TETRACYCLIC PYRIMIDINONE COMPOUND, PREPARATION METHOD THEREFOR, AND COMPOSITION AND USE THEREOF

Abstract

Disclosed is a tetracyclic pyrimidinone compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The compound having a structure represented by formula (I) is a new Lp-PLA2 inhibitor usable for treatment of neurodegenerative-related diseases, such as Alzheimer's disease (AD), glaucoma and age-related macular degeneration (AMD), or of cardiovascular diseases including atherosclerosis.

##STR00001##

Claims

1. A compound represented by formula (I): ##STR00037## or a pharmaceutically acceptable salt thereof, wherein: n.sub.1, n.sub.2, n.sub.3, and n.sub.4 are each independently 0, 1, or 2; R.sup.1 and R.sup.2 are each independently selected from: —H, hydroxyl, cyano, halogen, alkyl, deuteroalkyl, hydroxyalkyl, haloalkyl, cycloalkyl, and alkoxyl; X.sub.1, X.sub.2, and X.sub.3 are each independently selected from: alkylene, —O—, —S—, or —NR′—; R′ is selected from: —H, alkyl, deuteroalkyl, or cycloalkyl; Ar is arylene or heteroarylene; and hydrogen atoms in the arylene or heteroarylene are optionally substituted by one or more substituents that are each independently selected from: halogen, alkyl, haloalkyl, alkoxyl, haloalkoxyl, deuteroalkyl, deuteroalkoxyl, hydroxyl, hydroxyalkyl, cyano, amino, monoalkyl- or dialkyl-substituted amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl, or heteroaryl; Y is —H, halogen, alkyl, haloalkyl, haloalkoxyl, cycloalkyl, alkoxyl, deuteroalkyl, deuteroalkoxyl, —OAr′, —SAr′, —NH—Ar′, —NMe-Ar′, —NR″, or —R′″—Ar′; Ar′ is selected from aryl or heteroaryl; and hydrogen atoms in the aryl or heteroaryl are optionally substituted by one or more substituents that are each independently selected from: halogen, alkyl, haloalkyl, alkoxyl, hydroxyl, hydroxyalkyl, haloalkoxyl, deuteroalkyl, deuteroalkoxyl, cyano, amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl, or heteroaryl; R″ is alkyl; R′″ is alkylene; and Z is O or S; optionally, wherein: n.sub.1, n.sub.2, n.sub.3, and n.sub.4 are each independently 0, 1, or 2; R.sup.1 and R.sup.2 are each independently selected from: —H, hydroxyl, cyano, halogen, alkyl, deuteroalkyl, hydroxyalkyl, haloalkyl, cycloalkyl, and alkoxyl; X.sub.1, X.sub.2, and X.sub.3 are each independently selected from: alkylene, —O—, —S—, or —NR′—; R′ is selected from: —H, alkyl, deuteroalkyl, or cycloalkyl; Ar is arylene or heteroarylene; and hydrogen atoms in the arylene or heteroarylene are optionally substituted by one or more substituents that are each independently selected from: halogen, alkyl, haloalkyl, alkoxyl, haloalkoxyl, hydroxyl, hydroxyalkyl, cyano, amino, monoalkyl- or dialkyl-substituted amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl, or heteroaryl; Y is —H, halogen, alkyl, haloalkyl, haloalkoxyl, cycloalkyl, alkoxyl, —OAr′, —SAr′, —NH—Ar′, —NMe-Ar′, —NR″, or —R′″—Ar′; Ar′ is selected from aryl or heteroaryl; and hydrogen atoms in the aryl or heteroaryl are optionally substituted by one or more substituents that are each independently selected from: halogen, alkyl, haloalkyl, alkoxyl, hydroxyl, hydroxyalkyl, haloalkoxyl, cyano, amino, nitro, carboxyl, aldehyde, cycloalkyl, heterocyclyl, aryl, or heteroaryl; R″ is alkyl; R′″ is alkylene; and Z is O or S.

2. The compound of formula (I), or the pharmaceutically acceptable salt thereof according to claim 1, wherein halogen atoms in the “halogen”, “haloalkyl”, and “haloalkoxyl” are each independently selected from F, Cl, Br, or I; optionally, alkyls in the “alkyl”, “deuteroalkyl”, “deuteroalkoxyl”, “hydroxyalkyl”, “haloalkyl”, “haloalkoxyl”, “alkoxyl”, and “monoalkyl- or dialkyl-substituted amino” are each independently C.sub.1-C.sub.10 linear or branched alkyl; optionally, C.sub.1-C.sub.7 linear or branched alkyl; optionally, C.sub.1-C.sub.4 linear or branched alkyl; optionally, selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl, or 2,2,3-trimethylbutyl; optionally, the “alkylenes” are each independently C.sub.1-C.sub.10 linear or branched alkylene; optionally C.sub.1-C.sub.7 linear or branched alkylene; optionally, C.sub.1-C.sub.5 linear or branched alkylene; and optionally selected from methylene, ethylene, n-propylidene, isopropylidene, n-butylidene, isobutylidene, tert-butylidene, sec-butylidene, n-pentylidene, 1-methylbutylidene, 2-methylbutylidene, 3-methylbutylidene, isopentylidene, 1-ethylpropylidene, neopentylidene, n-hexylidene, 1-methylpentylidene, 2-methylpentylidene, 3-methylpentylidene, isohexylidene, 1,1-dimethylbutylidene, 2,2-dimethylbutylidene, 3,3-dimethylbutylidene, 1,2-dimethylbutylidene, 1,3-dimethylbutylidene, 2,3-dimethylbutylidene, 2-ethylbutylidene, n-heptylidene, 2-methylhexylidene, 3-methylhexylidene, 2,2-dimethylpentylidene, 3,3-dimethylpentylidene, 2,3-dimethylpentylidene, 2,4-dimethylpentylidene, 3-ethylpentylidene, or 2,2,3-trimethylbutylidene; optionally, the “cycloalkyl” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkyl, optionally C.sub.3-C.sub.7 monocyclic cycloalkyl, and optionally cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl; optionally, the “heterocyclyl” is 3- to 10-membered non-aromatic heterocyclic ring containing 1, 2, or 3 heteroatoms selected from N, O, and S on the ring, optionally, the heterocyclic ring is a 3- to 10-membered non-aromatic ring containing 1 or 2 heteroatoms selected from N and O on the ring; optionally, the heterocyclic ring is a 3- to 6-membered non-aromatic ring containing 1 or 2 heteroatoms selected from N and O on the ring; optionally, the heterocyclic ring is a 3- to 10-membered non-aromatic ring containing 1 or 2 heteroatoms selected from N and S on the ring; and optionally, the heterocyclic ring is a 3- to 6-membered non-aromatic ring containing 1 or 2 heteroatoms selected from N and S on the ring; optionally, the “aryl” is 6- to 10-membered aryl, optionally phenyl or naphthyl, and optionally phenyl, 1-naphthyl, or 2-naphthyl; optionally, the “arylene” is 6- to 10-membered arylene, and optionally phenylene or naphthylene; optionally, the “heteroaryl” is a 5- to 10-membered heteroaromatic ring containing 1 to 3 heteroatoms selected from N, O, and S on the ring, and optionally a 5- to 10-membered heteroaromatic ring containing 1 to 2 heteroatoms selected from N, O, and S on the ring; optionally, the heteroaromatic ring is selected from a pyridine ring, a pyrrole ring, a pyrimidine ring, a pyrazine ring, a pyridazine ring, a thiophene ring, and a furan ring; optionally, the heteroaryl is selected from pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyridazin-3-yl, pyridazin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrazin-2-yl, pyrazin-3-yl, indolyl, isoindolyl, indazolyl, indolizinyl, purinyl, quinolizinyl, quinolinyl, isoquinolinyl, cinolinyl, phthalazinyl, naphthyridinyl, quinazolinyl, quinoxalinyl, thieno[2,3-b]furyl, furo[3,2-b]-pyranyl, pyrido[2,3-d]oxazinyl, pyrazolo[4,3-d]oxazolyl, imidazo[4,5-d]thiazolyl, pyrazino[2,3-d]pyridazinyl, imidazo[2,1-b]thiazolyl, imidazo[],2-b][1,2,4]triazinyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzoxepinyl, benzoxazinyl, benzofuranyl, benzotriazolyl, pyrrolo[2,3-b]pyridyl, pyrrolo[3,2-c]pyridyl, pyrrolo[3,2-b]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrazolo[4,3-d]pyridyl, pyrazolo[4,3-c]pyridyl, pyrazolo[3,4-c]pyridyl, pyrazolo[3,4-d]pyridyl, pyrazolo[3,4-b]pyridyl, imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrrolo[1,2-b]pyridazinyl, imidazo[1,2-c]pyrimidinyl, pyrido[3,2-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[2,3-b]pyrazinyl, pyrido[3,4-b]pyrazinyl, pyrimido[5,4-d]pyrimidinyl, pyrazolo[2,3-b]pyrazinyl, and pyrimido[4,5-d]pyrimidinyl, and optionally pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, or pyrimidin-5-yl; and optionally, the “heteroarylene” is a 5- to 10-membered heteroarylene ring containing 1 to 3 heteroatoms selected from N, O, and S on the ring, and optionally a 5- to 10-membered heteroarylene ring containing 1 to 2 heteroatoms selected from N, O, and S on the ring; and optionally, the heteroarylene ring is selected from a pyridylidene ring, a pyrrylidene ring, a pyrimidylidene ring, a pyrazinylidene ring, a pyridazinylidene ring, a thienylidene ring, a furylidene ring.

3. The compound of formula (I), or the pharmaceutically acceptable salt thereof according to claim 1, wherein n.sub.1, n.sub.2, n.sub.3, and n.sub.4 are each independently 0, 1, or 2; optionally, n.sub.1 is 0, optionally, n.sub.2 is 0 or 1, optionally, n.sub.3 is 0, optionally, n.sub.4 is 1; optionally, R.sub.1, R.sub.2 are each independently selected from: —H, F, Cl, Br, hydroxyl, cyano, C.sub.1-C.sub.7 alkyl (methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl, or 2,2,3-trimethylbutyl), C.sub.1-C.sub.3 deuteroalkyl(—CD.sub.3, —C.sub.2D.sub.5, —C.sub.3D.sub.7), cyclopropanyl, cyclobutanyl, and cyclopentyl; optionally, R.sub.1 is —H, and optionally, R.sub.2 is —H; optionally, X.sub.1, X.sub.2, and X.sub.3 are each independently selected from: C.sub.1-C.sub.7 alkylene (optionally, —CH.sub.2—, ethylene, n-propylidene, isopropylidene, n-butylidene, or isobutylidene), —O—, —S—, or —NR′—; optionally, X.sub.1 is —CH.sub.2—; optionally, X.sub.2 is selected from —CH.sub.2— or —O—; optionally, X.sub.3 is —O—; optionally, R′ is selected from —H, C.sub.1-C.sub.7 alkyl (methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl, or 2,2,3-trimethylbutyl), deuteroalkyl (optionally —CD.sub.3, —C.sub.2D.sub.5, —C.sub.3D.sub.7), or C.sub.3-C.sub.6 cycloalkyl (cyclopropanyl, cyclobutanyl, and cyclopentyl, cyclohexyl); optionally, Ar is phenylene or pyridyl, and a hydrogen atom in the phenylene or pyridyl is optionally substituted by 1, 2, or 3 substituents that are each independently selected from: F, Cl, Br, I, —CN, -Me, —C.sub.2H.sub.5, cyclopropyl, —CD.sub.3, —OMe, or —OCF.sub.3; optionally, Ar is phenylene, and a hydrogen atom in the phenylene is optionally substituted by 2 substituents that are F; optionally, Y is —H, —F, —Cl, —Br, methyl, ethyl, n-propyl, isoproyl, —CD.sub.3, —CF.sub.3, —CH.sub.2CF.sub.3, —OCF.sub.3, —OCHF.sub.2, cyclopropyl, -cyclobutyl, -cyclopentyl, —OCH.sub.3, —OC.sub.2H.sub.5, —OC.sub.3H.sub.7, or —OAr′; optionally, Y is —H or —OAr′; optionally, Ar′ is selected from phenyl, pyridyl, pyrimidinyl, or quinolinyl, and hydrogen atoms in the phenyl, pyridyl, pyrimidinyl or quinolinyl ring are each independently optionally substituted with 1, 2 or 3 substituents that are each independently selected from: F, Cl, Br, —CN, C.sub.1-C.sub.7 alkyl (optionally selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, isopentyl, 1-ethylpropyl, neopentyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3-ethylpentyl, or 2,2,3-trimethylbutyl), —CD.sub.3, C.sub.1-C.sub.6haloalkyl, —OCH.sub.3, —OC.sub.2H.sub.7, —OC.sub.3H.sub.7, C.sub.1-C.sub.6 haloalkoxyl, or C.sub.3-C.sub.6 cycloalkyl (optionally cyclopropanyl, cyclobutanyl, cyclopentyl, or cyclohexyl); optionally, Ar′ is selected from phenyl, pyridin-3-yl, or pyridin-4-yl, or pyrimidin-5-yl, optionally, the Ar′ is substituted by 1 or 2 substituents that are selected from Cl, —CH.sub.3, —CF.sub.3, or —OCF.sub.3; and optionally, Z is O.

4. The compound of formula (I), or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) is selected from the following compounds: ##STR00038## ##STR00039## ##STR00040## ##STR00041##

5. The compound of formula (I), or the pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt includes an anionic salt and a cationic salt of the compound of formula (I); optionally, the pharmaceutically acceptable salt includes an alkali metal salt, an alkaline earth metal salt, and an ammonium salt of the compound of formula (I); optionally, the alkali metal includes sodium, potassium, lithium, and cesium, and the alkaline earth metal includes magnesium, calcium, and strontium; optionally, the pharmaceutically acceptable salt includes a salt formed by the compound of formula (I) and an organic base; optionally, the organic base includes trialkylamine, pyridine, quinoline, piperidine, imidazole, picoline, dimethylaminopyridine, dimethylaniline, N-alkylmorpholine, 1,5-diazabicyclo[4.3.0]nonene-5, 1,8-diazabicyclo[5.4.0]undecene-7, and 1,4-diazabicyclo[2.2.2]octane; optionally, the trialkylamine includes trimethylamine, triethylamine, and N-ethyldiisopropylamine; and optionally, the N-alkylmorpholine includes N-methylmorpholine; optionally, the pharmaceutically acceptable salt includes a salt formed by the compound of formula (I) and an acid; and optionally, the acid includes an inorganic acid and an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid; optionally, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, glutamic acid, and pamoic acid.

6. A method for preparing a compound of formula (I), or a pharmaceutically acceptable salt thereof according to claim 1, comprising the following synthetic route: ##STR00042## in respective formulas, n.sub.1, n.sub.2, n.sub.3, R.sub.1, R.sub.2, X.sub.1, X.sub.2, X.sub.3, Z, Ar, and Y are as defined; optionally, when X.sub.2 is —O— and n.sub.3 is 0, the following synthetic route is further adopted: ##STR00043##

7. A pharmaceutical composition, comprising one or more of a compound of formula (I), or a pharmaceutically acceptable salt thereof according to claim 1, and optionally a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, wherein a dosage form of the pharmaceutical composition includes an oral preparation, a rectally administered preparation, and a parenteral administered preparation; optionally, the oral preparation includes a solid preparation and a liquid preparation, optionally, the solid preparation includes tablets, powders, granules, and capsules; optionally, the liquid preparation includes aqueous or oily suspensions, and syrups; and optionally, the parenteral administered preparation includes solutions for injection, and aqueous or oily suspensions.

9. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a Lp-PLA2 inhibitor.

10. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for treating a neurodegenerative-related disease; optionally, the neurodegenerative-related disease includes Alzheimer's disease (AD), glaucoma, and age-related macular degeneration (AMD).

11. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for treating cardiovascular disease, diabetic macular edema (DME), or prostatic disease; and optionally, the cardiovascular disease includes atherosclerosis.

Description

DETAILED DESCRIPTION

[0072] The present disclosure will be further explained with reference to the following examples. It should be appreciated that the examples described herein are merely intended to exemplarily illustrate the present disclosure, and not limit the scope of the present disclosure in any way.

[0073] The starting materials of the present disclosure can be synthesized by or according to the methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Darui Chemicals, and other companies.

[0074] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0075] Unless otherwise specified in the examples, the reaction temperature is room temperature, e.g., 20° C. to 30° C.

Example 1

[0076] Preparation of Compound 1

##STR00009##

[0077] Step I: Preparation of Compound 1b

##STR00010##

[0078] At room temperature, 7-azabicyclo[2.2.1]heptane 1a (4 g, 41.1 mmol) was dissolved in 300 mL of dichloromethane, followed by addition of triethylamine (6.3 g, 61.8 mmol) and di-tert-butyl dicarbonate (13.5 g, 61.8 mmol). The materials were stirred at room temperature and reacted for 4 h, and the reactant was extracted with dichloromethane (100 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=20:1) by the silica gel column chromatography to give a colorless, oily product 1b (10 g, yield: 100%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.16 (s, 2H), 1.74-1.72 (m, 4H), 1.42 (s, 9H), 1.37-1.35 (m, 4H).

[0079] Step II: Preparation of Compound 1C

##STR00011##

[0080] At room temperature, tert-butyl 7-azabicyclo[2.2.1]heptane-7-carboxylate 1b (10 g, 50.8 mmol) and TMEDA (7.1 g, 60.9 mmol) were dissolved in 200 mL of dry ether, followed by adding dropwise a solution of s-BuLi (1.3 M, 60.9 mmol) in hexane at −65° C. under nitrogen protection. After the materials were stirred and reacted for 30 min, a solution of methyl formate (3.65 g, 60.9 mmol) in ether was added dropwise. This mixture was stirred and reacted at −65° C. for 30 min, heated to 0° C., and reacted for 2 h. To the reaction mixture, 100 mL of saturated aqueous ammonium chloride solution was added, and extracted with ether (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=40:1) by the silica gel column chromatography to give a colorless, oily product 1c (8.14 g, yield: 71%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.93 (s, 1H), 4.29 (s, 1H), 2.03-1.88 (m, 4H), 1.65-1.46 (m, 4H), 1.42 (s, 9H).

[0081] Step III: Preparation of Compound 1d

##STR00012##

[0082] Tert-butyl 1-formyl-7-azabicyclo[2.2.1]heptane-7-carboxylate 1c (8.14 g, 36.1 mmol) was dissolved in 300 mL of methanol, to which KBH.sub.4 (3.03 g, 54.2 mmol) was added in batch at 0° C. After the materials were stirred and reacted at room temperature for 1 h, the reactant was concentrated under reduced pressure, and extracted with dichloromethane (60 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=20/1) by the silica gel column chromatography to give a colorless, oily product 1d (6.53 g, yield: 79.5%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ4.96 (s, 1H), 4.24 (m, 1H), 3.90 (m, 2H), 1.89-1.77 (m, 4H), 1.48-1.35 (m, 13H).

[0083] Step IV: Preparation of Compound 1e

##STR00013##

[0084] Tert-butyl 1-(hydroxymethyl)-7-azabicyclo[2.2.1]heptane-7-carboxylate 1d (6.53 g, 28.7 mmol) was dissolved in 100 mL of hydrochloric acid/ethanol (2M). After the materials were stirred and reacted at room temperature for 4 h, the reactant was concentrated under reduced pressure to give a yellow solid product 1e (4.72 g, yield: 100%).

[0085] .sup.1H NMR (400 MHz, DMSO) δ 9.04 (s, 2H), 3.98 (m, 1H), 3.73 (s, 2H), 1.98-1.82 (m, 4H), 1.69-1.53 (m, 4H).

[0086] Step V: Preparation of Compound 1f

##STR00014##

[0087] At room temperature, (7-azabicyclo[2.2.1]heptane-1-yl)methanol 1e (4.5 g, 35.5 mmol), 2,4,6-trichloropyrimidine (7.79 g, 42.6 mmol), and diisopropylethylamine (13.7 g, 106.5 mmol) were dissolved in 200 mL of acetonitrile, stirred overnight, concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5:1) by the silica gel column chromatography to give a colorless solid product 1f (5.2 g, yield: 53%).

[0088] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.37 (s, 1H), 4.91 (m, 1H), 4.26 (m, 1H), 4.01 (m, 2H), 2.01-1.82 (m, 4H), 1.68-1.52 (m, 4H).

[0089] Step VI: Preparation of Compound 1g

##STR00015##

[0090] At room temperature, (7-(2,6-dichloropyrimidin-4-yl)-7-azabicyclo[2.2.1]heptan-1-yl)methanol 1f (5.46 g, 19.9 mmol) and triethylamine (6.0 g, 59.7 mmol) were dissolved in 100 mL of dry dichloromethane, to which MsCl (2.5 g, 21.9 mmol) was added dropwise at 0° C. After the materials were stirred and reacted at 0° C. for 1 h, the reactant was concentrated under reduced pressure, and dissolved in 100 mL of a mixed solvent of dioxane/water (1:1), to which potassium carbonate (8.2 g, 59.7 mmol) was added, and stirred at 90° C. overnight. Then the reactant was concentrated under reduced pressure, and purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a colorless solid product 1g (2.2 g, yield: 46%).

[0091] .sup.1H NMR (400 MHz, DMSO) δ 6.10 (s, 1H), 4.42 (m, 1H), 3.95 (s, 2H), 1.94-1.89 (m, 2H), 1.78-1.69 (m, 6H).

[0092] Step VII: Preparation of Compound 1

##STR00016##

[0093] (2,3-Difluorophenyl)methanol 1h (46 mg, 0.32 mmol) was dissolved in 2 mL of dry N,N-dimethylformamide for 1 h, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 1 (31 mg, yield: 43%).

[0094] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.24-7.03 (m, 3H), 5.45 (s, 2H), 5.15 (s, 1H), 4.09 (m, 1H), 3.99 (s, 2H), 2.14-1.99 (m, 2H), 1.78-1.66 (m, 6H). MS (ESI): m/z 346.0 [M+H].sup.+.

Example 2 Preparation of Compound 2

[0095] ##STR00017##

[0096] (4-Fluorophenyl)methanol (40 mg, 0.32 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 2 (40 mg, yield: 58%).

[0097] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.43-7.40 (m, 2H), 7.08-7.04 (m, 2H), 5.37 (s, 2H), 5.17 (s, 1H), 4.11 (m, 1H), 4.01 (s, 2H), 2.10-2.06 (m, 2H), 1.82-1.67 (m, 6H). MS (ESI): m/z 328.0 [M+H].sup.+.

Example 3 Preparation of Compound 3

[0098] ##STR00018##

[0099] (3,4-Dichlorophenyl)methanol (46 mg, 0.32 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 3 (17 mg, yield: 23%).

[0100] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.28-7.22 (m, 1H), 7.18-7.11 (m, 2H), 5.35 (s, 2H), 5.17 (s, 1H), 4.12 (m, 1H), 4.01 (s, 2H), 2.12-2.07 (m, 2H), 1.83-1.74 (m, 6H). MS (ESI): m/z 346.0 [M+H].sup.+.

Example 4 Preparation of Compound 4

[0101] ##STR00019##

[0102] Step I: Preparation of Compound 4c

[0103] At room temperature, 2-(trifluoromethyl)pyridin-4-ol 4b (0.85 g, 5.2 mmol), 3,4,5-trifluorobenzaldehyde (1 g, 6.2 mmol) and potassium carbonate (0.93 g, 6.76 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 1 h. The reactant was cooled to room temperature, then poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a yellow solid product 4c (1.47g, yield: 78.2%).

[0104] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.97 (s, 1H), 8.65 (m, 1H), 7.63 (m, 2H), 7.27 (m, 1H), 7.01 (m, 1H).

[0105] Step II: Preparation of Compound 4d

[0106] At room temperature, 3,5-difluoro-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)benzaldehyde 4c (1.47 g, 4.85 mmol) was dissolved in 50 mL of ethanol, to which NaBH.sub.4 (184 mg, 4.85 mmol) was added at 0° C. The materials were stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=2:1) by the silica gel column chromatography to give a white solid product 4d (1.04 g, yield: 70.3%).

[0107] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.59 (m, 1H), 7.24 (m, 1H), 7.11 (m, 2H), 6.99 (m, 1H), 4.75 (m, 2H), 2.19 (m, 1H).

[0108] Step III: Preparation of Compound 4

[0109] (3,5-Difluoro-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)phenyl)methanol (77 mg, 0.25 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 5 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 4 (35 mg, yield: 33%).

[0110] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.62 (m, 1H), 7.28 (s, 1H), 7.15 (m, 2H), 7.01 (m, 1H), 5.44 (s, 2H), 5.24 (s, 1H), 4.16 (m, 1H), 4.03 (s, 2H), 2.12-2.07 (m, 2H), 1.83-1.79 (m, 6H). MS (ESI): m/z 506.9 [M+H].sup.+.

Example 5 Preparation of Compound 5

[0111] ##STR00020##

[0112] Step I: Preparation of Compound 5b

[0113] At room temperature, 4-(trifluoromethyl)phenol 5a (0.84 g, 5.2 mmol), 3,4,5-trifluorobenzaldehyde 4a (1 g, 6.2 mmol) and potassium carbonate (0.93 g, 6.76 mmol) were dissolved in 30 mL N,N-dimethylformamide, and stirred and reacted at 90° C. for 1 h. The reactant was cooled to room temperature, then poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a yellow solid product 5b (1.33 g, yield: 71.0%).

[0114] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (m, 1H), 7.59 (m, 4H), 7.04 (m, 2H).

[0115] Step II: Preparation of Compound 5c

[0116] At room temperature, 3,5-difluoro-4-(4-(trifluoromethyl)phenoxy)benzaldehyde 5b (1.33 g, 4.4 mmol) was dissolved in 50 mL of methylethanol, to which NaBH.sub.4 (166 mg, 4.4 mmol) was added at 0° C. The materials were stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=2/1) by the silica gel column chromatography to give a colorless, oily product 5c (0.85g, yield: 63.6%).

[0117] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.57 (m, 2H), 7.09-7.00 (m, 4H), 4.72 (m, 2H), 2.03 (m, 1H).

[0118] Step III: Preparation of Compound 5

[0119] 3,5-Difluoro-4-(4-(trifluoromethyl)phenoxy)phenyl)methanol 5c (64 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 5 (17 mg, yield: 16%).

[0120] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.59 (m, 2H), 7.12-7.02 (m, 4H), 5.42 (s, 2H), 5.23 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.12 (m, 2H), 1.87-1.74 (m, 6H). MS (ESI): m/z 506.0 [M+H].sup.+.

Example 6 Preparation of Compound 6

[0121] ##STR00021##

[0122] Step I: Preparation of Compound 6b

[0123] At room temperature, 6-(trifluoromethyl)pyridin-3-ol 6a (0.85 g, 5.2 mmol), 3,4,5-trifluorobenzaldehyde 4a (1 g, 6.2 mmol) and potassium carbonate (0.93 g, 6.76 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 1 h. The reactant was cooled to room temperature, then poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a yellow solid product 6b (1.34 g, yield: 84.6%).

[0124] Step II: Preparation of Compound 6c

[0125] At room temperature, 3,5-difluoro-4-((6-(trifluoromethyl)pyridin-3-yl)oxy)benzaldehyde 6b (1.34 g, 4.4 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (167 mg, 4.4 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=2:1) by the silica gel column chromatography to give a colorless, oily product 6c (0.77 g, yield: 57.4%).

[0126] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.46 (s, 1H), 7.63 (m, 1H), 7.30 (m, 1H), 7.09 (m, 2H), 4.73 (m, 2H), 2.40 (m, 1H).

[0127] Step III: Preparation of Compound 6

[0128] (3,5-Difluoro-4-((6-(trifluoromethyl)pyridin-3-yl)oxy)phenyl)methanol 6c (50 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 6 (31 mg, yield: 29%).

[0129] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.51 (m, 1H), 7.65 (m, 1H), 7.31 (m, 1H), 7.13 (m, 2H), 5.42 (s, 2H), 5.23 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.12-2.06 (m, 2H), 1.85-1.70 (m, 6H). MS (ESI): m/z 507.0 [M+H].sup.+.

Example 7 Preparation of Compound 7

[0130] ##STR00022##

[0131] Step I: Preparation of Compound 7b

[0132] At room temperature, 6-methylpyridin-4-ol 7a (0.5 g, 4.6 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.88 g, 5.5 mmol) and potassium carbonate (0.823 g, 5.95 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. The reactant was cooled to room temperature, then poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 7b (0.4 g, yield: 33.8%).

[0133] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 8.39 (m, 1H), 7.62-7.56 (m, 2H), 6.70-6.66 (m, 2H), 2.52 (s, 3H).

[0134] Step II: Preparation of Compound 7c

[0135] At room temperature, 3,5-difluoro-4-((2-methylpyridin-4-yl)oxy)benzaldehyde 7b (0.4 g, 1.86 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (71 mg, 1.86 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 7c (0.40 g, yield: 85.7%).

[0136] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.29 (m, 1H), 7.07 (m, 2H), 6.70-6.64 (m, 2H), 4.73 (s, 2H), 3.20 (m, 1H), 2.50 (s, 3H).

[0137] Step III: Preparation of Compound 7

[0138] (3,5-Difluoro-4-((2-methylpyridin-4-yl)oxy)phenyl)methanol 7c (53 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 7 (17 mg, yield: 18%).

[0139] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.38 (m, 1H), 7.10 (m, 2H), 6.69 (m, 2H), 5.42 (s, 2H), 5.23 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.53 (s, 3H), 2.21-2.07 (m, 2H), 1.88-1.70 (m, 6H). MS (ESI): m/z 453.0 [M+H].sup.+.

Example 8 Preparation of Compound 8

[0140] ##STR00023##

[0141] Step I: Preparation of Compound 8b

[0142] At room temperature, 6-methylpyridin-3-ol 8a (0.57 g, 5.2 mmol), 3,4,5-trifluorobenzaldehyde 4a (1 g, 6.2 mmol) and potassium carbonate (0.93 g, 6.76 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 1 h. The reactant was cooled to room temperature, then poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 8b (0.91g, yield: 69.4%).

[0143] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.92 (s, 1H), 8.28 (s, 1H), 7.62-7.49 (m, 2H), 7.18-7.10 (m, 2H), 2.54 (s, 3H).

[0144] Step II: Preparation of Compound 8c

[0145] At room temperature, 3,5-difluoro-4-((2-methylpyridin-4-yl)oxy)benzaldehyde 8b (0.91 g, 4.3 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (161 mg, 4.3 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 8c (0.89 g, yield: 82.5%).

[0146] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.20 (m, 1H), 7.16-6.98 (m, 4H), 4.69 (m, 2H), 2.88 (m, 1H), 2.50 (s, 3H).

[0147] Step III: Preparation of Compound 8

[0148] (3,5-Difluoro-4-((2-methylpyridin-4-yl)oxy)phenyl)methanol 8c (53 mg, 0.21 mmol) was dissolved in 2 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 8 (20 mg, yield: 21%).

[0149] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.30 (m, 1H), 7.18-7.04 (m, 4H), 5.40 (s, 2H), 5.22 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.54 (s, 3H), 2.11 (m, 2H), 1.86-1.74 (m, 6H). MS (ESI): m/z 453.0 [M+H].sup.+.

Example 9 Preparation of Compound 9

[0150] ##STR00024##

[0151] Step I: Preparation of Compound 9b

[0152] At room temperature, 2-methylpyrimidin-5-ol 9a (0.25 g, 2.27 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.44 g, 2.72 mmol) and potassium carbonate (0.41 g, 2.95 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10:1) by the silica gel column chromatography to give a yellow solid product 9b (0.24 g, yield: 34.8%).

[0153] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.93 (s, 1H), 8.39 (s, 2H), 7.64-7.54 (m, 2H), 2.72 (s, 3H).

[0154] Step II: Preparation of Compound 9c

[0155] At room temperature, 3,5-difluoro-4-((2-methylpyrimidin-5-yl)oxy)benzaldehyde 9b (0.24 g, 0.79 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (30 mg, 0.79 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4:1) by the silica gel column chromatography to give a colorless, oily product 9c (0.17 g, yield: 85.4%).

[0156] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.33 (s, 2H), 7.04 (m, 2H), 4.71 (m, 2H), 2.70 (s, 3H).

[0157] Step III: Preparation of Compound 9

[0158] (3,5-Difluoro-4-((2-methylpyrimidin-5-yl)oxy)phenyl)methanol 9c (53 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 9 (18 mg, yield: 19%).

[0159] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 2H), 7.09 (m, 2H), 5.37 (s, 2H), 5.20 (s, 1H), 4.12 (m, 1H), 3.99 (s, 2H), 2.69 (s, 3H), 2.13-2.03 (m, 2H), 1.81-1.71 (m, 6H). MS (ESI): m/z 454.0 [M+H].sup.+.

Example 10 Preparation of Compound 10

[0160] ##STR00025##

[0161] Step I: Preparation of Compound 10b

[0162] At room temperature, 4-chloro-3-(trifluoromethyl)phenol 10a (0.5 g, 2.54 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.45 g, 2.8 mmol) and potassium carbonate (0.46 g, 3.3 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 10b (0.6 g, yield: 70.1%).

[0163] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 7.64-7.55 (m, 2H), 7.45 (m, 1H), 7.31 (m, 1H), 7.05 (m, 1H).

[0164] Step II: Preparation of Compound 10c

[0165] At room temperature, 4-(4-chloro-3-(trifluoromethyl)phenoxy)-3,5-difluorobenzaldehyde 10b (0.6 g, 1.78 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (67 mg, 1.78 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a white solid product 10c (0.28 g, yield: 46.4%).

[0166] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.41 (m, 1H), 7.28 (m, 1H), 7.08-7.00 (m, 3H), 4.73 (m, 2H), 1.94 (m, 1H).

[0167] Step III: Preparation of Compound 10

[0168] (4-(4-Chloro-3-(trifluoromethyl)phenoxy)-3,5-difluorophenyl)methanol 10c (71 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 10 (28 mg, yield: 25%).

[0169] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.41 (m, 1H), 7.29 (m, 1H), 7.08 (m, 2H), 7.01 (m, 1H), 5.38 (s, 2H), 5.20 (s, 1H), 4.13 (m, 1H), 4.00 (s, 2H), 2.19-2.08 (m, 2H), 1.84-1.74 (m, 6H). MS (ESI): m/z 539.9 [M+H].sup.+.

Example 11 Preparation of Compound 11

[0170] ##STR00026##

[0171] Step I: Preparation of Compound 11b

[0172] At room temperature, 3-(trifluoromethyl)phenol 11a (1 g, 6.17 mmol), 3,4,5-trifluorobenzaldehyde 4a (1.09 g, 6.79 mmol) and potassium carbonate (1.1 g, 8.02 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 11b (1.7 g, yield: 82.5%).

[0173] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 7.63-7.55 (m, 2H), 7.46 (m, 1H), 7.39 (m, 1H), 7.21 (s, 1H), 7.13 (m, 1H).

[0174] Step II: Preparation of Compound 11c

[0175] At room temperature, 3,5-difluoro-4-(3-(trifluoromethyl)phenoxy)benzaldehyde 11b (1.7 g, 5.6 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (213 mg, 5.6 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 11e (1.27 g, yield: 74.5%).

[0176] Step III: Preparation of Compound 11

[0177] (3,5-Difluoro-4-(3-(trifluoromethyl)phenoxy)phenyl)methanol 11e (71 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 11 (33 mg, yield: 31%).

[0178] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.44 (m, 1H), 7.36 (m, 1H), 7.22 (s, 1H), 7.11 (m, 3H), 5.42 (s, 2H), 5.23 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.12 (m, 2H), 1.83-1.77 (m, 6H). MS (ESI): m/z 505.9 [M+H].sup.+.

Example 12 Preparation of Compound 12

[0179] ##STR00027##

[0180] Step I: Preparation of Compound 12b

[0181] At room temperature, 4-chloro-3-methylphenol 12a (1 g, 7.0 mmol), 3,4,5-trifluorobenzaldehyde 4a (1.2 g, 7.7 mmol) and potassium carbonate (1.3 g, 9.1 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 12b (1.2 g, yield: 54.5%).

[0182] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.92 (s, 1H), 7.61-7.51 (m, 2H), 7.30-7.23 (m, 1H), 6.85 (m, 1H), 6.73 (m, 1H), 2.34 (s, 3H).

[0183] Step II: Preparation of Compound 12c

[0184] At room temperature, 4-(4-chloro-3-methylphenoxy)-3,5-difluorobenzaldehyde 12b (1.2 g, 4.2 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (161 mg, 4.2 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 12c (0.89 g, yield: 74.4%).

[0185] Step III: Preparation of Compound 12

[0186] 4-(4-Chloro-3-methylphenoxy)-3,5-difluorophenyl)methanol 12c (59 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 12 (29 mg, yield: 31%).

[0187] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.25 (m, 1H), 7.08 (m, 2H), 6.84 (m, 1H), 6.73 (m, 1H), 5.40 (s, 2H), 5.22 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.35 (s, 3H), 2.12 (m, 2H), 1.87-1.74 (m, 6H). MS (ESI): m/z 486.0 [M+H].sup.+.

Example 13: Preparation of Compound 13

[0188] ##STR00028##

[0189] Step I: Preparation of Compound 13b

[0190] At room temperature, 2-(trifluoromethyl)pyrimidin-5-ol 13a (0.25 g, 1.52 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.29 g, 1.83 mmol) and potassium carbonate (0.27 g, 1.98 mmol) were dissolved in 20 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 13b (0.24 g, yield: 52.0%).

[0191] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.97 (s, 1H), 8.59 (s, 2H), 7.69-7.54 (m, 2H).

[0192] Step II: Preparation of Compound 13c

[0193] At room temperature, 3,5-difluoro-4-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)benzaldehyde 13b (0.24 g, 0.79 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (30 mg, 0.79 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 13c (0.12 g, yield: 49.6%).

[0194] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.54 (s, 2H), 7.12 (m, 2H), 4.74 (m, 2H), 2.23 (m, 1H).

[0195] Step III: Preparation of Compound 13

[0196] (3,5-Difluoro-4-((2-(trifluoromethyl)pyrimidin-5-yl)oxy)phenyl)methanol 13c (64 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 13 (18 mg, yield: 17%).

[0197] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.58 (s, 2H), 7.17 (m, 2H), 5.43 (s, 2H), 5.24 (s, 1H), 4.16 (m, 1H), 4.03 (s, 2H), 2.13 (m, 2H), 1.88-1.76 (m, 6H).

[0198] MS (ESI): m/z 508.1 [M+H].sup.+.

Example 14: Preparation of Compound 14

[0199] ##STR00029##

[0200] Step I: Preparation of Compound 14b

[0201] At room temperature, 3-chloro-4-(trifluoromethyl)phenol 14a (0.25 g, 1.27 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.22 g, 1.4 mmol) and potassium carbonate (0.23 g, 1.65 mmol) were dissolved in 20 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 14b (0.32 g, yield: 74.0%).

[0202] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.95 (s, 1H), 7.69-7.56 (m, 3H), 7.10 (m, 1H), 6.92 (m, 1H).

[0203] Step II: Preparation of Compound 14c

[0204] At room temperature, 4-(3-chloro-4-(trifluoromethyl)phenoxy)-3,5-difluorobenzaldehyde 14b (0.32 g, 0.94 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (36 mg, 0.94 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a white solid product 14c (0.15 g, yield: 47.1%).

[0205] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.62 (m, 1H), 7.13-7.00 (m, 3H), 6.90 (m, 1H), 4.74 (m, 2H), 1.88 (m, 1H).

[0206] Step III: Preparation of Compound 14

[0207] (4-(3-Chloro-4-(trifluoromethyl)phenoxy)-3,5-difluorophenyl)methanol 14c (71 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 14 (12 mg, yield: 10%).

[0208] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.64 (m, 1H), 7.10 (m, 3H), 6.91 (m, 1H), 5.41 (s, 2H), 5.23 (s, 1H), 4.15 (m, 1H), 4.03 (s, 2H), 2.19-2.04 (m, 2H), 1.83-1.75 (m, 6H). MS (ESI): m/z 539.9 [M+H].sup.+.

Example 15 Preparation of Compound 15

[0209] ##STR00030##

[0210] Step I: Preparation of Compound 15b

[0211] Ethoxyethylene 15a (1 g, 13.9 mmol) and pyridine (1.65 g, 20.9 mmol) were dissolved in 20 mL of dichloromethane, and trifluoroacetic anhydride was added at 0° C. under nitrogen protection. The materials were stirred and reacted for 20 min, heated to room temperature, and reacted for 2 h. The reaction mixture was cooled to −20° C., to which dimethylamine was added dropwise within 10 min, heated again to room temperature, and reacted for 2 h. The reaction was quenched with water. The reactant was extracted with dichloromethane (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a white solid product 15b (540 mg, yield: 23.1%).

[0212] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.85 (m, 1H), 5.26 (m, 1H), 3.21 (s, 3H), 2.95 (s, 3H).

[0213] Step II: Preparation of Compound 15c

[0214] At room temperature, (E)-4-(dimethylamino)-1,1,1-trifluorobut-3-en-2-one 15b (0.54 g, 2.98 mmol) was dissolved in 20 mL of dichloroethane, and a solution of Tf.sub.2O in dichloroethane (5 mL) was added dropwise at 0° C., followed by adding a solution of 3-aminophenol in dichloroethane (10 mL). The materials were stirred and reacted at 40° C. for 2 h, and filtered to give a white solid product 15c (275 mg, 57.4%).

[0215] .sup.1H NMR (400 MHz, MeOD) δ 8.43 (m, 1H), 7.90 (m, 1H), 7.62 (m, 1H), 7.40 (m, 1H), 7.32 (m, 1H).

[0216] Step III: Preparation of Compound 15d

[0217] At room temperature, 2-(trifluoromethyl)quinolin-7-ol 15c (0.37 g, 1.71 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.28 g, 1.71 mmol) and potassium carbonate (0.71 g, 5.13 mmol) were dissolved in 20 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a white solid product 15d (310 mg, 51.5%).

[0218] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.97 (s, 1H), 8.36 (m, 1H), 7.96 (m, 1H), 7.69 (m, 1H), 7.62 (m, 3H), 7.40 (s, 1H).

[0219] Step IV: Preparation of Compound 15e

[0220] At room temperature, 3,5-difluoro-4-((2-(trifluoromethyl)quinolin-7-yl)oxy)benzaldehyde 15d (0.31 g, 0.88 mmol) was dissolved in 20 mL of methanol, to which NaBH.sub.4 (33 mg, 0.88 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 15e (230 mg, 74.6%).

[0221] Step V: Preparation of Compound 15

[0222] (3,5-Difluoro-4-((2-(trifluoromethyl)quinolin-7-yl)oxy)phenyl)methanol 15e (75 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. and purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 15 (18 mg, yield: 17%).

[0223] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.33 (m, 1H), 7.92 (m, 1H), 7.65 (m, 1H), 7.60 (m, 1H), 7.36 (s, 1H), 7.13 (m, 2H), 5.40 (s, 2H), 5.23 (s, 1H), 4.14 (m, 1H), 4.02 (s, 2H), 2.11 (m, 2H), 1.79 (m, 6H). MS (ESI): m/z 557.0 [M+H].sup.+.

Example 16 Preparation of Compound 16

[0224] ##STR00031##

[0225] Step I: Preparation of Compound 16b

[0226] At room temperature, 3-(trifluoromethoxy)phenol 16a (0.50 g, 2.8 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.5 g, 2.8 mmol) and potassium carbonate (0.5 g, 3.64 mmol) were dissolved in 30 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a white solid product 16b (0.73 g, yield: 81.4%).

[0227] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 7.64-7.54 (m, 2H), 7.34 (m, 1H), 7.00 (m, 1H), 6.87 (m, 2H).

[0228] Step II: Preparation of Compound 16c

[0229] At room temperature, 4-(3-(trifluoromethoxy)phenoxy)-3,5-difluorobenzaldehyde 16b (0.73 g, 2.28 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (86 mg, 2.28 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 16c (0.57g, yield: 71.2%).

[0230] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.30 (m, 1H), 7.06 (m, 2H), 6.94 (m, 1H), 6.85 (m, 1H), 6.81 (s, 1H), 4.72 (m, 2H), 1.94 (m, 1H).

[0231] Step III: Preparation of Compound 16

[0232] 4-(3-(Trifluoromethoxy)phenoxy)-3,5-difluorophenyl)methanol 16c (67 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 16 (25 mg, yield: 23%).

[0233] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.29 (m, 1H), 7.07 (m, 2H), 6.93 (m, 1H), 6.83 (m, 2H), 5.38 (s, 2H), 5.20 (s, 1H), 4.12 (m, 1H), 4.00 (s, 2H), 2.08 (m, 2H), 1.83-1.73 (m, 6H). MS (ESI): m/z 522.0 [M+H].sup.+.

Example 17 Preparation of Compound 17

[0234] ##STR00032##

[0235] Step I: Preparation of Compound 17b

[0236] At room temperature, 3-chloro-4-(trifluoromethoxy)phenol 17a (0.50 g, 2.34 mmol), 3,4,5-trifluorobenzaldehyde 4a (0.41 g, 2.58 mmol) and potassium carbonate (0.42 g, 3.04 mmol) were dissolved in 20 mL of N,N-dimethylformamide, and stirred and reacted at 90° C. for 2 h. After cooled, the reactant was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a yellow solid product 17b (0.62 g, yield: 74.4%).

[0237] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 7.63-7.54 (m, 2H), 7.29 (m, 1H), 7.07 (m, 1H), 6.90 (m, 1H).

[0238] Step II: Preparation of Compound 17c

[0239] At room temperature, 4-(3-chloro-4-(trifluoromethoxy)phenoxy)-3,5-difluorobenzaldehyde 17b (0.62 g, 1.94 mmol) was dissolved in 50 mL of methanol, to which NaBH.sub.4 (62 mg, 1.94 mmol) was added at 0° C., stirred and reacted at room temperature for 0.5 h, concentrated under reduced pressure, added with water, and extracted with ethyl acetate (100 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4/1) by the silica gel column chromatography to give a colorless, oily product 17c (0.53 g, yield: 77.0%).

[0240] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.25 (m, 1H), 7.06 (m, 2H), 7.01 (m, 1H), 6.87 (m, 1H), 4.72 (s, 2H), 2.04 (m, 1H).

[0241] Step III: Preparation of Compound 17

[0242] (4-(3-Chloro-4-(trifluoromethoxy)phenoxy)-3,5-difluorophenyl)methanol 17c (75 mg, 0.21 mmol) was dissolved in 5 mL of dry N,N-dimethylformamide, to which sodium hydride (60% in mineral oil, 17 mg, 0.42 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 1g (50 mg, 0.21 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. and purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 17 (23 mg, yield: 20%).

[0243] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.23 (s, 1H), 7.09 (m, 2H), 7.03 (m, 1H), 6.87 (m, 1H), 5.39 (s, 2H), 5.21 (s, 1H), 4.13 (m, 1H), 4.01 (s, 2H), 2.10 (m, 2H), 1.83-1.71 (m, 6H). MS (ESI): m/z 555.9 [M+H].sup.+.

Example 18 Preparation of Compound 18

[0244] ##STR00033##

[0245] Step I: Preparation of Compound 18b

[0246] At room temperature, 8-azabicyclo[3.2.1]octane 18a (5 g, 33.9 mmol) and triethylamine (5.2 g, 50.8 mmol) were dissolved in 300 mL of dichloromethane, and (Boc).sub.2O (11.1 g, 50.8 mmol) was added. The materials were stirred and reacted for 4 h, and extracted with dichloromethane (100 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a colorless liquid product 18b (9.4 g, 100%).

[0247] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.14 (m, 2H), 1.97-1.86 (m, 2H), 1.81-1.53 (m, 6H), 1.46 (s, 9H), 1.41-1.35 (m, 2H).

[0248] Step II: Preparation of Compound 18c

[0249] At room temperature, tert-butyl 8-azabicyclo[3.2.1]octane-8-carboxylate 18b (7.2 g, 34.1 mmol) and TMEDA (4.7 g, 40.9 mmol) were dissolved in 150 mL of dry ether, and s-BuLi (1.3 M in hexane, 31.5 mL, 40.9 mmol) was added dropwise at −65° C. under nitrogen protection. After stirring and reacting at −65° C. for 30 min, a solution of methyl formate (2.45 g, 40.9 mmol) in ether (20 mL) was added dropwise. After stirring and reacting at −65° C. for 30 min, the reactant was heated to 0° C., stirred and reacted for 2 h, added with a saturated aqueous ammonium chloride solution (100 mL), and extracted with ether (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10:1) by the silica gel column chromatography to give a colorless, liquid product 18c (3.13 g, 38.4%).

[0250] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.45 (s, 1H), 4.24 (m, 1H), 2.15 (s, 1H), 1.97 (m, 1H), 1.85 (m, 2H), 1.82-1.60 (m, 6H), 1.45 (s, 9H).

[0251] Step III: Preparation of Compound 18d

[0252] At room temperature, tert-butyl 1-formyl-8-azabicyclo[3.2.1]octane-8-carboxylate 18c (3.13 g, 13.1 mmol) was dissolved in 100 mL of dry methanol, and KBH.sub.4 (1.1 g, 19.6 mmol) was added in batch at 0° C. The materials were stirred and reacted at room temperature for 1 h, added with 40 mL of water, and extracted with dichloromethane (60 mL×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give a colorless liquid product 18d (3.45 g, 100%).

[0253] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.37 (m, 1H), 4.21 (m, 1H), 3.72-3.49 (m, 2H), 2.00-1.73 (m, 4H), 1.61 (m, 6H), 1.46 (s, 9H).

[0254] Step IV: Preparation of Compound 18e

[0255] At room temperature, tert-butyl 1-(hydroxymethyl)-8-azabicyclo[3.2.1]octane-8-carboxylate 18d (3.13 g, 13.1 mmol) was dissolved in 60 mL of a mixed solvent of HCl/ethanol, stirred and reacted for 4 h, and concentrated under reduced pressure to give a yellow solid product 18e (2.45 g, 100%).

[0256] .sup.1H NMR (400 MHz, DMSO) δ 8.94 (s, 2H), 5.55 (s, 1H), 3.80 (m, 1H), 3.58-3.48 (m, 2H), 2.04 (m, 1H), 1.86-1.40 (m, 9H).

[0257] Step V: Preparation of Compound 18f

[0258] At room temperature, (8-azabicyclo[3.2.1]oct-1-yl)methanol 18e (2.45 g, 17.35 mmol), 2,4,6-trichloropyrimidine (3.49 g, 19.1 mmol) and DIPEA (0.42 g, 52.05 mmol) were dissolved in 100 mL of acetonitrile, stirred and reacted overnight, concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=4:1) by the silica gel column chromatography to give a white solid product 18f (1 g, 20.1%).

[0259] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.41 (s, 1H), 5.43 (m, 1H), 4.30 (m, 1H), 3.83 (m, 1H), 3.72 (m, 1H), 2.13-1.52 (m, 10H).

[0260] Step VI: Preparation of Compound 18g

[0261] 18f (1 g, 3.1 mmol) and triethylamine (0.94 g, 1.3 mmol) were dissolved in 50 mL of dry dichloromethane, and MsCl (0.4 g, 3.4 mmol) was added dropwise at 0° C. After stirring and reacting at 0° C. for 1 h, the reactant was concentrated under reduced pressure to give a colorless, oily intermediate (1.1 g, 100%). This coarse product and potassium carbonate (1.28 g, 9.3 mmol) were dissolved in 60 mL of a mixed solution of dioxane/water (1/1), stirred and reacted at 90° C. overnight, concentrated under reduced pressure, and purified with an eluent system (dichloromethane/methanol=20/1) by the silica gel column chromatography to give a white solid product 18g (0.4 g, 51%).

[0262] .sup.1H NMR (400 MHz, .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.66 (s, 1H), 4.25 (m, 1H), 4.11 (m, 1H), 3.70 (m, 1H), 2.05-1.88 (m, 4H), 1.85-1.71 (m, 6H).

[0263] Step VII: Preparation of Compound 18

[0264] (3,5-Difluoro-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)phenyl)methanol 4d (92 mg, 0.3 mmol) was dissolved in 5 mL of dry DMF, to which sodium hydride (60% in mineral oil, 17 mg, 0.4 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 18g (50 mg, 0.2 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 18 (49 mg, yield: 47%).

[0265] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.60 (m, 1H), 7.26 (s, 1H), 7.13 (m, 2H), 6.98 (m, 1H), 5.41 (s, 2H), 5.19 (s, 1H), 4.24 (m, 1H), 4.07 (m, 1H), 3.70 (m, 1H), 2.05-1.87 (m, 3H), 1.73 (m, 7H). MS (ESI): m/z 521.2 [M+H].sup.+.

Example 19 Preparation of Compound 19

[0266] ##STR00034##

[0267] Step I: Preparation of Compound 19

[0268] (3,5-Difluoro-4-((2-methylpyridin-4-yl)oxy)phenyl)methanol 8c (53 mg, 0.21 mmol) was dissolved in 5 mL of dry DMF, to which sodium hydride (60% in mineral oil, 17 mg, 0.4 mmol) was added at 0° C. After the materials were stirred and reacted at room temperature for 20 min, Compound 18g (50 mg, 0.2 mmol) was added, and stirred and reacted for 1 h. Thereafter, a small amount of water was added to quench the reaction. The reactant was purified with an eluent system (dichloromethane/methanol=20:1) by the silica gel column chromatography to give a white solid product 19 (52 mg, yield: 55.5%).

[0269] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.38 (s, 2H), 7.10 (m, 2H), 5.41 (s, 2H), 5.20 (s, 1H), 4.23 (m, 1H), 4.06 (m, 1H), 3.68 (m, 1H), 2.70 (s, 3H), 2.00-1.90 (m, 3H), 1.85-1.69 (m, 7H). MS (ESI): m/z 468.2 [M+H].sup.+.

Example 20 Preparation of Compound 20

[0270] ##STR00035## ##STR00036##

[0271] Step I: Preparation of Compound 20b

[0272] At room temperature, the compound diethyl meso-2,5-dibromoadipate (10.0 g, 27.8 mmol) and potassium carbonate (4.60 g, 33.3 mmol) were dissolved in toluene and water (200 mL, v/v=4:1), and phenylmethylamine (2.99 g, 27.8 mmol) was added. The reaction mixture was heated to 80° C. and reacted for 24 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure to remove most of the solvent, added with water, and extracted with ethyl acetate (80 mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=10/1) by the silica gel column chromatography to give an oily product 20b (6.6 g, 77.7%).

[0273] .sup.1H NMR (400 MHz, DMSO) δ 7.31-7.18 (m, 5H), 3.96-3.86 (m, 4H), 3.83 (s, 2H), 3.38 (m, 2H), 2.08-1.98 (m, 2H), 1.94-1.83 (m, 2H), 1.08 (m, 6H).

[0274] Step II: Preparation of Compound 20c

[0275] At room temperature, the compound diethyl N-benzyl-2,5-pyrrole dicarboxylate (6.6 g, 21.6 mmol) was dissolved in methanol (100 mL), and palladium on carbon (20%, 1.3 g) was added. The reaction mixture was reacted for 7 h at room temperature under the H.sub.2 condition. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a colorless, oily Compound 20c (4.4 g, 94.4%).

[0276] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.23-4.11 (m, 4H), 3.81-3.71 (m, 2H), 2.18-2.06 (m, 2H), 1.97-1.83 (m, 2H), 1.26 (m, 6H).

[0277] Step III: Preparation of Compound 20d

[0278] At room temperature, the compound diethyl 2,5-pyrrolidinecarboxylate (4.4 g, 19.5 mmol) was dissolved in toluene, and di-tert-butyl dicarbonate (5.35 g, 24.5 mmol) was added. The reaction mixture was heated to 95° C. and reacted for 4 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a colorless, oily product 20d (4.3 g, 70.0%).

[0279] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.38 (s, 1H), 4.27 (s, 1H), 4.24-4.14 (m, 4H), 2.16 (m, 4H), 1.42 (s, 9H), 1.31-1.21 (m, 6H).

[0280] Step IV: Preparation of Compound 20e

[0281] At room temperature, the compound diethyl N-tert-butoxycarbonyl-2,5-pyrrole dicarboxylate (2.7 g, 8.6 mmol) was dissolved in dry tetrahydrofuran (80 mL). A solution of lithium diisopropylamide (2 M, 8.2 ml, 16.3 mmol) in tetrahydrofuran was added dropwise at −78° C. under nitrogen protection. Benzylchloromethyl ether (1.49g, 9.5 mmol) was added. The mixture was stirred and reacted at −78° C. for 1 h, and heated to the room temperature and reacted for 2 h. To the reaction mixture, 100 mL of saturated aqueous ammonium chloride solution was added, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=5/1) by the silica gel column chromatography to give a yellow, oily product 20e (2.3 g, 61.4%).

[0282] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.41-7.20 (m, 5H), 4.71-4.52 (m, 2H), 4.25-4.07 (m, 4H), 2.49 (m, 1H), 2.28 (m, 1H), 2.15-2.01 (m, 2H), 1.37 (s, 9H), 1.29-1.17 (m, 6H).

[0283] Step V: Preparation of Compound 20f

[0284] At room temperature, the compound diethyl N-tert-butoxycarbonyl-2,5-(2-phenoxymethyl)pyrrole dicarboxylate was dissolved in dioxane hydrochloride (4N, 20 mL), and stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure to give a brown oily coarse Compound 20f (1.44 g, 50.0%) for direct use in the next step without purification.

[0285] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39-7.27 (m, 5H), 4.60-4.46 (m, 2H), 3.71 (m, 1H), 3.55 (m, 2H), 3.44-3.27 (m, 3H), 1.89-1.76 (m, 1H), 1.76-1.45 (m, 3H).

[0286] Step VI: Preparation of Compound 20g

[0287] Dry tetrahydrofuran (40 mL) was added to a reaction flask, and lithium aluminum hydride (0.33 g, 8.6 mmol) was added under ice bath and nitrogen protection. Ten minutes later, a solution of [6-(benzyloxymethyl)-6-(hydroxymethyl)-2-piperidinyl]methanol in tetrahydrofuran was added dropwise, and then moved to room temperature and reacted for 1 h. The reaction was performed under ice-bath conditions again, and sodium hydroxide solution (2M, 2 mL) was added to produce a white solid. The white solid was filtered and washed with tetrahydrofuran. The filtrate was dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure to give a yellowish oily coarse Compound 20g (1.0 g, 92.5%) for direct use in the next step without purification.

[0288] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39-7.27 (m, 5H), 4.57-4.46 (m, 2H), 3.71 (m, 1H), 3.55 (m, 2H), 3.44-3.31 (m, 3H), 1.88-1.78 (m, 1H), 1.75-1.58 (m, 2H), 1.52 (m, 1H).

[0289] Step VII: Preparation of Compound 20h

[0290] At room temperature, the compound (2-((benzyloxy)methyl)pyrrolidine-2,5-diyl)dimethanol (1.0 g, 3.98 mmol) was dissolved in methanesulfonic acid under argon protection. The reaction mixture was heated to 140° C. and reacted for 8 h. After cooling to room temperature, the reaction solution was poured into ice water (20 ml), 24 ml of 50% sodium hydroxide was added at 0° C. After concentrated under reduced pressure, the mixture was dissolved in methanol and filtered, and the filtrate was concentrated under reduced pressure to give a yellowish oily coarse target Compound 20h (0.3 g, 52.8%).

[0291] Step VIII: Preparation of Compound 20i

[0292] At room temperature, the compound 3-oxa-8-azabicyclo[3.2.1]oct-5-ylmethanol (0.3 g, 2.1 mmol), 2,4,6-trichloropyrimidine (0.46 g, 2.5 mmol), and diisopropylethylamine (0.81 g, 6.3 mmol) were dissolved in acetonitrile (20 ml), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and purified with an eluent system (petroleum ether/ethyl acetate=2:1) by the silica gel column chromatography to give a white solid compound 20i (0.15g, 24.8%).

[0293] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.39 (s, 1H), 5.25 (m, 1H), 4.22 (m, 1H), 3.98-3.76 (m, 3H), 3.67 (m, 3H), 2.23-2.14 (m, 1H), 2.14-1.96 (m, 2H), 1.78 (m, 1H).

[0294] Steps IX & X: Preparation of Compound 20k

[0295] At room temperature, the compound (8-(2,6-dichloropyrimidin-4-yl)-3-oxa-8-azabicyclo[3.2.1]oct-1-yl)methanol (0.15, 24.8%) was dissolved in dry dichloromethane, triethylamine (158 mg, 1.56 mmol) was added, and placed in an ice bath. Methanesulfonyl chloride (66 mg, 0.57 mmol) was slowly added. The reaction mixture was reacted in the ice bath for 1 h. The reaction mixture was concentrated under reduced pressure to give a white solid compound, which was used directly in the next step without purification. The coarse product and potassium carbonate (215 mg, 1.56 mmol) were dissolved in dioxane and water (40 mL, 1/1). The reaction mixture was reacted at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and purified with an eluent system (dichloromethane/methanol=20/1) by the silica gel column chromatography to give a white solid Compound 20k (40 mg, 30.8%).

[0296] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.72 (s, 1H), 4.29 (m, 1H), 4.08 (m, 1H), 3.97 (m, 1H), 3.77 (m, 4H), 2.46-2.27 (m, 1H), 2.14 (m, 1H), 1.96 (m, 1H), 1.77 (m, 1H).

[0297] Step XI: Preparation of Compound 20

[0298] At room temperature, the compound (3,5-difluoro-4-((2-(trifluoromethyl)pyridin-4-yl)oxy)phenyl)methanol (58, 0.19 mmol) was dissolved in dry N,N-dimethylformamide (5 mL), and sodium hydride (60% in mineral oil, 13 mg, 0.32 mmol) was added in an ice bath. 7-Chloro-3,4-dihydro-1H-4,11a-ethanopyrimido[6′,1′:2,3]imidazo[5,1-c][1,4]azinyl-9(11H)-one (40 mg, 0.16 mol) was added 15 min later, and reacted at room temperature for 1 h. The reaction mixture was quenched by adding ice water, and extracted with ethyl acetate (20 ml×3). The organic layers were combined, dried over anhydrous sodium sulfate, and filtered to remove the drying agent. The filtrate was concentrated under reduced pressure, and purified with an eluent system (dichloromethane/methanol=20/1) by the silica gel column chromatography to give a white solid compound 20 (35 mg, 35.3%).

[0299] .sup.1H NMR (600 MHz, CDCl.sub.3) δ 8.61 (m, 1H), 7.26 (s, 1H), 7.14 (m, 2H), 6.99 (m, 1H), 5.43 (s, 2H), 5.22 (s, 1H), 4.26 (m, 1H), 4.00 (m, 1H), 3.93 (m, 1H), 3.78-3.68 (m, 4H), 2.33-2.26 (m, 1H), 2.08 (m, 1H), 1.99-1.92 (m, 1H), 1.73 (m, 1H). m/z 523.0 [M+H].sup.+.

[0300] Biological Evaluation

[0301] The bioactivity of a compound could be determined using any suitable assay as well as tissue and in vivo model for determining the activity of a compound as an LpPLA2 inhibitor.

[0302] (1) Recombinant Human Lp-PLA2 Assay (rhLp-PLA2), Also Referred to as PED6 Assay

[0303] PED6 was a fluorescently-labeled phospholipid that could be purchased dorectly from Invitogene or Molecular Probes. It had a fluorescence-quenching p-nitrophenyl group at the Sn3 position and a Bodipy fluorescein (FL) group at the sn2 position. Once cleaved by the Lp-PLA2 enzyme, it would release the FL group, resulting in enhanced fluorescence. However, the Lp-PLA2 inhibitor could prevent occurrence of such cleavage, so that no enhanced fluorescence could be observed.

[0304] Assay Method: The compound to be tested (as shown in Table 1) was mixed with a DMSO solution in a volume ratio of 1:3, and diluted to prepare a source plate of a 384-well microplate. Then 0.01 μl of the compound was transferred via an ECHO liquid dispenser from the source plate to a 384-well Greiner 784076 plate, and 5 μl of a buffer consisting of 50 mM HEPES, pH7.4, 150 mM NaCl, and 1 mM CHAPS (the buffer solution containing a recombinant human Lp-PLA2 enzyme at a concentration of 4 nM or 110 pM) was added to each well of the plate. The plate was centrifuged at 500 rpm for 10 seconds. After a 30-min pre-incubation, 5 μl of the above-mentioned buffer was added to a 384-well Greiner 784076 plate, the plate was centrifuged at 500 rpm for 10 seconds. After the plate was incubated at room temperature for 20 min in a dark place, the fluorescence intensity was read at ex 480/em 540 with a ViewLux microplate imager, and the XL fitting model in Excel was used to perform the curve analysis and QC analysis to calculate pIC50. The results were listed in Table 1.

TABLE-US-00001 TABLE 1 Compound No. rhLp-PLA2 (pIC.sub.50)  1 8.6  2 9.1  3 9.3  4 10.4  5 10.5  6 10.3  7 10.4  8 10.5  9 10.0 10 10.2 11 10.1 12 10.1 13 9.6 14 10.1 16 10.0 17 10.1 18 10.0 19 9.5 Positive compound Rilapladib 8.9

[0305] (2) Assay of Lp-PLA2 in Human Plasma (Also Referred to as Thio-PAF Assay)

[0306] The human plasma assay was conducted using the sulphatide analog of PAF (phosphatidylcholine). After hydrolysis, it would generate phospholipids containing free sulfhydryl groups, which would be subjected to Michael addition with CPM to generate fluorescence-enhancing maleimide. Continuous quantitative analysis of thiol could be conducted by detecting the fluorescence intensity. This assay could be used to detect the inhibitory activity of the Lp-PLA2 inhibitor on the Lp-PLA2 enzyme in human plasma.

[0307] Assay Method: The compound to be tested (as shown in Table 2) was mixed with a DMSO solution in a volume ratio of (1:3), and diluted to prepare a source plate of a 384-well microplate. Then 0.01 μl of the compound was transferred via an ECHO liquid dispenser from the source plate to a 384-well Greiner 784076 plate, and 8 μl of pre-aliquoted and frozen mixed human plasma was then added. The plate was centrifuged at 500 rpm for 10 seconds. After a 30-min pre-incubation, 2 μl of a substrate solution, and a buffer containing 2.5 mM 2-thio-PAF (a solution in ethanol), 32 μM CPM (a solution in DMSO) and 3.2 mM N-ethylmaleimide (NEM) (a buffer solution consisting of 50 mM HEPES, pH7.4, 150 mM NaCl, 1 mM CHAPS) was added by a BRAVO liquid handling station to a 384-well Greiner 784076 low-volume plate. Two minutes later, the reaction was quenched with 5 μl of 5% trifluoroacetic acid. After the plate was incubated at room temperature for 40 min in a dark place, the fluorescence intensity was read at ex 380/em 485 with an Envision microplate reader, and the XL fitting model in Excel was used to perform the curve analysis and QC analysis to calculate pIC50. The results were listed in Table 2.

TABLE-US-00002 TABLE 2 Compound No. Thio-PAF (pIC.sub.50)  1 7.0  2 7.6  3 7.8  4 8.4  5 8.2  6 8.1  7 8.5  8 8.1  9 7.9 10 8.1 11 7.9 12 8.0 13 7.7 14 8.1 16 7.9 17 8.0 18 8.1 19 7.8 Positive compound Rilapladib 7.8