Substituted pyrazoles as LRRK2 inhibitors

This application is a National Stage of International Application No. PCT/KR2018/015184 filed Dec. 3, 2018, claiming priority based on U.S. Provisional Patent Application No. 62/594,773 filed Dec. 5, 2017.

The present invention relates to a pyrrolo(pyrazolo)pyrimidine derivative having efficacy as an LRRK2 (leucine-rich repeat kinase 2) inhibitor and a pharmaceutical composition comprising the same for preventing or treating a degenerative brain disease.

LRRK2 (leucine-rich repeat kinase 2) is a member of the leucine-rich repeat kinase family. It consists of a sequence of 2,527 amino acids with highly conserved among interspecies. LRRK2 has dual functions with GTP hydrolase (GTPase) and serine-threonine kinase activities simultaneously.

LRRK2 is reported to be highly expressed in dopamine neurons of the cerebral cortex, substantia nigra, striatum, hippocampus, cerebellum associated with the pathogenesis of Parkinson's disease. In a postmortem study, expression of LRRK2 has been observed in abnormal protein aggregates called Lewy bodies, which result in specific dopamine neurons in Parkinson's disease patients.

In addition, it has been confirmed that a LRRK2 mutation with increased kinase activity (gain-of-functions' mutant) is closely associated with the occurrence of late-onset autosomal dominant Parkinson's disease. Thus, it is known that hyperactivity of LRRK2 has an important role on the initiation and progression of Parkinson's disease.

In particular, when normal LRRK2 is overexpressed in neurons, this leads to apoptosis induction. These physiological changes in LRRK2-derived cells are confirmed to be very similar to that of the cells from the neuronal cell death process in Parkinson's disease patients. The aberrantly activity of LRRK2 has a high clinical relationship with the initiation and progression of Parkinson's disease. It is expected that of the modulation of LRRK2 activity would be beneficial to control both neuronal apoptosis and neurocytic inflammatory responses, which are the most important pathological evidence in Parkinson's disease.

Meanwhile, 40% of the patients who are currently treated with the drugs for Parkinson's disease have experienced motor fluctuation. Furthermore, the rate of patients experiencing motor fluctuation increases by 10% per one year due to the occurrence of drug-resistance after a certain period of time (e.g., 4 to 6 years from onset).

Accordingly, there is a need for developing a novel small molecule drug which capable of oral administration and offer acceptable safety profile.

Technical Problem

Thus, as the results of the studies, the present inventors have developed and synthesized a novel pyrrolo(pyrazolo)pyrimidine derivative to selectively inhibit LRRK2. It has been confirmed that the derivatives had an excellent inhibitory activity to LRRK2, whereby the present invention has been completed.

Accordingly, an object of the present invention is to provide a novel compound excellent in the inhibitory effect on the activity of LRRK2.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating a degenerative brain disease, which comprises the compound as an active ingredient.

Solution to Problem

One aspect of the present invention provides a compound selected from the group consisting of a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, and isomer thereof.

In the above formula,

X is CH or N;

R 1 is hydrogen, halogen, or C 1-4 alkyl or C 1-4 alkoxy unsubstituted or substituted with one or more halogens;

R 2 is C 1-6 alkyl, C 1-3 alkoxy C 1-6 alkyl, C 3-6 cycloalkyl C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, 4- to 7-membered heterocycloalkyl, or C 6-10 aryl unsubstituted or substituted with one or more halogens;

wherein the 4- to 7-membered heterocycloalkyl may be substituted with C 1-4 alkyl, C 1-4 alkylcarbonyl unsubstituted or substituted with one or more halogens, C 3-6 cycloalkylcarbonyl, 4- to 7-membered heterocycloalkylcarbonyl unsubstituted or substituted with C 1-4 alkyl, C 1-4 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, or oxetanyl, and the heterocycloalkyl contains at least one atom selected from N, O, and S,

R 3 is hydrogen, halogen, or C 1-4 alkyl;

R 4 is C 1-6 alkyl or C 3-6 cycloalkyl unsubstituted or substituted with one or more halogens; and

R 5 is halogen or C 1-4 alkyl unsubstituted or substituted with one or more halogens.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a degenerative brain disease, which comprises the compound as an active ingredient.

Advantageous Effects

The pyrrolo(pyrazolo)pyrimidine derivative according to the present invention is excellent in the selective inhibitory activity to LRRK2 and is useful as a drug for preventing or treating a degenerative brain disease including Parkinson's disease.

Hereinafter, the invention will be described in detail with reference to embodiments. The embodiments are not limited to what is disclosed below. Rather, they may be modified in various forms as long as the gist of the invention is not altered.

In this specification, when a part is referred to as “comprising” a component, it means that the component can be further included rather than excluding other components, unless otherwise indicated.

In this specification, examples of halogen include fluorine, chlorine, bromine, and iodine.

One embodiment provides a compound selected from the group consisting of a compound represented by the following Formula 1, and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, and isomer thereof.

In the above formula,

X is CH or N;

R 1 is hydrogen, halogen, or C 1-4 alkyl or C 1-4 alkoxy unsubstituted or substituted with one or more halogens;

R 2 is C 1-6 alkyl, C 1-3 alkoxy C 1-6 alkyl, C 3-6 cycloalkyl C 1-6 alkyl, C 1-6 alkoxy, C 3-6 cycloalkyl, 4- to 7-membered heterocycloalkyl, or C 6-10 aryl unsubstituted or substituted with one or more halogens;

wherein the 4- to 7-membered heterocycloalkyl may be substituted with C 1-4 alkyl, C 1-4 alkylcarbonyl unsubstituted or substituted with one or more halogens, C 3-6 cycloalkylcarbonyl, 4- to 7-membered heterocycloalkylcarbonyl unsubstituted or substituted with C 1-4 alkyl, C 1-4 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, or oxetanyl, and

the heterocycloalkyl contains at least one atom selected from N, O, and S,

R 3 is hydrogen, halogen, or C 1-4 alkyl;

R 4 is C 1-6 alkyl or C 3-6 cycloalkyl unsubstituted or substituted with one or more halogens; and

R 5 is halogen or C 1-4 alkyl unsubstituted or substituted with one or more halogens.

According to an embodiment, R 1 is hydrogen, halogen, trifluoro C 1-4 alkyl, C 1-4 alkyl, or C 1-4 alkoxy.

According to an embodiment, R 4 is trifluoro C 1-4 alkyl, C 1-4 alkyl, or C 3-6 cycloalkyl.

According to an embodiment, R 5 is halogen, trifluoro C 1-4 alkyl, or C 1-4 alkyl.

According to an embodiment, R 2 is C 1-4 alkyl, C 1-3 alkoxy C 1-4 alkyl, C 3-6 cycloalkyl C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, 4- to 7-membered heterocycloalkyl, or C 6-10 aryl substituted with F or Cl, wherein the 4- to 7-membered heterocycloalkyl is substituted with C 1-4 alkyl, trifluoro C 1-4 alkylcarbonyl, C 1-4 alkylcarbonyl, C 3-6 cycloalkylcarbonyl, 4- to 7-membered heterocycloalkylcarbonyl unsubstituted or substituted with C 1-4 alkyl, C 1-4 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, or oxetanyl, and the heterocycloalkyl contains at least one atom selected from N and O.

According to an embodiment, R 2 may be selected from the group consisting of

In the above substituents,

is the point of attachment to the nitrogen atom of the pyrazolyl ring.

According to an embodiment, the compound represented by Formula 1 may be selected from the group consisting of the following compounds.

1) N-(5-chloro-1-methyl-1H-pyrazol-4-yl)-7-methyl-5-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 2) 5-chloro-N-(1-isopropyl-3-methoxy-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 3) 5-chloro-N-(3-chloro-1-isopropyl-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 4) 5-chloro-7-methyl-N-(1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 5) 5-chloro-N-(5-chloro-1-methyl-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 6) 5-chloro-N-(5-chloro-1-methyl-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 7) 5-chloro-N-(3-chloro-1-isopropyl-1H-pyrazol-4-yl)-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 8) 5-chloro-N-(3-chloro-1-isopropyl-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 9) 5-chloro-N-(1-isopropyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 10) 5-chloro-N-(1-(2-methoxyethyl)-3-(trifluoromethyl)-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 11) N-(1-butyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)-5-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 12) 5-chloro-N-(3-chloro-1-(2-methoxyethyl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 13) 5-chloro-N-(3-chloro-1-(cyclopropylmethyl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 14) 5-chloro-N-(3-chloro-1-cyclopropyl-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 15) 5-chloro-N-(3-chloro-1-cyclopropyl-1H-pyrazol-4-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 16) 5-chloro-N-(5-chloro-1-(oxetan-3-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 17) 5-chloro-N-(3-chloro-1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 18) 5-chloro-N-(3-chloro-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 19) 5-chloro-N-(3-chloro-1-(2,6-dimethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 20) 5-chloro-N-(3-chloro-1-(1-ethylpiperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 21) 5-chloro-N-(3-chloro-1-(4-fluorophenyl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 22) 5-chloro-N-(5-chloro-1-(4-chlorophenyl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 23) 1-(4-(5-chloro-4-(5-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)ethanone, 24) (4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone, 25) 5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 26) 1-(4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)-2-methylpropane-1-one, 27) 5-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 28) 5-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 29) 5-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 30) 5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 31) (4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)(morpholino)methanone, 32) (4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)(4-methylpiperazin-1-yl)methanone, 33) 1-(4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)-2,2-dimethylpropane-1-one, 34) 5-chloro-N-(3,5-dimethyl-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 35) 5-chloro-N-(3-chloro-1-(1-(cyclopropylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 36) (R)-5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 37) (S)-5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 38) (R)-(3-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)pyrrolidin-1-yl)(cyclopentyl)methanone, 39) (S)-(3-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)pyrrolidin-1-yl)(cyclopentyl)methanone, 40) 5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)azetidin-3-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 41) (3-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)azetidin-1-yl)(cyclopropyl)methanone, 42) 5-chloro-N-(3-chloro-1-(1-(methylsulfonyl)azepan-4-yl)-1H-pyrazol-4-yl)-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-amine, 43) (4-(3-chloro-4-(5-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-1H-pyrazol-1-yl)azepan-1-yl)(cyclopropyl)methanone, 44) 3-chloro-N-(3-chloro-1-(4-fluorophenyl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 45) 3-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 46) 3-chloro-N-(3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 47) 3-chloro-N-(3-chloro-1-(1-(cyclopropylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 48) 1-(4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)-2,2,2-trifluoroethanone, 49) 3-chloro-N-(3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 50) 3-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 51) 3-chloro-N-(3-chloro-1-(1-(methylsulfonyl)azepan-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 52) (4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)azepan-1-yl)(cyclopropyl)methanone, 53) 1-(4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)-2,2-dimethylpropane-1-one, 54) (4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)(morpholino)methanone, 55) 3-chloro-N-(3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 56) (4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-5-methyl-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone, 57) (4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone, 58) 1-(4-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)piperidin-1-yl)ethanone, 59) 3-chloro-N-(3-chloro-1-(1-(ethylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 60) (3-(3-chloro-4-(3-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)-1H-pyrazol-1-yl)azetidin-1-yl)(cyclopropyl)methanone, 61) 3-chloro-N-(5-chloro-1-(1-(oxetan-3-yl)azetidin-3-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine, 62) 3-chloro-N-(5-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-yl)-1-ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-amine.

According to an embodiment, the LRRK2 inhibition rate (IC 50 ) according to the substituent of the compound represented by Formula 1 is as shown in Table 1 below. (++++: greater than 0 to 10 nM, +++: greater than 10 nM to 100 nM, ++: greater than 100 nM to 1,000 nM, +: greater than 1,000 nM to 10,000 nM).

According to an embodiment, examples of the pharmaceutically acceptable salt of the compound represented by Formula 1 include alkaline metal salts, alkaline earth metal salts, ammonium salts, amine salts, acid addition salts, hydrate salts, and the like. They may be non-toxic and water-soluble. For example, they may be inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, and nitrate; and organic acid salts such as acetate, formate, lactate, tartrate, tannate, succinate, maleate, fumarate, benzoate, citrate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, isethionate, glucuronate, gluconate, and fatty acid salts.

According to an embodiment, the pharmaceutically acceptable salt may be one or more selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, methanesulfonate, ethanesulfonate, benzenesulfonate, and toluenesulfonate.

The hydrate and solvate of the compound represented by Formula 1 according to an embodiment may be prepared by a method well known in the field. Specifically, they may be a hydrate and a solvate to which 1 to 5 molecules of water and an alcohol-based solvent (especially, ethanol or the like) are bonded, respectively, and are preferably non-toxic and water-soluble.

The prodrug of the compound represented by Formula 1 according to an embodiment is a chemical derivative capable of being converted into the compound represented by Formula 1 in vivo when administered. It may be prepared by a method well known in the field.

The isomer of the compound represented by Formula 1 according to an embodiment include optical isomers (e.g., enantiomers, diastereomers, and mixtures thereof), as well as conformation isomers and position isomers.

The compound of Formula 1a (where X is CH) according to an embodiment may be prepared by the method shown in Reaction Scheme 1 below.

In Reaction Scheme 1, R 1 to R 5 are as defined in Formula 1 above.

Specifically, the compound of Formula 1a may be prepared by a process, which comprises (1) reacting a compound of Formula 2 with a compound of Formula 3 in an organic solvent to prepare a compound of Formula 4; (2) reacting the compound of Formula 4 with a compound of Formula 5 in an organic solvent to prepare a compound of Formula 6; and (3) reacting the compound of Formula 6 with a compound of Formula 7 in an organic solvent to prepare the compound of Formula Ta.

The organic solvent may be at least one selected from the group consisting of acetone, 1,4-dioxane, methyl ethyl ketone, N,N-dimethylformamide, N-methylpyrrolidone, and ethyl acetate, but it is not limited thereto.

In step (1), for example, a compound of Formula 2 may be reacted with N-chlorosuccinimide (NCS) in an N,N-dimethylformamide solvent for 1 to 2 hours at room temperature to prepare a compound of Formula 4.

In step (2), for example, the compound of formula 4 may be reacted with potassium hydroxide and a compound of Formula 5 in an N,N-dimethylformamide solvent for 2 hours at room temperature to prepare a compound of Formula 6.

Alternatively, unlike steps (1) and (2) above, after the compound of Formula 2 is reacted with iodine and potassium hydroxide in an N,N-dimethylformamide solvent at 0° C. to room temperature, iodomethane is added thereto, followed by reaction with methyl fluorosulfonyldifluoroacetate (MFSDA) at 100 to 130° C. for 4 hours to prepare 2-chloro-7-methyl-5-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidine.

In step (3), for example, after the compound of Formula 6 and a compound of Formula 7 are dissolved in an ethanol solvent, hydrochloric acid is added thereto, which is reacted at 80° C. for 12 hours to prepare a compound of Formula 1a.

Alternatively, palladium (II) acetate (0.1 to 0.5 eq.), Xantphos (0.5 to 1.0 eq.), potassium carbonate (2 to 3 eq.), compounds 6 and 7 are reacted in an organic solvent such as 1,4-dioxane at 80 to 110° C. for 10 to 12 hours to prepare the compound of Formula Ta.

The compound of Formula 1b (where X is N) according to an embodiment may be prepared by the method as shown in Reaction Scheme 2 below.

In Reaction Scheme 2, R 1 to R 5 are as defined in Formula 1 above.

Specifically, the compound of Formula 1b may be prepared by a process, which comprises (A) reacting a compound of Formula 8 with N 2 H 4 and acetonitrile (ACN) to prepare a compound of Formula 9; (B) reacting the compound of Formula 9 with KOH to prepare a compound of Formula 10; (C) reacting the compound of Formula 10 with phosphoryl chloride (POCl 3 ) to prepare a compound of Formula 11; (D) reacting the compound of Formula 11 with a compound of Formula 5 in an organic solvent to prepare a compound of Formula 12; (E) reacting the compound of Formula 12 with m-chloroperoxybenzoic acid (mCPBA) in an organic solvent to prepare a compound of Formula 13; and (F) reacting the compound of Formula 13 with p-toluenesulfonic acid and a compound of Formula 7 in an organic solvent to prepare the compound of Formula 1b.

The organic solvent may be at least one selected from the group consisting of acetone, dichloromethane, 1,4-dioxane, methyl ethyl ketone, N,N-dimethylformamide, N-methylpyrrolidone, and ethyl acetate, but it is not limited thereto.

In step (A), for example, a compound of Formula 8 may be reacted with N 2 H 4 and acetonitrile (ACN) for 1 to 2 hours at room temperature to prepare a compound of Formula 9.

In step (B), for example, the compound of Formula 9 may be reacted with 10% KOH at 90 to 110° C. for 2 to 3 hours to prepare a compound of Formula 10.

In step (C), for example, the compound of Formula 10 may be reacted with phosphoryl chloride (POCl 3 ) at 90 to 110° C. to prepare a compound of Formula 11.

In step (D), for example, the compound of formula 11 may be reacted with a compound of Formula 5 in an N,N-dimethylformamide solvent at 0° C. to room temperature for 2 hours to prepare a compound of Formula 12.

In step (E), for example, the compound of Formula 12 may be reacted with m-chloroperoxybenzoic acid (mCPBA) in a dichloromethane or N,N-dimethylformamide solvent at room temperature for 4 hours to prepare a compound of Formula 13.

In step (F), for example, the compound of formula 13 may be reacted with p-toluenesulfonic acid and a compound of Formula 7 in an N-methylpyrrolidone solvent at 100 to 120° C. for 6 hours to prepare the compound of Formula 1b.

For the synthesis of the compounds of Formulae 10 and 11, reference was made to the literature (Hauser, Martin. et al., J. Org, Chem. 1960, 25, 1570-1573; J. Org. Chem. 1961, 26, 451-455; and Liu, Jing. et al., ACS Comb. Sci. 2011, 13, 414-420).

The compound represented by Formula 1 according to an embodiment, and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, and isomer thereof have a high inhibitory activity to LRRK2, as well as excellent selectivity for other kinases. Thus, the compound represented by Formula 1 and a pharmaceutically acceptable salt, hydrate, solvate, prodrug, and isomer thereof are useful as a drug for preventing or treating a degenerative brain disease that requires inhibition of LRRK2 activity.

One embodiment provides a pharmaceutical composition for preventing or treating a degenerative brain disease, which comprises the compound as an active ingredient.

According to an embodiment, the degenerative brain disease may be Parkinson's disease.

According to an embodiment, the pharmaceutical composition has an inhibitory activity selective to LRRK2 (leucine-rich repeat kinase 2).

According to an embodiment, the pharmaceutical composition may comprise one or more pharmaceutically acceptable additives. For example, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc; excipients such as lactose, sodium citrate, calcium carbonate, and dicalcium phosphate; disintegrants such as starch, alginic acid, and specific complex silicates; and various carriers may be used for the preparation of tablets. In addition, the type of carrier may vary depending on the solubility, chemical properties, and mode of administration of the active ingredient.

The pharmaceutical composition may be prepared according to conventional methods using one or more pharmaceutically acceptable additives. The additives may include, in particular, diluents, sterilized aqueous media, and various non-toxic organic solvents. If necessary, sweeteners, fragrances, colorants, or stabilizers may be further comprised.

The pharmaceutical composition may be formulated in various dosage forms such as tablets, pills, granules, capsules, powders, aqueous solutions, or suspensions, injection solutions, elixirs, or syrups according to conventional formulation methods in the pharmaceutical field.

In order to prepare capsules, it is advantageous to use lactose and high molecular weight polyethylene glycol. If aqueous suspensions are used, they may contain emulsifiers or agents that facilitate suspension. Such diluents as sucrose, ethanol, polyethylene glycol, propylene glycol, glycerol, chloroform, or mixtures thereof may also be used.

As such, the pharmaceutical composition may be administered non-orally or orally as needed. It may be administered in single or multiple doses once a day. The non-oral dose range is from 0.5 mg/kg to 5 mg/kg per body weight, preferably 1 mg/kg to 4 mg/kg per body weight; of the oral dose range is form 5 mg/kg to 50 mg/kg per body weight, preferably 10 mg/kg to 40 mg/kg per body weight. The dosage for patients may vary depending on body weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of the disease for each patent.

According to an embodiment, there is provided a use of the compound for the preparation of a medicament for preventing or treating a degenerative brain disease.

According to an embodiment, there is provided a method of treating a degenerative brain disease, which comprises administering the compound to a patient.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are provided to illustrate the present invention with specific embodiments. It is not intended to limit the scope of the present invention to the those described therein.

Preparation Example 1: Synthesis of Intermediate 1 (2,5-dichloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine)

2,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (2.16 g, 11.48 mmol) was added to an N,N-dimethylformamide solvent (20 ml), which was cooled to 0° C. NaH (0.68 g, 17.23 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. Iodomethane (1.59 ml, 17.23 mmol) was slowly added thereto dropwise, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. Water (50 ml) was added to precipitate a solid. It was filtered with water under reduced pressure and washed with n-heptane to obtain 2.20 g (yield: 94%) of the target compound as a solid in beige color.

MS (ESI) m/z 201.9, 204.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.95 (s, 1H), 7.88 (s, 1H), 3.77 (s, 3H).

Preparation Example 2: Synthesis of Intermediate 2 (2,5-dichloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidine)

2,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (3.31 g, 17.60 mmol) was added to an N,N-dimethylformamide solvent (33 ml), which was cooled to 0° C. NaH (1.15 g, 26.40 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. Iodomethane (1.69 ml, 21.12 mmol) was slowly added thereto dropwise, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. Water (80 ml) was added to precipitate a solid. It was filtered with water under reduced pressure to obtain 3.15 g (yield: 82%) of the target compound.

MS (ESI) m/z 216.0, 217.8 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 8.92 (s, 1H), 7.96 (s, 1H), 4.21 (q, J=7.2 Hz, 2H), 1.38 (t, J=7.2 Hz, 3H).

Preparation Example 3: Synthesis of Intermediate 3 (2,5-dichloro-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidine)

2,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (900 mg, 4.79 mmol), cesium carbonate (Cs 2 CO 3 , 1.9 g, 5.7 mmol), and 1,1,1-trifluoro-2-iodoethane (9.58 mmol, 0.94 ml) were added to an N,N-dimethylformamide solvent (50 ml), which was stirred at 100° C. for 12 hours. It was extracted with water and dichloromethane (DCM), which was depressurized and subjected to column chromatography (EA:Hep=1:1) to obtain 1.0 g of the target compound (yield: 77%).

MS (ESI) m/z 269.8 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.89 (s, 1H), 7.279 (s, 1H), 4.84 (q, J=8.4 Hz, 2H).

Preparation Example 4: Synthesis of Intermediate 4 (2,5-dichloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine)

2,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (1.0 g, 5.31 mmol) was added to an N,N-dimethylformamide solvent (10 ml), which was cooled to 0° C. NaH (0.32 g, 7.97 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. 2-Iodopropane (0.79 ml, 7.97 mmol) was slowly added thereto dropwise, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. Water (30 ml) was added at room temperature to precipitate a solid. It was filtered with water under reduced pressure to obtain 1.05 g of the target compound (yield: 85%).

MS (ESI) m/z 230.0, 232.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.85 (s, 1H), 7.82 (s, 1H), 5.09 (p, J=6.9 Hz, 1H), 1.57 (s, 3H), 1.55 (s, 3H).

Preparation Example 5: Synthesis of Intermediate 5 (2,5-dichloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine)

2,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (0.2 g, 1.06 mmol) was added to an N,N-dimethylformamide solvent (10 ml), which was cooled to 0° C. NaH (69.6 mg, 1.59 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. Cyclopropanecarbonyl chloride (0.11 ml, 1.27 mmol) was slowly added thereto dropwise, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. Water (30 ml) was added at room temperature to precipitate a solid. It was filtered with water under reduced pressure to obtain 0.23 g of the target compound (yield: 87%).

MS (ESI) m/z 255.9, 258.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.90 (s, 1H), 7.98 (s, 1H), 3.85-3.79 (m, 1H), 1.43-1.40 (m, 2H), 1.33-1.30 (m, 2H).

Preparation Example 6: Synthesis of Intermediate 6 (2-chloro-7-methyl-5-(trifluoromethyl)-7H-pyrrolo[2,3-d]pyrimidine)

2-Chloro-7H-pyrrole[2,3-d]pyrimidine (1.0 g, 6.51 mmol) was dissolved in an N,N-dimethylformamide solvent (10 ml), which was cooled to 0° C. Potassium hydroxide (1.37 g, 24.41 mmol) and iodine (3.3 g, 13.02 mmol) were slowly added thereto, followed by gradual elevation of the temperature to room temperature. After it was stirred for 2 hours, water (80 ml) was added thereto to terminate the reaction, and a solid was precipitated. It was filtered with water under reduced pressure and washed with ethyl acetate (EtOAc) to obtain 1.68 g of the target compound as a solid in yellow color (yield: 89%).

MS (ESI) m/z 279.8, 281.8 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.37 (s, 1H), 7.64 (s, 1H).

2-Chloro-5-iodo-7H-pyrrole[2,3-d]pyrimidine (1 g, 3.57 mmol) was dissolved in an N,N-dimethylformamide solvent (5 ml), which was cooled to 0° C. NaH (0.21 g, 5.36 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. Iodomethane (0.49 ml, 5.36 mmol) was slowly added thereto dropwise, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. Water (10 ml) was added to precipitate a solid. It was filtered with water under reduced pressure and washed with n-heptane to obtain 0.60 g of the target compound as a solid in beige color (yield: 57%).

MS (ESI) m/z 236.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.60 (s, 1H), 7.29 (s, 1H), 3.88 (s, 3H).

2-Chloro-5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidine (0.5 g, 1.70 mmol) was dissolved in an N,N-dimethylformamide solvent (10 ml). Methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (0.53 ml, 4.25 mmol) and Cu(I)I (0.32 g, 1.70 mmol) were added thereto, which was stirred at 130° C. for 2 hours. The temperature was lowered to room temperature, ethyl acetate (50 ml) was added thereto, and water (30 ml) was added thereto to terminate the reaction. It was filtered with celite, extracted with ethyl acetate and water, and washed with brine. The organic layer was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.12 g of the target compound (yield: 310%).

MS (ESI) m/z 235.9, 238.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.96 (s, 1H), 7.57 (s, 1H), 3.90 (s, 3H).

Preparation Example 7: Synthesis of Intermediate 7 (3-chloro-1-methyl-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine)

Ethanol (5.5 ml) was added to ethyl 4-chloro-2-(methylthio)pyrimidin-5-carboxylate (1.3 g, 5.58 mmol), which was cooled to 0° C. Methyl hydrazine (0.58 ml, 11.17 mmol) dilute in ethanol (5.5 ml) was slowly added thereto, which was stirred at room temperature for 2 hours. Water (30 ml) was added thereto to terminate the reaction, and a white solid was precipitated. It was filtered under reduced pressure to obtain 0.86 g of the target compound (yield: 64%).

MS (ESI) m/z 243.3 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.33 (s, 1H), 4.33 (q, J=7.2 Hz, 2H), 4.17 (s, 2H), 3.40 (s, 3H), 2.53 (s, 3H), 1.37 (t, J=7.2 Hz, 3H).

Ethyl 4-hydrazinyl-2-(methylthio)pyrimidin-5-carboxylate (0.86 g, 3.54 mmol) was added to 10% KOH (9.0 ml), which was refluxed at 100° C. for 15 minutes. After the temperature was lowered to room temperature, a 25% aqueous acetic acid solution (8 ml) was added thereto for neutralization, thereby precipitating a yellow solid. It was filtered with water under reduced pressure to obtain 0.58 g of the target compound (yield: 84%).

MS (ESI) m/z 197.0 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 11.59 (s, 1H), 8.86 (s, 1H), 3.72 (s, 3H), 2.56 (s, 3H).

A suction flask was charged with 6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidin-3(2H)-one (0.58 g, 2.95 mmol), and POCl 3 (2.34 ml, 25.12 mmol) was added thereto, which was stirred at 140° C. for 5 hours. After the temperature was lowered to room temperature, ammonium hydroxide (20 ml) was slowly added thereto at 0° C. for neutralization, thereby precipitating a beige solid. It was filtered with water under reduced pressure and washed with a solution of ethanol and water (1:1) to obtain 0.50 g of the target compound (yield: 79%).

MS (ESI) m/z 214.9, 216.9 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.87 (s, 1H), 4.02 (s, 2H), 2.66 (s, 3H).

Preparation Example 8: Synthesis of Intermediate 8 (3-chloro-1-ethyl-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine)

3-Chloro-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine (12.7 g, 63.29 mmol) was dissolved in an N,N-dimethylformamide solvent (63 ml), which was cooled to 0° C. NaH (3.31 g, 75.95 mmol) was slowly added thereto, followed by stirring thereof at 0° C. for 30 minutes. Iodomethane (6.10 ml, 75.95 mmol) was slowly added thereto, followed by elevation of the temperature to room temperature and stirring thereof for 2 hours. After the reaction was terminated, water (100 ml) was added to precipitate a brown solid. It was filtered and dried under reduced pressure to obtain 12.26 g of the target compound (yield: 84%).

MS (ESI) m/z 228.70 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.86 (s, 1H), 4.44 (q, J=7.2 Hz, 2H), 2.65 (s, 3H), 1.53 (t, J=7.2 Hz, 3H).

Preparation Example 9: Synthesis of Intermediate 9 (3-chloro-6-(methylthio)-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin)

3-Chloro-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine (70 mg, 0.49 mmol) was dissolved in an N,N-dimethylformamide solvent (63 ml). Cesium carbonate (243 mg, 0.74 mmol) and 2,2,2-trifluoroethyl methanesulfonate (70.0 ml, 0.59 mmol) were added thereto, which was stirred at 80° C. for 5 hours. The temperature was lowered to room temperature, and it was extracted with water and ethyl acetate. The organic layer was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 48.3 mg of the target compound (yield: 48%).

MS (ESI) m/z 282.8, 285.0 [M+H] + ;

1 H NMR (300 MHz, Acetone-d 6 ) δ 9.09 (s, 1H), 5.27 (q, J=8.7 Hz, 2H), 2.66 (s, 3H).

Preparation Example 10: Synthesis of Intermediate 10 (3-chloro-1-cyclopropyl-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine)

A suction flask was charged with 3-chloro-6-(methylthio)-1H-pyrazolo[3,4-d]pyrimidine (0.2 g, 0.99 mmol), cycloboronic acid (0.17 g, 1.99 mmol), 2,2′-bipyridine (0.15 g, 0.99 mmol), and Na 2 CO 3 (0.21 g, 1.99 mmol). Dichloroethane (10 ml) was added thereto, and nitrogen (N 2 ) was bubbled for 10 minutes. Cu(OAc) 2 (0.18 g, 0.99 mmol) was added thereto, and N 2 was bubbled for 1 minute, which was stirred at 60° C. for 18 hours. The temperature was lowered to room temperature, and it was filtered with ethyl acetate and then concentrated. It was extracted with ethyl acetate and a 1N aqueous hydrochloric acid solution, dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.16 g of the target compound (yield: 67%).

MS (ESI) m/z 241.1, 243.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.84 (s, 1H), 3.82-3.75 (m, 1H), 2.67 (s, 3H), 1.36-1.33 (m, 2H), 1.18-1.14 (m, 2H).

Preparation Example 11: Synthesis of Intermediate 11 (1-isopropyl-3-methoxy-1H-pyrazol-4-amine)

HNO 3 (18 ml) was slowly added to acetic anhydride (42 ml) at 0° C. to prepare acetyl nitrate. Another flask was charged with 1H-pyrazole (10 g), and acetic acid (28 ml) was added thereto. The acetyl nitrate was slowly added to the pyrazole compound at 0° C., followed by stirring thereof for 1 hour and the addition of water to precipitate a solid. It was filtered with water under reduced pressure to obtain 16.0 g of the target compound (yield: 97%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.79 (t, J=3.3 Hz, 1H), 7.88 (d, J=0.6 Hz, 1H).

1-Nitro-1H-pyrazole (10 g) was dissolved in benzonitrile (70 ml), which was refluxed at 180° C. for 4 hours. The temperature was lowered to room temperature to precipitate a solid. Heptane (250 ml) was added thereto, which was stirred for 10 minutes, followed by filtration under reduced pressure to obtain 8.9 g of the target compound (yield: 89%).

1 H NMR (300 MHz, CDCl 3 ) δ 12.80 (br s, 1H), 7.87 (d, J=2.4 Hz, 1H), 7.04 (t, J=1.2 Hz, 1H).

3-Nitro-1H-pyrazole (2.5 g, 22.11 mmol) was dissolved in H 2 SO 4 (7.6 ml, 143.71 mmol), which was cooled to 0° C. Fuming nitric acid (fuming HNO 3 , 1.03 ml, 24.32 mmol) was slowly added thereto, followed by stirring thereof at 80° C. for 4 hours. The temperature was lowered to 0° C., and it was neutralized with NaHCO 3 . It was extracted with ethyl acetate, dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.80 g of the target compound (yield: 22%).

MS (ESI) m/z 159.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 9.12 (s, 1H).

3,4-Dinitro-1H-pyrazole (0.8 g, 5.06 mmol) was dissolved in dimethylformamide (DMF, 10 ml), which was cooled to 0° C. NaH (0.30 g, 7.59 mmol) was slowly added thereto, followed by stirring thereof for 30 minutes. 2-Iodopropane (0.75 ml, 7.59 mmol) was added thereto dropwise, followed by stirring thereof at room temperature for 1 hour. Water was added thereto to terminate the reaction, and it was extracted with ethyl acetate. It was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.31 g of the target compound (yield: 30%).

MS (ESI) m/z 201.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 4.58 (q, J=6.9 Hz, 1H), 1.64 (s, 3H), 1.61 (s, 3H).

1-Isopropyl-3,4-dinitro-1H-pyrazole (0.31 g, 1.54 mmol) was dissolved in MeOH, and NaOMe (0.78 g, 3.09 mmol) was added thereto. After it was refluxed for 6 hours, the temperature was lowered to room temperature, and it was extracted with water and ethyl acetate. It was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.26 g of the target compound (yield: 92%).

MS (ESI) m/z 186.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.03 (s, 1H), 4.32 (p, J=6.6 Hz, 1H), 4.06 (s, 3H), 1.53 (s, 3H), 1.51 (s, 3H).

1-Isopropyl-3-methoxy-4-nitro-1H-pyrazol (0.26 g, 1.40 mmol) was dissolved in MeOH, and 10 wt % Pd/C (37.75 mg, 0.03 mmol) was slowly added thereto. It was stirred for 14 hours under H 2 conditions. It was filtered with Celite, concentrated, and subjected to column chromatography to obtain 0.16 g of the target compound (yield: 75%).

MS (ESI) m/z 201.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 6.95 (s, 1H), 4.20 (q, J=6.6 Hz, 1H), 3.93 (s, 3H), 1.41 (s, 3H), 1.39 (s, 3H).

Preparation Example 12: Synthesis of Intermediate 12 (4-nitro-3-(trifluoromethyl)-1H-pyrazole)

3-(Trifluoromethyl)-1H-pyrazole (2 g, 14.69 mmol) and H 2 SO 4 (9.36 g, 95.53 mmol) were added, which was stirred in an ice bath. HNO 3 (1.01 g, 16.16 mmol) was added thereto, followed by stirring thereof at 80° C. for 4 hours. Upon termination of the reaction, ice was added thereto, followed by stirring thereof. The resulting solid compound was filtered first, and the filtrate was extracted again with ethyl acetate, dried over MgSO 4 , and depressurized to obtain 2.60 g of a white solid compound (yield: 97%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.46 (s, 1H).

Preparation Example 13: Synthesis of Intermediate 13 (1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazol-4-amine)

Dihydro-2H-pyran-4(3H)-one (1 g, 9.98 mmol) was added to tetrahydrofuran (THF, 10 ml), which was stirred for 10 minutes in an ice bath. LiAlH 4 (2 M in THF, 10 ml) was added thereto dropwise, followed by stirring thereof for 3 hours. Upon termination of the reaction, it was immersed in 2 N NaOH. NH 4 Cl was added thereto, which was stirred and then filtered. The solution was evaporated under reduced pressure, and isopropyl ether (IPE) was added thereto, which was stirred and then filtered. The filtrate was depressurized again to obtain 0.18 g of the target compound (yield: 18%).

1 H NMR (300 MHz, CDCl 3 ) δ 3.54-3.47 (m, 4H), 1.25-1.20 (m, 5H).

Tetrahydro-2H-pyran-4-ol (0.18 g, 1.82 mmol) and DCM (15 ml) were added, which was stirred for 10 minutes in an ice bath. Methanesulfonyl chloride (0.31 g, 2.73 mmol) and triethylamine (TEA, 0.27 g, 2.73 mmol) were added thereto, which was stirred for 7 hours. Upon termination of the reaction, it was extracted, dried over MgSO 4 , and depressurized to obtain 0.20 g of the target compound (yield: 62%).

1 H NMR (300 MHz, CDCl 3 ) δ 4.97-4.88 (m, 1H), 4.00-3.93 (m, 2H), 3.61-3.53 (m, 2H), 3.06 (s, 3H), 2.11-2.03 (m, 2H), 1.96-1.84 (m, 2H).

4-Nitro-3-(trifluoromethyl)-1H-pyrazole (0.1 g, 0.55 mmol, Intermediate 12), DMF (13 ml), and Cs 2 CO 3 (0.27 g, 0.82 mmol) were added and stirred for 30 minutes. Tetrahydro-2H-pyran-4-yl methanesulfonate (0.13 g, 0.82 mmol) was added thereto, which was refluxed for 2 hours. Upon termination of the reaction, the solution was evaporated under reduced pressure, and it was extracted with ethyl acetate. After it was dried over MgSO 4 , the solution was evaporated under reduced pressure, and it was subjected to column chromatography (MC:MeOH=10:1) to obtain 0.045 g of the target compound (yield: 31%).

4-Nitro-1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazole (0.045 g, 0.17 mmol) and THF:MeOH (7 ml, 10:1 v/v) were added, which was stirred for 10 minutes in an ice bath. Ni(OAc) 2 .4H 2 O (0.0042 g, 0.016 mmol) and NaBH 4 (0.025 g, 0.67 mmol) were added thereto, which was stirred for 30 minutes. Upon termination of the reaction, it was filtered, and the solution was evaporated under reduced pressure. It was subjected to column chromatography (MC:MeOH=10:1) to obtain 0.039 g of the target compound (yield: 98%).

MS (ESI) m/z 236.0 [M+H] +

Preparation Example 14: Synthesis of Intermediate 14 (1-isopropyl-3-(trifluoromethyl)-1H-pyrazol-4-amine)

4-Nitro-3-(trifluoromethyl)-1H-pyrazole (0.10 g, 0.58 mmol, Intermediate 12) and DMF (15 ml) were added and stirred for 10 minutes in an ice bath. NaH (0.038 g, 0.87 mmol) was added thereto, followed by stirring thereof for 30 minutes. 2-Iodopropane (0.13 g, 0.76 mmol) was added thereto, followed by refluxing thereof for 3 hours. Upon termination of the reaction, the solution was evaporated under reduced pressure, and it was extracted with ethyl acetate. After it was dried over MgSO 4 , the solution was depressurized, and it was subjected to column chromatography (Hep:EA=1:1) to obtain 0.09 g of the target compound (yield: 69%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.28 (s, 1H), 4.64-4.55 (m, 1H), 1.59 (s, 6H).

The same procedure as in steps 3 and 4 of Preparation Example 13 was carried out, except that 1-isopropyl-4-nitro-3-(trifluoromethyl)-1H-pyrazole was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.09 g of the target compound (yield: 69%).

1 H NMR (300 MHz, CDCl 3 ) δ 7.06 (s, 1H), 4.49-4.38 (m, 1H), 3.19 (bs, 2H), 1.49 (s, 3H), 1.47 (s, 3H).

Preparation Example 15: Synthesis of Intermediate 15 (1-(2-methoxyethyl)-3-(trifluoromethyl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 14 was carried out, except that 1-bromo-2-methoxyethane was used instead of 2-iodopropane, to obtain 0.14 g of the target compound (yield: 72%).

1 H NMR (300 MHz, CDCl 3 ) δ 7.14 (s, 1H), 4.22-4.19 (t, 2H, J=10.2 Hz), 3.73-3.71 (t, 2H, J=10.2 Hz), 3.36 (s, 3H), 3.19 (bs, 2H).

Preparation Example 16: Synthesis of Intermediate 16 (1-butyl-3-(trifluoromethyl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 14 was carried out, except that 1-bromobutane was used instead of 2-iodopropane, to obtain 0.17 g of the target compound (yield: 93%).

1 H NMR (300 MHz, CDCl 3 ) δ 7.01 (s, 1H), 4.06-4.01 (t, 2H, J=14.7 Hz), 3.19 (s, 2H), 1.87-1.77 (m, 2H), 1.40-1.28 (m, 2H), 0.97-0.93 (t, 3H, J=14.7 Hz).

Preparation Example 17: Synthesis of Intermediate 17 (5-chloro-1-methyl-1H-pyrazol-4-amine)

4-Nitropyrazole (1.0 g, 8.84 mmol) was dissolved in DMF (10 ml). K 2 CO 3 (3.6 g, 26.5 mmol) was added thereto, followed by stirring thereof for 30 minutes. Then, Mel (1.65 ml, 13.26 mmol) was added thereto. After overnight, it was filtered with K 2 CO 3 , and the solvent was concentrated. It was extracted with dichloromethane (DCM) and water, dried over MgSO 4 , and then concentrated to obtain 0.82 g of the target compound (yield: 73%).

MS (ESI) m/z 128.1 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 8.84 (s, 1H), 8.23 (s, 1H), 3.91 (s, 3H).

THF (20 ml) was added to 1-methyl-4-nitro-1H-pyrazole (1.0 g, 7.86 mmoles). After the temperature was lowered to −78° C., lithium bis(trimethylsilyl)amide (LiHMDS, 1 M in THF, 15.6 ml) was added thereto dropwise. After it was stirred for 30 minutes, hexachloroethane (2.79 g, 11.8 mmol) was added thereto. After it was stirred at room temperature for 1 hour, NaCl (10 ml) was added thereto to terminate the reaction. Then, it was extracted with ethyl acetate, dried over MgSO 4 , and concentrated to obtain 1.1 g of the target compound (yield: 87%).

MS (ESI) m/z 162.2 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 8.14 (s, 3H), 3.91 (s, 3H).

THF (50 ml) was added to 5-chloro-1-methyl-4-nitro-1H-pyrazole (1.1 g, 6.83 mmol). Ni(OAc) 2 .4H 2 O (169 mg, 0.68 mmol) was then added thereto. After the temperature was lowered to 0 to 4° C. using an ice bath, NaBH 4 (590 mg, 13.6 mmol) was added thereto in two portions. After it was stirred for 30 minutes and checked with TLC, aqueous NH 4 Cl was added thereto to terminate the reaction, followed by extraction with ethyl acetate. After it was dried over MgSO 4 , a solution (3.4 ml, 13.6 mmol) in which 4 M HCl was dissolved in dioxane was added thereto. After it was stirred for 30 minutes and concentrated, acetone was added for recrystallization to obtain 160 mg of the target compound (yield: 18%).

MS (ESI) m/z 132.0, 134.2 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 10.48 (brs, 2H), 7.68 (s, 1H), 3.81 (s, 3H).

Preparation Example 18: Synthesis of Intermediate 18 (5-chloro-1-(oxetan-3-yl)-1H-pyrazol-4-amine)

4-Nitro-1H-pyrazole (0.66 g, 5.88 mmol) and oxetan-3-yl methanesulfonate (0.94 g, 6.17 mmol) were dissolved in DMF. Cs 2 CO 3 (3.80 g, 11.67 mmol) was added thereto, followed by stirring thereof at 90° C. for 18 hours. The temperature was lowered to room temperature, and the reaction was then terminated with water. It was extracted with acetate and then dried over MgSO 4 . It was filtered under reduced pressure, concentrated, and subjected to column chromatography to obtain 0.83 g of the target compound (yield: 84%).

MS (ESI) m/z 170.0 [M+H] +

THF (10 ml) was added to 4-nitro-1-(oxetan-3-yl)-1H-pyrazole (0.83 g, 4.90 mmol). After the temperature was lowered to −78° C., LiHMDS (1 M in THF, 9.81 ml) was added thereto dropwise. After it was stirred for 30 minutes, hexachloroethane (1.39 g, 5.88 mmol) dissolved in THF (5 ml) was added thereto. After it was stirred at −78° C. for 1 hour, aqueous NaCl (10 ml) was added thereto to terminate the reaction, followed by extraction with ethyl acetate. It was dried over MgSO 4 , concentrated, and then subjected to column chromatography to obtain 0.42 g of the target compound (yield: 43%).

MS (ESI) m/z 204.1 [M+H] +

EtOH (20 ml) and water (2 ml) were added to 5-chloro-4-nitro-1-(oxetan-3-yl)-1H-pyrazole (0.42 g, 2.06 mmol). Fe (0.34 g, 6.18 mmol) and NH 4 Cl (0.33 g, 6.18 mmol) were added thereto, followed by refluxing thereof at 90° C. for 3 hours. After the temperature was lowered to room temperature, it was filtered and concentrated under reduced pressure. Ethyl acetate was added thereto, and it was subjected to sonication for 3 minutes. After the insoluble material was filtered out, it was concentrated to obtain 0.34 g of the target compound (yield: 95%).

MS (ESI) m/z 173.9, 176.1 [M+H] +

Preparation Example 19: Synthesis of Intermediate 19 (5-chloro-1-(4-chlorophenyl)-1H-pyrazol-4-amine)

4-Nitro-1H-pyrazole (1 g, 8.84 mmol), 4-chlorobenzeneboronic acid (2.76 g, 17.68 mmol) and Cu(OAc) 2 (2.40 g, 13.26 mmol) were added to DCM (40 ml). Pyridine (2.85 ml, 35.37 mmol) was added thereto, followed by stirring thereof at room temperature for 20 hours. Upon termination of the reaction, it was filtered and extracted with DCM and water. The organic layer was dried over MgSO 4 and filtered. It was concentrated under reduced pressure and subjected to column chromatography to obtain 0.88 g of the target compound (yield: 44%).

THF (6 ml) was added to 1-(4-chlorophenyl)-4-nitro-1H-pyrazole (0.64 g, 2.86 mmol). After the temperature was lowered to −78° C., LiHMDS (1 M in THF, 5.7 ml) was added thereto dropwise. After it was stirred for 30 minutes, hexachloroethane (0.81 g, 3.43 mmol) dissolved in THF (3 ml) was added thereto. After it was stirred at −78° C. for 1 hour, aqueous NaCl (10 ml) was added thereto to terminate the reaction. It was extracted with ethyl acetate, dried over MgSO 4 , and concentrated. It was subjected to column chromatography to obtain 0.20 g of the target compound (yield: 27%).

MS (ESI) m/z 258.2, 260.0 [M+H] +

EtOH (15 ml) and water (1.5 ml) were added to 5-chloro-1-(4-chlorophenyl)-4-nitro-1H-pyrazole (0.20 g, 0.77 mmol). Fe (0.12 g, 2.32 mmol) and NH 4 Cl (0.12 g, 2.32 mmol) were added thereto, followed by refluxing thereof at 90° C. for 3 hours. After the temperature was lowered to room temperature, it was filtered and concentrated under reduced pressure. Ethyl acetate was added thereto, and it was subjected to sonication for 3 minutes. After the insoluble material was filtered out, it was concentrated to obtain 0.17 g of the target compound (yield: 99%).

MS (ESI) m/z 228.0, 230.1 [M+H] +

Preparation Example 20: Synthesis of Intermediate 20 (tert-butyl 4-(methylsulfonyloxy)piperidin-1-carboxylate)

Tert-butyl-4-hydroxypiperidin-1-carboxylate (20.0 g, 99.42 mmol) was dissolved in DCM (200 ml). Methanesulfonyl chloride (10 ml, 1.3 eq.) was added thereto, followed by stirring thereof. Triethylamine (41.6 ml, 3.0 eq.) was added thereto, followed by stirring thereof for 1 hour. Upon termination of the reaction, it was extracted three times with brine and DCM, dried over MgSO 4 , and concentrated under reduced pressure to obtain 27.7 g of a solid compound in ivory color (yield: 99%).

MS (ESI) m/z 280.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.95-4.54 (m, 1H), 3.74-3.70 (m, 2H), 3.35-3.28 (m, 2H), 3.05 (s, 3H), 1.96-1.84 (m, 4H), 1.48 (s, 9H).

Preparation Example 21: Synthesis of Intermediate 21 (tert-butyl 3-(methylsulfonyloxy)azetidin-1-carboxylate)

Azetidin-3-ol hydrochloride (3.0 g, 27.38 mmol) was dissolved in H 2 O (6 ml). K 2 CO 3 (3.78 g, 1.0 eq.) dissolved in H 2 O (6 ml) was added thereto. Di-tert-butyl dicarbonate (5.98 g, 1.0 eq.) dissolved in THF (27 ml) was added thereto dropwise, followed by stirring thereof for 4 hours. Upon termination of the reaction, ethyl acetate was added thereto three times for extraction, it was dried and concentrated to obtain 4.7 g of the target compound in yellow color (yield: 99%).

MS (ESI) m/z 174.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.59-4.56 (m, 1H), 4.54-4.11 (m, 2H), 3.82-3.77 (m, 2H), 3.16 (s, 1H), 1.43 (s, 9H).

Tert-butyl-3-hydroxyazetidin-1-carboxylate (4.7 g, 27.13 mmol) was dissolved in DCM (50 ml). Methanesulfonyl chloride (2.7 ml, 1.3 eq.) was added thereto, followed by stirring thereof. Triethylamine (11.3 mL, 3.0 eq.) was added thereto, followed by stirring thereof for 1 hour. Upon termination of the reaction, it was extracted three times with brine and DCM, dried over MgSO 4 , and concentrated to obtain 6.8 g of the target compound in brown color (yield: 99%).

MS (ESI) m/z 252.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 5.24-5.17 (m, 1H), 4.31-4.25 (m, 2H), 4.13-4.08 (m, 2H), 3.07 (s, 3H), 1.45 (s, 9H).

Preparation Example 22: Synthesis of Intermediate 22 (tert-butyl 4-(methylsulfonyloxy)azepan-1-carboxylate)

The same procedure as in Preparation Example 20 was carried out, except that tert-butyl-4-hydroxyazepan-1-carboxylate (2.0 g, 9.29 mmol) was used instead of tert-butyl-4-hydroxypiperidin-1-carboxylate, to obtain 2.7 g of the target compound (yield: 99%).

MS (ESI) m/z 294.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.91 (s, 1H), 3.45-3.38 (m, 1H), 3.02 (s, 3H), 2.08-2.04 (m, 5H), 1.73-1.64 (m, 1H), 1.47 (s, 9H).

Preparation Example 23: Synthesis of Intermediate 23 ((R)-tert-butyl 3-(methylsulfonyloxy)pyrrolidin-1-carboxylate)

The same procedure as in Preparation Example 20 was carried out, except that (R)-tert-butyl-3-hydroxypyrrolidin-1-carboxylate (10.0 g, 53.41 mmol) was used instead of tert-butyl-4-hydroxypiperidin-1-carboxylate, to obtain 14.4 g of the target compound (yield: 99%).

MS (ESI) m/z 266.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 5.28-5.26 (m, 1H), 3.68-3.50 (m, 4H), 3.05 (s, 3H), 2.27-2.16 (m, 2H), 1.47 (s, 9H).

Preparation Example 24: Synthesis of Intermediate 24 ((S)-tert-butyl 3-(methylsulfonyloxy)pyrrolidin-1-carboxylate)

The same procedure as in Preparation Example 20 was carried out, except that (S)-tert-butyl-3-hydroxypyrrolidin-1-carboxylate (10.0 g, 53.41 mmol) was used instead of tert-butyl-4-hydroxypiperidin-1-carboxylate, to obtain 14.0 g of the target compound (yield: 98.8%).

MS (ESI) m/z 266.1 [M+H] +

Preparation Example 25: Synthesis of Intermediate 25 (4-(5-chloro-4-nitro-1H-pyrazol-1-yl)piperidine hydrochloride)

4-Nitro-1H-pyrazole (4.71 g, 41.66 mmol) was dissolved in DMF (42 ml). NaH (2.7 g, 1.5 eq.) was added thereto, followed by stirring thereof at 0° C. for 30 minutes. Tert-butyl 4-(methylsulfonyloxy)piperidin-1-carboxylate (12.8 g, 1.1 eq. Intermediate 20) was added thereto, followed by elevation of the temperature to 150° C. and stirring thereof for 12 hours. Upon termination of the reaction, it was extracted three times with H 2 O and DCM, dried over MgSO 4 , and concentrated. It was subjected to column chromatography (MeOH:MC=1:40) to obtain 10.2 g of the target compound in yellow color (yield: 83%).

MS (ESI) m/z 297.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.17 (s, 1H), 8.09 (s, 1H), 4.34-4.26 (m, 3H), 2.20-2.16 (m, 3H), 1.98-1.89 (m, 2H), 1.49 (s, 9H).

Tert-butyl-4-(4-nitro-1H-pyrazol-1-yl)piperidin-1-carboxylate (800 mg, 2.70 mmol) was dissolved in THF (9.0 ml). After the temperature was lowered to −78° C., LiHMDS (1 M in THF, 5.4 mL, 2.0 eq.) was added thereto, followed by stirring thereof for 30 minutes. Cl 3 CCCl 13 (765 mg, 1.2 eq.) dissolved in THF (8.0 ml) was added thereto, followed by stirring thereof for 1 hour. Water was added thereto to terminate the reaction. Ethyl acetate was added thereto three times for extraction, it was dried over MgSO 4 and concentrated to obtain 500 mg of the target compound in brown color (yield: 56%).

MS (ESI) m/z 331.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 7.28 (s, 1H), 4.54-4046 (m, 1H), 4.32 (d, J=12.3 Hz, 2H), 2.91 (t, J=12.6 Hz, 2H), 2.20-2.07 (m, 2H), 1.94 (d, J=12.3 Hz, 2H), 1.49 (s, 9H).

Tert-butyl-4-(5-chloro-4-nitro-1H-pyrazol-1-yl)piperidin-1-carboxylate (500 mg, 1.51 mmol) was dissolved in DCM (5 ml). HCl (4 M in dioxane, 1.13 ml, 3.0 eq.) was added thereto, followed by stirring thereof for 12 hours. Upon termination of the reaction, it was filtered to obtain 120 mg of the target compound in white color (yield: 34%).

MS (ESI) m/z 230.7 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 4.49-4.41 (m, 1H), 3.29 (d, J=12.9 Hz, 2H), 2.79 (t, J=10.8 Hz, 2H), 2.17-2.03 (m, 2H), 1.94 (d, J=11.7 Hz, 2H).

Preparation Example 26: Synthesis of Intermediate 26 (1-(4-(4-amino-5-chloro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone)

4-(5-Chloro-4-nitro-1H-pyrazol-1-yl)piperidine hydrochloride (120 mg, 0.52 mmol, Intermediate 25) was added to THF (1.0 ml). K 2 CO 3 (108 mg, 1.5 eq.) was added thereto, and acetyl chloride (0.04 ml, 1.1 eq.) was added thereto dropwise, followed by stirring thereof for 12 hours. Upon termination of the reaction, it was extracted with water and DCM and concentrated to obtain 110 mg of the target compound in white color (yield: 78%).

MS (ESI) m/z 273.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 4.82-4.78 (m, 1H), 4.63-4.53 (m, 1H), 4.07-4.01 (m, 1H), 3.44-3.27 (m, 2H), 2.83-2.75 (m, 1H), 2.16 (s, 3H), 2.06 (s, 3H).

1-(4-(5-chloro-4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone (110 mg, 0.403 mmol) was dissolved in EtOH (5 ml) and water (0.5 ml). NH 4 Cl (64 mg, 3.0 eq.) and Fe (67 mg, 3.0 eq.) were added thereto, followed by stirring thereof at 90° C. for 12 hours. Upon termination of the reaction, Fe was removed, and it was concentrated. It was extracted with water and ethyl acetate, dried, and concentrated to obtain 90 mg of the target compound in red color (yield: 92%).

MS (ESI) m/z 243.1 [M+H] + ;

1 H NMR (300 MHz. CDCl 3 ) δ 7.23 (s, 1H), 4.75-4.70 (m, 1H), 4.37-4.32 (m, 1H), 4.00-3.95 (m, 1H), 3.43-2.73 (m, 4H), 2.13 (s, 3H), 2.05-1.98 (m, 2H).

Preparation Example 27: Synthesis of Intermediate 27 (5-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 26 was carried out, except that methanesulfonyl chloride was used instead of acetyl chloride, to obtain 0.23 g of the target compound (yield: 73%).

MS (ESI) m/z 278.7, 280.4 [M+H] + ;

1 H NMR (300 MHz, DMSO-d 6 ) δ 7.21 (s, 1H) 4.13-4.05 (m, 1H), 3.90 (s, 2H), 3.65-3.59 (m, 2H), 2.92 (s, 3H), 2.90-2.83 (m, 2H), 2.02-1.88 (m, 2H), 1.88-1.82 (m, 2H).

Preparation Example 28: Synthesis of Intermediate 28 (4-(4-nitro-1H-pyrazol-1-yl)piperidine hydrochloride)

4-Nitro-1H-pyrazole (4.71 g, 41.66 mmol) was dissolved in DMF (42 ml). NaH (2.7 g, 1.5 eq.) was added thereto, followed by stirring thereof at 0° C. for 30 minutes. Tert-butyl 4-(methylsulfonyloxy)piperidin-1-carboxylate (12.8 g, 1.1 eq. Intermediate 20) was added thereto, followed by elevation of the temperature to 150° C. and stirring thereof for 12 hours. Upon termination of the reaction, it was extracted three times with H 2 O and DCM. It was dried over MgSO 4 , concentrated, and subjected to column chromatography (MeOH:MC=1:40) to obtain 10.2 g of the target compound in yellow color (yield: 83%).

MS (ESI) m/z 297.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.17 (s, 1H), 8.09 (s, 1H), 4.34-4.26 (m, 3H), 2.20-2.16 (m, 3H), 1.98-1.89 (m, 2H), 1.49 (s, 9H).

Tert-butyl-4-(4-nitro-1H-pyrazol-1-yl)piperidin-1-carboxylate (10.2 g, 34.42 mmol) was dissolved in DCM (68 ml). HCl (4 M in dioxane, 25.8 ml, 3.0 eq.) was added thereto, followed by stirring thereof for 12 hours. Upon termination of the reaction, it was filtered to obtain 6.0 g of the target compound in white color (yield: 75%).

MS (ESI) m/z 197.2 [M+H] + ;

1 H NMR (300 MHz. MeOD): δ 8.67 (s, 1H), 8.18 (s, 1H), 4.70-4.61 (m, 1H), 3.58 (d, J=13.2 Hz, 2H), 3.27-3.18 (m, 2H), 2.41-2.32 (m, 4H).

Preparation Example 29: Synthesis of Intermediate 29 (5-chloro-1-(1-(oxetan-3-yl)piperidin-4-yl)-1H-pyrazol-4-amine)

4-(4-Nitro-1H-pyrazol-1-yl)piperidine hydrochloride (0.40 g, 1.74 mmol, Intermediate 28), dichloroethane (DCE, 15 ml), diisopropylethylamine (DIPEA, 0.44 g, 3.48 mmol), and oxetan-3-one (0.31 g, 4.35 mmol) were added and stirred for 15 minutes. NaBH(OAc) 3 (1.10 g, 5.22 mmol) and AcOH (0.12 g, 2.08 mmol) were added thereto, followed by stirring thereof for 2 hours. Upon termination of the reaction, it was immersed in NaHCO 3 and then extracted with DCM. After it was dried, the solution was evaporated under reduced pressure to obtain 0.42 g of the target compound (yield: 96%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 8.10 (s, 1H), 4.72-4.62 (m, 4H), 4.19-4.17 (d, 1H, J=4.5 Hz), 3.60-3.52 (m, 1H), 2.92-2.90 (d, 2H, J=5.4 Hz), 2.231 (s, 2H), 2.11-2.05 (m, 4H).

4-(4-Nitro-1H-pyrazol-1-yl)-1-(oxetan-3-yl)piperidine (0.32 g, 1.26 mmol) was added to THF (15 ml), followed by stirring thereof at −78° C. LiHMDS (1 M in THF, 2.53 ml, 2.53 mmol) was added thereto, followed by stirring thereof for 30 minutes. Hexachloroethane (0.36 g, 1.52 mmol) dissolved in THF (3 ml) was added thereto dropwise, followed by stirring thereof for 30 minutes. Upon termination of the reaction, it was immersed in NH 4 Cl and then extracted with ethyl acetate. It was dried over MgSO 4 and depressurized to obtain 0.26 g of the target compound (yield: 72%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.20 (s, 1H), 4.72-4.62 (m, 4H), 4.42-4.31 (m, 1H), 3.61-3.52 (m, 1H), 2.95-2.91 (m, 2H), 2.38-2.25 (m, 2H), 2.09-1.96 (m, 4H).

The same procedure as in step 2 of Preparation Example 26 was carried out, except that 4-(5-chloro-4-nitro-1H-pyrazol-1-yl)-1-(oxetan-3-yl)piperidine was used instead of 1-(4-(5-chloro-4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone, to obtain 1.0 g of the target compound (yield: 86%).

MS (ESI) m/z 257.3, 259.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.24 (s, 1H), 4.70-4.62 (m, 4H), 4.19-4.09 (m, 1H), 3.59-3.51 (m, 1H), 2.90 (s, 4H), 2.28-2.16 (m, 2H), 2.05-1.92 (m, 4H).

Preparation Example 30: Synthesis of Intermediate 30 (1-(azetidin-3-yl)-4-nitro-1H-pyrazole hydrochloride)

4-Nitro-1H-pyrazole (2.9 g, 25.77 mmol) was dissolved in DMF (30 ml). Cs 2 CO 3 (17.6 g, 2.0 eq.) and tert-butyl-3-(methylsulfonyloxy)azetidin-1-carboxylate (6.8 g, 1.05 eq., Intermediate 21) were added thereto, followed by stirring thereof at 90° C. for 12 hours. Water was added thereto to terminate the reaction. It was extracted three times with ethyl acetate, dried over MgSO 4 , and concentrated. It was subjected to column chromatography (MeOH:MC=1:40) to obtain 6.45 g of the target compound in yellow color (yield: 93%).

MS (ESI) m/z: 268.9 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.30 (s, 1H), 8.14 (s, 1H), 5.09-5.03 (m, 1H), 4.44-4.29 (m, 4H), 1.46 (s, 9H).

Tert-butyl-3-(4-nitro-1H-pyrazol-1-yl)azetidin-1-carboxylate (6.45 g, 24.05 mmol) was dissolved in DCM (50 ml). HCl (4 M in dioxane, 20 ml, 3.0 eq.) was added thereto, followed by stirring thereof for 12 hours. Upon termination of the reaction, it was filtered to obtain 4.5 g of the target compound in white color (yield: 91%).

MS (ESI) m/z 429.9 [M+H] + ;

1 H NMR (300 MHz, MeOD) δ 8.68 (s, 1H), 8.32 (s, 1H), 5.54-5.50 (m, 1H), 4.62-4.55 (m, 4H).

Preparation Example 31: Synthesis of Intermediate 31 (5-chloro-1-(1-(oxetan-3-yl)azetidin-3-yl)-1H-pyrazol-4-amine)

1-(Azetidin-3-yl)-4-nitro-1H-pyrazole hydrochloride (1.0 g, 4.89 mmol, Intermediate 30) was added to DCE (20 ml). DIPEA (1.7 ml, 2.0 eq.) and oxetan-3-one (881 mg, 2.5 eq.) were added thereto, followed by stirring thereof at room temperature for 15 minutes. NaBH(OAc) 3 (3.1 g, 3.0 eq.) and AcOH (0.336 ml, 1.2 eq.) were added thereto, followed by stirring thereof at room temperature for 1.5 hours. Upon termination of the reaction, it was extracted with NaHCO 3 and DCM and concentrated. It was then subjected to column chromatography (MeOH:MC=1:40) to obtain 540 mg of the target compound in yellow color (yield: 49%).

MS (ESI) m/z 225.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.36 (s, 1H), 8.12 (s, 1H), 5.02-4.98 (m, 1H), 4.78-4.57 (m, 4H), 3.93-3.68 (m, 5H).

The same procedure as in step 2 of Preparation Example 29 was carried out, except that 4-nitro-1-(1-(oxetan-3-yl)azetidin-3-yl)-1H-pyrazole was used instead of 4-(4-nitro-1H-pyrazol-1-yl)-1-(oxetan-3-yl)piperidine, to obtain 0.43 g of the target compound (yield: 69%).

MS (ESI) m/z 259.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.25 (s, 1H), 5.28-5.19 (m, 1H), 4.79-4.74 (m, 2H), 4.64-4.60 (m, 2H), 3.79-3.76 (m, 5H).

The same procedure as in step 2 of Preparation Example 26 was carried out, except that 5-chloro-4-nitro-1-(1-(oxetan-3-yl)azetidin-3-yl)-1H-pyrazole was used instead of 1-(4-(5-chloro-4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone, to obtain 0.1 g of the target compound (yield: 26%).

MS (ESI) m/z 229.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.30 (s, 1H), 5.08-5.03 (m, 1H), 4.77-4.73 (m, 2H), 4.64-4.60 (m, 2H), 3.96-3.69 (m, 5H).

Preparation Example 32: Synthesis of Intermediate 32 (3,5-dimethyl-4-nitro-1H-pyrazole)

The same procedure as in Preparation Example 12 was carried out, except that 3,5-dimethyl-1H-pyrazole was used instead of 3-(trifluoromethyl)-1H-pyrazole, to obtain 7.0 g of the target compound (yield: 96%).

1 H NMR (300 MHz, CDCl 3 ) δ 11.01 (bs, 1H), 2.62 (s, 6H).

Preparation Example 33: Synthesis of Intermediate 33 (3-chloro-5-methyl-4-nitro-1H-pyrazole)

Ethyl 3-oxobutanoate (28.59 g, 219.73 mmol) was added to EtOH (50 ml), followed by stirring thereof for 10 minutes in an ice bath. Hydrazine hydrate (10 g, 199.76 mmol) was added thereto, followed by refluxing thereof overnight. Upon termination of the reaction, it was evaporated under reduced pressure. Ethyl acetate was added thereto, followed by stirring and filtering thereof to obtain 14.68 g of the target compound in white color (yield: 75%).

1 H NMR (300 MHz, DMSO-d 6 ) δ 5.21 (s, 1H), 2.08 (s, 3H)

A high-pressure reactor was charged with 5-methyl-1H-pyrazol-3(2H)-one (14 g, 101.93 mmol) and POCl 3 (42.20 g, 275.22 mmol), followed by refluxing thereof at 160° C. overnight. Upon termination of the reaction, it was evaporated and neutralized with NaHCO 3 and 5 N NaOH. It was extracted with DCM, dried over MgSO 4 , and evaporated under reduced pressure. It was subjected to column chromatography (Hep:EA=2:1) to obtain 2.90 g of the target compound (yield: 17%).

MS (ESI) m/z 117.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 11.58 (bs, 1H), 6.01 (s, 1H), 2.37 (s, 3H).

The same procedure as in Preparation Example 12 was carried out, except that 3-chloro-5-methyl-1H-pyrazole was used instead of 3-(trifluoromethyl)-1H-pyrazole, to obtain 3.68 g of the target compound (yield: 910%).

1 H NMR (300 MHz, CDCl 3 ) δ 2.72 (s, 3H).

Preparation Example 34: Synthesis of Intermediate 34 (3-chloro-1-isopropyl-1H-pyrazol-4-amine)

4-Nitro-1H-pyrazole (0.30 g, 2.65 mmol) and DMF (30 ml) were added and stirred for 10 minutes in an ice bath. NaH (0.17 g, 3.97 mmol) was added thereto, followed by stirring thereof for 30 minutes. 2-Iodopropane (0.58 g, 3.44 mmol) was then added thereto, followed by refluxing thereof for 3 hours. Upon termination of the reaction, it was evaporated under reduced pressure and extracted with ethyl acetate. It was dried over MgSO 4 and evaporated under reduced pressure. It was subjected to column chromatography (Hep:EA=1:1) to obtain 0.26 g of the target compound in liquid phase (yield: 65%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.17 (s, 1H), 8.09 (s, 1H), 4.58-4.49 (m, 1H), 1.56 (s, 6H).

4-Nitro-3-(trifluoromethyl)-1H-pyrazole (0.26 g, 1.72 mmol), EtOH (20 ml) and HCl (1.43 ml, 17.20 mmol) were added and stirred under N 2 . After Pd/Al (0.026 g) was added thereto, triethylsilane (1.20 g, 10.35 mmol) was added thereto dropwise, followed by reaction thereof overnight. Upon termination of the reaction, it was filtered with celite-545. It was extracted with ethyl acetate and evaporated under reduced pressure to obtain 0.17 g of the target compound (yield: 50%).

MS (ESI) m/z 160.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.59 (s, 1H), 4.67-4.58 (m, 1H), 1.38 (s, 3H), 1.36 (s, 3H).

Preparation Example 35: Synthesis of Intermediate 35 (3-chloro-1-cyclopropylmethyl-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 34 was carried out, except that (bromomethyl)cyclopropane was used instead of 2-iodopropane, to obtain 0.66 g of the target compound (yield: 88%).

MS (ESI) m/z 171.9, 174 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.78 (s, 1H), 3.99 (s, 2H), 1.27-1.23 (m, 1H), 0.62-0.60 (m, 2H), 0.41 (s, 2H).

Preparation Example 36: Synthesis of Intermediate 36 (3-chloro-1-cyclopropyl-1H-pyrazol-4-amine)

A suction flask was charged with 4-nitro-1H-pyrazole (0.5 g, 4.42 mmol), cyclopropylboronic acid (0.75 g, 8.84 mmol), 2,2′-bipyridine (0.69 g, 4.42 mmol), and Na 2 CO 3 (0.93 g, 8.84 mmol). Dichloroethane (22 ml) was added thereto, and nitrogen (N 2 ) was bubbled for 10 minutes. Cu(OAc) 2 (0.80 g, 4.42 mmol) was added thereto, and N 2 was bubbled for 1 minute, followed by stirring thereof at 70° C. for 18 hours. The temperature was lowered to room temperature, and it was filtered with ethyl acetate and then concentrated. It was extracted with ethyl acetate and a 1 N aqueous hydrochloric acid solution, dried over MgSO 4 , filtered under reduced pressure, and concentrated to obtain 0.67 g of the target compound (yield: 45%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.20 (s, 1H), 3.69 (p, J=1.8 Hz, 1H), 1.27-1.16 (m, 4H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 1-cyclopropyl-4-nitro-1H-pyrazole (0.3 g, 1.95 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 0.14 g of the target compound (yield: 47%).

MS (ESI) m/z 158.0, 160.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.16 (s, 1H), 4.37 (p, J=3.9 Hz, 1H), 2.89 (s, 2H), 1.18-1.14 (m, 2H), 1.07-1.04 (m, 2H).

Preparation Example 37: Synthesis of Intermediate 37 (3-chloro-1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-amine)

4-(4-Nitro-1H-pyrazol-1-yl)piperidine hydrochloride (0.4 g, 1.71 mmol) was dissolved in DMF (5 ml), and the temperature was lowered to 0° C. NaH (0.30 g, 6.87 mmol) was slowly added thereto, followed by stirring thereof for 30 minutes. Iodomethane (0.16 ml, 2.57 mmol) was added thereto dropwise, followed by stirring thereof at room temperature for 1 hour. Water was added thereto to terminate the reaction, and it was extracted with ethyl acetate. It was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.16 g of the target compound (yield: 45%).

MS (ESI) m/z 211.1 [M+H] + ;

1 H NMR (300 MHz, Acetone-d 6 ) δ 8.64 (s, 1H), 8.12 (s, 1H), 4.26 (p J=7.2 Hz, 1H), 2.95-2.91 (m, 2H), 2.25 (s, 3H), 2.12-2.05 (m, 6H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 1-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine (0.16 g, 0.76 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 0.18 g of the target compound (yield: 95%).

MS (ESI) m/z 215.0, 217.0 [M+H] +

Preparation Example 38: Synthesis of Intermediate 38 (3-chloro-1-(1-ethylpiperidin-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 37 was carried out, except that iodoethane was used instead of iodomethane, to obtain 0.05 g of the target compound (yield: 21%).

MS (ESI) m/z 229.1 [M+H] +

Preparation Example 39: Synthesis of Intermediate 39 (3-chloro-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine)

4-Nitro-1H-pyrazole (0.08 g, 0.76 mmol), DMF (10 ml), and Cs 2 CO 3 (0.37 g, 1.14 mmol) were added and stirred for 30 minutes. Tetrahydro-2H-pyran-4-yl methanesulfonate (0.20 g, 1.314 mmol) was added thereto, followed by refluxing thereof for 2 hours. Upon termination of the reaction, it was evaporated under reduced pressure and extracted with ethyl acetate. It was dried over MgSO 4 and evaporated under reduced pressure. It was subjected to column chromatography (MC:MeOH=10:1) to obtain 0.085 g of the target compound (yield: 57%).

1 H NMR (300 MHz, CDCl 3 ) δ 8.19 (s, 1H), 8.11 (s, 1H), 4.44-4.34 (m, 1H), 4.18-4.13 (m, 2H), 3.61-3.52 (m, 2H), 2.20-2.01 (m, 4H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 4-nitro-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 0.04 g of the target compound (yield: 99%).

MS (ESI) m/z 202.0 [M+H] +

Preparation Example 40: Synthesis of Intermediate 40 (3-chloro-1-(2,6-dimethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 39 was carried out, except that 2,6-dimethyltetrahydro-2H-pyran-4-yl methanesulfonate was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.09 g of the target compound (yield: 71%).

MS (ESI) m/z 230.1 [M+H] +

Preparation Example 41: Synthesis of Intermediate 41 (3-chloro-1-(4-fluorophenyl)-1H-pyrazol-4-amine)

4-Nitro-1H-pyrazole (1 g, 8.84 mmol), 4-fluorophenylboronic acid (2.47 g, 17.68 mmol), and Cu(OAc) 2 (2.41 g, 13.27 mmol) were added to DCM (40 ml). Pyridine (2.85 ml, 35.37 mmol) was added thereto, followed by stirring thereof at room temperature for 20 hours. Upon termination of the reaction, it was filtered and extracted with DCM and water. The organic layer was dried over MgSO 4 , filtered, and concentrated under reduced pressure. It was subjected to column chromatography to obtain 0.82 g of the target compound (yield: 45%).

MS (ESI) m/z 208.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 9.62 (s, 1H), 8.55 (s, 1H), 8.01-7.98 (m, 2H), 7.45-7.41 (m, 2H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 1-(4-fluorophenyl)-4-nitro-1H-pyrazole (0.82 g, 3.95 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 0.23 g of the target compound (yield: 27%).

MS (ESI) m/z 212.2, 214.1 [M+H] +

Preparation Example 42: Synthesis of Intermediate 42 ((4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone)

4-(4-Nitro-1H-pyrazol-1-yl)piperidine hydrochloride (0.25 g, 1.07 mmol, Intermediate 28) was dissolved in DCM. Triethylamine (0.37 ml, 2.68 mmol) was added thereto, and the temperature was lowered to 0° C. Cyclopropanecarbonyl chloride (0.11 ml, 1.28 mmol) was slowly added thereto dropwise, followed by stirring thereof at room temperature for 2 hours. Water was added thereto to terminate the reaction, and it was extracted with ethyl acetate. The organic layer was dried over MgSO 4 , filtered under reduced pressure, and concentrated. It was subjected to column chromatography to obtain 0.28 g of the target compound (yield: 99%).

MS (ESI) m/z 265.1 [M+H] + ;

1 H NMR (300 MHz, Acetone-d 6 ) δ 8.69 (s, 1H), 8.14 (s, 1H), 4.65 (m, 3H), 3.37 (s, 1H), 2.78-2.75 (m, 2H), 2.27-2.25 (m, 2H), 2.02-1.97 (m, 2H), 0.84-0.82 (m, 2H), 0.81-0.78 (m, 2H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that cyclopropyl(4-(4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)methanone (0.28 g, 1.07 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 0.10 g of the target compound (yield: 34%).

MS (ESI) m/z 269.1, 271.0 [M+H] + ;

1 H NMR (300 MHz, Acetone-d 6 ) δ 7.14 (s, 1H), 4.58-4.42 (m, 3H), 3.63 (s, 2H), 3.37 (m, 1H), 2.02-1.97 (m, 6H), 0.83-0.82 (m, 2H), 0.75-0.70 (m, 2H).

Preparation Example 43: Synthesis of Intermediate 43 (1-(4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)-2-methylpropane-1-one)

The same procedure as in Preparation Example 42 was carried out, except that isobutyryl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.35 g of the target compound (yield: 100%).

MS (ESI) m/z 271.1, 273.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.24 (s, 1H), 4.79-4.76 (d, 1H, J=11.4 Hz), 4.41-4.31 (m, 1H), 4.13-4.09 (d, 1H, J=12.6 Hz), 3.25-3.17 (t, 1H, J=24.6 Hz), 2.97-2.70 (m, 5H), 2.19-1.99 (m, 4H). 1.17-1.15 (d, 7H, J=6 Hz).

Preparation Example 44: Synthesis of Intermediate 44 (1-(4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)-2,2-dimethylpropane-1-one)

The same procedure as in Preparation Example 42 was carried out, except that pivaloyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.15 g of the target compound (yield: 22%).

MS (ESI) m/z 285.1, 287.0 [M+H] +

Preparation Example 45: Synthesis of Intermediate 45 ((4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)(morpholino)methanone)

The same procedure as in Preparation Example 42 was carried out, except that 4-morpholinecarbonyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.12 g of the target compound (yield: 48%).

MS (ESI) m/z 314.1, 316.0 [M+H] +

Preparation Example 46: Synthesis of Intermediate 46 ((4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)(4-methylpiperazin-1-yl)methanone)

The same procedure as in Preparation Example 42 was carried out, except that 4-methyl-1-piperazincarbonyl chloride hydrochloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.35 g of the target compound (yield: 95%).

MS (ESI) m/z 327.1, 329.0 [M+H] +

Preparation Example 47: Synthesis of Intermediate 47 (3-chloro-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 42 was carried out, except that methanesulfonyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 5.4 g of the target compound (yield: 53%).

MS (ESI) m/z 278.9 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.24 (s, 1H), 4.31-4.24 (m, 1H), 3.95-3.89 (m, 2H), 3.02-2.92 (m, 2H), 2.86 (s, 3H), 2.31-2.18 (m, 2H), 2.08-2.02 (m, 2H).

Preparation Example 48: Synthesis of Intermediate 48 (3-chloro-1-(1-(ethylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 42 was carried out, except that ethanesulfonyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.58 g of the target compound (yield: 46%).

MS (ESI) m/z 293.2, 295.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.11 (s, 1H), 4.29 (p, J=6.9 Hz, 1H), 4.00 (s, 2H), 3.72-3.66 (m, 2H), 3.08 (q, J=7.2 Hz, 2H), 1.94-1.86 (m, 4H), 1.22 (t, J=7.2 Hz, 3H).

Preparation Example 49: Synthesis of Intermediate 49 (3-chloro-1-(1-(cyclopropylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 42 was carried out, except that cyclopropylsulfonyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.56 g of the target compound (yield: 46%).

MS (ESI) m/z 305.1, 307.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.24 (s, 1H), 4.33-4.23 (m, 1H), 3.98-3.94 (m, 2H), 3.11-3.06 (m, 2H), 2.92 (s, 2H), 2.35-2.18 (m, 3H), 2.09-1.98 (m, 2H), 1.24-1.18 (m, 2H), 1.06-0.98 (m, 2H).

Preparation Example 50: Synthesis of Intermediate 50 (1-(4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)2,2,2-trifluoroethanone)

4-(5-Chloro-4-nitro-1H-pyrazol-1-yl)piperidine hydrochloride (500 mg, 2.15 mmol) was added to DCM (5 ml). TEA (0.66 ml, 2.2 eq.) and 2,2,2-trifluoroacetic anhydride (0.45 ml, 1.5 eq.) were added thereto, followed by stirring thereof at room temperature for 12 hours. Upon termination of the reaction, it was extracted with water and DCM and concentrated to obtain 680 mg of the target compound in white color (yield: 98%).

1 H NMR (300 MHz. CDCl 3 ) δ 8.21 (s, 1H), 8.10 (s, 1H), 4.70-4.64 (m, 1H), 4.52-4.43 (m, 1H), 4.21-4.17 (m, 1H), 3.42-3.32 (m, 1H), 3.11-3.03 (m, 1H), 2.34-2.30 (m, 2H), 2.16-2.07 (m, 2H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 2,2,2-trifluoro-1-(4-(4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone (680 mg, 2.33 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 200 mg of the target compound (yield: 29%).

MS (ESI) m/z: 297.1 [M+H] + ;

1 H NMR (300 MHz. CDCl 3 ) δ 7.24 (s, 1H), 4.61-4.57 (m, 1H), 4.48-4.40 (m, 1H), 4.19-4.15 (m, 1H), 3.40-3.30 (m, 1H), 3.13-3.04 (m, 1H), 2.93 (s, 2H), 2.22-2.06 (m, 4H).

Preparation Example 51: Synthesis of Intermediate 51 (3-chloro-1-(1-(methylsulfonyl)azetidin-3-yl)-1H-pyrazol-4-amine)

1-(Azetidin-3-yl)-4-nitro-1H-pyrazole hydrochloride (500 mg, 2.44 mmol, Intermediate 30) was added to DCM (12 ml). TEA (1.4 ml, 3.0 eq.) and methanesulfonyl chloride (0.266 ml, 1.2 eq.) were added thereto dropwise, followed by stirring thereof at room temperature for 1 hour. Upon termination of the reaction, it was extracted with water and DCM and concentrated to obtain 500 mg of the target compound in yellow color (yield: 83%).

MS (ESI) m/z 247.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.31 (s, 1H), 8.19 (s, 1H), 5.16-5.12 (m, 1H), 4.48-4.41 (m, 4H), 3.03 (s, 3H).

The same procedure as in step 2 of Preparation Example 34 was carried out, except that 21-(1-(methylsulfonyl)azetidin-3-yl)-4-nitro-1H-pyrazole (500 mg, 2.03 mmol) was used instead of 4-nitro-3-(trifluoromethyl)-1H-pyrazole, to obtain 240 mg of the target compound (yield: 47%).

MS (ESI) m/z 250.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.33 (s, 1H), 5.16-5.11 (m, 1H), 4.46-4.30 (m, 4H), 3.02 (s, 3H).

Preparation Example 52: Synthesis of Intermediate 52 ((3-(4-amino-3-chloro-1H-pyrazol-1-yl)azetidin-1-yl)(cyclopropyl)methanone)

The same procedure as in Preparation Example 51 was carried out, except that cyclopropanecarbonyl chloride was used instead of methanesulfonyl chloride, to obtain 0.21 g of the target compound (yield: 41%).

MS (ESI) m/z: 241.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.31 (s, 1H), 4.78-4.66 (m, 4H), 3.21-3.06 (m, 1H), 1.00-0.76 (m, 4H).

Preparation Example 53: Synthesis of Intermediate 53 (4-(4-nitro-1H-pyrazol-1-yl)azepane hydrochloride)

4-Nitro-1H-pyrazole (0.946 g, 8.37 mmol) was dissolved in DMF (9 ml). NaH (0.548 g, 1.5 eq.) was added thereto, followed by stirring thereof at 0° C. for 30 minutes. Tert-butyl 4-(methylsulfonyloxy)azepan-1-carboxylate (2.7 g, 1.1 eq.) was added thereto, followed by elevation of the temperature to 150° C. and stirring thereof for 12 hours. Upon termination of the reaction, it was extracted three times with H 2 O and DCM. It was dried over MgSO 4 , concentrated, and then subjected to column chromatography (MeOH:MC=1:40) to obtain 2.2 g of the target compound in yellow color (yield: 85%).

MS (ESI) m/z 311.3 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.15 (s, 1H), 8.07 (s, 1H), 4.33-4.26 (m, 1H), 3.78-3.33 (m, 4H), 2.29-1.72 (m, 6H), 1.49 (s, 9H).

Tert-butyl-4-(4-nitro-1H-pyrazol-1-yl)azepan-1-carboxylate (2.2 g, 7.09 mmol) was dissolved in DCM (14 ml). HCl (4 M in dioxane, 5.32 ml, 3.0 eq.) was added thereto, followed by stirring thereof for 12 hours. Upon termination of the reaction, it was filtered to obtain 1.4 g of the target compound in white color (yield: 80%).

MS (ESI) m/z 211.2 [M+H] + ;

1 H NMR (300 MHz, MeOD) δ 11.05 (s, 1H), 10.56 (s, 1H), 7.13-7.08 (m, 1H), 5.77-5.71 (m, 5H), 4.88-4.35 (m, 6H).

Preparation Example 54: Synthesis of Intermediate 54 ((R)-4-nitro-1-(pyrrolidin-3-yl)-1H-pyrazole hydrochloride)

The same procedure as in Preparation Example 53 was carried out, except that (R)-tert-butyl-3-hydroxypyrrolidin-1-carboxylate was used instead of tert-butyl-4-(methylsulfonyloxy)azepan-1-carboxylate, to obtain 3.5 g of the target compound (yield: 90%).

MS (ESI) m/z 183.3 [M+H] + ;

1 H NMR (300 MHz, MeOD) δ 8.73 (s, 1H), 8.21 (s, 1H), 5.39-5.32 (m, 1H), 3.80-3.53 (m, 4H), 2.69-2.47 (m, 2H).

Preparation Example 55: Synthesis of Intermediate 55 ((S)-4-nitro-1-(pyrrolidin-3-yl)-1H-pyrazole hydrochloride)

The same procedure as in Preparation Example 53 was carried out, except that (S)-tert-butyl-3-hydroxypyrrolidin-1-carboxylate was used instead of tert-butyl-4-(methylsulfonyloxy)azepan-1-carboxylate, to obtain 3.0 g of the target compound (yield: 77.4%).

MS (ESI) m/z 183.3 [M+H] +

Preparation Example 56: Synthesis of Intermediate 56 (3-chloro-1-(1-(methylsulfonyl)azepan-4-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 53 was carried out, except that 3-chloro-4-nitro-1H-pyrazole and tert-butyl-4-(methylsulfonyl)azepan-1-carboxylate were used instead of 4-nitro-1H-pyrazole and tert-butyl-4-(methylsulfonyloxy)azepan-1-carboxylate, to obtain 0.05 g of the target compound (yield: 11%).

MS (ESI) m/z 292.9 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.28 (s, 1H), 4.48-4.44 (m, 1H), 3.70-3.50 (m, 3H), 3.26-3.18 (m, 1H), 2.86 (s, 3H), 3.31-1.99 (m, 6H).

Preparation Example 57: Synthesis of Intermediate 57 ((4-(4-amino-3-chloro-1H-pyrazol-1-yl)azepan-1-yl)(cyclopropyl)methanone)

The same procedure as in Preparation Example 53 was carried out, except that 3-chloro-4-nitro-1H-pyrazole and tert-butyl-4-(cyclopropanecarbonyl)azepan-1-carboxylate were used instead of 4-nitro-1H-pyrazole and tert-butyl-4-(methylsulfonyloxy)azepan-1-carboxylate, to obtain 0.22 g of the target compound (yield: 53%).

MS (ESI) m/z 282.9 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.21 (s, 1H), 4.38-4.26 (m, 1H), 3.92-3.56 (m, 4H), 2.92 (s, 2H), 2.20-1.70 (m, 7H), 1.08-0.76 (m, 4H).

Preparation Example 58: Synthesis of Intermediate 58 ((R)-3-chloro-1-(1-(methylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 39 was carried out, except that (R)-1-(methylsulfonyl)pyrrolidin-3-yl methanesulfonate was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.22 g of the target compound (yield: 36%).

MS (ESI) m/z 264.9 [M+H] + ;

1 H NMR (300 MHz, MeOD) δ 7.25 (s, 1H), 5.11-5.07 (m, 1H), 4.82-3.50 (m, 4H), 2.90 (s, 3H), 2.45-2.35 (m, 2H).

Preparation Example 59: Synthesis of Intermediate 59 ((S)-3-chloro-1-(1-(methylsulfonyl)pyrrolidin-3-yl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 39 was carried out, except that (S)-1-(methylsulfonyl)pyrrolidin-3-yl methanesulfonate was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.2 g of the target compound (yield: 32.8%).

MS (ESI) m/z 264.9 [M+H] + ;

1 H NMR (300 MHz, CD 3 OD) δ 7.25 (s, TH), 5.11-5.07 (m, 1H), 3.81-3.52 (m, 4H), 2.90 (s, 3H), 2.39-2.36 (m, 2H).

Preparation Example 60: Synthesis of Intermediate 60 ((R)-(3-(4-amino-3-chloro-1H-pyrazol-1-yl)pyrrolidin-1-yl)(cyclopentyl)methanone)

The same procedure as in Preparation Example 39 was carried out, except that (R)-1-(cyclopentanecarbonyl)pyrrolidin-3-yl methanesulfonate was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.28 g of the target compound (yield: 46%).

MS (ESI) m/z 283.0 [M+H] +

Preparation Example 61: Synthesis of Intermediate 61 ((S)-(3-(4-amino-3-chloro-1H-pyrazol-1-yl)pyrrolidin-1-yl)(cyclopentyl)methanone)

The same procedure as in Preparation Example 39 was carried out, except that (S)-1-(cyclopentanecarbonyl)pyrrolidin-3-yl methanesulfonate was used instead of tetrahydro-2H-pyran-4-yl methanesulfonate, to obtain 0.27 g of the target compound (yield: 44.2%).

MS (ESI) m/z 283.0 [M+H] + ;

1 H NMR (300 MHz, CD 3 OD) δ 7.23 (s, 1H), 5.14-5.09 (m, 1H), 3.85-3.53 (m, 4H), 3.02-2.92 (m, 1H), 2.90 (s, 3H), 2.45-2.31 (m, 2H), 1.92-1.61 (m, 8H).

Preparation Example 62: Synthesis of Intermediate 62 ((4-(4-amino-3-chloro-5-methyl-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone)

3-Chloro-5-methyl-4-nitro-1H-pyrazole (1.84 g, 11.44 mmol, Intermediate 33) was dissolved in DMF (15 ml), which was stirred for 10 minutes in an ice bath. NaH (0.99 g, 22.89 mmol) was added thereto, followed by stirring thereof for 30 minutes. Tert-butyl 4-(methylsulfonyloxy)piperidin-1-carboxylate (4.79 g, 17.16 mmol) was added thereto, followed by stirring thereof at room temperature for 12 hours. Upon termination of the reaction, the solution was evaporated under reduced pressure, and it was extracted with ethyl acetate. It was dried over MgSO 4 and evaporated under reduced pressure. The filtrate was recrystallized with isopropyl ether to obtain 1.2 g of the target compound (yield: 30%).

Tert-butyl 4-(3-chloro-5-methyl-4-nitro-1H-pyrazol-1-yl)piperidin-1-carboxylate (0.90 g, 2.61 mmol) was added to DCM (dichloromethane, 30 ml). TFA (trifluoroacetic acid, 2.97 g, 26.10 mmol) was added thereto, followed by stirring thereof for 12 hours under reflux conditions. Upon termination of the reaction, the solution was evaporated under reduced pressure. Isopropyl ether was added to the residue, which was stirred to obtain 0.92 g of the target compound (yield: 98%).

Step 3: Synthesis of (4-(3-chloro-5-methyl-4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone)

4-(3-Chloro-5-methyl-4-nitro-1H-pyrazol-1-yl)piperidine (0.50 g, 1.39 mmol) was added to DCM (30 ml). TEA (0.42 g, 4.18 mmol) and cyclopropanecarbonyl chloride (0.21 g, 2.09 mmol) were added thereto, followed by stirring thereof at room temperature for 1 hour. Upon termination of the reaction, water was added for extraction, which was dried over MgSO 4 . The solution was evaporated under reduced pressure, and it was subjected to column chromatography (DCM:MeOH=10:1) to obtain 0.2 g of the target compound (yield: 46%).

(4-(3-Chloro-5-methyl-4-nitro-1H-pyrazol-1-yl)piperidin-1-yl)(cyclopropyl)methanone) (0.2 g, 0.63 mmol) was added to ethanol and water (20 ml, 3:1 v/v). NH 4 Cl (0.034 g, 0.63 mmol) and Fe (0.35 g, 6.39 mmol) were added thereto, followed by stirring at 70° C. for 1 hour. Upon termination of the reaction, it was filtered, and the solution was evaporated under reduced pressure. The filtrate was extracted again with ethyl acetate, dried over MgSO 4 , and the solution was evaporated under reduced pressure to obtain 0.14 g of the target compound in liquid phase (yield: 90%).

MS (ESI) m/z 283.1, 285.0 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.73-4.60 (m, 1H, 4.48-4.41 (m, 1H), 4.17-4.08 (m, 1H), 3.24 (s, 1H), 2.68 (s, 2H), 2.23 (s, 3H), 1.93-1.84 (m, 2H), 1.81-1.73 (m, 1H), 1.00 (s, 2H), 0.80-0.76 (m, 2H).

Preparation Example 63: Synthesis of Intermediate 63 (3,5-dimethyl-1-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazol-4-amine)

3,5-Dimethyl-4-nitro-1H-pyrazole (0.46 g, 3.25 mmol) was dissolved in DMF (4 ml). Tert-butyl 4-(methylsulfonyloxy)piperidin-1-carboxylate (1 g, 3.57 mmol) and Cs 2 CO 3 (2.12 g, 6.52 mmol) were added thereto, followed by stirring thereof at 80° C. for 15 hours. Upon termination of the reaction, it was extracted with ethyl acetate and dried over MgSO 4 . After the solution was evaporated under reduced pressure, ethyl acetate/heptane (1:3) was added to the residue to obtain 0.48 g of the target compound (yield: 46%).

MS (ESI) m/z 325.2 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.38-4.25 (m, 2H), 4.22-4.11 (m, 1H), 2.93-2.83 (m, 2H), 2.67 (s, 3H), 2.52 (s, 3H), 2.24-2.12 (m, 2H), 1.86-1.81 (m, 2H), 1.49 (s, 9H).

The same procedure as in step 3 of Preparation Example 25 was carried out to obtain 0.39 g of the target compound (yield: 99%).

MS (ESI) m/z 225.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 8.83 (br s, 2H), 4.68-4.59 (m, 1H), 3.44-3.37 (m, 2H), 3.10-3.00 (m, 2H), 2.62 (s, 3H), 2.41 (s, 3H), 2.27-2.12 (m, 2H), 2.02-1.98 (m, 2H).

The same procedure as in step 1 of Preparation Example 42 was carried out, except that methanesulfonyl chloride was used instead of cyclopropanecarbonyl chloride, to obtain 0.39 g of the target compound (yield: 97%).

MS (ESI) m/z 302.35 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.21-4.14 (m, 1H), 4.01-3.96 (m, 2H), 3.00-2.91 (m, 2H), 2.88 (s, 3H), 2.66 (s, 3H), 2.51 (s, 3H), 2.39-2.30 (m, 4H), 2.02-1.97 (m, 2H).

The same procedure as in step 3 of Preparation Example 17 was carried out, except that 4-(3,5-dimethyl-4-nitro-1H-pyrazol-1-yl)-1-(methylsulfonyl)piperidine was used instead of 5-chloro-1-methyl-4-nitro-1H-pyrazole, to obtain 0.21 g of the target compound (yield: 67%).

MS (ESI) m/z 273.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 4.05-3.89 (m, 3H), 2.96-2.91 (m, 2H), 2.86 (s, 3H), 2.30-2.24 (m, 2H), 2.17 (s, 6H), 1.99-1.94 (m, 2H).

Preparation Example 64: Synthesis of Intermediate 64 (3-chloro-1-(2-methoxyethyl)-1H-pyrazol-4-amine)

The same procedure as in Preparation Example 34 was carried out, except that 2-bromomethyl methyl ether was used instead of 2-iodopropane, to obtain 0.46 g of the target compound (yield: 59%).

MS (ESI) m/z 176.2, 178.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.25 (s, 1H), 4.23 (t, J=5.7 Hz, 2H), 3.74 (t, J=5.7 Hz, 2H), 3.35 (s, 3H), 2.85 (s, 2H).

Preparation Example 65: Synthesis of Intermediate 65 (1-(4-(4-amino-3-chloro-1H-pyrazol-1-yl)piperidin-1-yl)ethanone)

The same procedure as in Preparation Example 37 was carried out, except that acetyl chloride was used instead of iodomethane, to obtain 0.22 g of the target compound (yield: 29%).

MS (ESI) m/z 243.2, 245.1 [M+H] + ;

1 H NMR (300 MHz, CDCl 3 ) δ 7.24 (s, 1H), 4.77-4.74 (m, 1H), 4.39-4.30 (m, 1H), 4.01-3.97 (m, 1H), 3.28-3.19 (m, 1H), 2.92 (br s, 2H), 2.81-2.72 (m, 1H), 2.14 (s, 3H), 2.02-1.95 (m, 4H).

After the compound was dissolved in 100% DMSO at 10 mM, it was serially diluted to the range of 1 μM to 10 μM using biochemical LRRK2 assay buffer (50 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) pH 7.5, 10 mM MgCl 2 , 1 mM EGTA (1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid), 2 mM DTT (dithiothreitol), and 0.01% TWEEN-20 (Aldrich)). Purified LRRK2 (CARNA BIOSCIENCES) was added to a black U-bottom 96-well microtiter plate containing 6 μl of the serially diluted compound, followed by incubation at room temperature for 30 minutes. For the kinase reaction, ATP and a substrate solution specific urea-polypeptide (ULIGHT™-poly TK, PerkinElimer) were added. The reaction was carried out at room temperature for 1 hour, and it was detected with an detection solution (brand name: LANCE™) containing EDTA. A LANCE detection solution containing europium-labeled antibody (LRRK2 specific PT66) was added, followed by incubation at room temperature for 50 minutes, to terminate the kinase experiment. The phosphorylated substrate was detected by 665 nm emission measurement. IC 50 values were calculated by a non-linear regression analysis using a Prism program (PRISM™ software) with reference to the absence of a kinase inhibitor. The results are shown in Table 6 below.

As a comparative assay, a compound LRR2-IN-1, that known to be a reference compound for LRRK2 inhibitor, have also tested an inhibitory activity.

As shown in Table 6 above, the compounds of the present invention had an inhibitory activity to LRRK2 with an single digit nM IC 50 values and had superior to those of LRRK2-IN-1. Specifically, six tyrosine kinase panel tests were carried out on Compounds 25, 28, 30, 31, 32, 33, 35, 42, 43, 46, 47, 51, 52, 55, 57, 58, and 59 in order to evaluate the selectivity.

As a result, they selectively showed high inhibitory activity to LRRK2, indicating that the selectivity to the LRRK2 inhibitory activity was excellent.