EA043797B1 - PYRIMIDINE-2-YLAMINO-1H-PYRAZOLES AS LRRK2 INHIBITORS FOR USE IN THE TREATMENT OF NEURODEGENERATIVE DISEASES - Google Patents

Description

Cross references to related applications

This application claims priority under 35 U.S.C. §119(e) by US Provisional Application No. 62/350876 filed June 16, 2016, No. 62/417151 filed November 3, 2016, No. 62/476581 filed March 24, 2017, and No. 6 2/510711 filed May 24, 2017, each of which is incorporated herein by reference in its entirety.

Field of technology

This document generally introduces new heteroaryl-substituted pyrimidines and their use as therapeutic agents, for example, as LRRK2 inhibitors.

State of the art

Neurodegenerative diseases such as Parkinson's disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Lewy body dementia, and Huntington's disease affect millions of people. Parkinson's disease is a chronic, progressive disorder of the motor system characterized by selective degeneration and cell death of dopaminergic neurons in the substantia nigra pars compacta of the brain. As a result, patients' ability to direct and control their movements is impaired. The cause of the disease was generally considered sporadic and unknown, but significant advances in understanding have been made over the past 15 years.

The genetic basis of the disease and the associated development of pathological processes led to the study of the gene encoding the protein leucine-rich repeat kinase 2 (LRRK2) in the study of the inherited form of Parkinson's disease (Paisan-Ruiz et al., Neuron, Vol. 44(4), 2004, 601-607). LRRK2 is a member of the ROCO protein family and shares five domains conserved with all other family members. Many missense mutations of the LRRK2 gene have been associated with autosomal dominant Parkinson's disease in familial studies (Trinh and Farrar, Nature Reviews in Neurology, Vol. 9, 2013, 445-454; Paisan-Ruiz et al., J. Parkinson's Disease, Vol. 3, 2013, 85-103). The most common pathogenic mutation, G2019S, is present in the highly conserved kinase domain of LRRK2 (See Gilks et al., Lancet, Vol. 365, 2005, 415-416). In vitro studies have shown that Parkinson's disease-associated mutations lead to increased LRRK2 kinase activity and a decreased rate of GTP hydrolysis (Guo et al., Experimental Cell Research, Vol. 313(16), 2007, 3658-3670). These data suggest that the kinase and GTPase activities of LRRK2 are important for pathogenesis, and the LRRK2 kinase domain may regulate the overall function of LRRK2 (see Cookson, Nat. Rev. Neurosci., Vol. 11, 2010, 791-797).

Despite progress in this field, there remains a need for improved LRRK2 inhibitors that are useful for the treatment of various neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis.

Description of the invention

Provided herein are compounds that are useful as LRRK2 inhibitors. Also provided herein are compositions, including pharmaceutical compositions that include the compounds, and methods of use (or administration). This document further provides compounds or compositions thereof for use in a method of treating a disease, disorder or condition that is mediated at least in part by LRRK2 kinase. Further provided herein are the uses of the compounds or compositions thereof for the manufacture of a medicament for the treatment of a disease, disorder or condition that is mediated at least in part by LRRK2 kinase. In one embodiment of the present invention, a compound of formula Ia is provided:

or a pharmaceutically acceptable salt, deuterated analogue, prodrug, stereoisomer or mixture of stereoisomers thereof, wherein:

R ¹ represents C ₃ . ₆ cycloalkyl, optionally substituted with one or more halogen, hydroxyl or cyano atoms, or C ₁ . ₆ alkyl, optionally substituted with halogen;

R ² represents halogen, cyano or C ₁ . ₆ alkyl, optionally substituted with halogen;

R ³ represents C ₁ . ₆ alkoxy, C ₃ . ₆ cycloalkyl, C ₃ . ₆ cycloalkoxy or -N(R11)(R ¹² );

R ⁴ represents hydrogen or halogen;

R ⁶ and R ⁷ are each independently hydrogen or C ₁ . ₆ alkyl, optionally substituted with halogen; And

R ⁸ and R ⁹ are each independently hydrogen, cyano, halogen, C1.6alkyl or C1. 6alkoxy; and R ¹¹ and R ¹² are each independently hydrogen, C1. ₆ alkyl or C ₃ . ₆ cycloalkyl.

In some embodiments of this invention, the compound is a compound presented in table. 1, or is a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof.

- 1 043797

In other embodiments of the present invention, a pharmaceutical composition is provided comprising the compound illustrated in Table 1, or a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In other embodiments, the present invention provides a method of treating a disease or condition mediated at least in part by LRRK2, the method comprising administering an effective amount of a pharmaceutical composition containing a compound illustrated in Table 1. 1, or a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier, diluent or excipient, to a subject in need thereof. In other embodiments of the present invention, a pharmaceutical composition is provided containing the compound illustrated in table. 1, or a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In other embodiments, the present invention provides a method of treating a disease or condition mediated at least in part by LRRK2, the method comprising administering an effective amount of a pharmaceutical composition containing a compound illustrated in Table 1. 1, or a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier, diluent or excipient, to a subject in need thereof. In other embodiments, the present invention provides a method of treating a disease or condition mediated at least in part by LRRK2, the method comprising administering an effective amount of a pharmaceutical composition containing a compound illustrated in Table 1. 1, or a pharmaceutically acceptable salt, deuterated analogue, prodrug, tautomer, stereoisomer or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier, diluent or excipient, to a subject in need thereof.

This document provides illustrative embodiments of the present invention. However, it should be recognized that this description is not intended to limit the scope of the present disclosure, but is instead presented as a description of illustrative embodiments of the present invention.

1. Definitions.

As used herein, the following words, phrases and symbols are generally intended to have the meanings set forth below, except that the context in which they are used indicates otherwise. A dash (-) that is not between two letters or symbols is used to indicate the point of attachment of a substituent. For example, -C(O)NH ₂ is attached via a carbon atom. The dash at the front or end of a chemical group is a matter of convenience; chemical groups may be shown with or without one or more dashes without losing their normal meaning. A wavy line or a dotted line drawn through a line in the structure indicates the indicated location of the group attachment. Unless chemically or structurally required, the direction or stereochemistry is not indicated or implied by the order in which the chemical group is written or named.

The prefix C _u - _v indicates that the following group has from and to v carbon atoms. For example, i-balkyl indicates that the alkyl group has 1 to 6 carbon atoms.

Reference to a value about or parameter herein includes (and describes) embodiments of the invention that are directed to the value or parameter itself. In some embodiments, the term about includes a specified amount of ±10%. In other embodiments of the invention, the term about includes the specified amount ±5%. In other embodiments of the invention, the term about includes the specified amount ±1%. Also, the term about X includes a description of X. In addition, singular forms include plural forms unless the context clearly indicates otherwise. Thus, for example, a reference to a compound includes a plurality of such compounds, and a reference to an assay includes a reference to one or more assays and their equivalents known to those skilled in the art. Alkyl refers to a straight or branched saturated hydrocarbon chain. As used herein, _{alkyl has 1 to 20 carbon atoms (i.e. C1-20 alkyl), 1 to 8 carbon atoms (i.e. C1-8 alkyl )} , 1 to 6 carbon atoms (t i.e. _C1.6 alkyl) or from 1 to 4 carbon atoms _{(i.e. C1.4 alkyl )} . Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl and 3-methylpentyl. In the event that an alkyl moiety having a specific number of carbon atoms is named by its chemical name or identified by its molecular formula, all positional isomers having that number of carbon atoms may be included; thus, for example, butyl includes n-butyl (i.e. -(CH ₂ ) ₃ CH ₃ ), sec-butyl (i.e. CH(CH ₃ )CH ₂ CH ₃ ), isobutyl (i.e. -CH ₂ CH(CH ₃ ) ₂ ) and tert-butyl (i.e. -C(CH ₃ ) ₃ ); and propyl includes n-propyl (ie -(CH ₂ ) ₂ CH ₃ ) and isopropyl (ie -CH(CH ₃ ) ₂ ).

- 2 043797

Some common alternative chemical names may be used. For example, a divalent group such as a divalent alkyl group, a divalent aryl group, etc. may also be referred to as an alkylene group or alkylenyl group, an arylene group or an arylenyl group, respectively. In addition, unless explicitly stated otherwise, where combinations of groups are referred to herein as a single moiety, such as arylalkyl or aralkyl, the last mentioned group contains the atom through which the moiety is attached to the rest of the molecule.

Alkenyl refers to an alkyl group containing at least one carbon-carbon double bond and having from 2 to 20 carbon atoms (i.e. C ₂ - ₂₀ alkenyl), from 2 to 8 carbon atoms (i.e. C ₂ - ₈ alkenyl), 2 to 6 carbon atoms (ie C ₂ - ₆ alkenyl) or 2 to 4 carbon atoms (ie C ₂ - ₄ alkenyl). Examples of alkenyl groups include ethenyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

Alkynyl refers to an alkyl group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e. _C2-20 alkynyl), 2 to 8 carbon atoms (i.e. _{C2_ 8} alkynyl), from 2 to _{6 carbon atoms (i.e. C 2_6 alkynyl ) or from 2 to 4 carbon atoms (i.e. C 2_4 alkynyl} ). The term alkynyl also includes groups having one triple bond and one double bond.

Alkoxy belongs to the alkyl-O- group. Examples of alkoxy groups include methoxy, ethoxy, npropoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, n-hexoxy and 1,2dimethylbutoxy.

Alkoxyalkyl belongs to the alkyl-O-alkyl group.

Alkylthio belongs to the alkyl-S- group.

Alkylsulfinyl belongs to the alkyl-S(O)- group.

Alkylsulfonyl belongs to the alkyl-S(O) ₂ - group.

Alkylsulfonylalkyl refers to -alkyl-S(O) ₂ -alkyl.

Acyl refers to the group -C(O)R ^y , where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Examples of acyl include formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl and benzoyl.

Amido refers to both the C-amido group, which refers to the -C(O)NR ^y R ^z group, and the N-amido group, which refers to the -NR ^y C(O)R ^z group, where R ^y and R ^z are independent are hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted, as defined herein, or R ^y and R ^z taken together to form a cycloalkyl or heterocyclyl; each of which may be optionally substituted as defined herein.

Amidoalkyl refers to an alkyl group, as defined above, where one or more hydrogen atoms are replaced by an amido group.

Amino refers to the group -NR ^y R ^z where R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Aminoalkyl refers to the group -alkyl-NR ^y R ^z where R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Amidino refers to -C(NR ^y )(NR ^z 2), wherein R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Aryl refers to an aromatic carbocyclic group having one ring (eg, monocyclic) or multiple rings (eg, bicyclic or tricyclic), including fused systems. As used herein, aryl has 6 to 20 _{ring carbon atoms (i.e., C6_20 aryl), 6 to 12 ring carbon atoms (i.e., C6_12 aryl ) ,} or 6 to 10 ring carbon atoms carbon atoms (i.e. C ₆ _ ₁₀ aryl). Examples of aryl groups include phenyl, naphthyl, fluorenyl and anthryl.

Aryl, however, does not encompass or overlap in any way with heteroaryl, as defined below. If one or more aryl groups are fused to a heteroaryl, the resulting ring system is heteroaryl. If one or more aryl groups are fused to a heterocyclyl, the resulting ring system is a heterocyclyl.

Arilalkyl or Aralkyl refers to the aryl-alkyl group.

Carbamoyl refers to both the O-carbamoyl group, which refers to the -OC(O)NR ^y R ^z group, and the N-carbamoyl group, which refers to the -NR ^y C(O)OR ^z group, where R ^y and R ^z is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

- 3 043797

A carboxylic acid ester or ester refers to both -OC(O)R ^x and -C(O)OR ^x , where R ^x is alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Cyanoalkyl refers to an alkyl group, as defined above, where one or more hydrogen atoms are replaced by a cyano group.

Cycloalkyl refers to a saturated or partially unsaturated cyclic alkyl group having one or more rings, including fused, bridged and spiro ring systems. The term cycloalkyl includes cycloalkenyl groups (ie, a cyclic group having at least one double bond) and carbocyclic fused ring systems having at least one sp ³ carbon atom (ie, at least one non-aromatic ring). As used herein, a cycloalkyl has 3 to 20 ring carbon atoms (i.e., _C3 to ₂₀ cycloalkyl), 3 to 12 ring carbon atoms (i.e., _C3 to ₁₂ cycloalkyl), 3 to 10 ring carbons carbon atoms (i.e. C ₃ - ₁₀ cycloalkyl), 3 _to 8 ring carbon atoms (i.e. C ₃ - ₈ cycloalkyl), or 3 to 6 ring carbon atoms (i.e. C _{3_6} cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

Polycyclic groups include, for example, bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, adamantyl, norbornyl, decalinyl, 7,7-dimethylbicyclo[2.2.1]heptanyl and the like. Additionally, the term cycloalkyl is intended to cover any non-aromatic ring that can be fused to an aryl ring, regardless of attachment to the rest of the molecule. In addition, cycloalkyl also includes spirocycloalkyl when there are two positions for substitution on the same carbon atom, for example spiro[2.5]octanyl, spiro[4.5]decanyl or spiro[5.5]undecanyl.

Cycloalkoxy refers to -O-cycloalkyl.

Cycloalkylalkyl belongs to the cycloalkyl-alkyl group.

Cycloalkylalkoxy refers to -O-alkyl-cycloalkyl.

Guanidino refers to -NR ^y C(=NR ^z )(NR ^y R ^z ), wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Hydrazino belongs to -NHNH2.

Imino refers to the group -C(NR ^y )R ^z wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Imido refers to the group -C(O)NR ^y C(O)R ^z wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Halogen refers to atoms occupying group VIIA of the periodic table, such as fluorine, chlorine, bromine or iodine.

Haloalkyl refers to a straight or branched alkyl group, as defined above, where one or more hydrogen atoms are replaced by halogen.

For example, where a moiety is substituted with more than one halogen, it may be referred to using a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyl substituted with two (di) or three (three) halogen-containing groups, which may, but are not necessarily, the same halogen atoms. Examples of haloalkyl include trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl and the like.

Haloalkoxy refers to an alkoxy group, as defined above, where one or more hydrogen atoms are replaced by halogen.

Hydroxyalkyl refers to an alkyl group, as defined above, where one or more hydrogen atoms are replaced by a hydroxy group.

Heteroalkyl refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are each independently substituted by the same or another heteroatom group, provided that the attachment to the rest of the molecule is through a carbon atom. The term heteroalkyl includes a straight or branched saturated chain containing a carbon atom and heteroatoms. As an example, 1, 2 or 3 carbon atoms may be independently replaced by the same or a different heteroatom group. Heteroatomic groups include, but are not limited to, -NR ^y -, -O-, -S-, -S(O)-, -S(O) ₂ -, and the like, where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of heteroalkyl groups include ethers (for example, -CH2OCH3, -CH(CH3)OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3, etc.), thioethers (for example, -CH2SCH3, -CH(CH3)SCH3, -CH2CH2SCH3, - CH2CH2SCH2CH2SCH3, etc.), sulfones (for example, -CH2S(O)2CH3, -CH(CH3)S(O)2CH3,

- 4 043797

-CH2CH2S(O)2CH ₃ , -CH2CH2S(O)2CH2CH2OCH3, etc.) and amines (for example, -CH2NR ^y CH3, -CH(CH3)NR ^y CH3, -CH2CH2NR ^y CH3, -CH2CH2NR ^y CH2CH2NR ^y CH ₃ , etc., wherein R ^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may be optionally substituted as defined herein). As used herein, heteroalkyl includes 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.

Heteroaryl refers to an aromatic group having a single ring, multiple rings, or multiple fused rings, with one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. As used herein, heteroaryl includes 1 to 20 ring carbon atoms (i.e., _{C1_20} heteroaryl), 3 to 12 ring carbon atoms (i.e., _C3 to ₁₂ heteroaryl), or 3 to 8 ring atoms carbon (ie C ₃ - ₈ heteroaryl); and 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, oxygen, and sulfur atoms. In some instances, heteroaryl includes 5-10 membered ring systems, 5-7 membered ring systems, or 5-6 membered ring systems, each independently having 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms or 1 ring heteroatom independently selected from nitrogen, oxygen and sulfur atoms. Examples of heteroaryl groups include acridinyl, benzimidazolyl, benzothiazolyl, benzidolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothiophenyl), benzothriazolyl, benzo[4,6]imidazo[1,2-a]pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl , dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, phenazinyl, phthalazine yl, pteridinyl, purinyl, pyrrolyl , pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl and triazinyl. Examples of fused heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl and imidazo[1,5 -a]pyridinyl, where heteroaryl can be linked through any ring of the fused system. Any aromatic ring having one or more fused rings containing at least one heteroatom is considered a heteroaryl regardless of attachment to the rest of the molecule (ie, through any of the fused rings). Heteroaryl does not span or overlap with aryl as defined above. Heteroarylalkyl belongs to the heteroaryl-alkyl group. Heterocyclyl refers to a saturated or partially unsaturated cyclic alkyl group with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur atoms. The term heterocyclyl includes heterocycloalkenyl groups (ie, a heterocyclyl group having at least one double bond), bridged heterocyclyl groups, fused heterocyclyl groups, and spiro-heterocyclicline groups.

The heterocyclyl may be a single ring or multiple rings, in which the multiple rings may be fused, bridged or spiro, and may contain one or more oxo (=O) or N-oxide (-O-) moieties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of attachment (i.e., can be bonded through a carbon atom or a heteroatom). In addition, the term heterocyclyl is intended to cover any non-aromatic ring containing at least one heteroatom, which ring can be fused to an aryl or heteroaryl ring independent of attachment to the rest of the molecule. As used herein, a heterocyclyl has 2 to 20 ring carbon atoms (i.e., _C2 to ₂₀ heterocyclyl), 2 to 12 ring carbon atoms (i.e., _C2 to ₁₂ heterocyclyl), 2 to 10 ring carbons carbon atoms (i.e. C ₂ - ₁₀ heterocyclyl), from 2 to 8 ring carbon atoms (i.e. C ₂ - ₈ heterocyclyl), from 3 to 12 ring carbon atoms (i.e. C ₃ - ₁₂ heterocyclyl ), from 3 to ₈ ring carbon atoms (i.e. C _{3_8} heterocyclyl _{) or from 3 to 6 ring carbon atoms (i.e. C 3_6 heterocyclyl} ); has 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 heteroatom independently selected from nitrogen, sulfur or oxygen. Examples of heterocyclyl groups include azetidinyl, azepinyl, benzodioxolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzopyranyl, benzodioxinyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[1,3]dithianyl, decahydroisoquinolyl , furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, oxetanil, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4-piperidonyl Pirrolidinil, pyrazolidinil, chinuclidinyl, thiazolidinyl, tetrahydrofiril, tetrahydropiralil, tritianil, tetrahydrochinolinil, thiophenil (i.e. thyenil), tetrahydrine, thiomorpholinyl, thiamorpholinyl, 1,1,1-oxotiomorpholinyl. The term heterocyclyl also includes spiroheterocyclyl when there are two substitution positions on the same carbon atom. Examples of spiro

- 5 043 797 heterocyclyl rings include bicyclic and tricyclic ring systems such as 2-oxa-7-azaspiro[3,5]nonanyl, 2-oxa-6-azaspiro[3,4]octanyl and 6-oxa-1-azaspiro[3, 3]heptanyl. Examples of fused-heterocyclyl rings include, but are not limited to, 1,2,3,4tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl and isoindolinyl, where the heterocyclyl may be linked through any ring condensed system.

Heterocyclylalkyl belongs to the heterocyclyl-alkyl group.

The term leaving group refers to an atom or group of atoms that is displaced in a chemical reaction as a stable species, carrying bonding electrons with it. Non-limiting examples of the leaving group include halogen, methanesulfonylnoxy, p-toluenesulfonyloxy, trifluoromethanesulfonyloxy, nonafluorobutanesulfonyloxy, (4-bromobenzene)sulfonyloxy, (4-nitrobenzene)sulfonyloxy, (2-nitrobenzene)sulfonyloxy, (4-isopropylbenzene)sulfonyloxy, (2,4, 6-triisopropylbenzene)sulfonyloxy, (2,4,6-trimethylbenzene)sulfonyloxy, (4-tert-butylbenzene)sulfonyloxy, benzenesulfonyloxy, (4-methoxybenzene)sulfonyloxy, and the like.

Oxime refers to the group -CR ^y (=NOH), where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Sulfonyl refers to the group -S(O)2R ^y , where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of sulfonyl are methylsulfonyl, ethylsulfonyl, phenylsulfonyl and toluenesulfonyl.

Sulfinyl refers to the group -S(O)R ^y , where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of sulfinyl are methylsulfinyl, ethylsulfinyl, phenylsulfinyl and toluenesulfinyl.

Sulfonamido refers to the groups -SO2NR ^y R ^z and -NR ^y SO2R ^z , where R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

The terms optional or optional mean that the subsequently described event or circumstance may or may not occur, and that the description includes cases in which the event or circumstance has occurred and cases in which it has not. In addition, the term optionally substituted refers to one or more hydrogen atoms on the designated atom or group, which may or may not be replaced by the remainder other than hydrogen.

As used herein, substituted means any of the above groups (e.g., alkyl, alkenyl, alkynyl, alkylene, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl and/or heteroalkyl) wherein at least one hydrogen atom is replaced by a non with a hydrogen atom, such as, but not limited to, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, amido, amino, amidino, aryl, aralkyl, azido, carbamoyl, carboxyl, carboxyl ether, cyano, cycloalkyl, cycloalkylalkyl, guanadine , halogen, haloalkyl, haloalkoxy, hydroxyalkyl, heteroalkyl, heteroaclyl, heteroaklylkyl, heterocyclyl, heterocyclylalkyl, hydrazine, hydrazone, imino, imido, hydroxy, oxo, oxime, nitro, sulfonyl, sulfinyl, alkylsulfonyl, alkylsulfinyl, thiocyanate, sulfinic acid, sulf onic acid , sulfonamide, thiol, thioxo, N-oxide or -Si(R ^y )3, where each R ^y independently represents hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, aryl, heteroaryl or heterocyclyl. In one embodiment of the invention, substituted includes any of the above groups (for example, alkyl, alkenyl, alkynyl, alkylene, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl and/or heteroalkyl) in which one or more hydrogen atoms are substituted -NR ^g R ^h , -NR ^g C(=O)R ^h , -NR ^g C(=O)NR ^g R ^h , -NR ^g C(=O)OR ^h , -NR ^g SO2R ^h , -OC( =O)NR ^g R ^h , -OR ^g , -SR ^g , -SOR ^g , -SO2R ^g , -OSO2R ^g , -SO2OR ^g , =NSO2R ^g and -SO2NR ^g R ^h . Substituted also means any of the above groups in which one or more hydrogen atoms are substituted -C(=O)R ^g , -C(=O)OR ^g , -C(=O)NR ^g R ^h , -CH2SO ₂ R ^g , -CH ₂ SO ₂ NR ^g R ^h . In the previous case, R ^g and R ^h are the same or different, and independently represent hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl and/or heteroarylalkyl. Substituted further means any of the above groups in which one or more hydrogen atoms are replaced by a bond with amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl and/or heteroarylalkyl group. In addition, each of the above substituents may also be optionally replaced by one or more of the above substituents.

Polymers or similar undefined structures described by defining substituents with additional substituents, extrapolating to infinity (e.g., substituted

- 6 043797 aryl having a substituted alkyl which is itself substituted by a substituted aryl group which is further substituted by a substituted heteroalkyl group, etc.) are not intended to be included herein. Unless otherwise stated, the maximum number of consecutive substitutions in the compounds described herein is three. For example, successive substitutions of substituted aryl groups with two other substituted aryl groups are limited to ((substituted aryl) substituted aryl) substituted aryl. Likewise, the above definitions are not intended to include unacceptable substitution schemes (eg, methyl substituted with 5 fluorine atoms or heteroaryl groups having two adjacent ring oxygen atoms). Such unacceptable substitution patterns are well known to one skilled in the art. When used to modify a chemical group, the term substituted may describe other chemical groups as defined herein. Unless otherwise indicated, if a group is described as optionally substituted, then any substituents on the group are themselves unsubstituted. For example, in some embodiments of this invention, the term substituted alkyl refers to an alkyl group having one or more substituents, including hydroxy, halogen, alkoxy, acyl, oxo, amino, cycloalkyl, heterocyclyl, aryl, and heteroaryl. In other embodiments, one or more substituents may be further substituted with halogen, alkyl, haloalkyl, hydroxy, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is substituted. In other embodiments of the invention, the substituents may be further substituted with halogen, alkyl, haloalkyl, alkoxy, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is unsubstituted.

Any compound or structure given herein is also intended to represent the unlabeled forms as well as the isotopically labeled forms of the compounds. Isotopically labeled compounds have the structures depicted herein, except that one or more atoms are replaced by an atom having a specified atomic mass or mass number. Examples of isotopes that may be included in the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as ^2H , ^3H , ^nC , ^13C , ^14C , ^13N , ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²³ I and ¹²⁵ I, respectively. Included herein are various isotopically labeled compounds of the present invention, which include radioactive isotopes such as ³ H, ¹³ C and ¹⁴ C. Such isotopically labeled compounds may be useful in metabolic studies, reaction kinetic studies, detection methods or imaging such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including tests of substrate or drug tissue distribution, or when treating patients with radioactive drugs.

This invention also includes deuterated analogues of the compounds described herein, in which 1 to n hydrogen atoms attached to the carbon atom are replaced by deuterium, in which n represents the number of hydrogen atoms in the molecule. Such compounds exhibit increased resistance to metabolism and are therefore useful for increasing the half-life of any compound when administered to a mammal, especially a human. See, for example, Foster, Deuterium Isotope Effects in Studies of Drug Metabolism, Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are synthesized using methods known in the art, for example, using starting materials in which one or more hydrogen atoms are replaced by deuterium.

Deuterium-labeled or substituted therapeutic compounds of this invention may have improved DMPK (drug metabolism and pharmacokinetics) properties related to absorption, distribution, metabolism and elimination (ADME). Substitution with heavier isotopes such as deuterium may confer certain therapeutic benefits associated with greater metabolic stability, such as increased in vivo half-life, reduced dosage requirements, and/or improved therapeutic index. The ¹⁸ F, ³ H, ^P S labeled compound may be useful for PET or SPECT, or other imaging studies. The isotopically labeled compounds of this invention and their prodrugs can generally be prepared by the procedures described in the Schemes or in the Examples and Procedures described below by replacing the non-isotopically labeled reagent with a readily available isotopically labeled reagent. It is understood that deuterium in this context is considered as a substituent in the compound described herein.

The concentration of such a heavier isotope, in particular deuterium, can be determined using the isotope enrichment factor. In the compounds of this invention, any atom not specifically designated as a particular isotope is intended to be any stable isotope of that atom. Unless otherwise noted, when an item is designated specifically as H or hydrogen, the item is considered to have hydrogen in the isotopic composition of the natural isotope. Accordingly, in the compounds of this invention, any atom specifically designated as deuterium (D) is assumed to be deuterium. In many cases, the compounds of this invention are capable of forming acidic and/or basic salts due to the presence of an amino group and/or carboxyl groups or groups similar thereto.

Pharmaceutically acceptable salts, hydrates, solvates, tautomeric forms, stereoisomers, and prodrugs of the compounds described herein are also provided. Pharmaceutically at

- 7 043797 acceptable or physiologically acceptable refers to compounds, salts, compositions, dosage forms and other substances that can be used for the preparation of a pharmaceutical composition suitable for use in veterinary medicine or in the pharmaceutical industry.

The term pharmaceutically acceptable salt of a given compound refers to salts that retain the biological effectiveness and properties of the compound and that are not biologically or otherwise undesirable. Pharmaceutically acceptable salts or physiologically acceptable salts include, for example, inorganic acid salts and organic acid salts. In addition, when the compounds described herein are prepared as an acid addition salt, the free base can be prepared by making the acid salt solution alkaline. Conversely, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid in accordance with conventional methods for preparing acid addition salts from basic compounds. Those skilled in the art will appreciate the various synthetic approaches that can be used to prepare non-toxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Salts derived from organic acids include acetic acid, propionic acid, gluconic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. Likewise, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, aluminum, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines such as alkylamines (i.e. NH ₂ (alkyl)), dialkylamines (i.e. HN (alkyl) ₂ ), trialkylamines (i.e. N(alkyl) ₃ ), substituted alkylamines (i.e. _NH2 (substituted alkyl)), di(substituted alkyl)amines (i.e. HN(substituted alkyl) ₂ ), tri(substituted alkyl)amines (i.e. N(substituted alkyl) ₃ ), alkenyl amines (i.e. NH2 (alkenyl)), dialkenyl amines (i.e. HN(alkenyl) ₂ ), trialkenyl amines (i.e. N(alkenyl) ) ₃ ), substituted alkenyl amines (i.e. NH ₂ (substituted alkenyl)), di(substituted alkenyl) amines (i.e. HN (substituted alkenyl) ₂ ), tri(substituted alkenyl) amines (i.e. N (substituted alkenyl) ₃ , mono-, di- or tricycloalkylamines (i.e. NH ₂ (cycloalkyl), HN(cycloalkyl) ₂ , N(cycloalkyl) ₃ ), mono-, di- or triarylamines (i.e. NH ₂ (aryl), HN(aryl) ₂ , N(aryl) ₃ ), or mixed amines, etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri( npropyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine and the like.

The term hydrate refers to the complex formed by combining a compound described herein and water.

Solvate refers to an associate or complex of one or more solvent molecules and a compound of this invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid and ethanolamine. Some of the compounds exist as tautomers. Tautomers are in equilibrium with each other. For example, amide-containing compounds can be in equilibrium with imido acid tautomers. Regardless of which tautomer is shown and regardless of the nature of the equilibrium between tautomers, those skilled in the art will understand that these include both amide and imido acid tautomers. Thus, amide-containing compounds are understood as their imido acid tautomers. Likewise, imido acid-containing compounds are intended to include their amide tautomers.

The compounds of this invention or their pharmaceutically acceptable salts include an asymmetric center and thus can form enantiomers, diastereomers and other stereoisomeric forms, which can be defined in terms of absolute stereochemistry as (R)- or (S)- or, as ( D)- or (L)- for amino acids. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or separated using conventional methods, e.g. , chromatography and fractional crystallization. Conventional methods for preparing/isolating individual enantiomers include chiral synthesis from a suitable optically pure starting material or resolution of the racemate (or racemate salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). In the event that the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are intended to include geometric isomers of E,

- 8 043797 and Z.

The term stereoisomer refers to a compound consisting of the same atoms linked by the same bonds, but having different three-dimensional structures that are not interchangeable. The present invention relates to various stereoisomers and mixtures thereof and includes enantiomers, which refer to two stereoisomers whose molecules are incomparable mirror images of each other.

Diastereomers are stereoisomers that have at least two asymmetric atoms but that are not mirror images of each other.

Prodrugs mean any compound that releases the active parent drug according to the structure described herein in vivo when such prodrug is administered to a mammalian subject. Prodrugs of a compound described herein are prepared by modifying functional groups present in a compound described herein such that the modifications can be cleaved in vivo to release the parent compound. Prodrugs can be prepared by modifying functional groups present in compounds such that the modifications are cleaved either by routine manipulation or in vivo into the parent compounds. Prodrugs include compounds described herein, wherein the hydroxy, amino, carboxyl or sulfhydryl group in the compound described herein is linked to any group that can be cleaved in vivo to release the free hydroxy, amino or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, esters (eg, acetate, formate and benzoate derivatives), amides, guanidines, carbamates (eg, N,N-dimethylaminocarbonyl) of the hydroxy functional groups in the compounds described herein, and the like. The preparation, selection and use of prodrugs are discussed in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series; Design of Prodrug, ed. H. Bundgaard, Elsevier, 1985; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, each of which is incorporated herein by reference in its entirety.

As used herein, the term pharmaceutically acceptable carrier or pharmaceutically acceptable excipient or excipient includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorbent retardants, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except that any conventional vehicle or agent is incompatible with the active ingredient, their use is contemplated in therapeutic compositions. Additional ingredients may also be included in the compositions.

2. Connections.

The compounds provided herein are compounds that are useful as LRRK2 inhibitors.

In one embodiment of the invention, there is provided a compound of formula I represented by formula Ia

NR ³ or a pharmaceutically acceptable salt, deuterated analogue, prodrug, stereoisomer, or a mixture of stereoisomers thereof, where

R ¹ represents C _3-6 cycloalkyl, optionally substituted with one or more halogen, hydroxyl or cyano atoms, or C 1 _-6 alkyl, optionally substituted with halogen;

R ² represents halogen, cyano or C _1-6 alkyl, optionally substituted with halogen;

R ³ represents C _1-6 alkoxy, C _3-6 cycloalkyl, C _3-6 cycloalkoxy or -N(R11)(R ¹² );

R ⁴ represents hydrogen or halogen;

R ⁶ and R ⁷ are each independently hydrogen or C _1-6 alkyl, optionally substituted with halogen; And

R ⁸ and R ⁹ are each independently hydrogen, cyano, halogen, C _1-6 alkyl or C _1-6 alkoxy; And

R ¹¹ and R ¹² are each independently hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl.

In some embodiments, R ⁶ and R ⁷ are methyl.

In some embodiments, R ⁸ and R ⁹ are hydrogen.

In some embodiments, at least one of R ⁸ and R ⁹ is hydrogen.

- 9 043797

In some embodiments, R ¹ is optionally substituted cyclopropyl or optionally substituted cyclobutyl. In some embodiments, R ¹ is cyclopropyl, cyclobutyl, hydroxycylobut-3-yl, cyanocylobut-3-yl, or fluorocyclobut-3-yl.

In some embodiments, R ¹ is CD3, ethyl or prop-2yl.

In some embodiments, R ² is bromine. In some embodiments, R ² is -CF3. In some embodiments, R ³ is cyclopropyl, methoxy, cyclopropylamino, -NH(CH ₃ ) or -NH(CH ₂ CH ₃ ). In some embodiments, R ⁴ is hydrogen.

In some embodiments, R ¹ is C ₃ . ₆ cycloalkyl, independently substituted with one or more hydroxy or cyano; R ² represents a halogen atom or Scbfluoroalkyl; R ³ represents -N(R ¹¹ )(R ¹² ) or C1.balkoxy; and R ⁴ represents hydrogen.

In some embodiments of the invention, certain compounds provided herein are unexpectedly brain-penetrating. In some embodiments, the compounds further have an MDR1-MDCK efflux ratio of less than or equal to about five. In one embodiment of the invention, the compound illustrated in table is provided. 1 or a pharmaceutically acceptable salt, deuterated analogue, prodrug, stereoisomer or mixture of stereoisomers thereof.

Table 1

- 10 043797

- 11 043797

- 12 043797

- 13 043797

or a pharmaceutically acceptable salt, deuterated analogue, prodrug, stereoisomer or mixture of stereoisomers thereof.

In one embodiment of the invention, the compound may be selected from the compounds of the table. 1. Also included in the disclosure are pharmaceutically acceptable salts, prodrugs, stereoisomers, or a mixture of stereoisomers.

3. Methods of treatment and application.

Treatment is an approach to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results may include one or more of the following: a) inhibiting a disease or condition (eg, reducing one or more symptoms caused by the disease or condition and/or reducing the severity of the disease or condition); b) slowing or stopping the development of one or more clinical symptoms associated with a disease or condition (eg, stabilizing the disease or condition, preventing or slowing the worsening or progression of the disease or condition, and/or preventing or slowing the spread (eg, metastasis) of the disease or condition ); and/or c) alleviation of the disease, that is, regression of clinical symptoms (for example, improving the status of the disease, providing partial or complete remission of the disease or condition, enhancing the effect of another drug, delaying the progression of the disease, increasing quality of life and/or increasing survival.

Prevention means any treatment of a disease or condition that does not result in clinical symptoms of the disease or condition. The compounds may, in some embodiments of the present invention, be administered to a subject (including a human) who is at risk for or has a family history of a disease or condition.

Subject refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experiment. The methods described herein may be useful in human treatment and/or veterinary applications. In some embodiments of the present invention, the subject is a mammal. In one embodiment of the invention, the subject is a human. The term therapeutically effective amount or effective amount of a compound described herein or a pharmaceutically acceptable salt, deuterated analogue, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue means an amount sufficient to treat when administered to a subject to provide a therapeutic benefit such as improvement symptoms or slowing the progression of the disease. For example, a therapeutically effective amount may be an amount sufficient to reduce a symptom of a disease or condition, as described herein. The therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the route of administration, which can be readily determined by one skilled in the art.

The methods described herein can be applied to a cell population in vivo or ex vivo. In vivo means within a living individual, either animal or human. IN

- 14 043797 in this context, the methods described herein can be applied therapeutically to an individual. Ex vivo means outside the living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from individuals. Such samples can be prepared by methods well known in the art. Examples of body fluids include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used ex vivo to determine the optimal schedule and/or dosage of administration of a compound of the present invention for a given indicator, cell type, individual, and other parameters. Information obtained from such applications can be used for experimental purposes or in the clinic to establish protocols for in vivo treatment. Other ex vivo uses for which the compounds and compositions described herein may be suitable are described below or will become apparent to those skilled in the art. Selected compounds may be further characterized to examine safe or tolerable dosage in human or non-human subjects. Such properties can be examined using methods well known to those skilled in the art. LRRK2 is involved in the transition from mild cognitive impairment to Alzheimer's disease; levodopa-induced dyskinesia (Hurley et al., Eur. J, Neurosci., Vol. 26, 2007, 171-177); disorders of the CNS associated with the proliferation and migration of neuroprogenitor cells, and regulation of LRRK2 activity may be beneficial in improving neurological outcome after ischemic injury and promoting recovery of CNS function after neuronal injury such as ischemic stroke, traumatic brain injury, or spinal cord injury (Milosevic et al., Neurodegen., Vol. 4, 2009, 25; see Zhang et al., J. Neurosci. Res. Vol. 88, 2010, 3275-3281); Parkinson's disease, Alzheimer's disease, multiple sclerosis and HIV-induced dementia (see Milosevic et al., Mol. Neurodegen., Vol. 4, 2009, 25); kidney cancer, breast cancer, prostate cancer (eg, solid tumor), blood and lung cancer, acute myeloid leukemia (AML); lymphomas and leukemias (see Ray et al., J. Immunolo., Vol. 230, 2011, 109); multiple myeloma (Chapman et al., Nature, Vol. 471, 2011, 467-472); papillary renal cancer and thyrocarcinoma; multiple myeloma (Chapman et al., Nature, Vol. 471, 2011, 467-472); diseases of the immune system, including rheumatoid arthritis, autoimmune hemolytic anemia of systemic lupus erythematosus, true red cell aplasia, idiopathic thrombocytopenic purpura (ITP), Evans syndrome, vasculitis, bullous skin inflammation, type 1 diabetes mellitus, Sjögren's syndrome, Delvik's disease and inflammatory myopathies (Nakamura et al., DNA Res. Vol. 13(4), 2006, 169-183; see Engeletal., Pharmacol. Rev. Vol. 63, 2011, 127-156; Homametal., J. Clin. Neuromuscular Disease , Vol. 12, 2010, 91-102); ankylosing spondylitis and leprosy infection (DAnoyetal., PLoSGenetics, Vol. 6(12), 2010, e1001195, 1-5; see, Zhangetal., N. Eng. J. Med. Vol. 361, 2009, 2609-2618) ; alpha-synucleinopathies, tauopathy (see Lietal., 2010 Neurodegen. Dis. Vol. 7, 2010, 265-271); Gaucher disease (see, Westbroeketal., Trends. Mol. Med. Vol. 17, 2011, 485-493); tauopathies characterized by hyperphosphorylation of tau protein, such as argyrophilic grain disease, Pick's disease, corticobasal degeneration, progressive supranuclear palsy and inherited frontotemporal dementia and parkinsonism associated with chromosome 17 (see, Goedert, M and Jakes, R, Biochemica et Biophysica Acta, Vol. 1739, 2005, 240-250); diseases characterized by decreased dopamine levels, such as withdrawal/relapse symptoms associated with drug addiction (see Rothman et al., og. Brain Res., Vol. 172, 2008, 385); microglial proinflammatory responses (see Moehle et al., J. Neuroscience, Vol. 32, 2012, 1602-1611); pathogenesis of Crohn's disease (see Barrett et al., Nature Genetics, Vol. 40, 2008, 955-962); and amyotrophic lateral sclerosis (ALS).

It is suggested that increased LRRK2 activity may be characteristic of ALS. Significantly increased levels of LRRK2 mRNA were observed in fibroblasts from patients with NiemannPick disease type C (NPC), indicating abnormal LRRK2 function that may play a role in lysosomal disorders.

In another aspect, the present invention provides a method of treating a disease or condition mediated at least in part by LRRK2. In particular, the present invention provides methods for preventing or treating a disorder associated with LRRK2 in a mammal, comprising the step of administering to said mammal a therapeutically effective amount of a compound from Table 1. 1, or a therapeutic agent according to this invention. In some embodiments of the present invention, the disease or condition mediated at least in part by LRRK2 is a neurodegenerative disease, for example, a central nervous system (CNS) disorder such as Parkinson's disease (PD), Alzheimer's disease (AD), dementia ( including diffuse Lewy body disease and vascular dementia), amyotrophic lateral sclerosis (ALS), age-related memory dysfunction, mild cognitive impairment (eg, transition from mild cognitive impairment to Alzheimer's disease), argyrophilic granule disease, lysosomal disorder (eg, Niemann's disease) Pica type C, Gaucher disease), corticobasal degeneration, progressive supranuclear palsy, inherited frontotemporal dementia with parkinsonism, associated

- 15 043797 diagnosed with a disorder in chromosome 17 (FTDP-17), withdrawal/relapse syndromes associated with drug dependence, levodopa-induced dyskinesia, Huntington's disease (HD) and HIV-associated dementia (HAD). In other embodiments, the disorder is an ischemic disease of organs including, but not limited to, the brain, heart, kidneys, and liver. In some other embodiments, the disease or condition mediated at least in part by LRRK2 is cancer. In some specific embodiments, the cancer is thyroid, kidney (including papillary renal cancer), breast, lung, blood and prostate cancer (eg, solid tumor), leukemia (including acute myeloid leukemia (AML)), or lymphoma. In some embodiments of the present invention, the cancer is kidney cancer, breast cancer, prostate cancer, blood cancer, papillary thyroid cancer, lung cancer, acute myeloid leukemia, or multiple myeloma.

In other embodiments, the compounds described herein are used in methods for treating inflammatory diseases. In some embodiments of the present invention, the disease is an inflammatory bowel disease, such as Crohn's disease or ulcerative colitis (commonly known collectively as inflammatory bowel disease). In other embodiments, the inflammatory disease is leprosy, amyotrophic lateral sclerosis, rheumatoid arthritis, or ankylosing spondylitis. In some embodiments of the present invention, the inflammatory disease is leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis, or ankylosing spondylitis. In other embodiments, the compounds disclosed herein are used in methods of treating multiple sclerosis, systemic lupus erythematosus, autoimmune hemolytic anemia, pure red cell aplasia, idiopathic thrombocytopenic purpura (ITP), Evans syndrome, vasculitis, bullous cutaneous inflammation, type 1 diabetes mellitus , Sjogren's syndrome, Devic's disease and inflammatory myopathy.

Other embodiments include methods of enhancing the cognitive memory of a subject, the method comprising administering an effective amount of a composition containing a compound of Table 1. 1 subject in need. Other embodiments include the use of compounds disclosed herein for treatment. Some embodiments include their use for treating a neurodegenerative disease, cancer, or inflammatory disease.

In other embodiments, compounds provided herein for use in the treatment of Alzheimer's disease, levodopa-induced dyskinesia, Parkinson's disease, dementia, ALS, kidney cancer, breast cancer, prostate cancer, blood cancer, papillary cancer, lung cancer, acute myeloid leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis or ankylosing spondylitis. In other embodiments, the invention provides for the use of compounds disclosed herein for the manufacture of a medicament for the treatment of a neurodegenerative disease, cancer or inflammatory disease. In other embodiments, the invention provides the use of compounds disclosed herein for the manufacture of a medicament for the treatment of Alzheimer's disease, levodopa-induced dyskinesia, Parkinson's disease, dementia, amyotrophic lateral sclerosis, kidney cancer, breast cancer, prostate cancer, blood cancer, papillary cancer, lung cancer, acute myeloid leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis or ankylosing spondylitis. In the context of this document, the term injury refers to physical damage to the body caused by violence, accident, fracture, etc. The term ischemia refers to cardiovascular disorders characterized by low oxygen levels, usually due to obstruction of arterial blood supply or insufficient blood flow leading to tissue hypoxia. The term stroke refers to a cardiovascular disorder caused by a blood clot or bleeding in the brain, most often caused by an interruption of blood flow in the brain due to a clot blocking a blood vessel, and in some embodiments, the disclosure of the term stroke refers to ischemic stroke or hemorrhagic stroke. The term myocardial infarction refers to cardiovascular disorders characterized by localized necrosis resulting from obstruction of the blood supply.

In some embodiments, the present invention provides compounds for inhibiting cell death, the compounds being illustrated in Table. 1. In some embodiments, the compounds of the present invention are cell death inhibitors. In any case, the compounds of the present invention preferably exert their cell death inhibitory effects at a concentration of less than about 50 micromolar, more preferably at a concentration of less than about 10 micromolar, and most preferably at a concentration of less than about 1 micromolar.

- 16 043797

4. Pharmaceutical compositions and methods of administration.

The compounds presented herein are typically administered in the form of pharmaceutical compositions. Thus, also provided herein are pharmaceutical compositions that contain one or more of the compounds described herein or a pharmaceutically acceptable salt, deuterated analogue, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue thereof, and one or more pharmaceutically acceptable carriers. , selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable carriers may include, for example, inert solid diluents and excipients, diluents including sterile aqueous solution and various organic solvents, penetration enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical field. See, for example, Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G. S. Banker & C. T. Rhodes, Eds.).

Pharmaceutical compositions may be administered as a single or multiple dose. The pharmaceutical composition can be administered in a variety of ways, including, for example, rectally, buccally, intranasally and transdermally. In some embodiments of the invention, the pharmaceutical composition may be administered by intra-arterial injection, intravenous, intraperitoneal, parenteral, intramuscular, subcutaneous, oral, topical, or by inhaler. One route of administration is parenteral administration, for example by injection. The forms in which the pharmaceutical compositions described herein may be formulated for administration by injection include, for example, aqueous or oily suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil, as well as elixirs, mannitol, dextrose or sterile aqueous solution, and similar pharmaceutical carriers.

Oral administration may be another method of administering the compounds described herein. Administration may be via, for example, enteric-coated capsules or tablets. In the manufacture of pharmaceutical compositions that include at least one compound described herein, or a pharmaceutically acceptable salt, deuterated analogue, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue thereof, the active ingredient is typically diluted with an excipient and/or contained within such carrier, which may be in the form of a capsule, sachet, paper or other container. In case the excipient serves as a diluent, it may be in the form of a solid, semi-solid or liquid material which acts as a carrier or vehicle for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, starch capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions and sterile packaged powders. Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup and methylcellulose.

The compositions may further include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; and flavors.

Compositions that include at least one compound described herein, or a pharmaceutically acceptable salt, deuterated analogue, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue thereof may be formulated to provide rapid, sustained or sustained release of the active ingredient after administration to a subject using procedures known in the art. Controlled-release drug delivery systems for oral administration include osmotic pump systems and dissolution systems containing polymer-coated reservoirs or drug polymer matrix formulations. Transdermal patches can be used to provide continuous or intermittent infusion of the compounds described herein in controlled amounts. Transdermal patches can be designed for continuous, pulsatile, or just-in-time delivery of pharmaceutical agents.

For the preparation of solid compositions, such as tablets, the main active ingredient can be mixed with a pharmaceutical excipient to form a solid composition to form a dosage form containing a homogeneous mixture of the compound described herein or a pharmaceutically acceptable salt, deuterated analogue, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue. When these compositions are formulated into a homogeneous dosage form, the active ingredient may be uniformly dispersed throughout the composition so that the composition can be readily divided into equally effective unit dosage forms such as tablets, pills and capsules.

- 17 043797

Tablets or pills of the compounds described herein may be coated or otherwise formulated to provide a dosage form that provides the benefit of long-lasting action or to protect against the acidic environment of the stomach. For example, a tablet or pill may include an internal dosage component and an external dosage component, the latter being in the form of a shell over the former. The two components may be separated by an enteric layer, which serves to resist disintegration in the stomach and allows the internal component to pass intact into the duodenum or be delayed in release. Various materials can be used for such enteric layers or coatings, including a number of polymer acids and mixtures of polymer acids with substances such as shellac, cetyl alcohol and cellulose acetate.

Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures and powders thereof. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In some embodiments of the present invention, the compositions are administered orally or through the respiratory tract for local or systemic action. In other embodiments of the invention, compositions in pharmaceutically acceptable solvents can be nebulized using inert gases. Inhalation solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a breathing mask or to an intermittent positive pressure breathing apparatus. The solution, suspension or powder compositions can be administered, preferably orally or nasally, from devices that deliver the composition in an appropriate manner.

5. Dosing.

The specific dosage level of a compound of the present invention for any particular subject will depend on a variety of factors, including the potency of the particular compound used, age, body weight, general health, sex, diet, time of administration, route of administration and rate of elimination, drug combination, and severity specific disease in the subject being treated. For example, the dose may be expressed as the number of milligrams of a compound described herein per kilogram of subject body weight (mg/kg). A dose in the range of about 0.1 and 150 mg/kg may be acceptable. In some embodiments of the present invention, a dose in the range of about 0.1 and 100 mg/kg may be appropriate. In other embodiments of the invention, a dose in the range of 0.5 and 60 mg/kg may be appropriate. In some embodiments of the present invention, a dosage of about 0.0001 to about 100 mg per kg of body weight per day, about 0.001 to about 50 mg of the compound per kg of body weight, or from about 0.01 to about 10 mg of the compound may be appropriate. per kg body weight. Normalization according to subject body weight is particularly useful when adjusting doses between subjects of very different sizes, for example when administering a drug to both children and adults, or when converting the effective dose for a non-human subject such as dog, at a dose suitable for humans. A daily dose may also be described as the total amount of a compound described herein to be administered as a single dose or per day. The daily dose of a compound from Table 1 may be from about 1 to 4000 mg, from about 2000 to 4000 mg/day, from about 1 to 2000 mg/day, from about 1 to 1000 mg/day, from about 10 to 500 mg/day , from about 20 to 500 mg/day, from about 50 to 300 mg/day, from about 75 to 200 mg/day, or from about 15 to 150 mg/day.

When administered orally, the total daily dose for humans may range from 1 mg to 1000 mg, between about 1000-2000 mg/day, between about 10-500 mg/day, between about 50-300 mg/day, between about 75-200 mg/day , or between about 100-150 mg/day. The compounds of the present invention or compositions thereof can be administered once, twice, three, four or more times per day using any suitable method described above.

In a specific embodiment, the method comprises administering to a subject an initial daily dose of about 1 to 800 mg of a compound described herein and then increasing the dose in increments until clinical efficacy is achieved. Increases of about 5, 10, 25, 50 or 100 mg can be used to increase the dose in steps. The dosage may be increased daily, every other day, twice a week or once a week.

6. Synthesis of compounds.

The compounds can be prepared using the methods described herein and conventional modifications thereof, as will be apparent in view of this disclosure and methods well known in the art. In addition to the present methods, conventional and known synthetic methods can be used. The synthesis of typical compounds described herein can be carried out as described in the following examples. If possible, reagents can be purchased commercially, for example from Sigma Aldrich or other chemical suppliers.

The compounds of this invention can be prepared using the methods described herein, and their usual modifications, which will be obvious in view of this disclosure and methods well known in the art. In addition to the present methods, conventional and known synthetic methods can be used. Synthesis of those described in this document

- 18 043797 those connections can be made as described in the following examples. If possible, reagents can be purchased commercially, for example from Sigma Aldrich or other chemical suppliers.

The compounds of this invention can be prepared from readily available starting compounds using, for example, the following general methods and techniques. It is understood that when typical or preferred process conditions are indicated (i.e., reaction temperatures, times, reactant molar ratios, solvents, pressures, etc.), other process conditions may be used unless otherwise indicated. Optimal reaction conditions may vary depending on the specific reagents or solvent used, but such conditions can be determined by one of ordinary skill in the art using routine optimization procedures. In addition, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent unwanted reactions of certain functional groups. Suitable protecting groups for various functional groups, as well as suitable conditions for protecting and deprotecting certain functional groups are well known in the art. For example, numerous protecting groups are described in Wuts, P. G. M., Greene, T. W., & Greene, T. W. (2006). Greene's protective groups in organic synthesis. Hoboken, N.J., Wiley-Interscience and references cited in this book. In addition, the compounds of this invention may contain one or more chiral centers. Accordingly, if desired, such compounds can be prepared or isolated as pure stereoisomers, that is, as individual enantiomers or diastereomers or as stereoisomer-enriched mixtures. All such stereoisomers (and enriched mixtures) are included within the scope of the present disclosure unless otherwise indicated. Pure stereoisomers (or enriched mixtures) can be prepared using, for example, optically active starting materials or stereoselective reagents well known in the art. Alternatively, racemic mixtures of such compounds can be separated, for example, using chiral column chromatography, chiral solvents, and the like. The starting materials for the following reactions are generally known compounds or can be prepared by known methods or obvious modifications. For example, many of the starting materials are available from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, WI, USA), Bachem (Torrance, CA, USA), Emka-Chemce or Sigma (St. Louis, MO, USA). Others can be prepared by methods or obvious modifications described in standard references such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15 (John Wiley, and Sons, 1991), Rodd's Chemistry of Carbon Compounds, Volumes 1-5 , and Supplementals (Elsevier Science Publishers, 1989), Organic Reactions, Volumes 1-40 (John Wiley, and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley, and Sons, 5th Edition, 2001), and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

The terms solvent, inert organic solvent, or inert solvent refer to a solvent that is inert under the reaction conditions described with it (including, for example, benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, methylene chloride (or dichloromethane) , diethyl ether, methanol, pyridine and the like). Unless otherwise specified, the solvents used in the reactions of the present disclosure are inert organic solvents and the reactions are carried out under an inert gas atmosphere, preferably nitrogen.

The term q.s. means adding an amount sufficient to achieve the stated function, for example, to bring the solution to the desired volume (i.e. 100%).

It will also be appreciated that in each of the above schemes, the addition of any substituent can result in a number of isomeric products (including, but not limited to, enantiomers or one or more diastereomers), any or all of which can be isolated and purified using conventional ways. When enantiomerically pure or enriched compounds are desired, chiral chromatography and/or enantiomerically pure or enriched starting materials can be used as commonly used in the art or as described in the examples.

General synthesis.

The following General Reaction Scheme I illustrates the general method for preparing the compounds described herein.

Scheme I

Referring to general reaction scheme I, compounds of formula (X) are prepared by coupling a substituted pyrimidine of formula (Y) with an amine of formula (Z) wherein R ² , R ³ , ring B and m are defined as in any of the formulas herein or as specific compounds listed in table. 1, and X represents a leaving group. In some embodiments, X is a halogen. The corresponding compounds of formula (Y) or (Z) can be prepared in accordance with the more specific methods described in the following examples, or methods known

- 19 043797 knowledgeable to a specialist in this field of technology. The combination of compounds of formula (Y) and (Z) in the presence of an acid results in a compound of formula (X). In some embodiments, the acid is toluenesulfonic acid or trifluoroacetic acid. In some embodiments, the combination of compounds of formula (Y) and (Z) in the presence of a base results in a compound of formula (X). In some embodiments, the base is triethylamine.

In one embodiment, a method for preparing a compound of formula (X) is provided, comprising combining a compound of formula (Y) with a compound of formula (Z) under conditions for preparing a compound of formula (X), wherein R ¹ , R ² , R ³ , ring B and m are defined as in any of the formulas given herein, or as the specific compounds listed in Table. 1, and X represents a leaving group. In some embodiments, X is a halogen. When amines of formula (Z) are not commercially available, they can be prepared from commercially available starting materials. For example, in some embodiments, amines of formula (Z) can be prepared by reducing the corresponding nitro compound. Amines of formula (Z) are typically functionalized to couple with a substituted pyrimidine of formula (Y). When a particular stereoisomer is desired (eg, a cis or trans stereoisomer of formula III, IIIA or IIIB), a single stereoisomer of the corresponding amine can be prepared prior to coupling with the substituted pyrimidine of formula (Y). Each of the cis and trans stereoisomers can be obtained by selectively inverting the stereochemistry before installing a cyano group on the cyclobutyl ring. In some embodiments, amines of formula (Z) are prepared via 1,3-dipolar cycloaddition reactions using suitably functionalized starting materials. Further functionalization or interconversion of functional groups can be performed before or after the cycloaddition reaction.

In some embodiments, compounds of Formula Ia can be prepared in accordance with Scheme II.

Scheme II

Referring to the general reaction scheme II, compounds of formula (2-3) can be prepared by coupling an appropriately substituted triazole (2-1) with an appropriately substituted ester (2-2). Conversion of the ester of compound (2-3) to α-cyanoketone (24) can be accomplished under displacement reaction conditions using a strong base (eg butyllithium) and acetonitrile. Contacting compound (2-4) with suitably substituted hydrazine (2-5) or a salt thereof results in the amine of formula (2-6). The coupling of an amine of formula (2-6) with a suitably substituted pyrimidine of formula (Y) can be carried out according to Scheme I, thereby leading to a compound of formula Ia.

Examples

The following examples are included to demonstrate specific embodiments of the disclosure. Those skilled in the art will appreciate that the methods described in the following examples are methods that function well in the present disclosure and, therefore, can be considered to represent specific methods of the present disclosure. However, those skilled in the art will, in light of the present disclosure, understand that specific embodiments disclosed herein that achieve the same or a similar result do not depart from the spirit and scope of the disclosure. General experimental methods:

All moisture-sensitive reactions were carried out in glassware previously dried in an oven, or using a burner flame, and under a nitrogen atmosphere. All chemicals were purchased from commercial suppliers and used unchanged unless otherwise stated. Reactions were magnetically stirred and monitored by thin layer chromatography (TLC) on 250 μm silica gel precoated plates and visualized in either UV or an iodine chamber. Flash column chromatography was performed using silica gel (100-200 mesh). Chemical shifts are reported relative to chloroform (δ 7.26), methanol (δ 3.31) or DMSO (52.50) for 1H NMR. HPLC analysis was carried out on a Shimadzu 20AB HPLC system with detector

- 20 043797 rum of a photodiode array and a Luna-C18 column (2) 2.0x50 mm, 5 µm with a flow rate of 1.2 ml/min and a solvent gradient. Mobile phase A (MFA, H ₂ O + 0.037% (v/v) TFA). Mobile phase B (MFV, ACN + 0.018% (v/v) TFA) (0.01 min, 10% MFV; 4 min, 80% MFV; 4.9 min, 80% MFV; 4.92 min, 10% MFV; 5.5 min, 10% MFV). LCMS was detected at 220 and 254 nm or by evaporative light scattering detection (ELSD) and positive electrospray ionization (MS). Semipreparative HPLC was performed using either acidic or neutral conditions. Acidic conditions: Luna C18 100x30 mm, 5 µm; MFA: HCl/H ₂ O = 0.04%, or formic acid/H ₂ O = 0.2% (v/v); IMF: ACN. Neutral conditions: Waters Xbridge 150x25.5 µm; MFA: 10 mM NH4HCO3 in H2O; IMF: ACN. Gradient for both conditions: 10% MPV to 80% MPV over 12 minutes at a flow rate of 20 ml/min, then 100% MPV over 2 min, 10% MPV over 2 min, UV detection. SFC analysis was carried out on a Thar analytical SFC system with a UV/visible light detector and a series of 4.6x100 mm, 3 µm chiral columns, including AD-3, AS-H, OJ-3, OD-3, AY-3 and IC- 3, with a flow rate of 4 ml/min and a solvent gradient. Mobile phase A (MPA, CO ₂ ). Mobile phase B (MFV, MeOH + 0.05% (v/v) IPS) (0.01 min, 10% MFV; 3 min, 40% MFV; 3.5 min, 40% MFV; 3.56 -5 min, 10% MFV). Preparative SFC was performed on a Thar 80 preparative SFC system with a UV/vis light detector and a series of 30x250 mm, 5 µm chiral preparative columns, including AD-H, AS-H, OJ-H, OD-H, AY-H and IC -H, with a flow rate of 65 ml/min and a solvent gradient. Mobile phase A (MPA, CO ₂ ). Mobile phase B (MFV, MeOH + 0.1% (v/v) NH3H2O) (0.01 min, 10% MFV; 5 min, 40% MFV; 6 min, 40% MFV; 6.1-10 min, 10% MFV).

Compounds were named using ChemBioDraw Ultra 13.0 or Chemaxon.

Receiving connections.

Where preparation of starting materials is not described, they are commercially available, known in the literature, or readily obtainable by those skilled in the art using standard techniques. When compounds are stated to be prepared similarly to earlier examples or intermediates, it will be appreciated by one skilled in the art that reaction times, reagent equivalents, and temperatures may be modified for each particular reaction and that it may be necessary or desirable to use different work-up methods or cleaning. When reactions are carried out using microwave irradiation, the Biotage Initiator microwave instrument is used. The actual power supplied varies during the reaction to maintain a constant temperature.

Example 1. Synthesis of N4-ethyl-N2-[1-(3-isocyanocyclobutyl)-5-methylpyrazol-4-yl]-5-(trifluoromethyl)pyrimidine-2,4-diamine (26).

-(Benzyloxy)cyclobutanol.

To a stirred solution of 3-benzyloxycyclobutanone (125 g, 709.38 mmol) in MeOH (1.5 L) was added NaBH ₄ (26.84 g, 709.38 mmol) in portions at -20°C under N2 for 4 hours . After addition, the mixture was allowed to warm to 25°C and stirred for 30 minutes. Water (50 ml) was added to the mixture and stirred for 30 minutes. The mixture was concentrated under reduced pressure to obtain a residue. (Two batches of the same size were combined for workup.) The residue was purified by silica gel column chromatography (PE:EtOAc=6:1) to give (1S,3S)-3(benzyloxy)cyclobutanol as a colorless oil.

1-(3-(Benzyloxy)cyclobutyl)-4-nitro-1H-pyrazole.

To a mixture of (1S,3S)-3-(benzyloxy)cyclobutanol (250 g, 1.40 mol) and 4-nitro-1H-pyrazole (158.3 g, 1.40 mol) in THF (5 L) was added PPh ₃ (477.37 g, 1.82 mol) and DIAD (36.82 g, 1.82 mol, 353.87 ml) dropwise at 0°C under N2. After addition, the mixture was stirred at 25°C for 16 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was triturated with PE:EtOAc = 2:1 (2 l) and filtered. The filtered cake was washed with PE:EtOAc=2:1 (2x1 L) and the combined filtrate was concentrated to give the crude product. The crude product was purified by silica gel column chromatography (PE:EtOAc = 6:1) to give 1-((1R,3R)-3(benzyloxy)cyclobutyl)-4-nitro-1H-pyrazole as a white solid. LCMS: RT 0.851 min, mz = 274.2 [M + H] ⁺ . 1H NMR (400 MHz, CDCI3) δ 8.15 (s, 1H), 8.12 (s, 1H), 7.29-7.41 (m, 5H), 4.92-4.99 (m, 1H), 4.49 (s, 2H), 4.41-4.47 (m, 1H), 2.63-2.84 (m, 4H).

1-(3-(Benzyloxy)cyclobutyl)-5-chloro-4-nitro-1H-pyrazole.

LiHMDS (1 M, 567 .90 ml) dropwise at -75°C in an N2 atmosphere for 1 hour. After addition, the mixture was stirred for 1 hour, then a solution of 1,1,1,2,2,2hexachloroethane (83.16 g, 351.28 mmol) in THF (200 ml) was added dropwise at -78°C, the mixture was stirred at -78 °C and stirred for 1 hour. The mixture was poured into an aqueous NH4Cl solution (1.5 L). The organic phase was separated and the aqueous phase was extracted with EtOAc (2x500 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (1 L), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1) to give 1-((1R,3R)-3-(benzyloxy)cyclobutyl)-5-chloro-4-nitro-1H-pyrazole in

- 21 043797 in the form of a white solid. 1H NMR (400 MHz, CDCl ₃ ) δ 8.21 (s, 1H), 7.29-7.41 (m, 5H), 5.16-5.24 (m, 1H), 4.50 (s , 2H), 4.42-4.47 (m, 1H) 2.81-2.89 (m, 2H), 2.61-2.70 (m, 2H).

1-(3-(Benzyloxy)cyclobutyl)-5-methyl-4-nitro-1H-pyrazole.

To a mixture of 1-((1R,3R)-3-(benzyloxy)cyclobutyl)-5-chloro-4-nitro-1H-pyrazole (65 g, 211.22 mmol), 2,4,6-trimethyl-1, 3,5,2,4,6-trioxatriborinane (212.12 g, 844.90 mmol, 235.69 ml) and Na ₂ CO ₃ (44.78 g, 422.45 mmol) in 1,4-dioxane ( 1.5 L) and H2O (150 ml) was added Pd(dppf)Cl ₂ -CH ₂ Cl ₂ (27.6 g, 33.80 mmol) at 25°C under N2 atmosphere. The mixture was then heated to 100°C and stirred for 40 hours. The mixture was cooled to 25°C and concentrated under reduced pressure to dryness. The residue was dissolved in PE:EtOAc = 2:1 (2 l), then anhydrous Na ₂ SO ₄ (100 g), celite (100 g) were added and stirred for 30 minutes. The mixture was filtered through a pad of celite. The filtered cake was washed with PE:EtOAc = 2:1 (2x 1 L) and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1) to give 1-((1R,3R)-3(benzyloxy)cyclobutyl)-5-methyl-4-nitro-1H-pyrazole as a white solid substances. LCMS: RT 0.844 min, m/z = 288.2 [M+H]+.

3-(5-Methyl-4-nitro-1H-pyrazol-1-yl)cyclobutanol.

To a solution of 1-((1R,3R)-3-(benzyloxy)cyclobutyl)-5-methyl-4-nitro-1H-pyrazole (59.5 g, 207.09 mmol) in DCM (1.2 L) BCl ₃ (1 M, 621.27 ml) dropwise at 0°C under N2 atmosphere for 2 hours. The mixture was then stirred at 0°C for 1 hour. The mixture was poured into ice water (600 ml). The aqueous phase was extracted with DCM (2x600 ml). The combined organic phase was washed with aqueous NaHCO3 (500 ml), aqueous saturated sodium chloride (500 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. (Four batches of the same size were combined for processing). The residue was purified by silica gel column chromatography (PE:EtOAc =1:1) to give (1R,3R)-3-(5-methyl-4-nitro-1H-pyrazol-1-yl)cyclobutanol as a white solid. . 'll NMR (400 MHz, CDCl ₃ ) δ 8.10 (br.d, J=4.63 Hz, 1H), 4.98-5.03 (m, 1H), 4.70-4.82 ( m, 1H), 2.85-2.97 (m, 2H), 2.59-2.66 (m, 3H), 2.47-2.58 (m, 2H), 2.38 (br. s, 1H).

1-(3-Iodocyclobutyl)-5-methyl-4-nitro-1H-pyrazole.

To a mixture of (1R,3R)-3-(5-methyl-4-nitro-1H-pyrazol-1-yl)cyclobutanol (70 g, 354.99 mmol), PPh ₃ (139.66 g, 532.49 mmol ) and imidazole (36.25 g, 532.49 mmol) in THF (1.2 L), a solution of I ₂ (135.15 g, 532.49 mmol) in THF (200 ml) was added dropwise at 0°C in atmosphere N2. After this, the mixture was stirred at 25°C for 16 hours. The mixture was poured into ice water (500 ml). The aqueous phase was extracted with EtOAc (2x300 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (500 ml), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1) to give 1-((1R,3R)-3-iodocyclobutyl, CDCl ₃ ) δ 8.14 (s, 1H), 4.61-4 .83 (m, 1H), 4.12-4.34 (m, 1H), 3.09-3.36 (m, 4H), 2.61 (s, 3H).

3-(5-Methyl-4-nitropyrazol-1-yl)cyclobutanecarbonitrile.

To a solution of 1-(3-iodocyclobutyl)-5-methyl-4-nitropyrazole (2 g, 6.51 mmol) in DMF (30 ml) was added KCN (2.5 g, 39.06 mmol) at 0°C. The mixture was then stirred at 70°C for 2 days. The mixture was diluted with water (60 ml), extracted with EtOAc (4x20 ml). The combined organic layers were washed with water (2x50 ml), aqueous saturated sodium chloride solution (50 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 1:1) to give 3-(5-methyl-4-nitropyrazol-1-yl)cyclobutanecarbonitrile as a yellow solid. LCMS: RT 1.066 min, m/z = 207.3 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) δ ppm 8.16 (s, 1H), 5.11 (quin, J=7.81 Hz, 1H), 3.32-3.47 (m, 1H), 3.08-3.21 (m, 2H) , 2.85-2.95 (m, 2H), 2.67 (s, 3H), 1.59 (s, 1H).

3-(4-Amino-5-methylpyrazol-1-yl)cyclobutanecarbonitrile.

To a mixture of 3-(5-methyl-4-nitropyrazol-1-yl)cyclobutanecarbonitrile (200 mg, 969.93 µmol), NH4C1 (259 mg, 4.85 mmol) in a mixture of EtOH (4.8 ml) and H2O ( 1.2 ml) Fe (270 mg, 4.85 mmol) was added at 15°C. The mixture was stirred at 80°C for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was diluted with water (10 ml), extracted with EtOAc (10x5 ml). The combined organic layers were dried over Na2SO4, filtered and concentrated to give 3-(4-amino-5methylpyrazol-1-yl)cyclobutanecarbonitrile. LCMS: RT 0.101 min, m/z = 177.2 [M+H] ⁺ .

2-Chloro-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine.

To a solution of 2,4-dichloro-5-(trifluoromethyl)pyrimidine (70 g, 322.61 mmol) in THF (1.4 L) was added a solution of ethylamine (32 g, 709.74 mmol, 46.37 ml) in THF (100 ml) dropwise at 0°C under N2 atmosphere for 1 hour. After addition, the mixture was stirred at 25°C for 1 hour. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was triturated with DCM (200 ml) and filtered. The filtrate was recrystallized from n-heptane (600 ml) and MTBE (400 ml). The precipitated phase was a syrupy substance. The liquid was discarded. The syrupy material was purified by silica gel column chromatography (PE:EtOAc = 20:1) to give 2-chloro-N-ethyl-5(trifluoromethyl)pyrimidin-4-amine as a white solid. ^1H NMR (400 MHz, CDCl ₃ ): δ 8.228.27 (m, 1H), 5.40 (br.s, 1H), 3.56-3.65 (m, 2H), 1.29 (t , J=7.22 Hz, 3H). HPLC: RT: 2.68 min.

- 22 043797

N4-Ethyl-N2-[1-(3-isocyanocyclic'lobutyl)-5-methylpyrazol-4-yl]-5-(trifluoromethyl)pyrimidine-2,4diamine.

Mixture of 1-(3-isocyanocyclobutyl)-5-methylpyrazol-4-amine (170 mg, 964.70 µmol), 2-chloro-M-ethyl5-(trifluoromethyl)pyrimidin-4-amine (217 mg, 964.70 µmol ), p-TsOHH ₂ O (55 mg, 289.41 µmol) in 1,4dioxane (10 ml) was stirred at 90°C for 2 hours. The mixture was concentrated under reduced pressure. The residue was diluted with water (20 ml), extracted with EtOAc (3x10 ml). The combined organic layers were washed with water (30 ml), aqueous saturated sodium chloride solution (30 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by preparative TLC (DCM:MeOH = 15:1) to give N4-ethyl-N2-[1-(3-isocyanocyclobutyl)-5-methylpyrazol-4-yl]-5(trifluoromethyl)pyrimidine-2, 4-diamine. 1H-NMR (400 MHz, CDC13) δ ppm 8.10 (s, 1H), 7.65-7.93 (m, 1H), 6.15-6.60 (m, 1H), 4.91-5.15 (m, 2H) 3.44 -3.55 (m, 2H), 3.23-3.35 (m, 1H), 3.07-3.21 (m, 2H), 2.75-2.89 (m, 2H), 2 .20 (s, 3H), 1.61 (br.s, 1H), 1.25 (t, J=7.1 Hz, 3H). HPLC: RT: 1.73 min. MS: m/z = 366.2 [M+H]+.

Example 2 Synthesis [9] N4-ethyl-N2-[1-( ² H ₃ methyl-3-[2-(2H-1,2,3-triazol-2-yl)propan-2-yl]-1Hpyrazol- 5-yl]-5-(trifluoromethyl)pyrimidine-2,4-diamine (34).

Ethyl 2-methyl-2-(2H-1,2,3-triazol-2-yl)propanoate.

To a mixture of 2H-triazole (20 g, 289.56 mmol) in DMF (200 ml) was added t-BuOK (48.74 g, 434.34 mmol) at 0°C. After the addition, ethyl 2-bromo-2-methylpropanoate (78.63 g, 434.34 mmol) was added dropwise at 0° C., then the mixture was stirred at 25° C. for 3 hours. The mixture was poured into ice water (70 ml) and stirred for 5 minutes. The aqueous phase was extracted with EtOAc (3x300 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (2x200 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1) to give ethyl 2-methyl-2-(1H-1,2,3-triazol-1-yl)propanoate and the isomer ethyl 2-methyl- 2-(2H-1,2,3-triazol-1yl)propanoate. LCMS: RT 0.565 min, m/z = 184.1 [M+H]+. 1H-NMR (400 MHz, CDCI3): δ ppm 7.64 (s, 2H), 4.12-4.18 (m, 2H), 1.95 (s, 6H), 1.18 (t, J=7.28 Hz, 3H). An undesired isomer, ethyl 2-methyl-2-(1H-1,2,3-triazol-1yl)propanoate. 1H NMR (400 MHz, CDCI3): δ ppm 7.70 (d, J=6.40 Hz, 2H), 4.14-4.19 (m, 2H), 1.94 (s, 6H), 1.20 (t, J=7.28 Hz , 3H).

4-Methyl-3-oxo-4-(2H-1,2,3-triazol-2-yl)pentanenitrile.

To a mixture of MeCN (96.88 mg, 2.36 mmol) in THF (10 ml) was added n-BuLi (2.5 M, 0.94 ml) dropwise at -78°C under N2 atmosphere. After 0.5 h, ethyl 2-methyl-2-(2H-1,2,3-triazol-1yl)propanoate (200 mg, 2.36 mmol) was added dropwise over 1 h at -78°C, stirred at -78°C for 2 hours. The mixture was poured into ice water (20 ml) and stirred for 5 minutes. The pH of the mixture was adjusted to pH 5-6 using HCl (1 M). The aqueous phase was extracted with ethyl acetate EtOAc (3x10 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (10 ml), dried with anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 1:1) to give 4-methyl-3-oxo-4-(2H-1,2,3-triazol-2yl)pentanenitrile. LCMS: RT 0.945 min, m/z = 179.1 [M+H] ⁺ . 1H NMR (400 MHz, CDC13): δ ppm 7.76 (s, 1H), 3.11 (s, 2H), 1.90 (s, 6H).

1-( ² H3)Methyl-3-[2-(2H-1,2,3-triazol-2-yl)propan-2-yl]-1H-pyrazol-5-amine.

TEA (992 mg, 9.8 mmol, 1 .36 ml, 7 eq.) at 0°C. After addition, the mixture was stirred at 95°C for 4 hours. The reaction mixture was concentrated to give a residue which was diluted with H2O (5 ml) and extracted with EtOAc (3x5 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 1-( ² H ₃ )methyl-3-[2 -(2H-1,2,3-triazol-2yl)propan-2-yl]-1H-pyrazol-5-amine. LCMS: RT 0.236 min, m/z = 210.2 [M+H] ⁺ . 1H NMR (400 MHz, CDCl·,): δ ppm 7.61 (s, 1H), 5.25 (s, 1H), 3.39 (br.s, 1H), 2.05 (s, 3H).

_N4 -Ethyl-N2-[1-( ^2H3 )methyl-3-[2-(2H-1,2,3-triazol-2-yl)propan-2-yl]-1H-pyrazol-5-yl ]-5-(trifluoromethyl)pyrimidine-2,4-diamine.

To a solution of 1-( ² H3)methyl-3-[2-(2H-1,2,3-triazol-2-yl)propan-2-yl]-1H-pyrazol-5-amine (100 mg, 477, 85 µmol) and 2-chloro-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine (107.8 mg, 477.85 µmol) in 1,4dioxane (10 ml) p-TsOH (24.69 mg) was added , 143.36 µmol). The mixture was stirred at 90°C for 3 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (5 ml) and adjusted to pH 8-9 with aqueous NaHCO3 and extracted with EtOAc (3x8 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (10 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (PE:EtOAc = 1:1) and trituration with n-heptane to give N4-ethyl-N2-[1( ^2H3 )methyl-3-[2-(2H-1,2,3 -triazol-2-yl)propan-2-yl]-1H-pyrazol-5-yl]-5-(trifluoromethyl)pyrimidin-2,4diamine. ¹ H-NMR (400 MHz, CDCI3): δ ppm 8.10 (s, 1H), 7.62 (s, 2H), 6.73 (br.s., 1H), 6.03 (s, 1H), 5.15 (br.s. H), 3 .35-3.44 (m, 2H), 2.11 (s, 6H), 1.18-1.21 (t, J=7.28 Hz, 3H). HPLC: RT 2.24 min, m/z: 399.2 [M+H] ⁺ .

- 23 043797

Example 3. Synthesis of N-[2-cyclopropyl-5-[1-methyl-1-(triazol-2-yl) ethyl]pyrazol-3-yl]-N4-ethyl-5(trifluoromethyl)pyrimidin-2,4- diamine (78).

4-Methyl-3-oxo-4-(triazol-2-yl)pentanenitrile.

To a mixture of 2H-triazole (20 g, 289.56 mmol) in DMF (200 ml) was added t-BuOK (48.74 g, 434.34 mmol) in one portion at 0°C under N2. After addition, methyl 2-bromo-2-methylpropanoate (78.63 g, 434.34 mmol, 56.16 ml) was added dropwise. The mixture was stirred at 25°C for 3 hours. The residue was poured into ice water (700 ml) and stirred for 5 minutes. The aqueous phase was extracted with EtOAc (3x300 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (2x100 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give methyl 2-methyl-2-(triazol-2yl)propanoate as a yellow oil. 1H NMR (400 MHz, CDC13): δ ppm 7.649 (s, 2H), 3.701 (s, 3H), 1.963 (s, 6H).

4-Methyl-3-oxo-4-(triazol-2-yl)pentanenitrile.

To a solution of CH3CN (485.21 mg, 11.82 mmol) in THF (20 mL), n-BuLi (2.5 M, 4.73 mL) was added dropwise at -78°C over 10 min. After addition, the mixture was stirred at this temperature for 50 minutes and then methyl 2-methyl-2-(triazol-2-yl)propanoate (1 g, 5.91 mmol) was added dropwise at -78°C. The resulting mixture was stirred at -78°C for 2 hours. The reaction mixture was poured into ice water (50 ml), adjusted to pH 5-6 with HCl (1N) and extracted with EtOAc (3x20 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (10 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =10:1 to 1:1) to give 4-methyl-3-oxo-4-(triazol-2-yl)pentanenitrile as a yellow solid. 1H NMR (400 MHz, CDCI3): δ ppm 7.761 (s, 2H), 3.106 (s, 2H), 1.904 (s, 6H).

2-Cyclopropyl-5-[1-methyl-1-(triazol-2-yl)ethyl]pyrazol-3-amine.

To a mixture of 4-methyl-3-oxo-4-(triazol-2-yl)pentanenitrile (400 mg, 2.24 mmol) and cyclopropylhydrazine dihydrochloride (974.6 mg, 6.72 mmol) in EtOH (10 ml) was added HCl (12 M, 560 µl) at 25°C in an N2 atmosphere. The mixture was stirred at 90°C for 12 hours. The mixture was concentrated. The residue was poured into aqueous NaHCO ₃ solution (10 ml) and stirred for 5 minutes. The aqueous phase was extracted with EtOAc (3x5 ml). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (PE:EtOAc = 1/1) to give 2-cyclopropyl-5-[1-methyl-1-(triazol-2-yl)ethyl]pyrazol-3-amine as a yellow oil. 1H NMR: (400 MHz, CDC13): δ ppm 7.756-7.722 (d, J=13.6 Hz, 1H), 7.616 (s, 1H), 2.041 (s, 6H), 1.1391.100 (m, 2H) 1.022-1.004 (m, 2H).

N2-[2-Cyclopropyl-5-[1-methyl-1-(triαzol-2-yl)ethyl]pyrazol-3-yl]-N4-ethyl-5-(trifluoromethyl)pyrimidine2,4-diamine.

Mixture of 2-cyclopropyl-5-[1-methyl-1-(triazol-2-yl)ethyl]pyrazol-3-amine (77 mg, 331.5 µmol), 2chloro-N-ethyl-5-(trifluoromethyl)pyrimidine -4-amine (74.79 mg, 331.5 µmol) and p-TsOHH ₂ O (31.53 mg, 165.75 µmol) in 1,4-dioxane (10 ml) were stirred at 90°C for 3 h in N2 atmosphere. The reaction mixture was quenched with saturated aqueous NaHCO3 (10 ml) and extracted with EtOAc (2x10 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (10 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO ₂ , PE:EtOAc = 1:1) and further purified by preparative HPLC (FA) to give N2-[2-cyclopropyl-5-[1-methyl-1-(triazole-2 -yl)ethyl]pyrazol3-yl]-N4-ethyl-5-(trifluoromethyl)pyrimidin-2,4-diamine. 1H NMR (400 MHz, CDC13): δ 8.13 (s, 1H), 7.62 (s, 2H), 7.30 (br.s, 1H), 6.13 (s, 1H), 5. 18 (br.s, 1H), 3.38-3.47 (m, 2H), 3.24 (tt, J=3.59, 6.95 Hz, 1H), 2.10 (s, 6H) , 1.24 (t, J=7.22 Hz, 3H), 1.09-1.21 (m, 4H). HPLC: RT 2.61 min. MS: m/z: 422.3 [M+H]+.

Example 4. Synthesis of (3S)- and (3R)-3-[1-cyclopropyl-5-[[4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2yl]amino]pyrazol-3-yl]-3- methyl-tetrahydrofuran-2-one (143 and 144).

tert-Butyl N-(1-methylcyclopropyl)carbamate.

To a mixture of sodium (5.34 g, 232.32 mmol) in diethyl carbonate (50 ml) was added a solution of tetrahydrofuran-2-one (20 g, 232.32 mmol) in diethyl carbonate (25 ml) at 100°C for period 3 h. The mixture was cooled to 20°C and quenched with an ice-cold saturated aqueous solution of NH4Cl, then the pH was adjusted to pH 5 by adding 1N. HCl. The aqueous phase was extracted with EtOAc (3x30 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (2x20 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give ethyl 2-oxotetrahydrofuran-3-carboxylate as a light yellow oil. 1H NMR (400 MHz, CDC13): δ ppm 4.49 (td, J=8.47, 5.52 Hz, 1H), 4.34 (dt, J=8.69, 7.45 Hz, 1H), 4.244.30 (m, 2H), 3 .55 (dd, J=9.35, 7.59 Hz, 1H), 2.69 (dav, J=13.07, 7.57 Hz, 1H), 2.51 (dddd, J=13.08 , 9.32, 7.59, 5.52 Hz, 1H) 1.32 (t, J=7.09 Hz, 3H).

Ethyl 3-methyl-2-oxo-tetrahydrofuran-3-carboxylate.

To a solution of ethyl 2-oxotetrahydrofuran-3-carboxylate (6.9 g, 43.63 mmol) in THF (150 ml) was added NaH (1.92 g, 47.99 mmol, 60% purity) at 0°C for 30 min. After addition, the mixture was stirred at 20°C for 30 min and then MeI (9.29 g, 65.45 mmol, 4.07 ml) was added dropwise at

- 24 043797

0°C for 30 min. The resulting mixture was stirred at 20°C for 10.5 hours. The mixture was poured into an aqueous saturated solution of NH ₄ Cl at 0°C and extracted with EtOAc (3x50 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (30 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 20:1 to 1:1) to give ethyl 3-methyl-2-oxotetrahydrofuran-3-carboxylate as a yellow oil. 1H NMR (400 MHz, CDC13): δ ppm 4.32-4.44 (m, 2H) 4.24 (kv, J=7.20 Hz, 2H), 2.76 (ddd, J=13.01, 7.06, 4.19 Hz, 1H ) 2.20 (dt, J=13.23, 8.38 Hz, 1H), 1.54 (s, 3H), 1.30 (t, J=7.17 Hz, 3H).

3-(3-Methyl-2-oxo-tetrahydrofuran-3-yl)-3-oxopropanenitrile.

To a solution of CH3CN (1.2 g, 30.03 mmol, 1.58 ml) in THF (50 ml) was added dropwise n-BuLi (12.01 ml, 2.5 M) at -78°C for 30 min in N2 atmosphere. After addition, the mixture was stirred at this temperature for 30 minutes. The suspension mixture was added dropwise to a solution of ethyl 3-methyl-2oxo-tetrahydrofuran-3-carboxylate (4.7 g, 27.30 mmol) in THF (50 ml) at -78°C for 30 minutes. The resulting mixture was heated to -40°C and stirred at -40°C for 1.5 hours. The reaction mixture was quenched using a saturated aqueous solution of NH ₄ Cl at 0°C, and then the pH was adjusted to pH = 4-5 using 1n. HCl and extracted with EtOAc (3x50 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (20 ml), dried over Na2SO4, filtered and concentrated under reduced pressure to give 3-(3-methyl-2-oxo-tetrahydrofuran-3-yl)-3oxopropanenitrile as a yellow solid. , which was used in the next step without further purification. 1H NMR (400 MHz, CDCl ₃ ): δ ppm 4.29-4.46 (m, 2H), 3.79-4.12 (m, 2H), 3.03 (ddd, J=13.40, 7.55, 6.17 Hz, 1H), 2.10 (dt, J=13.73, 7.14 Hz, 1H), 1.60 (s, 3H).

-(5-Amino-1-cyclopropylpyrazol-3-yl)-3-methyl-tetrahydrofuran-2-one.

A mixture of 3-(3-methyl-2-oxo-tetrahydrofuran-3-yl)-3-oxopropanenitrile (200 mg, 1.2 mmol) and cyclopropylhydrazine dihydrochloride salt (174 mg, 1.2 mmol) in i-PrOH (5 ml) was stirred at 50°C for 16 hours under N ₂ atmosphere. The reaction solution was adjusted to pH 7 with saturated aqueous NaHCO3 and extracted with EtOAc (3x5 ml). The organic layers were combined, washed with aqueous saturated sodium chloride solution (5 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by preparative TLC (DCM:MeOH = 10:1) to give 3-(5-amino-1-cyclopropylpyrazol-3-yl)-3-methyltetrahydrofuran-2-one as a yellow oil. 1H NMR (400 MHz, CDCl ₃ ): δ ppm 5.47 (s, 1H), 4.24-4.41 (m, 2H), 3.76-3.94 (br.s, 2H), 3.04-3.14 (m, 1H), 2.89-3.02 (m, 1H), 2.12-2.28 (m, 1H), 1.53 (s, 3H), 0.95-1.04 (m, 4H).

(3S) and (3R) N2-[5-Cyclopropyl-1-[3-(triazol-2-yl)cyclobutyl]pyrazol-4-yl]-N4-ethyl-5(trifluoromethyl)pyrimidine-2,4-diamine .

To a solution of 3-(5-amino-1-cyclopropylpyrazol-3-yl)-3-methyl-tetrahydrofuran-2-one (90 mg, 406.76 μmol) in 1,4-dioxane (5 ml) was added 2-chloro -M-ethyl-5-(trifluoromethyl)pyrimidin-4-amine (91.77 mg, 406.76 µmol) and p-TsOH (14.01 mg, 81.35 µmol). The mixture was stirred at 90°C for 10 hours. The pH of the reaction solution was adjusted to pH 7 using saturated aqueous NaHCO ₃ solution, extracted with EtOAc (3x5 ml). The organic layers were combined, washed with aqueous saturated sodium chloride solution (5 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by preparative TLC (PE:EtOAc = 1:1) to obtain enantiomer mixtures, which were separated by SFC.

First eluting isomer: 1H NMR (400 MHz, CDC13): δ ppm 8.09 (s, 1H), 7.16 (br.s, 1H), 6.55 (s, 1H), 5.17 (br.s, 1H), 4.20-4.32 (m, 2H), 3.48-3.57 (m, 2H), 3.12-3.20 (m, 1H), 2.93 (ddd, J=12.58, 6.49, 4.02 Hz, 1H), 2.19 (dt, J=12.58, 8.52 Hz, 1H), 1.53 (s, 3H), 1.24 (t, J=7.22 Hz, 3H), 1, 02-1.13 (m, 4H). HPLC: RT: 2.33 min. MS: m/z: 411.2 [M+H]+. Second eluting isomer: 1H NMR (400 MHz, CDC13): δ ppm. 8.17 (d, J=0.75 Hz, 1H), 7.28 (br.s, 1H), 6.62 (s, 1H), 5.26 (br.s, 1H) 4.28- 4.42 (m, 2H), 3.55-3.66 (m, 2H), 3.173.28 (m, 1H), 3.01 (ddd, J=12.61, 6.46, 4.02 Hz, 1H), 2.26 (dt, J=12.55, 8.47 Hz, 1H), 1.53-1.64 (m, 3H), 1.32 (t, J=7.22 Hz , 3H), 1.10-1.21 (m, 5H). HPLC: RT: 2.33 min. MC: m/z: 411.2[M+H] ⁺ .

Example 5. Synthesis of 1-(1-cyclopropyl-5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)1H-pyrazol-3-yl)pyrrolidin-2-one (153 ).

-Cyclopropyl-1 H-pyrazole-3,5-diamine.

A mixture of propane dinitrile (6.15 g, 93.09 mmol) and cyclopropylhydrazine (9 g, 62.06 mmol, 2HC1 salt) in i-PrOH (10 ml) was heated at 105°C for 5 hours. The reaction solution was cooled to 0°C, the pH was adjusted to pH 7 using a saturated aqueous NaHCO ₃ solution, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH = 30:1 to 10:1) to give 1-cyclopropylpyrazole-3,5-diamine as a brown syrup. 1H NMR (400 MHz, CDC13): δ ppm 4.88 (s, 1H), 3.80 (br.s, 2H), 2.98 (tt, J=6.89, 3.47 Hz, 1H), 2.84 (br.s, 2H) , 1.05 (dkv, J=7.86, 3.70 Hz, 2H), 0.93-1.00 (m, 2H).

N-(5-Amino-1-cyclopropyl-1H-pyrazol-3-yl)-4-chlorobutαnamide.

To a solution of 1-cyclopropylpyrazole-3,5-diamine (2.25 g, 16.28 mmol) and TEA (3.29 g, 32.56 mmol) in DCM (200 ml) was added dropwise 4-chlorobutanoyl chloride (2, 07 g, 14.65 mmol) at 0°C for 30

- 25 043797 min. The mixture was stirred at 0°C for 30 minutes and stirred at 15°C for 1 hour. The reaction mixture was diluted with H2O (50 ml) and extracted with DCM: i-PrOH (v:v = 3:1, 3x30 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give N-(5-amino-1-cyclopropylpyrazol-3-yl)-4-chlorobutanamide as a yellow oil. 1H NMR (400 MHz, CDCl3): δ ppm 7.97 (br.s, 1H), 5.94 (s, 1H), 3.90 (br.s, 2H), 3.63 (t, J=6.21 Hz, 2H), 3.08 (tt, J=6.82, 3.59 Hz, 1H), 2.49 (t, J= 7.09 Hz, 2H), 2.16 (quin., J= 6.62 Hz, 2H), 0.93-1.13 (m, 4H).

-(5-Amino-1-cyclopropyl-1H-pyrazol-3-yl)pyrrolidin-2-one.

To a solution of N-(5-amino-1-cyclopropylpyrazol-3-yl)-4-chlorobutanamide (1.3 g, 5.36 mmol) in THF (390 ml) was added NaH (536 mg, 13.40 mmol, 60 % purity) at 0°C for 10 minutes. After addition, the mixture was stirred at 0°C for 20 minutes and then stirred at 15°C for 1.5 hours. The reaction mixture was quenched with saturated aqueous NH ₄ Cl (100 ml) at 0°C and then extracted with DCM: i-PrOH (v:v = 3:1.3 x 100 ml). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give 1-(5-amino-1cyclopropylpyrazol-3-yl)pyrrolidin-2-one as an off-white solid. 1H NMR (400 MHz, CDCl ₃ ): δ ppm 6.10 (s, 1H), 3.89 (t, J=7.06 Hz, 4H), 3.10 (tt, J=6.86, 3.61 Hz, 1H), 2.52 (t , J=8.05 Hz, 2H), 2.11 (quin, J=7.61 Hz, 2H), 1.06-1.12 (m, 2H), 1.00-1.06 (m, 2H).

1-(1-Cyclopropyl-5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-3-yl)pyrrolidin-2-one.

To a solution of 1-(5-amino-1-cyclopropylpyrazol-3-yl)pyrrolidin-2-one (180 mg, 872.77 µmol) and 2chloro-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine (197 mg, 872.77 µmol) in 1,4-dioxane (10 ml) p-TsOH was added. H ₂ O (45 mg, 261.83 µmol). The mixture was stirred at 90°C for 12 hours. The reaction mixture was diluted with H2O (30 ml) and adjusted to pH 8-9 with aqueous NaHCO ₃ (10 ml) at 0°C and extracted with EtOAc (3x30 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (10 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (FA) to give 1-(1-cyclopropyl-5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-3-yl) pyrrolidin-2-one. 1H NMR (400 MHz, CDCl ₃ ): δ ppm 8.16 (s, 1H), 7.35 (br.s, 1H), 7.22 (s, 1H), 5.27 (br.s, 1H), 3.93 (t, J=7, 06 Hz, 2H), 3.62-3.73 (m, 2H), 3.19-3.27 (m, 1H), 2.56 (t, J=8.16 Hz, 2H), 2, 09-2.20 (m, 2H), 1.34 (t, J=7.28 Hz, 3H), 1.15-1.20 (m, 2H), 1.09-1.15 (m, 2H). HPLC: RT 2.11 min. MS: m/z: 396.2 [M+H]+.

Example 6. Synthesis of N2-[3-cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-yl]-N4-ethyl-5(trifluoromethyl)pyrimidine-2,4-diamine (110).

3-(3-Cyclopropyl-4-nitropyrazol-1-yl)thietane 1,1-dioxide.

To a mixture of 3-cyclopropyl-4-nitro-1H-pyrazole (500 mg, 3.26 mmol) in DMF (15 ml) was added NaH (156 mg, 3.91 mmol, 60% purity) at 0°C in an N2 atmosphere . The mixture was stirred at 20°C for 30 minutes, then treated with 1,1-dioxane-3-bromothiethane (1.01 g, 3.26 mmol) and stirred at 20°C for 15.5 hours. The mixture was poured into ice water (30 ml) and extracted with EtOAc (3x15 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (3x 15 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 3:1) to give 3-(3-cyclopropyl-4-nitropyrazol-1yl)thietane 1,1-dioxide as a yellow oil.

3-Cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-amine.

Fe (174 mg, 3.11 mmol) and NH4C1 (166 mg, 3.11 mmol, 108.71 µl) at 20°C. The reaction mixture was heated at 70°C for 2 hours, then concentrated under reduced pressure. The residue was washed with a solvent mixture of DCM and MeOH (10 ml, 10:1), filtered and the filtrate was concentrated under reduced pressure to give 3-cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-amine as brown oils

N2-[3-Cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-yl]-N4-ethyl-5-(trifluoromethyl)pyrimidine2,4-diamine.

To a solution of 3-cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-amine (100 mg, 439.99 µmol) and 2-chloro-methyl-5-(trifluoromethyl)pyrimidin-4-amine (99 mg, 439.99 µmol) in 1,4-dioxane (5 ml) p-TsOH (15 mg, 88 µmol) was added. The reaction mixture was stirred at 80°C for 1 hour. The pH of the mixture was adjusted to pH 7 using saturated aqueous NaHCO ₃ solution, extracted with EtOAc (3x5 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (5 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC (neutral) to give N2-[3-cyclopropyl-1-(1,1-dioxothietan-3-yl)pyrazol-4-yl]-N4-ethyl-5(trifluoromethyl)pyrimidine-2 ,4-diamine. 1H NMR (400 MHz, CDC13): δ 8.16 (s, 1H), 8.12 (br.s, 1H), 6.62-7.03 (m, 1H), 5.15 (br.s , 1H), 5.07 (br.s, 1H), 4.66 (br.s, 2H), 4.58 (br.s, 2H), 3.58 (br.s, J=5.90 Hz, 2H), 1.67-1.78 (m, 1H), 1.32 (br.t, J=6.78 Hz, 3H), 0.91-0.98 (m, 2H), 0 .81-0.90 (m, 2H). HPLC: RT: 1.92 min. MC: m/z = 417.2 [M+H]+.

- 26 043797

Example 7 Synthesis of (1R,5S)-1-[1-cyclopropyl-5-[[4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2yl]amino]pyrazol-3-yl]-3-oxabicyclo[3.1 .0]hexan-2-one (162).

Methyl (1R,5S)-2-oxo-3-oxabicyclo[3.1.0]hexane-1-carboxylate.

Na (8.27 g, 359.52 mmol) was added to MeOH (500 ml) and the mixture was stirred at 20°C for 3 hours until the Na dissolved. Dimethylpropanedioate (50 g, 378.44 mmol) was added at 0°C, then after 30 min (2S)-2-(chloromethyl)oxirane (31.51 g, 340.6 mmol) was added at 20°C under N2. The mixture was stirred at 90°C for 12 hours. The mixture was concentrated under reduced pressure at 45°C. The residue was poured into ice water (100 ml) and stirred for 5 minutes. The aqueous phase was extracted with EtOAc (3x300 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:1-5:1) to give methyl (1R,5S)-2-oxo-3-oxabicyclo[3.1.0]hexane-1-carboxylate as an oil . 1H NMR (400 MHz, CDCl ₃ ): δ ppm 4.35 (dd, J=9.37, 4.74 Hz, 1H), 4.18 (d, J=9.48 Hz, 1H), 3.79 (s, 3H), 3.33-3 .40 (m, 1H), 2.74 (dt, J=7.94, 5.18 Hz, 1H), 2.07 (dd, J=7.94, 4.85 Hz, 1H), 1, 39 (t, J=5.07 Hz, 1H).

-Oxo-3-[(1 R, 5 S)-2-okco-3 -oxabicyclo[3.1.0]hexan-1 -yl]propanenitrile.

To a mixture of MeCN (1.45 g, 35.22 mmol) in THF (20 ml) was added n-BuLi (2.5 M, 14.09 ml) at -78°C under N2. After 1 h, the mixture was added to a solution of methyl (1R,5S)-2-oxo-3-oxabicyclo[3.1.0]hexane-1carboxylate (5 g, 32.02 mmol) in THF (30 ml) at 78 °C, then the mixture stirred at -78°C for 2 hours. The mixture was poured into aqueous NH ₄ Cl (30 ml) and stirred for 5 min and adjusted to pH 3 with dilute HCl (1N). The aqueous phase was extracted with EtOAc (3x30 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (30 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:MTBE = 50:1 to 0:1) to give 3-okco-3-[(1R,5S)-2-oxo-3-oxabicyclo[3.1.0]hexane- 1-yl]propanenitrile as a white solid. 1H NMR (400 MHz, CDCl ₃ ): δ ppm 4.25-4.47 (m, 3H), 4.03-4.15 (m, 1H), 3.02 (dt, J=7.99, 5.26 Hz, 1H), 2.19 ( dd, J=8.16, 4.41 Hz, 1H), 1.58-1.65 (m, 1H).

(1R,5S)-1-(5-Amino-1-cyclopropylpyrazol-3-yl)-3-oxabicyclo[3.1.0]hexan-2-one.

To a mixture of 3-okco-3-[(1R,5S)-2-oxo-3-oxabicyclo[3.1.0]hexan-1-yl]propanenitrile (800 mg, 4.84 mmol) in i-PrOH (20 ml ) cyclopropylhydrazine dihydrochloride (632.28 mg, 4.36 mmol) was added in one portion at 25°C under N2. The mixture was stirred at 50°C for 12 hours. The mixture was poured into aqueous NaHCO3 solution (50 ml) and stirred for 10 min. The aqueous phase was extracted with DCM/MeOH (3:1, 3x20 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (20 ml), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO ₂ , DCM:MeOH = 20:1) to give (1R,5S)-1-(5-amino-1-cyclopropylpyrazol-3-yl)-3-oxabicyclo[3.1.0] hexan-2-one as a brown oil. LCMS: RT 0.370 min, m/z = 220.2 [M+H]+. 1H NMR (400 MHz, CDCI3): δ ppm 5.76 (s, 1H), 4.38 (dd, J=9.15, 4.74 Hz, 1H), 4.20 (d, J=9.26 Hz, 1H) 3.83 (br. s, 2H), 3.06 (tt, J=6.89, 3.58 Hz, 1H), 2.61 (dt, J=7.72, 4.63 Hz, 1H), 1.81 (dd , J=7.72, 4.41 Hz, 1H), 1.24 (t, J=4.74 Hz, 1H), 0.94-1.11 (m, 4H).

(1R,5S)-1-[1-Cyclopropyl-5-[[4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-3yl]-3-oxabicyclo[3.1.0]hexane -2-on.

To a mixture of (1R,5S)-1-(5-amino-1-cyclopropylpyrazol-3-yl)-3-oxabicyclo[3.1.0]hexan-2-one (150 mg, 684.18 µmol) and 2-chloro -N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine (154.35 mg, 684.18 µmol) in 1,4dioxane (5 ml) was added p-TsOH* _H2O (26.03 mg, 136. 84 µmol) in one portion at 20°C in an N2 atmosphere. The mixture was stirred at 90°C for 12 hours. The mixture was poured into aqueous NaHCO ₃ solution (30 ml) and stirred for 10 minutes. The aqueous phase was extracted with EtOAc (3x20 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (30 ml), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (neutral conditions) to give (1R,5S)-1-[1-cyclopropyl-5-[[4(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-3 -yl]-3-oxabicyclo[3.1.0]hexan-2-one. 1H NMR (400 MHz, CDCl·,): δ ppm 8.10 (s, 1H), 7.13 (br.s, 1H), 6.87 (s, 1H), 5.16 (br.s, 1H), 4.35 (dd, J=9, 22, 4.71 Hz, 1H), 4.18 (d, J=9.29 Hz, 1H), 3.49-3.64 (m, 2H), 3.07-3.19 (m, 1H ), 2.57-2.68 (m, 1H), 1.81 (dd, J=7.72, 4.45 Hz, 1H), 1.22-1.30 (m, 4H), 0 .98-1.12 (m, 3H), 1.09 (br.s, 1H). HPLC: RT: 2.17 min. MS: m/z: 409 [M+H]+.

Example 8. Synthesis of (1R,3R)-3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1Hpyrazol-1-yl)cyclobutanecarbonitrile (181).

3-[2-(3-Benzyloxycyclobutylidene)hydrazine]propanenitrile.

A mixture of 3-benzyloxycyclobutanone (10 g, 56.75 mmol) and 3-hydrazinopropanenitrile (4.83 g, 56.75 mmol) in EtOH (150 ml) was stirred at 20°C for 16 hours. The mixture was concentrated under reduced pressure to give 3-[2-(3-benzyloxycyclobutylidene)hydrazino]propanenitrile (13.81 g, crude) as a yellow oil. LCMS: RT 0.686 min, m/z = 244.2 [M+H] ⁺ .

- 27 043797

2-(3-Benzyloxycyclobutyl)pyrazol-3-amine.

To a mixture of 3-[2-(3-benzyloxycyclobutylidene)hydrazino]propanenitrile (13.81 g, 56.76 mmol) in tBuOH (130 ml) was added t-BuONa (5.45 g, 56.76 mmol) under N2 atmosphere . The mixture was stirred at 110°C for 3 hours. The mixture was poured into ice water (100 ml) and extracted with EtOAc (2x00 ml). The pH of the organic phase was adjusted to pH 3 with 2N. HCl and washed with water (3x100 ml). The pH of the aqueous phase was adjusted to pH 8 with 6 N. NaOH, extracted with EtOAc (3x100 ml), washed with aqueous saturated sodium chloride solution (100 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give 2-(3-benzyloxycyclobutyl)pyrazole-3-amine as a yellow oil. LCMS: RT 0.625 min, m/z = 244.2 [M+H]+.

3-(5-Aminopyrazol-1-yl)cyclobutanol.

To a solution of 2-(3-benzyloxycyclobutyl)pyrazol-3-amine (5 g, 20.55 mmol) in DCM (200 mL) was added BCl ₃ (1 M, 8.02 mL) at 0° C. under N2. The mixture was stirred at 20°C for 2 hours. The mixture was poured into saturated aqueous NaHCO ₃ solution (200 ml) and the aqueous phase was concentrated under reduced pressure. The residue was washed with DCM:MeOH (v:v = 10:1, 100 ml), filtered and the filtrate was concentrated under reduced pressure to give 3-(5-aminopyrazol-1-yl)cyclobutanol as a yellow oil. LCMS: RT 0.096 min, m/z = 154.1 [M+H] ⁺ .

(1S,3 S)-3-(5-((4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1 H-pyrazol-1 -yl)cyclobutanol (1R,3R)- 3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)cyclobutanol.

To a mixture of 3-(5-aminopyrazol-1-yl)cyclobutanol (2.2 g, 14.36 mmol) in NMP (22 ml) was added 2chloro-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine (2 .59 g, 11.49 mmol) and p-TsOHH ₂ O (819.59 mg, 4.31 mmol) in one portion at 20°C under N2 atmosphere. The mixture was then heated to 100°C and stirred for 16 hours. The mixture was cooled to 20°C, poured into water (150 ml) and adjusted to pH 7-8 with an aqueous solution of NaHCO ₃ . The aqueous phase was extracted with EtOAc (3x50 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (50 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (DCM:MeOH = 30:1) to give a mixture of (1S,3S)-3(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino )-1H-pyrazol-1-yl)cyclobutanol and (1R,3R)3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1- yl)cyclobutanol in the form of a yellow resin.

(1S,3 S)-3-(5-((4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1 H-pyrazol-1 -yl)cyclobutyl methanesulfonate, (1 R, 3R)-3-(5-((4-(ethylamino)-5 -(trifluoromethyl)pyrimidin-2-yl)amino)-1 Npyrazol-1-yl)cyclobutyl methanesulfonate.

To a mixture of (1S,3S)-3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1yl)cyclobutanol and (1R,3R)-3 -(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol1-yl)cyclobutanol (2 g, 5.84 mmol) in DCM (40 ml) added TEA (709.14 mg, 7.01 mmol) and MsCl (802.77 mg, 7.01 mmol) at 0°C in N2 atmosphere. The mixture was then stirred at 0°C for another 1 hour. The mixture was diluted with water (10 ml) and stirred for 3 minutes. The organic phase was separated, washed with aqueous saturated sodium chloride solution (10 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (DCM:MeOH = 30:1) to give a mixture of (1S,3S)-3-(5-((4(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino )-1H-pyrazol-1-yl)cyclobutyl methanesulfonate and (1R,3R)-3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)1H-pyrazol-1 -yl)cyclobutyl methanesulfonate as a yellow oil.

(1R,3R)-3-(5-((4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)cyclobutanecarbonitrile.

To a mixture of (1S,3S)-3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1yl)cyclobutyl methanesulfonate and (1R,3R)- 3-(5-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1Hpyrazol-1-yl)cyclobutyl methanesulfonate (200 mg, 475.73 µmol) in DMSO (4 ml) 18-crown-6 (12 mg, 47.57 µmol) and NaCN (140 mg, 2.85 mmol) were added at 20°C under N2. The mixture was then heated to 120°C and stirred for 8 hours. The mixture was cooled to 20°C and poured into water (50 ml). The aqueous phase was extracted with EtOAc (3 x 20 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (20 ml), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (FA) to give the product, which was further purified by preparative TLC (PE:EtOAc = 1:1) to give (1R,3R)-3-(5-((4-(ethylamino)- 5(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)cyclobutanecarbonitrile and by-product 2(6,7-dihydro-5,7-methanopyrazolo[1,5-a]pyrimidin-4(5H )-yl)-N-ethyl-5-(trifluoromethyl)pyrimidin-4-amine.

(1R,3R)-3-(5-((4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)cyclobutanecarbonitrile. 1H NMR (400 MHz, CDCI3): δ 8.08 (s, 1H), 7.57 (d, J=1.76 Hz, 1H), 7.08 (br.s, 1H), 6.19 ( d, J=1.76 Hz, 1H), 5.16 (br.s, 1H), 5.06 (quin., J=7.87 Hz, 1H), 3.36-3.48 (m, 2H), 3.24-3.36 (m, 1H), 3.06-3.18 (m, 2H), 2.73-2.84 (m, 2H), 1.20 (t, J= 7.22 Hz, 3H). LCMS: RT: 0.652 min. MS: m/z: 352.1 [M+H] ⁺ .

- 28 043797

Example 9. Synthesis of N2-(1-((1R,3R)-3-(2H-1,2,3-triazol-2-yl)cyclobutyl)-1H-pyrazol-5-yl)-N4-ethyl5-( trifluoromethyl)pyrimidin-2,4-diamine (182) and N2-(1-((1 R,3R-3-(1 H-1,2,3-triazol-1 -yl)cyclobutyl)1H-pyrazole-5 -yl)-M4-ethyl-5-(trifluoromethyl)pyrimidine-2,4-diamine (183).

N2-(1-((1R,3R)-3-(2H-1,2,3-Triazol-2-yl)cyclobutyl)-1H-pyrazol-5-yl)-N4-ethyl-5-(grifromethyl) pyrimidine-2,4-diamine and N2-(1-((1R,3R)-3-(1H-1,2,3-triazol-1-yl)cyclobutyl)-1H-pyrazol-5-yl)-N4 ethyl 5(trifluoromethyl)pyrimidine-2,4-diamine.

To a mixture of (1S,3S)-3-(5-((4-(ethylamino)-5-(mrifluoromethyl)pyrimidin-2-yl)amino)-1H-pyrazol-1yl)cyclobutylmethanesulfonate and (1R,3R)-3 -(5-((4-(ethylamino)-5-(grifformethyl)pyrimidin-2-yl)amino)-1Hpyrazol-1-yl)cyclobutylmethanesulfonate (300 mg, 713.59 µmol) in DMF (5 ml) was added K2CO3 (148 mg, 1.07 mmol) and 2H-triazole (74 mg, 1.07 mmol) in one portion at 20°C under N2 atmosphere. The mixture was then heated to 120°C and stirred for 8 hours. The mixture was cooled to 20°C and poured into water (50 ml). The aqueous phase was extracted with EtOAc (3x20 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (20 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was separated by HPLC (FA conditions) to give N2-(1-((1R,3R)-3-(2H-1,2,3-mriazol-2yl)cyclobugyl)-1H-pyrazol-5-yl)- N4-ethyl-5-(gryfluoromethyl)pyrimidin-2,4-diamine and N2-(1-((1R,3R)-3-(1H-1,2,3triazol-1-yl)cyclobugyl)-1H-pyrazole -5-yl)-N4-ethyl-5-(mrifluoromethyl)pyrimidine-2,4-diamine.

N2-(1-((1R,3R)-3-(2H-1,2,3-Triazol-2-yl)cyclobutyl)-1H-pyrazol-5-yl)-N4-ethyl-5-(grifromethyl) pyrimidine-2,4-diamine (182). 1H NMR (400 MHz, CDCI3): δ 8.11 (s, 1H), 7.64 (s, 2H), 7.61 (d, J=1.88 Hz, 1H), 6.83 (br. s, 1H), 6.29 (d, J=1.76 Hz, 1H), 5.50 (tt, J=4.49, 8.69 Hz, 1H), 5.17-5.27 (m , 1H), 5.13 (br.s, 1H), 3.39-3.51 (m, 2H), 3.25-3.36, 2H), 3.01-3.14 (m, 2H ), 1.21 (t, J=7.22 Hz, 3H).LCMS: VU: 0.706 min. MS: m/z: 394.3 [M+H]+.

N2-(1-((1R,3R)-3-(1H-1,2,3-Triazol-1-yl)cyclobutyl)-1H-pyrazol-5-yl)-N4-ethyl-5-(grifromethyl) pyrimidine-2,4-diamine (183). 1H NMR (400 MHz, CDCl): δ 8.10 (s, 1H), 7.74 (s, 1H), 7.61 (d, J=1.63 Hz, 1H), 7.60 ( s, 1H), 6.70 (br.s, 1H) 6.28 (d, J=1.76 Hz, 1H), 5.36-5.45 (m, 1H), 5.18-5, 27 (m, 1H), 5.14 (br.s, 1H), 3.37-3.53 (m, 2H), 3.31 (ddd, J=5.77, 8.31, 13.65 Hz, 2H), 3.11-3.23 (m, 2H), 1.22 (t, J=7.22 Hz, 3H).LCMS: VU: 0.660 min. MS: m/z: 394.2 [M+H] ⁺ .

Example 10. Synthesis of N2-(5-cyclopropyl-1-pyrazin-2-yl-pyrazol-4-yl)-N4-ethyl-5-(trifluoromethyl)pyrimidine-2,4-diamine (105).

2-(4-Nitropyrazol-1-yl)pyrazine.

To a solution of 4-nitro-1H-pyrazole (1 g, 8.84 mmol) in DMF (20 ml) was added NaH (424 mg, 10.61 mmol, 60% purity) at 0°C under N2 atmosphere. The mixture was stirred at 0°C for 1 hour. 2-chloropyrazine (1.01 g, 8.84 mmol, 790.99 μl) was then added at 0°C and the mixture was heated to 80°C and stirred for 12 hours. The mixture was cooled to 20°C and quenched with a cold aqueous saturated solution of NH4C1 (60 ml). The aqueous phase was extracted with EtOAc (3x20 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (3x15 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give 2-(4-nitropyrazol-1yl)pyrazine as a light yellow solid. 1H NMR (400 MHz, DMSO-d ₆ ): δ ppm 9.54 (s, 1H), 9.31 (d, J=1.13 Hz, 1H), 8.81 (d, J=2.51 Hz, 1H), 8.70-8.74 (m , 1H), 8.71 (s, 1H), 8.69 (dd, J=2.45, 1.32 Hz, 1H).

2-(5-Chloro-4-nitropyrazol-1-yl)pyrazine.

To a solution of 2-(4-nitropyrazol-1-yl)pyrazine (0.78 g, 4.08 mmol) in THF (15 ml) was added LiHMDS (1M, 4.49 mmol, 4.49 ml) at -78° C in N2 atmosphere. The mixture was stirred at -78°C for 30 min, then a solution of 1,1,1,2,2,2-hexachloroethane (1.06 g, 4.49 mmol, 508.45 μl) in THF (10 ml) was added. at -78°C under N2 atmosphere and the mixture was stirred for 3.5 hours. The mixture was quenched with cold aqueous saturated NH4C1 solution (30 ml). The aqueous phase was extracted with EtOAc (3x10 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (10 ml), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 1:1) to give 2-(5-chloro-4-nitropyrazol-1yl)pyrazine as a white solid. LCMS: RT 1.066 min. MS m/z = 226.0 [M+H] ⁺ .

2-(5-Cyclopropyl-4-nitropyrazol-1-yl)pyrazine.

To a mixture of 2-(5-chloro-4-nitropyrazol-1-yl)pyrazine (200 mg, 886.56 µmol) and cyclopropylboronic acid (380 mg, 4.43 mmol) in 1,4-dioxane (10 ml) was added KF (154 mg, 2.66 mmol) and Pd (dppf)Cl ₂ -CH ₂ Cl ₂ (145 mg, 177.31 µmol) at 20°C in a N2 atmosphere. The mixture was heated to 110°C and stirred for 12 hours. The mixture was cooled to 20°C and filtered. The residue was diluted with water (15 ml). The aqueous phase was extracted with EtOAc (3x8 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (8 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give 2-(5-cyclopropyl-4-nitropyrazol-1-yl)pyrazine. 1H NMR (400 MHz, CDC13): δ ppm 9.08 (s, 1H), 8.71 (d, J=2.38 Hz, 1H), 8.56-8.61 (m, 1H), 8.29 (s, 1H) 2.36 ( tt, J=8.52, 5.79 Hz, 1H), 1.07-1.17 (m, 2H), -0.17 (tt, J=8.96, 5.91 Hz, 2H).

5-Cyclopropyl-1-pyrazin-2-yl-pyrazol-4-amine.

To a solution of 2-(5-cyclopropyl-4-nitropyrazol-1-yl)pyrazine (240 mg, 1.04 mmol) in EtOH (16 ml) and H2O (4 ml) was added NH4C1 (277 mg, 5.19 mmol) and Fe (290 mg, 5.19 mmol) at 20°C. The mixture was heated

- 29 043797 to 80°C and stirred for 2 hours. The mixture was cooled to 20°C, filtered and concentrated under reduced pressure. The residue was washed with DCM:MeOH (10 ml, v:v = 10:1), filtered and concentrated under reduced pressure to give 5-cyclopropyl-1-pyrazin-2-yl-pyrazol-4-amine as a brown oil. LCMS: RT 0.711 min. MS m/z = 202.1 [M+H]+.

N2-(5-Cyclopropyl-1-pyrazin-2-yl-pyrazol-4-yl)-N4-ethyl-5-(trifluoromethyl)pyrimidin-2,4-diamine.

To a mixture of 5-cyclopropyl-1-pyrazin-2-yl-pyrazol-4-amine (100 mg, 496.95 µmol) and 2-chloro-Nethyl5-(trifluoromethyl)pyrimidin-4-amine (112 mg, 496. 95 µmol) in 1,4-dioxane (5 ml) pTsOH.H ₂ O (34 mg, 198.78 µmol) was added at 20°C. The mixture was heated to 90°C and stirred for 2 hours. The mixture was cooled to 20°C, water (10 ml) was added and the pH was adjusted to pH 7-8 using saturated aqueous NaHCO ₃ solution. The aqueous phase was extracted with EtOAc (3x8 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (2x5 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 0:1) to give N2-(5-cyclopropyl-1-pyrazin-2-yl-pyrazol-4-yl)-N4ethyl-5-( trifluoromethyl)pyrimidine-2,4-diamine. 1H NMR (400 MHz, MeOD): δ ppm 9.08 (s, 1H), 8.55 (s, 2H), 8.04 (br.s, 2H), 3.53 (kv, J=6.82 Hz, 2H), 2.16-2 .34 (m, 1H), 1.20 (br.t, J=7.03 Hz, 3H), 0.91 (br.d, J=6.90 Hz, 2H), 0.55 (br.d., J=6.90 Hz, 2H). d, J=4.77 Hz, 2H). HPLC: RT: 2.06 min. MS: m/z: 391.2 [M+H] ⁺ .

Example 11 Synthesis of (3S)-3-[3-cyclopropyl-4-[[4-(methylamino)-5-(trifluoromethyl)pyrimidin-2yl]amino]pyrazol-1-yl]-3-methyltetrahydrofuran-2-one and (3R)-3-[3-cyclopropyl-4-[[4-(methylamino)-5(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-1-yl]-3-methyltetrahydrofuran-2-one (113 and 122).

3-(3-Cyclopropyl-4-nitropyrazol-1-yl) tetrahydrofuran-2-one.

To a solution of 3-cyclopropyl-4-nitro-1H-pyrazole (1 g, 6.53 mmol) in DMF (10 ml) was added NaH (313 mg, 7.84 mmol, 60% purity) at 0°C under N2 atmosphere . The mixture was stirred at 20°C for 30 minutes, then treated with 3-bromotetrahydrofuran-2-one (1.19 g, 7.18 mmol, 670 μl) and stirred for 15.5 hours. The mixture was poured into ice water (20 ml) and EtOAc extract (3x10 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (3x10 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 1:1) to give 3-(3cyclopropyl-4-nitropyrazol-1-yl)tetrahydrofuran-2-one as a yellow oil. 1H NMR (400 MHz, CDC13): δ 8.31 (s, 1H), 4.96 (t, J=9.16 Hz, 1H), 4.65 (td, J=8.88, 3.45 Hz, 1H), 4.39-4.51 (m, 1H), 2.95 (dd, J= 13.25, 8.92 Hz, 1H), 2.77-2.87 (m, 1H) , 2.56-2.65 (m, 1H), 1.01-1.09 (m, 2H), 0.93-1.01 (m, 2H). LCMS: RT 0.746 min, m/z = 252.1 [M+H] ⁺ .

-(3-Cyclopropyl-4-nitropyrazol-1-yl)-3-methyl-tetrahydrofuran-2-one.

To a solution of 3-(3-cyclopropyl-4-nitropyrazol-1-yl) tetrahydrofuran-2-one (780 mg, 3.29 mmol) in THF (15 ml) was added LDA (4.93 mmol, 2 M, 2. 47 ml) at -78°C in an N2 atmosphere. The mixture was stirred at -78°C for 30 min, then treated with MeI (700 mg, 4.93 mmol, 307 μl) at -78°C, warmed to 0°C and stirred for 1.5 hours. The mixture was poured into saturated aqueous NH4CI (15 ml) and extracted with EtOAc (3 x 5 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (5 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 1:1) to give 3-(3-cyclopropyl-4-nitropyrazol-1-yl)-3methyltetrahydrofuran-2-one as a colorless oil. 1H NMR (400 MHz, CDC13): δ 8.40 (s, 1H), 7.27 (s, 1H), 4.55 (td, J=8.53, 5.77 Hz, 1H), 4, 38-4.48 (m, 1H), 3.12-3.22 (m, 1H), 2.56-2.65 (m, 1H), 2.49 (ddd, J=13.49, 7 .59, 5.90 Hz, 1H), 1.84 (s, 3H), 1.00-1.09 (m, 2H), 0.90-1.00 (m, 3H). LCMS: RT 0.746 min, m/z = 252.1 [M+H] ⁺ .

-(4-Amino-3-cyclopropylpyrazol-1-yl)-3-methyltetrahydrofuran-2-one.

To a solution of 3-(3-cyclopropyl-4-nitropyrazol-1-yl)-3-methyltetrahydrofuran-2-one (555 mg, 2.21 mmol) in MeOH (15 ml) was added Pd-C (10%, 220 mg ) in an N2 atmosphere. The suspension was degassed under reduced pressure and purged with H ² three times. The mixture was stirred under H2 (15 psi) at 20°C for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give 3-(4-amino-3-cyclopropylpyrazol-1-yl)-3-methyltetrahydrofuran-2-one as a yellow oil. 1H NMR (400 MHz, CDCI3): δ 7.18 (s, 1H), 4.43-4.51 (m, 1H), 4.30-4.39 (m, 1H), 3.25 (ddd , J=13.05, 7.53, 5.02 Hz, 1H), 2.91 (br.s, 2H), 2.36 (dt, J=13.43, 7.47 Hz, 1H), 1.72 (s, 3H), 1.62-1.70 (m, 1H), 0.82-0.90 (m, 2H), 0.79 (ddd, J=7.81, 4.99 , 2.38 Hz, 2H).

(3S)-3-[3-Cyclopropyl-4-[[4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-1-yl]3-methyltetrahydrofuran-2-one and (3R )-3-[3-cyclopropyl-4-[[4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-1-yl]-3-methyltetrahydrofuran-2-one.

To a mixture of 2-chloro-N-methyl-5-(trifluoromethyl)pyrimidin-4-amine (143 mg, 677.95 µmol) and 3-(4amino-3-cyclopropylpyrazol-1-yl)-3-methyl-tetrahydrofuran- 2-one (150 mg, 677.95 µmol) in 1,4dioxane (10 ml), p-TsOHH ₂ O (40 mg, 203.39 µmol) was added at 20°C under nitrogen and stirred at 90°C for 4 hours. The mixture was poured into ice water (10 ml) and EtOAc extracts (3x8

- 30 043797 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (8 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1) to give 3-[3-cyclopropyl-4[[4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol- 1-yl]-3-methyl-tetrahydrofuran-2one. Enantiomers were separated by SFC to give (3S)-3-[3-cyclopropyl-4-[[4-(methylamino)5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-1-yl]-3-methyltetrahydrofuran -2-one and (3R)-3-[3cyclopropyl-4-[[4-(methylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]pyrazol-1-yl]-3-methyltetrahydrofuran-2- she.

The isomer eluting first is 1H NMR (400 MHz, CDCl ₃ ): δ 8.28 (br.s, 1H), 8.13 (br.s, 1H), 7.08 (br.s, 1H), 5, 25 (br.s, 1H) 4.47 (br.d, J=7.53 Hz, 1H), 4.38 (td, J=8.38, 4.83 Hz, 1h), 3.31 ( br.s, 1h), 3.11 (br.s, 3H), 2.43 (dt, J=13.52, 7.48 Hz, 1H), 1.78 (s, 3H), 1.67 -1.75 (m, 1H), 0.77-0.95 (m, 4H). HPLC: RT: 2.00 min. MS: m/z = 397.2 [M+H]+.

The second isomer eluting - 1H NMR (400 MHz, CDCl ₃ ): δ 8.28 (br.s, 1H), 8.13 (br.s, 1H), 7.08 (br.s, 1H), 5, 25 (br.s, 1H) 4.47 (br.d, J=7.40 Hz, 1H), 4.33-4.42 (m, 1H), 3.32 (br.s, 1H), 3.11 (br.s, 3H), 2.37-2.49 (m, 1H), 1.78 (s, 3H), 1.67-1.76 (m, 1H), 0.79- 0.94 (m, 4H). HPLC: RT: 2.00 min. MS: m/z = 397.2 [M+H] ⁺ .

Example 12. Synthesis of 2-[4-[[4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3-methylpyrazol1 -yl]-2-methylcyclopentanone (194).

2-(4-Bromo-3-methylpyrazol-1-yl)cyclopentanone and 2-(4-bromo-5-methylpyrazol-1-yl)cyclopentanone.

To a solution of 4-bromo-3-methyl-1H-pyrazole (10 g, 62.11 mmol) in DMF (60 ml) was added NaH (3.23 g, 80.75 mmol, 60% purity) at 0°C and stirred at 15°C for 1 hour. Then, 2-chlorocyclopentanone (8.84 g, 74.53 mmol, 7.43 ml) was added to the mixture and stirred at 15°C for 15 hours. The reaction mixture was quenched by adding saturated aqueous NH ₄ Cl (300 ml) at 0°C and then extracted with EtOAc (3x100 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (200 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:MTBE = 2:1 to 1:1) to give a mixture of 2-(4-bromo-3-methylpyrazol-1-yl)cyclopentanone and 2-(4-bromo-5 -methylpyrazol-1yl)cyclopentanone in the form of a yellow resin. LCMS: RT 2.119 min, m/z = 243.1 [M+H] ⁺ .

2-(4-Bromo-3-methylpyrazol-1-yl)-2-methylcyclopentanone and 2-(4-bromo-5-methylpyrazol-1-yl)-2methylcyclopentanone.

To a mixture of 2-(4-bromo-3-methylpyrazol-1-yl)cyclopentanone and 2-(4-bromo-5-methylpyrazol-1yl)cyclopentanone (6.5 g, 26.74 mmol) in THF (30 ml) LiHMDS (1 M, 34.76 ml) was added and stirred at -78°C for 1 hour. MeI (4.93 g, 34.76 mmol, 2.16 ml) was added at -78°C and stirred at 15°C for 15 hours. The reaction mixture was quenched by adding saturated aqueous NH ₄ Cl (200 ml) at 0°C and then extracted with EtOAc (3x70 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (100 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 4:1 to 2:1) to give a mixture of 2-(4-bromo-3-methylpyrazol-1-yl)-2methylcyclopentanone and 2-(4-bromo-5 -methylpyrazol-1-yl)-2-methylcyclopentanone in the form of a yellow resin. LCMS: RT 0.747 min, m/z = 257.1 [M+H] ⁺ .

tert-Butyl N-[3-methyl-1-(1-methyl-2-oxocyclopentyl)pyrazol-4-yl]carbamate and tert-butyl N-[5methyl-1-(1-methyl-2-oxocyclopentyl)pyrazol- 4-yl] carbamate.

Mixture of 2-(4-bromo-3-methylpyrazol-1-yl)-2-methylcyclopentanone and 2-(4-bromo-5-methylpyrazol-1-yl)2-methylcyclopentanone (160 mg,

622.26 µmol), NH ₂ Boc (437 mg, 3.73 mmol), t-BuONa (120 mg, 1.24 mmol) and [2-(2aminoethyl)phenyl]chloropalladium-ditert-butyl-[2-( 2,4,6-triisopropylphenyl)phenyl]phosphane (107 mg, 155.57 µmol) in THF (2 ml) was degassed and purged with N2 3 times, and then the mixture was stirred at 90°C for 2 hours under an _N2 atmosphere. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (neutral conditions) to give tert-butyl N-[3-methyl-1-(1-methyl-2-oxocyclopentyl)pyrazol-4-yl]carbamate and tert-butyl N-[5methyl-1 -(1-methyl-2-oxocyclopentyl)pyrazol-4-yl]carbamate in the form of a yellow resin. LCMS: RT 1.203 min, m/z = 294.3 [M+H] ⁺ .

2-(4-Amino-3-methylpyrazol-1-yl)-2-methylcyclopentanone.

A solution of tert-butyl-N-[3-methyl-1-(1-methyl-2-oxocyclopentyl)pyrazol-4-yl]carbamate (80 mg, 272.7 µmol) in HCl/EtOAc (3 ml) was stirred at 0 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain 2-(4-amino-3-methylpyrazol-1-yl)-2methylcyclopentanone as a yellow solid. LCMS: RT 1.032 min, m/z = 194.2 [M+H] ⁺ .

2-[4-[[4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino]-3-methylpyrazol-1-yl]-2-methylcyclopentanone.

2-(4-Amino-3-methylpyrazol-1-yl)-2-methylcyclopentanone (55 mg, 284.61 µmol), 2-chloro-M-ethyl-5(trifluoromethyl)pyrimidin-4-amine (64 mg, 284.61 µmol) and TEA (86 mg, 853.84 µmol,

- 31 043797

118.84 μL) vn-BuOH (1 mL) was transferred into a microwave reactor vial. The hermetically sealed ampoule was heated at 110°C for 1 hour in a microwave reactor. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (neutral conditions) and preparative TLC (PE:EtOAc = 1:1) to give 2-[4-[[4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl]amino] -3-methylpyrazol-1-yl]-2methylcyclopentanone. 1H NMR (400 MHz, CDC13): δ ppm 8.12 (br.s, 2H), 6.66 (br.s, 1H), 5.15 (br.s, 1H), 3.58 (br.s, 2H), 2.90-3, 07 (m, 1H), 2.38-2.58 (m, 2H), 2.24 (s, 3H), 2.04-2.19 (m, 2H), 1.88-2.00 ( m, 1H), 1.58 (s, 3H), 1.31 (br.t., J=7.09 Hz, 3H). HPLC: retention time: 2.557 min. MS: (M+H)+ m/z: 383.2.

Example 13. Synthesis of (S)-3-(4-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1Hpyrazol-1-yl)-3-(fluoromethyl )dihydrofuran-2(3H)-one and (R)-3-(4-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1 -yl)-3-(fluoromethyl)dihydrofuran-2(3H)-one (216 and 217).

3-(G hydroxymethyl)-3 -(3 -methyl-4-nitro-1H-pyrazol-1 -yl)dihydrofuran-2(3H)-one.

To a mixture of 3-(3-methyl-4-nitropyrazol-1-yl)tetrahydrofuran-2-one (2 g, 9.47 mmol) in THF (25 ml) was added LiHMDS (1M, 12.31 ml) at -78 °C under N2, and then the mixture was stirred at -78 °C for 0.5 hour. A solution of paraformaldehyde (1.02 g, 11.37 mmol) in THF (1 ml) was then added to the reaction mixture, and the mixture was stirred at 10° C. for 2.5 hours. The reaction mixture was quenched by adding saturated aqueous NH4C1 (150 ml) at 0°C and then extracted with EtOAc (3x50 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (50 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 2:1 to 1:1) to give 3-(hydroxymethyl)-3-(3-methyl-4-nitropyrazol-1-yl)tetrahydrofuran-2-one in as a white solid. LCMS: RT 0.497 min, m/z = 242.1 [M+H] ⁺ . 1H NMR (400 MHz, CDC13): δ 8.54 (s, 1H), 4.59-4.48 (m, 2H), 4.20-4.07 (m, 2H), 3.07-2 .98 (m, 1H), 2.95-2.86 (m, 2H), 2.55 (s, 3H).

3-(Fluoromethyl)-3-(3-methyl-4-nitro-1H-pyrazol-1-yl)dihydrofuran-2(3H)-one.

DAST (5, 88 g, 36.48 mmol, 4.82 ml) at 0°C, then the mixture was stirred at 20°C for 15 hours. The reaction mixture was quenched by adding saturated aqueous NaHCO ₃ solution (200 ml) at 0°C and extracted with EtOAc (3x70 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (70 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (TE:EtOAc = 3:1 to 1:1) to give 3-(fluoromethyl)-3-(3-methyl-4-nitropyrazol-1-yl)tetrahydrofuran-2-one in as a white solid. LCMS: RT 0.576 min, m/z = 244.1 [M+H] ⁺ . 1H NMR (400 MHz, CDC13): δ 8.56 (s, 1H), 4.96-4.74 (m, 2H), 4.59-4.49 (m, 2H), 3.30-3 .20 (m, 1H), 2.95-2.86 (m, 1H), 2.55 (s, 3H).

-(4-Amino-3 -methyl-1 H-pyrazol-1 -yl)-3-(fluoromethyl)dihydrofuran-2 (3H)-one.

A mixture of 3-(fluoromethyl)-3-(3-methyl-4-nitropyrazol-1-yl)tetrahydrofuran-2-one (0.7 g, 2.88 mmol), Fe (804 mg, 14.39 mmol) and NH4C1 (770 mg, 14.39 mmol) in EtOH (8 ml) and H ₂ O (2 ml) was stirred at 70°C for 2 hours. The reaction mixture was concentrated under reduced pressure, the residue was diluted with DCM:MeOH (50 ml, ratio = 10:1), filtered and concentrated under reduced pressure to give 3-(4-amino-3methylpyrazol-1-yl)-3-(fluoromethyl) tetrahydrofuran-2-one as a brown solid. LCMS: RT 0.087 min, m/z = 214.1 [M+H] ⁺ . 1H NMR (400 MHz, CDCI3): δ 7.30 (s, 1H), 4.89-4.66 (m, 2H), 4.52-4.40 (m, 2H), 3.31 (br .dd, J=6.2, 13.2 Hz, 1H), 2.87-2.80 (m, 1H), 2.21-2.15 (m, 3H).

(I)-3-(4-((4-(Ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1H-pyrazol-1-yl)-3-(fluoromethyl)dihydrofuran -2(3H)-one and (S)-3-(4-((4-(ethylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)-3-methyl-1Hpyrazol-1 -yl)- 3 -(fluoromethyl)dahydrofuran-2(3H)-one.

Mixture of 3-(4-amino-3-methylpyrazol-1-yl)-3-(fluoromethyl)tetrahydrofuran-2-one (0.2 g, 938.05 µmol), 2chloro-N-ethyl-5-(trifluoromethyl) pyrimidin-4-amine (190 mg, 844.24 µmol) and p-TsOHH ₂ O (71 mg, 375.22 µmol) in 1,4-dioxane (3 ml) were stirred at 90°C for 6 hours under N2 . The reaction mixture was quenched by adding saturated aqueous NaHCO ₃ (60 ml) at 0°C and then extracted with EtOAc (3x20 ml). The combined organic layers were washed with aqueous saturated sodium chloride solution (20 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (TE:EtOAc = 3:1 to 1:1) to give the title compound as a brown oil, which was separated by SFC. SPC, first eluting isomer: 1H NMR (400 MHz, CDCl·): δ 8.30 (br.s, 1H), 8.12 (s, 1H), 7.01-6.61 (m, 1H) , 5.32-5.06 (m, 1H), 4.91-4.68, 2H), 4.54-4.37 (m, 2H), 3.643.53 (m, 2H), 3.32 (br.s, 1H), 2.92-2.79 (m, 1H), 2.26 (s, 3H), 1.33 (br.t, J=7.0 Hz, 3H). HPLC: retention time: 2.02 min. MS: (M+H+) m/z = 403.3.

SFC, second eluting isomer: 1H NMR (400 MHz, CDC13): δ 8.30 (br s, 1H), 8.12 (s, 1H), 7.01-6.61 (m, 1H), 5. 32-5.06 (m, 1H), 4.91-4.68, 2H), 4.54-4.37 (m, 2H), 3.64-3.53 (m, 2H), 3, 32 (br.s, 1H), 2.92-2.79 (m, 1H), 2.26 (s, 3H), 1.33 (br.t, J=7.0 Hz, 3H). HPLC: retention time: 1.99 min. mS: (M+H) ⁺ m/z = 403.3.

- 32 043797

Example 14. Synthesis of N2-[5-chloro-1-[(3S)-1-ethyl-4,4-difluoro-3-piperidyl]pyrazol-4-yl]-N4-ethyl-5(trifluoromethyl)pyrimidin-2 ,4-diamine and N2-[5-chloro-1-[(3R)-1-ethyl-4,4-difluoro-3-piperidyl]pyrazol-4yl]-N4-ethyl-5-(trifluoromethyl)pyrimidine-2 ,4-diamine (204 and 205).

tert-Butyl-3-(4-nitropyrazol-1-yl)-4-oxopiperidine-1-carboxylate.

To a solution of tert-butyl-3-bromo-4-oxopiperidine-1-carboxylate (20 g, 71.91 mmol) and 4-nitro-Spirazole (8.94 g, 79.10 mmol) in DMF (100 ml) was added K ₂ CO ₃ (19.88 g, 143.81 mmol) at 20°C in an N ₂ atmosphere. The mixture was stirred at 20°C for 16 hours. The mixture was poured into ice water (300 ml) and extracted with EtOAc (3x100 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (3x100 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 3:1) to give tert-butyl 3-(4-nitropyrazol-1-yl)-4-oxopiperidine-1-carboxylate as a yellow oil. LCMS: RT 1.306 min, m/z = 255.2 [M-56] ⁺ . 1H NMR (400 MHz, CDCl ₃ ): δ 8.22-8.27 (m, 1H), 8.12 (s, 1H), 4.97 (dd, J=10.85, 6.34 Hz, 1H), 4.75 (br.s, 1H), 4.43 (br.s, 1H), 3.64 (br.t, J=11.86 Hz, 1H), 3.30 (br.d , J=5.77 Hz, 1H), 1.41-1.58 (m, 9H), 1.41-1.58 (m, 2H).

tert-Butyl-4,4-difluoro-3-(4-nitropyrazol-1-yl)piperidine-1-carboxylate.

To a solution of tert-butyl-3-(4-nitropyrazol-1-yl)-4-oxopiperidin-1-carboxylate (1 g, 3.22 mmol) in DCM (10 mL) was added DAST (2.6 g, 16. 11 mmol, 2.13 ml) at -78°C in an N2 atmosphere. The mixture was stirred at 20°C for 16 hours. The mixture was poured into saturated aqueous NaHCO ₃ (15 ml) and extracted with EtOAc (3x5 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (5 ml), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 3:1) to give tert-butyl 4,4-difluoro-3-(4-nitropyrazol-1-yl)piperidine-1-carboxylate as a white solid. LCMS: RT 1.335 min, m/z = 277.1 [M-56] ⁺ . 1H NMR (400 MHz, CDCl ₃ ): δ 8.30 (s, 1H), 8.13 (s, 1H), 4.52 (ddq, J=14.23, 9.60, 4.65, 4 .65, 4.65 Hz, 1H), 4.39 (br.s, 1H), 4.10 (br.s, 1H), 3.66 (br.t, J=11.36 Hz, 1H) , 3.30 (br.t, J=11.42 Hz, 1H), 2.26-2.42 (m, 1H), 1.95-2.18 (m, 1H), 1.37-1 .57 (m, 9H).

tert-Butyl-3-(5-chloro-4-nitropyrazol-1-yl)-4,4-difluoropiperidine-1-carboxylate.

LiHMDS (1M, 3.34 mmol, 3.34 ml) at -78°C in an N2 atmosphere. The reaction mixture was stirred at -78°C for 1 hour. 1,1,1,2,2,2-hexachloroethane (1.05 g, 4.45 mmol, 504.49 µl) in THF (5 ml) was then added dropwise and the mixture was stirred at -78°C for 1 hours. The mixture was poured into saturated aqueous NH4Cl (15 ml) and extracted with EtOAc (3x5 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (5 ml), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:0 to 3:1) to give tert-butyl-3-(5-chloro-4-nitropyrazol-1-yl)-4,4-difluoropiperidin- 1-carboxylate as a yellow oil. LCMS: RT 1.352 min, m/z = 311.2 [M-56] ⁺ . 1H NMR (400 MHz, CDCl ₃ ): δ 8.22 (s, 1H), 4.59-4.72 (m, 1H), 4.00-4.16 (m, 2H), 3.81- 3.90 (m, 1H) 3.55 (br.d, J=9.03 Hz, 1H), 2.38-2.54 (m, 1H), 1.96-2.15 (m, 1H ), 1.39-1.56 (m, 9H).

3-(5-Chloro-4-nitropyrazol-1-yl)-4,4-difluoro-piperidine.

A mixture of tert-butyl 3-(5-chloro-4-nitropyrazol-1-yl)-4,4-difluoropiperidine-1-carboxylate (1.8 g, 4.91 mmol) in HCl/EtOAc (40 ml) was stirred at 20°C for 2 hours. The reaction mixture was concentrated under reduced pressure and the pH of the mixture was adjusted to pH 7-8 using saturated aqueous NaHCO ₃ solution. The aqueous phase was then extracted with EtOAc (3x15 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give 3-(5-chloro-4-nitropyrazol-1yl)-4,4-difluoropiperidine as light yellow solid matter. 1H NMR (400 MHz, CDCl ₃ ): δ 8.178.32 (m, 1H), 4.57-4.81 (m, 1H), 3.59 (br.dd, J=13.68, 4.89 Hz, 1H), 3.36 (br.dd, J=13.93, 4.02 Hz, 1H), 3.143.27 (m, 1H), 2.98-3.11 (m, 1H), 2 .37 (br.s, 1H), 2.14-2.34 (m, 1H).

3-(5-Chloro-4-nitropyrazol-1-yl)-1-ethyl-4,4-difluoropiperidine.

To a mixture of 3-(5-chloro-4-nitropyrazol-1-yl)-4,4-difluoropiperidine (0.5 g) and acetaldehyde (2.07 g, 18.75 mmol, 2.63 ml) in MeOH ( 10 ml) NaBH ₃ CN (589 mg, 9.38 mmol) was added and stirred for 15 minutes. CH3COOH (1.13 g, 18.75 mmol, 1.07 mL) was then added to the solution at 20°C and the mixture was stirred at 20°C for 1 hour. The pH of the mixture was adjusted to pH 7-8 with saturated aqueous NaHCO ₃ solution and the aqueous phase was extracted with EtOAc (3x15 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (10 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 0:1) to give 3-(5-chloro-4-nitropyrazol-1-yl)-1ethyl-4,4-difluoropiperidine as a yellow oil . LCMS: RT 0.939 min, m/z = 295.1 [M+H] ⁺ . 1H NMR (400 MHz, CDCl ₃ ) δ: 8.19-8.33 (m, 1H), 4.78-4.95 (m, 1H), 3.10-3.22 (m, 2H), 2.97-3.06 (m, 1H), 2.57-2.67 (m, 2H) 2.39-2.51 (m, 1H), 2.22-2.36 (m, 1H) , 2.12-2.21 (m, 1H), 1.13 (t, J=7.22 Hz, 3H).

- 33 043797

5-Chloro-1 -(1-ethyl-4,4-difluoro-3-piperidyl)pyrazol-4-amine.

To a mixture of 3-(5-chloro-4-nitropyrazol-1-yl)-1-ethyl-4,4-difluoropiperidine (0.15 g, 509.02 µmol) in EtOH (4 ml) and H ₂ O (1 ml) Fe (142 mg, 2.55 mmol) and NH ₄ Cl (136 mg, 2.55 mmol, 88.98 μl) were added at 20°C. The mixture was then stirred at 80°C for 1 hour. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was washed with DCM:MeOH (v:v = 10:1) (30 ml), filtered and the filtrate was concentrated under reduced pressure to give 5chloro-1-(1-ethyl-4,4-difluoro-3-piperidyl) pyrazol-4-amine as a red solid. LCMS: RT 1.150 min, m/z = 265.1 [M+H]+.

N2-[5-Chloro-1-[(3S)-1-ethyl-4,4-difluoro-3-piperidyl]pyrazol-4-yl]-N4-ethyl-5-(trifluoromethyl)pyrimidine-2,4diamine and N2-[5-chloro-1-[(3R)-1-ethyl-4,4-difluoro-3-piperidyl]pyrazol-4-yl]-N4-ethyl-5-(trifluoromethyl)pyrimidine2,4-diamine.

To a mixture of 5-chloro-1-(1-ethyl-4,4-difluoro-3-piperidyl)pyrazol-4-amine (0.13 g, 491.12 μmol) and 2-chloro-N-ethyl5-(trifluoromethyl )pyrimidin-4-amine (110 mg, 491.12 µmol) in 1,4-dioxane (3 ml), p-TsOHH ₂ O (25 mg, 147.33 µmol) was added at 20°C and the mixture was stirred at 90° Glow 5 hours. The pH of the mixture was adjusted to pH 7-8 with saturated aqueous NaHCO ₃ solution and the aqueous phase was extracted with EtOAc (3x5 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (8 ml), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO ₂ , EtOAc) to give the desired compound as a white syrup, which was further separated by SFC to give N2-[5-chloro-1-[(3S)-1-ethyl-4. 4-difluoro-3-piperidyl]pyrazol-4-yl]-N4ethyl-5-(trifluoromethyl)pyrimidine-2,4-diamine in the form of a white syrupy substance and N2-[5-chloro-1-[(3R)-1ethyl -4,4-difluoro-3-piperidyl]pyrazol-4-yl]-M4-ethyl-5-(trifluoromethyl)pyrimidine-2,4-diamine.

SFC, first eluting isomer: 1H NMR (400 MHz, CdC1 ₃ ): 8.23 (br.s, 1H), 8.13 (s, 1H), 6.72 (br.s, 1H), 5.14 (br.s, 1H), 4.64-4.79 (m, 1H), 3.48-3.64 (m, 2H), 3.14 (br.d, J=8.41 Hz, 2H ), 2.99 (br.d, J=10.67 Hz, 1H), 2.60 (kv, J=7.15 Hz, 2H), 2.35-2.50 (m, 1H), 2 .04-2.34 (m, 2H), 1.27 (t, J=7.22 Hz, 3H), 1.13 (t, J=7.15 Hz, 3H). HPLC:VU: 1.116 min. MS: m/z = 454.4 [M+H] ⁺ . SFC: retention time: 1.621 min.

SPC, second eluting isomer: 1H NMR (400 MHz, CDC13): 8.23 (br s, 1H), 8.14 (s, 1H), 6.71 (br s, 1H), 5.13 (br .s, 1H), 4.60-4.81 (m, 1H), 3.49-3.61 (m, 2H), 3.15 (br.d, J=8.28 Hz, 2H), 2.99 (br.d., J=11.80 Hz, 1H), 2.60 (kv, J=7.15 Hz, 2H), 2.43 (b.t., J=12.05 Hz, 1H), 2.06-2.33 (m, 2H), 1.27 (t, J=7.22 Hz, 3H), 1.13 (t, J=7.15 Hz, 3H). HPLC: retention time: 1.108 min. MS: m/z = 454.4 [M+H] ⁺ . SFC: retention time: 1.785 min.

Example 15. Synthesis of (1S,2R)-2-[4-[(5-bromo-4-methoxypyrimidin-2-yl)amino]-3-cyclopropylpyrazol1-yl]cyclopropanecarbonitrile and (1R,2S)-2-[4 -[(5-bromo-4-methoxypyrimidin-2-yl)amino]-3cyclopropylpyrazol-1-yl]cyclopropanecarbonitrile (213 and 214).

-Cyclopropyl-4-nitro-1-vinylpyrazole.

NaOH (9 .14 g, 228.55 mmol) and water (9 ml) at 20°C in an N2 atmosphere. The mixture was stirred at 80°C for 8 hours. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 1:1) to give 3-cyclopropyl-4-nitro-1-vinylpyrazole as a yellow solid. 1H NMR (400 MHz, CDC13): δ ppm 8.23 (s, 1H), 6.87 (dd, J=15.55, 8.71 Hz, 1H), 5.70 (d, J=15.66 Hz, 1H) 5.06 (d, J=8.60 Hz, 1H), 2.53-2.68 (m, 1H), 0.97-1.11 (m, 4H).

Ethyl (1S,2R)-2-(3-cyclopropyl-4-nitropyrazol-1-yl)cyclopropanαnαrboxylate and ethyl (1S,2S)-2-(3cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarboxylate.

To a mixture of 3-cyclopropyl-4-nitro-1-vinylpyrazole (4.7 g, 26.23 mmol) and bis-3-[3-(2-carboxy-2methylpropyl)phenyl]-2,2-dimethylpropanoic acid dirhodium (200 mg, 262.31 µmol) in DCM (100 ml) was added dropwise ethyl 2-diazoacetate (17.96 g, 157.39 mmol) in DCM (30 ml) at 20°C under N2 for 3 hours. The mixture was stirred at 20°C for 12 hours. The mixture was concentrated. The residue was purified by silica gel column chromatography (PE:EEAc = 100:1 to 1:1) to give ethyl (1S*,2R*)-2-(3-cyclopropyl-4nitropyrazol-1-yl)cyclopropanecarboxylate and ethyl (1S *,2S*)-2-(3-cyclopropyl-4-nitropyrazol-1yl)cyclopropanecarboxylate as a brown oil. (1S*,2R*)-2-(3-cyclopropyl-4-nitropyrazol-1yl)cyclopropanecarboxylate: 1H NMR (400 MHz, CDC13): δ 8.15 (s, 1H), 4.12-4.37 ( m,1H), 3.97-4.07 (m, 2H), 3.90 (td, J=7.50, 5.71 Hz, 1H), 2.43-2.71 (m, 1H) , 2.13-2.37 (m, 1H) 1.88-2.07 (m, 1H), 1.59 (td, J=8.06, 6.46 Hz, 1H), 1.23- 1.36 (m, 1H), 1.17 (t, J=7.15 Hz, 3H), 0.84-1.06 (m, 4H).

(1S*,2S*)-2-(3-Cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarboxylate: 1H NMR (400 MHz, CDC13): δ 8.18 (s, 1H), 4.08-4, 32 (m, 3H), 3.98 (ddd, J=7.97, 4.89, 3.07 Hz, 1H), 2.50-2.65 (m, 1H), 2.30 (ddd, J=9.54, 6.27, 3.01 Hz, 1H), 1.79 (dt, J=9.91, 5.21 Hz, 1H), 1.65 (dt, J=8.03, 5.96 Hz, 1H), 1.24-1.36 (m, 4H), 0.92-1.10 (m, 4H).

(1 S,2R)-2-(3-Cyclopropyl-4-nitropyrazol-1 -yl)cyclopropanecarboxylic acid.

HCl (2 M, 20 ml) at 20°C in an N2 atmosphere. The mixture was stirred at 60°C for 12 hours. The mixture was concentrated to give (1S,2R)-2-(3-cyclopropyl-4

- 34 043797 nitropyrazol-1-yl)cyclopropanecarboxylic acid in the form of a brown solid. 1H NMR (400 MHz, DMSO): δ 8.84 (s, 1H), 4.01-4.10 (m, 1H), 2.39-2.46 (m, 1H), 2.02-2 .10 (m, 1H), 1.98 (sq., J=6.03 Hz, 1H), 1.46-1.55 (m, 1H), 0.93-1.07 (m, 2H) , 0.76-0.89 (m, 2H).

(1 S,2R)-2-(3-Cyclopropyl-4-nitropyrazol-1 -yl)cyclopropanecarboxamide.

To a mixture of (1S,2R)-2-(3-cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarboxylic acid (2 g, 8.43 mmol), NH ₄ Cl (2.71 g, 50.59 mmol) and DIPEA (6.54 g, 50.59 mmol) in DMF (20 ml) was added to HATU (6.41 g, 16.86 mmol) at 20°C under N2 atmosphere. The mixture was stirred at 20°C for 4 hours. The mixture was poured into ice water (100 ml). The aqueous phase was extracted with EtOAc (3x50 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (3x50 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give (1S,2R)-2-(3cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarboxamide in as a brown solid. 1H NMR (400 MHz, DMSO): δ 8.67 (s, 1H), 7.65 (br.s, 1H), 6.87 (br.s, 1H), 3.81-3.98 (m , 1H), 2.38-2.47 (m, 1H), 2.04 (kv, J=7.57 Hz, 1H), 1.93 (kv, J=5.73 Hz, 1H), 1 .37 (td, J=8.05, 5.95 Hz, 1H), 1.21-1.29 (m, 1H), 0.941.01 (m, 2H), 0.78-0.84 (m , 1H).

(1S, 2R)-2-(3-Cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarbonitrile.

T3P (18.32 g, 28 .79 mmol, 17.12 ml, 50% purity) at 20°C in an N2 atmosphere. The mixture was stirred at 75°C for 12 hours. The mixture was poured into aqueous NaHCO3 solution (200 ml). The aqueous phase was extracted with EtOAc (3x50 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (150 ml), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 1:1) to give (1S,2R)-2-(3-cyclopropyl-4nitropyrazol-1-yl)cyclopropanecarbonitrile as a white solid. LCMS: RT 1.20 min, m/z = 219.2 [M+H]+. 1H NMR (400 MHz, CDCI3): δ 8.26 (s, 1H), 3.90-4.09 (m, 1H), 2.62 (tt, J=8.05, 5.29 Hz, 1H ), 2.10-2.20 (m, 1H), 2.01 (dt, J=9.43, 6.64 Hz, 1H), 1.75 (dt, J=9.26, 7.39 Hz, 1H), 1.00-1.11 (m, 4H).

(1S,2R)-2-(4-Amino-3-cyclopropylpyrazol-1-yl)cyclopropanecarbonitrile.

To a mixture of (1S,2R)-2-(3-cyclopropyl-4-nitropyrazol-1-yl)cyclopropanecarbonitrile (0.8 g, 3.67 mmol) and Fe (1.02 g, 18.33 mmol) in EtOH (20 ml) and water (5 ml), NH4C1 (981 mg, 18.33 mmol) was added at 20°C under N2 atmosphere. The mixture was stirred at 75°C for 1 hour. The mixture was filtered and the filtrate was concentrated. The residue was washed with DCM:MeOH (10:1.3x10 ml), filtered and the filtrate was concentrated under reduced pressure to give (1S,2R)-2-(4-amino-3-cyclopropylpyrazol-1yl)cyclopropanecarbonitrile (0.75 g crude) as brown butter. LCMS: RT 0.81 min, m/z = 189.3 [M+H] ⁺ . 1H NMR (400 MHz, CDC13): δ 6.97-7.15 (m, 1H), 3.74-3.91 (m, 1H), 2.03 (q, J=6.25 Hz, 1H ), 1.80 (dt, J=9.43, 6.42 Hz, 1H), 1.64-1.74 (m, 1H), 1.54-1.63 (m, 1H), 0, 78-0.93 (m, 4H).

(1S,2R)-2-[4-[(5-Bromo-4-methoxypyrimidin-2-yl)αmino]-3-cyclopropylpyrazol-1-yl]cyclopropαnkarbonitrile and (1R,2S)-2-[4-[ (5-bromo-4-methoxypyrimidin-2-yl)amino]-3-cyclopropylpyrazol-1-yl]cyclopropanecarbonitrile.

To a mixture of (1S,2R)-2-(4-amino-3-cyclopropylpyrazol-1-yl)cyclopropanecarbonitrile (0.1 g, 531.27 µmol) and 5-bromo-2-chloro-4-methoxypyrimidine (119 mg , 531.27 µmol) in 1,4-dioxane (2 ml) was added p-TsOH.H ₂ O (30 mg, 159.38 µmol) at 20°C under N2 atmosphere. The mixture was stirred at 85°C for 4 hours. The mixture was poured into aqueous NaHCO3 (5 ml) and extracted with EtOAc (3x5 ml). The combined organic phase was washed with aqueous saturated sodium chloride solution (10 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 1:1) and separated by SFC to give (1S,2R)-2-[4-[(5-bromo-4-methoxypyrimidine-2 -yl)amino]-3cyclopropylpyrazol-1-yl]cyclopropanecarbonitrile and (1R,2S)-2-[4-[(5-bromo-4-methoxypyrimidin-2-yl)amino]3-cyclopropylpyrazol-1-yl]cyclopropanecarbonitrile . SPC, first eluting isomer: 1H NMR (400 MHz, CDC13): δ 8.23 (s, 1H), 8.01 (s, 1H), 6.76 (br.s, 1H), 4.05 (s , 3H), 3.85-3.97 (m, 1H), 2.11 (kv, J=6.27 Hz, 1H), 1.88 (dt, J=9.29, 6.46 Hz , 1H), 1.60-1.78 (m, 2H), 0.84-0.97 (m, 4H). LCMS: retention time: 1.475 min. MS: [M + H] ⁺ m/z: 375.2.

SPC, first eluting isomer: 1H NMR (400 MHz, CDC13): δ 8.22 (s, 1H), 8.01 (s, 1H), 6.77 (br.s, 1H), 4.05 (s , 3H), 3.86-3.96 (m, 1H), 2.11 (kv, J=6.27 Hz, 1H), 1.88 (dt, J=9.29, 6.53 Hz , 1H), 1.61-1.77 (m, 2H), 0.85-0.97 (m, 4H), LCMS: retention time: 1.465 min. MS: [M+H] ⁺ m/z: 375.2.

Other compounds from the table. 1 were or can be prepared in accordance with the examples above and/or the general procedures described herein using appropriate starting materials.

Example 16. Biochemical analysis of compounds.

Materials:

enzyme LRRK2 G2019S, substrate (LRRKtide),

ATP,

- 35 043797 TR-FRET dilution buffer, pLRRKtide antibody,

384 well plate,

DMSO.

Enzyme reaction conditions:

mMTris, pH 7.5, 10 mM _MgCl3 , 1 mM EGTA, 0.01% Brij-35, 2 mM DTT, nM LRRK2,

134 µM ATP, reaction time 60 minutes, reaction temperature 23°C, total reaction volume 10 µl, detection reaction conditions, 1 x TR-FRET dilution buffer, 10 mM EDTA, nM antibody, reaction temperature 23°C, total reaction volume 10 µl

Compounds were prepared by first diluting to 1 mM solution in DMSO. 35 μl of reference compound solution, 35 μl of test compound solution and 35 μl of HPE were sequentially added to the original plate (384-well plate, Labcyte). The plates were centrifuged at 2500 rpm for 1 minute and covered with foil. The POD was used to perform a 3.162-fold serial dilution, and 100 nL of reference compound solution, test compound solution, HPE, and ZPE were transferred to assay plates. The assay plate was centrifuged at 2500 rpm for 1 minute and covered with foil. To carry out the enzymatic reaction, 5 μl of LRRKtide substrate and kinase mixture in assay buffer was added to each well of the plate. The plate was centrifuged to concentrate the mixture at the bottom of the wells. The assay plate was incubated at 23°C for 20 minutes. After incubation, 5 μl of 2xATP in assay buffer was added to each well, and the plates were centrifuged to concentrate the mixture at the bottom of the wells. The plate was incubated at 23°C for 60 minutes.

To perform the detection reaction, EDTA, fully mixed in TR-FRET dilution buffer, was added to the antibody reagent. 10 μl of detection reagent was added to all wells of each well of the assay plate and the plate was centrifuged to concentrate the mixture at the bottom of the wells. The plate was then incubated at 23°C for 60 minutes. The plates were read on a Perkin Elmer Envision 2104 in TR-FRET mode using a 340 nm excitation filter, a 520 nm fluorescence filter, and a 490 or 495 terbium emission filter. Some of the compounds described herein were tested in accordance with the above methods and found to have the ICR LRRK2 G2019S IC ₅₀ as listed in Table 1. 2. In the table below, the activity is shown as follows: In the table below, the activity is shown as follows: +++ = IC ₅₀ less than 30 nM; ++ = IC ₅₀ between 30 nM and 60 nM; + = IC ₅₀ more than 60 nM.

table 2

Number LRRK2 TR-FRET IC50 (nM) ms [M+1] ⁺ 8 +++ 325.1 9 +++ 409 10 + 409.1 AND +++ 389.1 12 +++ 389.1 13 +++ 389.1 14 +++ 396.2

- 36 043797

15 +++ 395.2 16 +++ 341.2 17 +++ 341.1 18 +++ 409.2 19 +++ 350.2 20 +++ 355.2 21 +++ 378.1 22 +++ 404.2 23 +++ 404.3 24 +++ 404.2 25 +++ 387.3 26 +++ 366.2 27 +++ 366.1 28 +++ 468.2 29 +++ 468.2 thirty ++ 371.2 31 ++ 371.2 32 +++ 352.1 33 +++ 410.1 34 +++ 399.2 35A +++ 366.2 35V +++ 366.2 37 +++ 349.0 38 +++ 366.1 39A +++ 384.2 40 +++ 352.2 41 +++ 352.1 42 +++ 350.1 43 +++ 364.1 44 +++ 363.3 45 +++ 349.2

46 +++ 399.2 48 +++ 375.2 50 +++ 331.1 52A +++ 338.1 52V +++ 338.2 54A +++ 380.2 54V +++ 380.1 57 +++ 396.2 58 +++ 362.1 59 +++ 390.2 60 +++ 333.1 61 +++ 380.2 62 +++ 343.2 63 +++ 346.1 64A +++ 381.1 64V +++ 381.1 65 +++ 381.1 66 +++ 376.1 67 +++ 343.2 68 +++ 411.3 69 +++ 408.2 70 +++ 398.2 71 +++ 410.2 72 +++ 332.1 73 +++ 400.1 74 +++ 368.3 75 +++ 409.1 76 +++ 404.1 77 +++ 408.2 78 +++ 422.3 79 +++ 421.1

- 37 043797

80 +++ 346.2 81 +++ 423.0 82 +++ 399.2 83 +++ 399.2 84 +++ 380.2 85 +++ 413.1 86 +++ 424.3 87 +++ 354.2 88 +++ 415.2 89 +++ 359.2 90 +++ 382.1, 384.0 91 +++ 374.3 92 +++ 434.4 93 +++ 390.1, 392.1 94 +++ 394.2 95 +++ 390.1, 392.1 96 +++ 429.1 97 +++ 434.2 98 +++ 385.1 99 +++ 391.3 100 +++ 385.2 101 +++ 434.2 102 +++ 382.1, 384 103 +++ 382.2 104 +++ 411.2 105 +++ 391.2 106 +++ 489.3 107 +++ 378.3

108 +++ 411.2 109 +++ 273.2 BY +++ 417.25 111 +++ 401.1 112 +++ 391.1 FROM +++ 397.2 114 +++ 423.1 115 +++ 423.1 116 +++ 434.4 117 +++ 405.3 118 +++ 380.2 119 +++ 371.2 120 +++ 382.2 121 +++ 489.3 122 +++ 397.2 123 +++ 391.2 124 +++ 371.2 125 +++ 385.2 126 +++ 433.8 127 +++ 436.3 128 +++ 419.2 129 +++ 410.2 130 +++ 384.2 131 +++ 380.2 132 +++ 391.1 133 + 338.1 134 +++ 371.2 135 + 355.2 136 ++ 410.2 137 +++ 408.2 138 +++ 408.2

- 38 043797

139 +++ 433.1 140 +++ 443.1, 445.2 141 +++ 392.2 142 +++ 394.2 143 +++ 411.2 144 +++ 411.2 145 +++ 383.3 146 +++ 418.2,420.2 147 +++ 418.2, 420.2 148 +++ 393.1 149 ++ 410.2 150 +++ 421.1, 423.1 151 +++ 421.1, 423.1 152 +++ 378.2 153 +++ 396.2 154 +++ 432.2 155 +++ 397.2 156 +++ 397.2 157 ++ 419.2, 421.2 158 +++ 408.1 159 +++ 442.1, 444.1 160 ++ 408.2, 410.1 161 +++ 408.1, 410.1 192 +++ 399.3 193 ++ 374.3 194 +++ 383.2 195 +++ 369.2 196 +++ 434.4 197 +++ 434.4 198 +++ 420.4 199 ++ 420.4 200 +++ 406.4 201 +++ 406.4 202 +++ 440.4 203 +++ 440.4 204 +++ 454.4 205 +++ 454.4

162 +++ 409 163 +++ 395.1 164 +++ 378.3 165 +++ 409.3 166 + 433.2 167 +++ 433.2 168 +++ 395.2 169 +++ 425.3 170 +++ 396.3 171 + 434.3 172 +++ 434.3 173 +++ 388.2 174 +++ 388.3 175 +++ 374.2 176 +++ 374.3 177 + 419.3 178 +++ 419.3 179 +++ 343.3 180 ++ 343.2 181 +++ 352.1 182 +++ 394.3 183 +++ 394.2 184 +++ 386.2 185 +++ 386.2 186 +++ 374.3 187 +++ 390.1, 392.1 188 ++ 394.2 189 ++ 394.2 190 +++ 345.1 191 +++ 385.3 206 +++ 372.3 207 +++ 372.3 208 +++ 364.3 209 +++ 364.3 210 +++ 414.3 211 +++ 414.3 212 +++ 376.2 213 +++ 375.2 214 +++ 388.3 215 +++ 388.3 216 +++ 403.3 217 +++ 403.3 218 +++ 396.2

Example 17 Metabolic stability.

Metabolic stability of the compounds was assessed using human liver microsomes (from Corning or XenoTech, LLC) using a 96-well plate. Compounds were incubated at 37°C at 1 μM final concentration in the microsomal matrix (0.5 mg/ml total protein) in the presence or absence of the cofactor NADPH. The analysis used a NADPH regeneration system consisting of NADP, MgCl2, isocitric acid and isocitrate dehydrogenase. Enzymatic reactions were carried out for 0, 5, 10, 20, 30 or 60 minutes until the reactions were terminated by the addition of acetonitrile containing tolbutamide and labetalol internal standards (100 ng/ml). After shaking for 10 min, the plates were centrifuged (4000 rpm at 4°C) for 20 min and the supernatants were mixed 1:3 with HPLC grade water. Samples were analyzed by LCMS/MS using the appropriate MRM transitions for each analyte and internal standard (IS). Analyte/BC peak area ratios were used to determine the percentage of compound remaining at each time point.

Internal clearance (Clint, expressed in ml*min ^-1 -mg ^-1 ) was calculated from the first order elimination constant (k, min ^-1 ) of the breakdown of the test product and the incubation volume. These values were scaled to internal organ clearance (Clint) using specific scales.

- 39 043797 human factors (48.8 mg of microsomal protein per 1 g of liver, 25.7 g of liver per kg of body weight). Organ Cl _int was then converted to liver clearance (CLhep, ml-min ^-1 -kg ^-1 ) using a thoroughly mixed hepatic elimination model, where Qh represents human hepatic blood flow (20.7 ml-min ^-1 -kg ^-1 ) .

_ Qh* CLi _nt ^LLhep ~ (Qh + CL _int )

CLhep is the predicted human hepatic clearance based on the above in vitro assay. A lower value indicates that less of the compound is removed by the liver. Surprisingly, it has been found that compounds containing a C5-pyrazole attached to an aminopyrimidine core provide lower clearance (ie, improved stability) compared to compounds containing a C4-pyrazole attached to an aminopyrimidine core, without a significant change in activity.

Table 3

No. Connections Structure LRRK2 TR-FRET ICso (nM) Human liver microsomes CLhep (ml/min/kg) 122 7f A L HN N NH /\ 1 . N-N ,0 V (First eluting isomer) 0.72 9.856 155 Lf νΑΆ A L HN N NH M o ¹ A (First eluting isomer) 1.95 5,519 114 AG HN N NH AS NN O A ⁰ (First eluting isomer) 1.05 16,508 150 dT HN N NH O A (First eluting isomer) 1.22 11,077 104 % A L HN N NH N-N O Ύ (First eluting isomer) 0.96 11.59

- 40 043797

143 a N^fT A A HN N NH O % (First eluting isomer) 1.21 7.81 93 dT HN N NH N-N 1.87 17,663 141 HN N NH nA V—=Ν 3.94 13,508 118 A N^A^F A Λ HN Ν NH N-N 1.55 7.81 131 A N^fT A A HN N NH nA\—SN 4.98 2.237

Example 18: MDR1-MDCK permeability.

The blood-brain barrier (BBB) separates circulating blood from the extracellular fluid of the central nervous system (CNS). Passive membrane permeability (Papp) and efflux potential of the MDR1 substrate (P-glycoprotein) were determined using the MDR1-MDCK cell line as a model for efficient permeabilization of the junction through the BBB. The bidirectional assay was performed in pretreated MDR1-MDCK cells using a 12- or 96-well plate in the absence or presence of an MDR1 inhibitor (GF120918 or VBACpodar). Assays were performed in duplicate in transport buffer (HBSS, pH 7.4) for 90 or 120 minutes at 37°C using a test compound concentration of 1 μM. Monolayer integrity was confirmed using Luciferin yellow and appropriate positive controls for passive permeation and MDR1 were included in each experiment. After incubation, samples from the donor and receiving compartments were removed and quenched with acetonitrile containing the appropriate internal standard (IS). Protein was pelleted by centrifugation for 10 min at 3220 g and supernatants were diluted in ultrapure water (if necessary) before analysis by LCMS/MS using appropriate MRM transitions for analytes and VS. Papp (apparent permeability expressed in cm/s [centimeter/second]) was calculated using the following equation:

_R . / dCo V _R V _R C _R (cm/sec) - —— x —--------—— or —------------x — ^pp dt (Area x C _A ) Area x Time _Where V _R is the volume of solution in the receiving chamber (apical or basolateral side), area is the surface area of the insert membrane), time is the incubation time expressed in seconds, C _R is the peak area ratio (analyte/BC) in the receiver compartment, CA is the average of the initial and final concentrations in the donor chamber, and Co is the initial peak area ratio in the donor chamber. P _arr was determined in both the apical-basolateral (A^B) and basolateral-apical (B^A) directions.

Monolayer efflux ratios (ER) were obtained using the following equation:

_ER _ \P _arr (B^A^1 [P _arr (L - V)]

Compounds with an MDR1-MDCK efflux ratio of less than or equal to five are likely to demonstrate the ability to cross the blood-brain barrier. Compounds having 1,2,3-triazole

- 41 043797 substituent were surprisingly brain permeable compared to molecules having a 1,2,4 triazole moiety.

Table 4

No. Connected Structure LRRK2 TR- FRET IC50(hM) Human liver microsomes CLhep (ml/min/kg) MDR1 ER 172 A n^A^t D Λ ΗΝ Ν ΝΗ A (Second eluting isomer) 1.47 19.9 42 34 ρ AND ^F _n HN^N NH ^D AO 2.31 0.75 4.8 68 μ aT ^f _n HN^N NH φιΑ λ oo 2.23 3.18 4.01 78 p aT ^f HN^ N NH 3.69 4.81 2 27 A n^A^t HN^N NH N-N / N 3.41 7.81 0.88 118 A ^F aT ^f HN^N NH ⁿ -n^n 1.55 7.81 0.83 218 y _F Λ Λ ΗΝ N NH 1 1 AiO NN ΑΛ 5.91 0 86.02

Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which the present invention relates. The inventions illustratively described herein may be suitably practiced in the absence of any of the elements or elements, limitations or restrictions not specifically disclosed herein. So, for example, the terms including, comprising, containing, etc. must be perceived expansively and without restrictions. Furthermore, the terms and expressions used herein are used as terms of description and not limitation, and there is no further intention to use such terms and expressions for

-

Claims (35)

excluding any equivalents of the features shown and described or parts thereof, but various modifications are possible within the scope of the claimed invention. Thus, it is to be understood that while the present invention has been specifically disclosed in preferred embodiments and optional aspects, modifications, improvements and variations of the inventions embodied herein may be employed by those skilled in the art and that such modifications, improvements and variations are contemplated within the scope of the present invention. The materials, methods and examples presented herein represent preferred embodiments and are illustrative and not intended to limit the scope of the invention. The invention is described herein broadly and generally. Each of the narrower types and subgeneric groups included in the general description is also part of the invention. This includes a general description of the invention with a qualification or negative limitation that removes any subject matter from the class, whether or not the omitted material is specifically stated herein. Moreover, when features or aspects of the invention are described in terms of Markush groups, those skilled in the art will understand that the invention is also so described in terms of any individual member or subgroup of members of the Markush group. All publications, patent applications, patents and other references mentioned herein are expressly incorporated by reference in their entirety to the same extent as if each had been individually incorporated herein by reference. In the event of a conflict, this specification, including definitions, shall prevail. It should be understood that while the disclosure has been described in connection with the above embodiments, the above description and examples are intended to be illustrative only and do not limit the scope of the present disclosure. Other aspects, advantages and modifications will be apparent to those skilled in the art within the scope of the disclosure of the present invention. CLAIM 1. Compound of formula Ia or a pharmaceutically acceptable salt, deuterated analogue, stereoisomer or mixture of stereoisomers thereof, wherein R ¹ is C ₃ . ₆ cycloalkyl, optionally substituted with one or more halogen, hydroxyl or cyano atoms, or C ₁ . ₆ αlkyl, optionally substituted with halogen; R ² represents halogen, cyano or C ₁ . ₆ alkyl, optionally substituted with halogen; R ³ represents C ₁ . ₆ alkoxy, C ₃ . ₆ cycloalkyl, C _{3 .6} cycloalkoxy or -N(R11)(R ¹² ); R ⁴ represents hydrogen or halogen; R ⁶ and R ⁷ are each independently hydrogen or C ₁ . ₆ αlkyl, optionally substituted with halogen; And R ⁸ and R ⁹ are each independently hydrogen, cyano, halogen, C ₁ . ₆ alkyl or Cl. ₆ alkox; And R ^{1 1} and R ¹² are each independently hydrogen, C ₁ . ₆ alkyl or C ₃ . ₆ cycloalkyl. 2. The compound according to claim 1, where R ⁶ and R ⁷ represent methyl. 3. A compound according to claim 1, wherein at least one of R ⁸ and R ⁹ is hydrogen. 4. A compound according to claim 3, wherein both R ⁸ and R ⁹ are hydrogen. 5. A compound according to any one of claims 1 to 4, wherein R ¹ represents optionally substituted cyclopropyl or optionally substituted cyclobutyl. 6. A compound according to claim 1, wherein R ¹ is cyclopropyl, cyclobutyl, hydroxycyclobut-3-yl, cyanocyclobut-3-yl or fluorocyclobut-3-yl. 7. A compound according to any one of claims 1 to 4, where R ¹ represents CD ₃ , ethyl or prop-2-yl. 8. A compound according to any one of claims 1 to 7, where R ² represents bromine. 9. A compound according to any one of claims 1 to 7, where R ² represents -CF ₃ . 10. A compound as claimed in any one of the preceding claims, wherein R ³ is cyclopropyl, methoxy, cyclopropylamino, -NH(CH ₃ ) or -NH(CH ₂ CH ₃ ). 11. A compound according to any of the preceding claims, wherein R ⁴ represents hydrogen. 12. A compound according to any of the preceding claims, wherein R ¹ represents C3.6 cycloalkyl independently substituted with one or more hydroxy or cyano; R ² represents halogen or C1. ₆ fluoroalkyl; R ³ is -N(R ¹¹ )(R ¹² ) or C ₁ . ₆ alkoxy and R ⁴ is H. - 43 043797 13. A compound or a pharmaceutically acceptable salt thereof selected from No. Structure 34 HA HN N NH ^d 3C'n^% nA A ^N 57 n ^CFa HN N NH N=< ,% Δ— NJ / ΪΓ 68 Lf AA _n HN N NH “M / nA A Av 69 Aq 4V I Vz / 75 aT _D HN N NH aL a W 77 A ^F AA HN N NH A 78 A ^F n^A^AAA HN N NH Ao - 44 043797 79 η ^βΓ _η ΗΝ Ν ΝΗ Μ / 81 η ΗΝ Ν 0 Μ V ^Ν >5 86 ρ N^<b l Λ ΗΝ Ν ΝΗ V.V. 88 f _f N^fT l Λ D _D HN Ν ΝΗ 126 1Ύ ^Γ ΗΝ Ν ΝΗ Vo / Ν^ 127 % Λ Λ ΗΝ Ν ΝΗ <5 - 45 043797 129 A A Λ HN N NH ^A < nA n^ AA uh 137 A n^At A Λ HN N NH AS < AO /N^ (First eluting isomer) 138 Qf N^Y^F A A HN N NH Ao (Second eluting isomer) 146 dT HN N NH (First eluting isomer) 147 A HN N NH A / ΪΤ (Second eluting isomer) 157 H' Q H^A T - 46 043797 14. The connection according to claim 13, selected from 15. A compound having the structure or a pharmaceutically acceptable salt thereof. 16. A compound having the structure or a pharmaceutically acceptable salt thereof. 17. A compound having the structure or a pharmaceutically acceptable salt thereof. 18. A compound having the structure or a pharmaceutically acceptable salt thereof. 19. A compound having the structure or a pharmaceutically acceptable salt thereof. - 47 043797 20. A pharmaceutical composition for the treatment of a disease or condition mediated at least in part by leucine-rich repeat kinase 2 (LRRK2), containing a compound according to claim 1 or a pharmaceutically acceptable salt, a deuterated analogue, a tautomer, a stereoisomer or a mixture of stereoisomers and a pharmaceutically acceptable carrier, diluent or excipient. 21. A method of treating a disease or condition mediated at least in part by LRRK2, the method comprising administering an effective amount of the pharmaceutical composition of claim 20 to a subject in need thereof. 22. The method of claim 21, wherein the disease or condition is a neurodegenerative disease. 23. The method according to claim 22, characterized in that the neurodegenerative disease is Parkinson's disease or dementia. 24. The method of claim 21, wherein the disease or condition is a disorder of the central nervous system (CNS). 25. The method according to claim 24, characterized in that the CNS disorder is Alzheimer's disease or levodopa-induced dyskinesia. 26. The method of claim 21, wherein the disease or condition is cancer. 27. The method of claim 26, wherein the cancer is kidney cancer, breast cancer, prostate cancer, blood cancer, papillary cancer, lung cancer, acute myelogenous leukemia or multiple myeloma. 28. The method according to claim 21, characterized in that the disease or condition is an inflammatory disease. 29. The method of claim 28, wherein the inflammatory disease is leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis or ankylosing spondylitis. 30. A method for enhancing cognitive memory, comprising administering an effective amount of the pharmaceutical composition according to claim 20 to a subject in need thereof. 31. Use of a compound according to claim 1 for the treatment of a disease or condition mediated at least in part by LRRK2 kinase. 32. Use according to claim 31, wherein the disease or condition mediated at least in part by LRRK2 kinase is a neurodegenerative disease, cancer or inflammatory disease. 33. The use of claim 31, wherein the disease or condition mediated at least in part by LRRK2 kinase is Alzheimer's disease, levodopa-induced dyskinesia, Parkinson's disease, dementia, amyotrophic lateral sclerosis (ALS), kidney cancer, breast cancer glands, prostate cancer, blood cancer, papillary cancer, lung cancer, acute myelogenous leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis or ankylosing spondylitis. 34. Use of a compound according to claim 1 for the manufacture of a medicament for the treatment of a neurodegenerative disease, cancer or inflammatory disease. 35. Use of a compound according to claim 1 for the manufacture of a medicinal product for the treatment of Alzheimer's disease, levodopa-induced dyskinesia, Parkinson's disease, dementia, amyotrophic lateral sclerosis, kidney cancer, breast cancer, prostate cancer, blood cancer, papillary cancer, lung cancer , acute myelogenous leukemia, multiple myeloma, leprosy, Crohn's disease, inflammatory bowel disease, ulcerative colitis, amyotrophic lateral sclerosis, rheumatoid arthritis or ankylosing spondylitis. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky lane, 2