mmol) in MeOH (10 mL) was added Et3N (120 mg, 1.2 mmol) and CH3NH2 (1.0 mL, 4M in MeOH). The solution was stirred at 70° C. for 4 h. The solvent was evaporated in vacuo, and the residue was purified by silica gel-column (eluting with PE/EA=10:1) to afford 4-methoxy-2-methyl-5-nitroisoindolin-1-one as a yellow solid (110 mg, yield 50%). m/z (ES+APCI)+: [M+H]+223.1.
Step 5: 5-Amino-4-methoxy-2-methylisoindolin-1-one




To a solution of 4-methoxy-2-methyl-5-nitroisoindolin-1-one (155 mg, 0.7 mmol) in MeOH (10 mL) was added Pd/C (60 mg). The mixture was stirred at 50° C. under H2 for 6 h. The resulting suspension was filtered. The filtrate was concentrated under reduced pressure to afford 5-amino-4-methoxy-2-methylisoindolin-1-one (70 mg, yield 52%). m/z (ES+APCI)+: [M+H]+193.1.
Example 1
7-Chloro-4-methyl-8-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one
Step 1: 2-((tert-Butyldimethylsilyl)oxy)-N-methylethanamine




A solution of tert-butyldimethylchlorosilane (4.2 g, 27.9 mmol) in dichloromethane (10 mL) was added dropwise over 3 min to a stirred solution of 2-(methylamino)ethanol (2.0 g, 26.6 mmol) and imidazole in dichloromethane (20 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. Water (20 mL) was added and the phases were separated. The aqueous phase was extracted with dichloromethane then the combined organic phases were passed through a phase separation cartridge and the solvent removed under reduced pressure to afford a colorless oil (5.10 g, 95%). This was taken through to the next step with no further purification.
Step 2: 4-Amino-N-2-tert-butyldimethylsilyl)oxy)ethyl)-5-chloro-2-methoxy-N-methylbenzamide




4-Amino-5-chloro-2-methoxybenzoic acid (3.0 g, 14.9 mmol), DIPEA (3.84 g, 29.8 mmol) and HATU (6.80 g, 17.9 mmol) were stirred at room temperature for 20 min in dichloromethane (180 mL). 2-((Tert-butyldimethylsilyl)oxy)-N-methylethanamine (3.12 g, 16.4 mmol) was added, then the reaction was stirred for 1 h after which time analysis by LCMS indicated the reaction was complete. The dichloromethane was removed and the resulting residue partitioned between ethyl acetate and water. The layers were separated and the aqueous phase extracted with ethyl acetate. The combined organic extracts were concentrated and the resulting residue was triturated with ethyl acetate-petroleum ether (1:1). The solid was collected by filtration and discarded. The supernatant was retained and concentrated to afford a crude product that was used directly in the next step (2.0 g, 36%).
Step 3: 4-Amino-5-chloro-2-hydroxy-N-(2-hydroxyethyl)-N-methylbenzamide




4-Amino-N-(2-((tert-butyldimethylsilyl)oxy)ethyl)-5-chloro-2-methoxy-N-methylbenzamide (2.0 g, 5.3 mmol) was stirred in dichloromethane (100 mL) at −10° C. under an atmosphere of nitrogen and a 1M solution of boron trichloride in dichloromethane (10.68 mL, 10.7 mmol) was added dropwise. The reaction was allowed to warm to room temperature then left to stir at this temperature for 18 h. The reaction was cooled to −10° C. and a further 1 equivalent of boron trichloride solution was added dropwise, then the reaction was allowed to warm to room temperature and stirred for 3 h. After this time analysis by LCMS indicated that the reaction was complete. The reaction was cooled in a ice bath and methanol (20 mL) was added drop-wise to the reaction followed by 7N methanolic ammonia solution (17 mL) and then a further portion of MeOH ( ̃80 mL) until all the solids had dissolved. The solution was stirred for 1 h at room temperature then concentrated. The resulting residue was stirred with saturated aqueous NaHCO3 (150 mL) for 30 min then the aqueous phase was extracted with ethyl acetate. The combined extracts were washed with brine then passed through a phase separation cartridge and the solvent was removed under reduced pressure to afford a green gum (1.00 g, 77%). This material was taken through to the next step with no further purification.
Step 4: 8-Amino-7-chloro-4-methyl-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one




4-Amino-5-chloro-2-hydroxy-N-(2-hydroxyethyl)-N-methylbenzamide (300 mg, 1.22 mmol) was stirred in dichloromethane at 0° C. under an atmosphere of nitrogen then TEA (247 mg, 2.4 mmol), triphenylphosphine (320 mg, 1.2 mmol) and DEAD (212 mg, 1.2 mmol) were added. The reaction was allowed to stir for 1 h at 0° C., after which time LCMS indicated the formation of a peak consistent with the target material. The volatiles were removed under reduced pressure and the resulting gum was dissolved in dichloromethane then passed through a SCX-2 20 g cartridge. The cartridge was washed with dichloromethane-methanol 10% and the fractions were collected in 100 mL flasks. TLC indicated that the fractions were not identical in composition which was verified by LCMS. The purest fraction was concentrated to afford the target material (18 mg, 5%).
Step 5: 7-Chloro-4-methyl-8-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one




A mixture of 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (0.5 g, 2.36 mmol), 8-amino-7-chloro-4-methyl-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one (0.38 mmol) and p-toluene sulphonic acid (0.49 g, 2.6 mmol) in dioxane (10 mL) was heated at 100° C. for 2 h. The mixture was cooled, filtered and the solid washed with dioxane. The solid was dried in a vacuum oven and purified via silica gel column chromatography (20-100% ethyl acetate/isohexane) gave 7-chloro-4-methyl-8-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one as a white solid (10 mg, 28%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=402 at 3.04 min. 1H NMR δ (ppm)(400 MHz, CDCl3): 8.41 (1H, s), 8.22 (1H, s), 7.97 (1H, s), 7.77-7.67 (1H, m), 5.29 (1H, s), 4.47-4.39 (2H, m), 3.61-3.51 (2H, m), 3.21-3.18 (3H, m), 3.20-3.07 (3H, m).
Example 2
7-((5-Chloro-4-(methylamino)pyrimidin-2-yl)amino)-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one
Step 1: 6-Chloro-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one




To a suspension of anhydrous potassium fluoride (4.71 g, 81 mmol) in DMF (40 mL) was added 2-amino-5-nitrophenol (5.00 g, 26.5 mmol) and the mixture was stirred at room temperature for 1 h. To the suspension was added a solution of ethyl-α-bromoisobutyrate (6.33 g, 32.5 mmol) in DMF (10 mL) dropwise over a period of 20 min. The mixture was stirred at 60° C. for 20 h. The reaction was allowed to cool, then water (150 mL) was added then the aqueous was extracted with ethyl acetate. The combined organic extracts were washed successively with 2M aqueous HCl (100 mL), water (100 mL) and brine (100 mL) then passed through a phase separation cartridge. The solvent was removed under reduced pressure and the resulting residue was suspended in ethyl acetate. The precipitate was collected by filtration and washed with ethyl acetate to afford a pale yellow solid (2.29 g, 34%) which was taken through to the next step with no further purification.
Step 2: 6-Chloro-2,2,4-trimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one




6-Chloro-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (2.29 g, 8.9 mmol) was stirred in DMF at 0° C. under an atmosphere of nitrogen then sodium hydride (60% dispersion in oil) (392 mg, 9.79 mmol) was added. The reaction was allowed to stir at room temperature for 30 min then iodomethane (1.89 g, 13.4 mmol) was added and the reaction was stirred at room temperature for 18 h. Analysis by LCMS indicated that the reaction was complete. Water (10 mL) was added carefully then the DMF was removed under reduced pressure. The residue was partitioned between ethyl acetate and water. The layers were separated and the aqueous phase was extracted with ethyl acetate. The organic portions were combined then passed through a phase separation cartridge. The solvents were evaporated to afford an orange solid (2.4 g, quant.) which was taken through to the next step with no further purification.
Step 3: 6-Methoxy-2,2,4-trimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one




Toluene-methanol (4:1) (50 mL) was degassed, then cesium carbonate (2.85 g, 7.4 ml) was added followed by rac-2-(di-tert-butylphosphine)1,1′-binaphthyl (44 mg, 0.11 mmol) and 6-chloro-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (1.0 g, 3.7 mmol) with stirring under an atmosphere of nitrogen. Palladium (II) acetate (20 mg, 0.074 mmol) was added then the reaction was heated to 75° C. for 18 h with stirring under nitrogen. The reaction was allowed to cool, then a further aliquot of palladium (II) acetate were added and the reaction heated at 75° C. for 18 h to complete the conversion. The reaction was allowed to cool, water (100 mL) was added then the whole was extracted with ethyl acetate. The combined organic extracts were washed with brine, passed through a phase separation cartridge and evaporated to afford a brown solid ( ̃800 mg). Purification via silica gel column chromatography (0-100% ethyl acetate/isohexane) gave 6-methoxy-2,2,4-trimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (0.47 g, 48%).
Step 4: 7-Amino-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one




6-Methoxy-2,2,4-trimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (250 mg, 0.94 mmol) was stirred in ethanol (15 mL) then tin (II) chloride was added. The reaction was heated to 80° C. with stirring for 2 h. The volatiles were removed under reduced pressure and the residue was slurried with 2M sodium hydroxide aqueous solution. The aqueous was extracted with ethyl acetate and the combined organic extracts were passed through a phase separation cartridge and evaporated, to afford a brown crystalline solid (220 mg, 99%) which was taken through to the next step with no further purification.
Step 5: 7-((5-Chloro-4-(methylamino)pyrimidin-2-yl)amino)-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one




A mixture of 2,5-dichloro-N-methylpyrimidin-4-amine (1 mmol), 7-amino-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (222 mg, 0.94 mmol), cesium carbonate (0.65 g, 2 mmol), XantPhos (17 mg, 0.03 mmol) and Pd2(dba)3 (5 mg, 0.02 mmol) in dioxane (3 mL) was sonnicated in an ultrasonic bath for 1 min. The mixture was then degassed under a stream of nitrogen for 5 min. The tube was sealed and the reaction was heated at 100° C. for 18 h. The reaction mixture was cooled and diluted with ethyl acetate (15 mL). The organic layer was washed with water and the combined aqueous extracts were further washed with ethyl acetate. The combined organics were passed through a phase separation cartridge and the solvent removed under reduced pressure. Purification of the residue via silica gel column chromatography (0-100% ethyl acetate/isohexane) gave 7-((5-chloro-4-(methylamino)pyrimidin-2-yl)amino)-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (30 mg, 15%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=378 at 2.77 min. 1H NMR δ (ppm)(400 MHz, d6-DMSO): 8.05 (1H, s), 7.98 (1H, s), 7.54 (1H, s), 7.28 (1H, d, J=5.2), 6.85 (1H, s), 3.93 (3H, s), 3.66 (3H, s), 2.91 (3H, d, J=4.6), 1.41 (6H, s).
Example 3
5-Methoxy-N,N-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzofuran-2-carboxamide
Step 1: Methyl 5-methoxybenzofuran-2-carboxylate




2-Hydroxy-5-methoxy-benzaldehyde (10 g, 65.8 mmol) was stirred in DMF (90 mL) with potassium carbonate (23 g, 0.17 mol) under nitrogen, then methyl bromoacetate was added dropwise. The mixture was stirred at room temperature for 4 h. The reaction was then heated to 80° C. for 18 h. The reaction was allowed to cool then poured onto ice water (200 mL) and the resulting brown suspension was allowed to stir for 30 min. After this time the solid was collected by filtration and washed with water. The solid was dried to afford (5.22 g, 39%), which was taken through to the next step with no further purification.
Step 2: Methyl 4-chloro-5-methoxybenzofuran-2-carboxylate




Methyl 5-methoxybenzofuran-2-carboxylate (5.22 g, 25.3 mmol) was dissolved in dichloromethane, and sulfuryl chloride (3.76 g, 27.9 mmol) was added at such rate to maintain a temperature of 20° C. After stirring for 5 h the solution was dripped slowly onto ice water (70 mL). The organic phase was then washed with water, aqueous saturated NaHCO3 and again with water. The layers were separated and the organic portion was passed through a phase separation cartridge and evaporated to afford a solid (5.81 g), which was taken through to the next step with no further purification.
Step 3: Methyl 4-chloro-5-methoxy-6-nitrobenzofuran-2-carboxylate




Methyl 4-chloro-5-methoxybenzofuran-2-carboxylate was dissolved in conc. sulphuric acid (58 mL) with stirring in an ice bath then conc. nitric acid was added dropwise, maintaining the temperature below 5° C. The reaction was then stirred at room temperature for 2.5 h. The reaction was added to ice water (500 mL) over 2 h. The resulting solid was filtered and washed with water until neutral. The solid was dried to afford methyl 4-chloro-5-methoxy-6-nitrobenzofuran-2-carboxylate (6.3 g, 91%), which was taken through to the next step with no further purification.
Step 4: 4-Chloro-5-methoxy-6-nitrobenzofuran-2-carboxylic acid




Lithium hydroxide (0.9 g, 21.0 mmol) was added to a stirred suspension of methyl 4-chloro-5-methoxy-6-nitrobenzofuran-2-carboxylate (2.0 g, 7.0 mmol) in methanol-water (3:1) (40 mL). The reaction was allowed to stir for 3 h at room temperature. The reaction was then concentrated under reduced pressure and the resulting residue was dissolved in water and washed with diethyl ether. The aqueous phase was acidified by the addition of 2M aqueous hydrochloric acid then extracted with ethyl acetate. The combined organic extracts were passed through a phase separation cartridge and evaporated to afford a white solid (1.17 g, 61%), which was taken through to the next step with no further purification.
Step 5: 4-Chloro-5-methoxy-N,N-dimethyl-6-nitrobenzofuran-2-carboxamide




4-Chloro-5-methoxy-N,N-dimethyl-6-nitrobenzofuran-2-carboxamide was prepared by the procedure described in Example 1, step 2 starting from 4-chloro-5-methoxy-6-nitrobenzofuran-2-carboxylic acid (1.17 g, 4.31 mmol) an dimethylamine hydrochloride (386 mg, 4.74 mmol). Purification via silica gel column chromatography (20-100% ethyl acetate/isohexane) gave 4-chloro-5-methoxy-N,N-dimethyl-6-nitrobenzofuran-2-carboxamide as a orange solid (850 mg, 66%).
Step 6: 6-Amino-5-methoxy-N,N-dimethylbenzofuran-2-carboxamide




4-Chloro-5-methoxy-N,N-dimethyl-6-nitrobenzofuran-2-carboxamide (500 mg, 1.68 mmol), TEA (184 mg, 1.83 mmol) and 10% palladium on charcoal (90 mg) were stirred at room temperature in THF-DMF (2:1) (30 mL) under an atmosphere of hydrogen for 18 h. The reaction was recharged with hydrogen then allowed to stir for an additional 18 h. The catalyst was removed by filtration and the filtrate was concentrated under reduced pressure to afford a brown solid (400 mg, quant.), which was taken through to the next step with no further purification.
Step 7: 5-Methoxy-N,N-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzofuran-2-carboxamide




5-Methoxy-N,N-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzofuran-2-carboxamide was prepared by the procedure described in Example 2, starting from 4-chloro-5-methoxy-N,N-dimethyl-6-nitrobenzofuran-2-carboxamide (220 mg, 0.94 mmol). The crude material was submitted to preparative HLPC for purification to give 5-methoxy-N,N-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzofuran-2-carboxamide (40 mg, 21%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=410 at 3.01 min. 1H NMR δ(ppm) (400 MHz, d6-DMSO): 8.70 (1H, s), 8.28 (1H, s), 8.17 (1H, s), 7.40-7.32 (3H, m), 3.98 (3H, s), 3.41-3.28 (6H, m), 3.01 (3H, s).
Example 4
5-Chloro-1,3-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-benzo[d]imidazol-2(3H)-one
Step 1: 5-Chloro-6-nitro-1H-benzo[d]imidazol-2(3H)-one




A stirred solution of 4-chloro-5-nitro-1,2-phenylenediamine (2.0 g, 10.66 mmol) and CDI (2.59 g, 16 mmol) in anhydrous THF (50 mL) was heated to 85° C. for 4 h. The reaction was allowed to cool and the solvents were removed under reduced pressure. The resulting residue was washed with 2M aqueous hydrochloric acid and recrystallised from methanol to afford 5-chloro-6-nitro-1H-benzo[d]imidazol-2(3H)-one as a brown solid (1.9 g, 84%), which was taken through to the next step with no further purification.
Step 2: 5-Chloro-1,3-dimethyl-6-nitro-1H-benzo[d]imidazol-2(3H)-one




5-Chloro-6-nitro-1H-benzo[d]imidazol-2(3H)-one (1.00 g, 4.7 mmol) was stirred in DMF (50 mL) then potassium carbonate (1.29 g, 9.4 mmol) and iodomethane (2.66 g, 18.7 mmol) were added. The reaction was stirred at 75° C. for 1.5 h. The reaction was allowed to cool and solvents were removed under reduced pressure. The residue was partitioned between water and ethyl acetate and the phases were separated. The aqueous phase was extracted with ethyl acetate and the combined organic extracts were through a phase separation cartridge and evaporated to afford a brown solid (1.07 g, 95%), which was taken through to the next step with no further purification.
Step 3: 5-Amino-6-chloro-1,3-dimethyl-1H-benzo[d]imidazol-2(3H)-one




5-Amino-6-chloro-1,3-dimethyl-1H-benzo[d]imidazol-2(3H)-one was prepared by the procedure described in Example 2, step 4 starting from 5-chloro-6-nitro-1H-benzo[d]imidazol-2(3H)-one (400 mg, 1.64 mmol) to afford a brown solid (314 mg, 90%), which was taken through to the next step with no further purification.
Step 4: 5-Chloro-1,3-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-benzo[d]imidazol-2(3H)-one




5-Chloro-1,3-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-benzo[d]imidazol-2(3H)-one was prepared by the procedure described in Example 2, starting from 5-amino-6-chloro-1,3-dimethyl-1H-benzo[d]imidazol-2(3H)-one (200 mg, 0.94 mmol). Purification via silica gel column chromatography (0-100% ethyl acetate/isohexane) afforded the title compound (20 mg, 11%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=387 at 3.10 min. 1H NMR δ (ppm)(400 MHz, DMSO-d6): 8.83 (1H, s), 8.13 (1H, s), 7.58 (1H, s), 7.38 (1H, s), 7.08 (1H, s), 3.36 (6H, s), 2.84 (3H, d, J=4.2).
Example 5
5-Chloro-3-methyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzo[d]oxazol-2(3H)-one
Step 1: 5-Chloro-6-nitrobenzo[d]oxazol-2(3H)-one




5-Chloro-6-nitrobenzo[d]oxazol-2(3H)-one was prepared by the procedure described in Example 4, step 1 starting from 2-amino-4-chloro-5-nitrophenol (5.53 g, 29.3 mmol), to afford a brown solid (5.25 g, 83%), which was taken through to the next step with no further purification.
Step 2: 5-Chloro-3-methyl-6-nitrobenzo[d]oxazol-2(3H)-one




5-Chloro-3-methyl-6-nitrobenzo[d]oxazol-2(3H)-one was prepared by the procedure described in Example 2, step 2 starting from 5-chloro-6-nitrobenzo[d]oxazol-2(3H)-one (1.75 g, 8.1 mmol) substituting iodomethane for dimethylsulfate. The crude product was recrystallised from methanol to afford an orange solid (0.6 g, 32%), which was taken through to the next step with no further purification.
Step 3: 6-Amino-5-chloro-3-methylbenzo[d]oxazol-2(3H)-one




6-Amino-5-chloro-3-methylbenzo[d]oxazol-2(3H)-one was prepared by the procedure described in Example 2, step 4 starting from 5-chloro-3-methyl-6-nitrobenzo[d]oxazol-2(3H)-one (520 mg, 2.3 mmol) to afford a solid (400 mg, 88%), which was taken through to the next step with no further purification.
Step 4: 5-Chloro-3-methyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzo[d]oxazol-2(3H)-one




5-Chloro-3-methyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzo[d]oxazol-2(3H)-one was prepared by the procedure described in Example 2, step 5 starting from 6-amino-5-chloro-3-methylbenzo[d]oxazol-2(3H)-one (224 mg, 1.1 mmol). Purification via silica gel column chromatography (0-100% ethyl acetate/isohexane) gave a solid which was triturated with petroleum ether-diethyl ether (1:1) to afford a white solid (15 mg, 7%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=374 at 3.58 min. 1H NMR δ (ppm) (400 MHz, CDCl3): 8.57 (1H, s), 8.19 (1H, s), 7.48 (1H, s), 7.01 (1H, s), 5.28 (1H, s), 3.39 (3H, s), 3.09 (3H, d, J=4.67).
Example 6
N2-(6-chloro-2-methylbenzo[d]oxazol-5-yl)-N4-methyl-5-(trifluoromethyl)pyrimidine-2,4-diamine
Step 1: 6-Chloro-2-methylbenzo[d]oxazol-5-amine




6-Chloro-2-methylbenzo[d]oxazol-5-amine was prepared by the procedure described in Example 2, step 4 starting from 6-chloro-2-methyl-5-nitrobenzo[d]oxazole (493 mg, 2.3 mmol) to afford a solid (200 mg, 47%). This was taken through to the next step with no further purification.
Step 2: N2-(6-chloro-2-methylbenzo[d]oxazol-5-yl)-N4-methyl-5-(trifluoromethyl)pyrimidine-2,4-diamine




N2-(6-chloro-2-methylbenzo[d]oxazol-5-yl)-N4-methyl-5-(trifluoromethyl)pyrimidine-2,4-diamine was prepared by the procedure described in Example 2, step 5 starting from 6-chloro-2-methylbenzo[d]oxazol-5-amine (205 mg, 1.1 mmol). Purification via silica gel column chromatography (0-100% ethyl acetate/isohexane) gave a solid which was triturated with petroleum ether-diethyl ether (1:1) to afford a white solid (15 mg, 7%). LCMS (10 cm_ESCI_Formic_MeCN): [MH+]=358 at 3.49 min. 1H NMR δ (ppm) (400 MHz, d6-DMSO): 8.90 (1H, s), 8.19 (2H, d, J=11.5), 7.87 (1H, s), 7.19 (1H, d, J=5.2), 2.86 (3H, d, J=4.3), 2.65 (3H, s).
Example 7
5-Methoxy-2-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one




A mixture of impure 6-amino-5-methoxy-2-methylisoindolin-1-one (<0.57 mmol), 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (66 mg, 0.31 mmol) and pTSA (59 mg, 0.31 mmol) in dioxane (4 mL) was heated to 100° C. for 4 h. DCM (10 mL) and saturated aqueous NaHCO3 (10 mL) were added and the organic phase was passed through a hydrophobic frit and the solvent was removed in vacuo. Purification of the residue via reversed phase preparative HPLC gave 5-methoxy-2-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one (15 mg, 13%) as an off-white solid. LCMS (10 cm_ESCI_Bicarb_MeCN): [M+H]+=368 at 2.89 min. 1H NMR (400 MHz, CDCl3): δ 9.04 (s, 1H), 8.17 (s, 1H), 7.80 (s, 1H), 6.93 (s, 1H), 5.26 (br s, 1H), 4.31 (s, 2H), 3.97 (s, 3H), 3.20 (d, J 4.7, 3H), 3.18 (s, 3H).
Example 8
6-Chloro-5-(5-chloro-4-(methylamino)pyrimidin-2-ylamino)-2-methylisoindolin-1-one




A mixture of 5-amino-6-chloro-2-methylisoindolin-1-one (61 mg, 0.31 mmol), 2,5-dichloro-4-methoxypyrimidine (55 mg, 0.31 mmol) and pTSA (59 mg, 0.31 mmol) in dioxane (4 mL) was heated to 100° C. for 18 h. DCM (10 mL) and saturated aqueous NaHCO3 (10 mL) were added and the mixture was passed through a hydrophobic frit. The solvent was removed in vacuo and purification of the residue via reversed phase preparative HPLC gave 6-chloro-5-(5-chloro-4-(methylamino)pyrimidin-2-ylamino)-2-methylisoindolin-1-one (26 mg, 25%) as an off-white solid. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=338 at 2.83 min. 1H NMR (400 MHz, d6-DMSO): δ 8.46 (s, 1H), 8.29 (s, 1H), 8.01 (s, 1H), 7.73 (s, 1H), 7.41 (q, J 4.6, 1H), 4.48 (s, 2H), 3.08 (s, 3H), 2.91 (d, J 4.6, 3H).
Example 9
6-Chloro-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one




6-Chloro-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one was prepared according to the procedure described for Example 8, using 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine. LCMS (10 cm_ESCI_Formic_MeCN): [M+H]+=372 at 3.48 min. 1H NMR (400 MHz, d6-DMSO): δ 8.91 (s, 1H), 8.23 (s, 1H), 8.23 (s, 1H), 7.75 (s, 1H), 7.26 (q, J 4.5, 1H), 4.49 (s, 2H), 3.09 (s, 3H), 2.88 (d, J 4.5, 3H).
Example 10
4-methoxy-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one




To a solution of 5-amino-4-methoxy-2-methylisoindolin-1-one (81 mg, 0.42 mmol) in t-BuOH (3 mL) was added 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (89 mg, 0.42 mmol). The mixture was heated at 100° C. for 1 h under microwave irradiation. The solvent was evaporated in vacuo and the residue was purified by reverse-phase prep-HPLC to afford the title compound as a white solid (35 mg, yield 23%). m/z (ES+APCI)+: [M+H]+368.1. 1H-NMR (Bruker, 500 MHz, MeOD) δ 8.65 (d, J=8.5 Hz, 1H), 8.13 (s, 1H), 7.43 (d, J=8.5 Hz, 1H), 4.69 (s, 2H), 4.06 (s, 3H), 3.19 (s, 3H), 3.05 (s, 3H)
The above compounds, together with additional compounds made using the above procedures, are shown below in Table 3 together with LRRK2 Ki values (micromolar).











TABLE 3






Structure
Name
H1 NMR
Ki







 1




7-chloro-4-methyl-8- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)- 3,4-dihydrobenzo[f][1,4] oxazepin-5(2H)-one
1H NMR δ (ppm)(400 MHz, CDCl3): 8.41 (1 H, s), 8.22 (1 H, s), 7.97 (1 H, s), 7.77- 7.67 (1 H, m), 5.29 (1 H, s), 4.47-4.39 (2 H, m), 3.61- 3.51 (2 H, m), 3.21-3.18 (3 H, m), 3.20-3.07 (3 H, m).
0.003


 


 2




7-((5-chloro-4- (methylamino) pyrimidin-2-yl)amino)-6- methoxy-2,2,4- trimethyl-2H- benzo[b][1,4]oxazin- 3(4H)-one
1H NMR δ (ppm)(400 MHz, DMSO): 8.05 (1 H, s), 7.98 (1 H, s), 7.54 (1 H, s), 7.28 (1 H, d, J = 5.19 Hz), 6.85 (1 H, s), 3.93 (3 H, s), 3.66 (3 H, s), 2.91 (3 H, d, J = 4.55 Hz), 1.41 (6 H, s).
0.001


 


 3




5-methoxy-N,N- dimethyl-6-((4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- yl)amino)benzofuran- 2-carboxamide
1H NMR δ (ppm)(400 MHz, DMSO): 8.70 (1 H, s), 8.28 (1 H, s), 8.17 (1 H, s), 7.40- 7.32 (3 H, m), 3.98 (3 H, s), 3.41-3.28 (6 H, m), 3.01 (3 H, d).
0.001


 


 4




5-chloro-1,3-dimethyl- 6-((4-(methylamino)- 5- (trifluoromethyl) pyrimidin-2-yl)amino)-1H- benzo[d]imidazol- 2(3H)-one
1H NMR δ (ppm)(400 MHz, DMSO-d6): 8.83 (1 H, s), 8.13 (1 H, s), 7.58 (1 H, s), 7.38 (1 H, s), 7.08 (1 H, s), 3.36 (6 H, s), 2.84 (3 H, d, J = 4.21 Hz).
0.002


 


 5




5-chloro-3-methyl-6- ((4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- yl)amino)benzo[d] oxazol-2(3H)-one
1H NMR δ (ppm)(400 MHz, CDCl3): 8.57 (1 H, s), 8.19 (1 H, s), 7.48 (1 H, s), 7.01 (1 H, s), 5.28 (1 H, s), 3.39 (3 H, s), 3.09 (3 H, s).
0.004


 


 6




N2-(6-chloro-2- methylbenzo[d]oxazol- 5-yl)-N4-methyl-5- (trifluoromethyl) pyrimidine-2,4-diamine
1H NMR δ (ppm)(400 MHz, DMSO): 8.90 (1 H, s), 8.19 (2 H, d, J = 11.50 Hz), 7.87 (1 H, s), 7.19 (1 H, d, J = 5.17 Hz), 2.86 (3 H, d, J = 4.33 Hz), 2.65 (3 H, s).
0.005


 


 7




5-methoxy-2-methyl- 6-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one
1H NMR (400 MHz, CDCl3): δ 9.04 (s, 1H), 8.17 (s, 1H), 7.80 (s, 1H), 6.93 (s, 1H), 5.26 (br s, 1H), 4.31 (s, 2H), 3.97 (s, 3H), 3.20 (d, J = 4.7 Hz, 3H), 3.18 (s, 3H).
0.005


 


 8




6-chloro-5-(5-chloro- 4- (methylamino) pyrimidin-2-ylamino)-2- methylisoindolin-1- one
1H NMR (400 MHz, DMSO): δ 8.46 (s, 1H), 8.29 (s, 1H), 8.01 (s, 1H), 7.73 (s, 1H), 7.41 (q, J = 4.6 Hz, 1H), 4.48 (s, 2H), 3.08 (s, 3H), 2.91 (d, J = 4.6 Hz, 3H).
0.003


 


 9




6-chloro-2-methyl-5- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one
1H NMR (400 MHz, DMSO): δ 8.91 (s, 1H), 8.23 (s, 1H), 8.23 (s, 1H), 7.75 (s, 1H), 7.26 (q, J = 4.5 Hz, 1H), 4.49 (s, 2H), 3.09 (s, 3H), 2.88 (d, J = 4.5 Hz, 3H).
0.001


 


10




2-cyclopropyl-4- methoxy-5-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.007


 


11




4-methoxy-2-methyl- 5-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.001


 


12




6-methoxy-2,2,4- trimethyl-7-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-2H- benzo[b][1,4]oxazin- 3(4H)-one
1H NMR δ (ppm)(DMSO- d6): 8.21 (1 H, s), 8.00 (1 H, s), 7.90 (1 H, s), 7.18 (1 H, d, J = 5.15 Hz), 6.86 (1 H, s), 3.35 (3H, s), 3.92 (3 H, s), 2.92 (3 H, s), 1.41 (6 H, s).
0.008


 


13




5-chloro-6-(5-chloro- 4- (methylamino) pyrimidin-2- ylamino)benzo[d] oxazol-2(3H)-one
1H NMR δ (ppm)(400 MHz, CDCl3): 8.53 (1 H, s), 7.90 (1 H, s), 7.06-7.02 (1 H, m), 5.47 (1 H, s), 3.09 (4 H, m)
0.004


 


14




5-chloro-6-(5-chloro- 4- (methylamino) pyrimidin-2-ylamino)-3- methylbenzo[d]oxazol- 2(3H)-one
1H NMR δ (ppm)(DMSO- d6): 8.30 (1 H, s), 7.90 (2 H, d, J = 4.87 Hz), 7.51 (1 H, s), 7.29-7.22 (1 H, m), 3.36 (3 H, s), 2.86 (3 H, d, J = 4.52 Hz).
0.002


 


15




7-methoxy-2-methyl- 6-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-3,4- dihydroisoquinolin- 1(2H)-one

0.007


 


16




2-(7-methoxy-2- methyl-1-oxo-1,2,3,4- tetrahydroisoquinolin- 6-ylamino)-4- (methylamino) pyrimidine-5-carbonitrile

0.044


 


17




7-methoxy-2-methyl- 6-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)phthalazin- 1(2H)-one

0.027


 


18




2-ethyl-4-methoxy-5- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.009


 


19




2-(2-hydroxypropan- 2-yl)-4-methoxy-5-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.004


 


20




4-methoxy-5-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-2- (oxetan-3- yl)isoindolin-1-one

0.009


 


21




6-methoxy-2-methyl- 5-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.005


 


22




(5-chloro-1-methyl-6- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-1H- indol-3- yl)(morpholino) methanone

0.003


 


23




2-(2-hydroxy-2- methylpropyl)-6- methoxy-5-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one

0.01


 


24




5-(4-(ethylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-2-(2- hydroxy-2- methylpropyl)-6- methoxyisoindolin-1- one

0.003


 


25




6-methoxy-5-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-2- (oxetan-3- yl)isoindolin-1-one

0.006


 


26




5-(4-(ethylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-6- methoxy-2-(oxetan-3- yl)isoindolin-1-one

0.002


 


27




5-chloro-N,N,1- trimethyl-6-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-1H- indole-3-carboxamide

0.006


 


28




(5-chloro-1-methyl-6- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-1H- indol-3-yl)(piperazin- 1-yl)methanone

0.004


 


29




(5-chloro-6-(4- (ethylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-1- methyl-1H-indol-3- yl)morpholino) methanone

0.001


 


30




(5-methoxy-1-methyl- 6-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-1H- indol-3- yl)(morpholino) methanone

0.001


 


31




(5-chloro-1-methyl-6- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)-1H- indol-3-yl)(4- methylpiperazin-1- yl)methanone

0.001


 


32




(5-chloro-1-methyl-6- (4-(methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)- 1H-indol-3-yl)(4- ethylpiperazin-1- yl)methanone

0.001


 


33




(1S,4S)-2-oxa-5- azabicyclo[2.2.1] heptan-5-yl(5-chloro-1- methyl-6-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2-ylamino)-1H- indol-3-yl)methanone

0.003


 


34




4-methoxy-2-methyl- 5-(4-(methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)isoindolin-1- one




 


35




(5-methoxy-6-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)benzofuran- 3- yl)(morpholino) methanone




 


36




5-methoxy-N,N- dimethyl-6-(4- (methylamino)-5- (trifluoromethyl) pyrimidin-2- ylamino)benzofuran- 3-carboxamide









Example 10
In Vitro LRRK2 LabChip Assay
This assay was used to determine a compound's potency in inhibiting activity of LRRK2 by determining, Kiapp, IC50, or percent inhibition values. In a polypropylene plate, LRRK2, fluorescently-labeled peptide substrate, ATP and test compound were incubated together. Using a LabChip 3000 (Caliper Life Sciences), after the reaction the substrate was separated by capillary electrophoresis into two populations: phosphorylated and unphosphorylated. The relative amounts of each were quantitated by fluorescence intensity. LRRK2 Ki was determined according to the 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 μM.

Assay conditions and materials used were as follows:
Final Assay Conditions:
LRRK2 G2019S in 5 mM MgCl2: 5.2 nM (Invitrogen lot #567054A)
LRRK2 G2019S in 1 mM MnCl2: 11 nM (Invitrogen lot #567054A)
LRRK2 Wild type in 5 mM MgCl2: 15 nM (Invitrogen lot #500607F)
LRRK2 I2020T in 5 mM MgCl2: 25 nM (Invitrogen lot #43594)
Substrate: 1 μM
ATP: 130 μM
Kinase reaction time: 2 hours
Temperature: ambient
Total volume: 20 μl
ATPapp Kms:
G2019S in 5 mM MgCl2: 130 μM
G2019S in 1 mM MnCl2: 1 μM
Wild type in 5 mM MgCl2: 80 μM
I2020T in 5 mM MgCl2: 14 μM
Materials:
Solid Support: Black 50 μL volume polypropylene 384 well plate (MatriCal cat #MP101-1-PP)
Kinase: LRRK2 G2019S (Invitrogen cat #PV4882).

    
    
        LRRK2 Wild type (Invitrogen cat #PV4874).
    
    


Substrate: 5FAM-GAGRLGRDKYKTLRQIRQ-CONH2 
Non-binding plate: 384 well clear V-bottom polypropylene plates (Greiner cat #781280).
ATP: 10 mM 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 #34869-100 mL).
Complete Reaction Buffer: H2O/25 mM Tris, pH 8.0/5 mM MgCl2/2 mM DTT/0.01% Triton X-100.
Stop Solution: H2O/100 mM HEPES, pH 7.2/0.015% Brij-35/0.2% Coating Reagent #3/20 mM EDTA.
Separation Buffer: H2O/100 mM HEPES, pH 7.2/0.015% Brij-35/0.1% Coating Reagent #3/1:200 Coating Reagent #8/10 mM EDTA/5% DMSO.
Compound Plate Preparation:
For serial dilutions, 34.6 μl DMSO was added to columns 3-24. For the assay controls, 37.5 μl DMSO was added to columns 1 and 2 of rows A and P. a,d and 50 μl 25 μM G-028831 (Staurosporine) was added to columns 1 and 2, row B. For the samples: to start at 100 μM, 37.5 μl DMSO was to columns 1 and 2, then 12.5 μl 10 mM compound; to start at 10 μM, 78 μl DMSO was added to columns 1 & 2, then 2 μl 10 mM compound; and to start at 1 μM, 25 μM compound (2 μl 10 mM cmpd+798 μl DMSO) was added to empty columns 1 and 2. A Precision instrument was used to perform 1:3.16 serial dilutions (“PLK_BM_serial_halflog”).
ATP Preparation:
ATP was diluted to 282.1 μM in Complete Kinase Buffer (final concentration was 130 μM).
Total and Blank Preparation:
In Complete Reaction Buffer, substrate was diluted to 4 μM. Equal volumes of Complete Reaction Buffer and 4 μM substrate were combined to obtain the blank. Equal volumes of Complete Reaction Buffer and 4 μM substrate were combined and to the combined solution was added 2× final LRRK2 concentration.
Assay Procedure:
To a 50 μl polypropylene plate, 5 μl/well buffer/substrate was added by hand to Blank wells. A Biomek FX was used to start the kinase reaction (“PLK SAR 23 ATP”). The following were added to the appropriate wells:
2 μA compound+23 μA ATP;
5 μl/well compound/ATP in Assay Plate;
5 μl/well kinase/substrate in Assay Plate;
The plate was incubated for 2 hours in the dark. Biomek FX was used to stop the kinase reaction (“PLK Stop”), and 10 μl/well Stop solution was added to the Assay Plate.
Results were read on the LabChip 3000.
Lab Chip 3000 Protocol:
The LabChip 3000 was run using the job “LRRK2 IC50” with the following job settings:
Pressure: −1.4 psi
Downstream voltage: −500 V
Upstream voltage: −2350 V
Post sample buffer sip time: 75 seconds
Post dye buffer sip time: 75 seconds
Final delay time: 200 seconds
Example 11
In Vitro LRRK2 Lanthascreen Binding Assay
This assay was used to determine a compound's potency in inhibiting activity of LRRK2 by determining, Kiapp, IC50, or percent inhibition values. In 384 well proxiplates F black, shallow well plates LRRK2, Eu-anti-GST-antibody, Alexa Fluor® Kinase tracer 236 and test compound were incubated together.
Binding of the Alexa Fluor® “tracer” to a kinase was detected by addition of a Eu-labeled anti-GST antibody. Binding of the tracer and antibody to a kinase results in a high degree of FRET, whereas displacement of the tracer with a kinase inhibitor results in a loss of FRET.
Assay conditions and materials used were as follows:
Final Assay Conditions:
GST-LRRK2 G2019S 10 nM
Eu-anti-GST-antibody 2 nM
Kinase tracer 236 8.5 nM
Kinase reaction time: 1 hour
Temperature: ambient
Total volume: 15 μl
DMSO 1%
Materials:
384 well proxiplates F black shallow well Perkin Elmer cat#6008260
Kinase: LRRK2 G2019S Invitrogen cat #PV4882(LOT 567054A).
Eu-labeled anti-GST antibody Invitrogen cat #PV5594
Alexa Fluor® Kinase tracer 236 Invitrogen cat #PV5592
TRIS-HCl Sigma cat #T3253
EGTA Sigma cat #E3889
Brij-35: Sigma cat #B4184(30% w/v)
DMSO: Sigma cat #D8418
MgCl2 Sigma cat #M9272
Reaction Buffer: H2O/50 mM Tris, pH 7.4/10 mM MgCl2/1 mM EGTA/0.01% Brij 35.
Compound Plate Preparation:
Serially dilute test compounds (10 mM stock) 1:3.16 (20 ul+43.2 ul) in 100% DMSO. 12 pt curve. Dilute each concentration 1:33.3 (3 ul+97 ul) in reaction buffer. Stamp 5 ul to assay plate. Final top test concentration 100 uM.
Total and Blank Preparation:
In Reaction Buffer, 5 ul of DMSO (3%) was added to total and blank wells and 5 ul of Eu-labeled anti-GST antibody (6 nM) was added to blank wells.
Assay Procedure:
Add 5 ul LRRK2(30 nM)/Eu-labeled anti-GST antibody (6 nM) mix to compound and total wells. Add 5 ul kinase tracer (25.5 nM) to all wells. Incubate plates at room temperature for 1 hour on a plate shaker (gentle shaking). Read on Perkin Elmer EnVision reader HTRF protocol.
Data Handling:
Calculate ratio: (665/620)*10000. Substract mean background values from all data points. Calculate % of control for each test value. Plot % of control vs Compound concentration. Calculate Ki Value (xlfit curve fitting—Morrison equation).
Results expressed as a Ki in μM. The equation for Ki: 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)))
Where Et=4 nM
kd (Tracer)=8.5 nM
Tracer concentration (S)=8.5 nM.
Example 12
Parkinson's Disease Animal Model
Parkinson's disease can be replicated in mice and in primates by administration of 1-methyl-4-phenyl tetrahydropyridine (MPTP), a selective nigrostriatal dopaminergic neurotoxin that produces a loss of striatal dopamine (DA) nerve terminal markers. Compounds of the invention may be evaluated for effectiveness in treatment of Parkinson's disease using MPTP induced neurodegeneration following generally the protocol described by Saporito et al., J. Pharmacology (1999) Vol. 288, pp. 421-427.
Briefly, MPTP is dissolved in PBS at concentrations of 2-4 mg/ml, and mice (male C57 weighing 20-25 g) are given a subcutaneous injection of 20 to 40 mg/kg. Compounds of the invention are solubilized with polyethylene glycol hydroxystearate and dissolved in PBS. Mice are administered 10 ml/kg of compound solution by subcutaneous injection 4 to 6 h before MPTP administration, and then daily for 7 days. On the day of the last injection, mice are sacrificed and the midbrain blocked and postfixed in paraformaldehyde. Striata are dissected free, weighed, and stored at −70° C.
The striata thus collected are evaluated for 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 striata may also be evaluated using the tyrosine hydroxylase assay of Okunu et al., Anal Biochem (1987) Vol. 129, pp. 405-411 by measuring 14CO2 evolution associated with tyrosine hydroxylase-mediated conversion of labeled tyrosine to L-dopa. The striata may further be evaluated using the Monoamine oxidase-B assay as described by White et al., Life Sci. (1984), Vol. 35, pp. 827-833, and by monitoring dopamine uptake as described by Saporito et al., (1992) Vol. 260, pp. 1400-1409.
While the present invention has been described with reference to the specific embodiments thereof, it should 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, 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 claims appended hereto.

## Claims

1. A compound of formula I:



or pharmaceutically acceptable salts thereof,
wherein:
A is a five- or six-membered saturated or unsaturated ring that includes one or two heteroatoms selected from O, N and S, which is substituted once with R5, and which is optionally substituted one, two or three times with R6;
X is: —NRa—; —O—; or —S(O)r— wherein r is from 0 to 2 and Ra is hydrogen or C1-6alkyl;
R1 is: C1-6alkyl; C1-6alkenyl; C1-6alkynyl; halo-C1-6alkyl; C1-6alkoxy-C1-6alkyl; hydroxy-C1-6alkyl; amino-C1-6alkyl; C1-6alkylsulfonyl-C1-6alkyl; C3-6cycloalkyl optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; oxetanyl; or oxetan-C1-6alkyl;
or R1 and Ra together with the atoms to which they are attached may form a three to six membered ring that may optionally include an additional heteroatom selected from O, N and S, and which is substituted with oxo, halo or C1-6alkyl;
R2 is: halo; C1-6alkoxy; cyano; C1-6alkynyl; C1-6alkenyl; halo-C1-6alkyl; halo-C1-6alkoxy; C3-6cycloalkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; C3-6cycloalkyl-C1-6alkyl wherein the C3-6cycloalkyl portion is optionally substituted with C1-6alkyl; tetrahydrofuranyl; tetrahydrofuranyl-C1-6alkyl; acetyl; oxetanyl; or oxetan-C1-6alkyl;
one of R3 and R4 is: halo; or C1-6alkoxy, and the other is hydrogen;
R5 is; oxo; C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; or —C(O)—NRbRc wherein Rb and Rc each independently is hydrogen or —C1-6alkyl, or Rb and Rc together with the atoms to which they are attached may form a heterocyclyl group that optionally includes an additional heteroatom selected from 0, N and S and which is optionally substituted one or more times with R6; and
each R6 is independently: C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; oxo; or —C(O)—NRbRc.

2. The compound of claim 1, wherein X is —NH— or —O—.

3. The compound of claim 1, wherein R1 is C1-6alkyl.

4. The compound of claim 1, wherein R2 is: halo; halo-C1-6alkyl; or cyano.

5. The compound of claim 1, wherein R2 is chloro, trifluoromethyl or cyano.

6. The compound of claim 1, wherein R2 is trifluoromethyl.

7. The compound of claim 1, wherein R3 is halo or C1-6alkoxy.

8. The compound of claim 1, wherein R3 is halo or methoxy.

9. The compound of claim 1, wherein R4 is halo or C1-6alkoxy.

10. The compound of claim 1, wherein R4 is halo or methoxy.

11. The compound of claim 1, wherein R5 is: oxo; C1-6alkyl; or —C(O)—NRbRc.

12. The compound of claim 1, wherein each R6 is independently: C1-6alkyl, hydroxy-C1-6alkyl, C1-6alkoxy-C1-6alkyl, heterocyclyl, oxo, or —C(O)—NRbRc.

13. The compound of claim 1, wherein said compound is of formula IIa or IIb:



wherein:
m is: 0; 1 or 2;
Z is; —C(R7)2—; —NR8— or —O—; or Z is absent;
each R7 is independently: hydrogen; or C1-6alkyl; and
each R8 is independently: hydrogen; C1-6alkyl; C3-6cycloalkyl; C3-6cycloalkyl-C1-6alkyl; halo; halo-C1-6alkyl; hydroxy-C1-6alkyl; C1-6alkoxy-C1-6alkyl; heterocyclyl; or —C(O)—NRbRc.

14. The compound of claim 13, wherein m is 0 or 1.

15. The compound of claim 13, wherein Z is —C(R7)2—.

16. The compound of claim 13, wherein Z is —NR8—.

17. The compound of claim 13, wherein Z is —O—.

18. The compound of claim 13, wherein Z is absent.

19. A composition comprising:
(a) a pharmaceutically acceptable carrier; and
(b) a compound of claim 1.

20. A method for treating Parkinson's disease, said method comprising administering to a subject in need thereof an effective amount of a compound of claim 1.

21. The compound of claim 1, wherein said compound is selected from:
7-chloro-4-methyl-8-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one;
7-((5-chloro-4-(methylamino)pyrimidin-2-yl)amino)-6-methoxy-2,2,4-trimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one;
5-methoxy-N,N-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzofuran-2-carboxamide
5-chloro-1,3-dimethyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-benzo[d]imidazol-2(3H)-one;
5-chloro-3-methyl-6-((4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzo[d]oxazol-2(3H)-one;
N2-(6-chloro-2-methylbenzo[d]oxazol-5-yl)-N4-methyl-5-(trifluoromethyl)pyrimidine-2,4-diamine;
5-methoxy-2-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
6-chloro-5-(5-chloro-4-(methylamino)pyrimidin-2-ylamino)-2-methylisoindolin-1-one;
6-chloro-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
2-cyclopropyl-4-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
4-methoxy-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
6-methoxy-2,2,4-trimethyl-7-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2H-benzo[b][1,4]oxazin-3(4H)-one;
5-chloro-6-(5-chloro-4-(methylamino)pyrimidin-2-ylamino)benzo[d]oxazol-2(3H)-one;
5-chloro-6-(5-chloro-4-(methylamino)pyrimidin-2-ylamino)-3-methylbenzo[d]oxazol-2(3H)-one;
7-methoxy-2-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroisoquinolin-1(2H)-one;
2-(7-methoxy-2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-ylamino)-4-(methylamino)pyrimidine-5-carbonitrile;
7-methoxy-2-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)phthalazin-1(2H)-one;
2-ethyl-4-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one
2-(2-hydroxypropan-2-yl)-4-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
4-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-(oxetan-3-yl)isoindolin-1-one;
6-methoxy-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
(5-chloro-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)(morpholino)methanone;
2-(2-hydroxy-2-methylpropyl)-6-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
5-(4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-(2-hydroxy-2-methylpropyl)-6-methoxyisoindolin-1-one;
6-methoxy-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-2-(oxetan-3-yl)isoindolin-1-one;
5-(4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-6-methoxy-2-(oxetan-3-yl)isoindolin-1-one;
5-chloro-N,N,1-trimethyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indole-3-carboxamide;
(5-chloro-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)(piperazin-1-yl)methanone;
(5-chloro-6-(4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1-methyl-1H-indol-3-yl)(morpholino)methanone;
(5-methoxy-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)(morpholino)methanone;
(5-chloro-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)(4-methylpiperazin-1-yl)methanone;
(5-chloro-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)(4-ethylpiperazin-1-yl)methanone;
(1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl(5-chloro-1-methyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-1H-indol-3-yl)methanone;
4-methoxy-2-methyl-5-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)isoindolin-1-one;
(5-methoxy-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)benzofuran-3-yl)(morpholino)methanone; and
5-methoxy-N,N-dimethyl-6-(4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)benzofuran-3-carboxamide.