CN110669044A

CN110669044A - Donepezil-oxadiazole fusion compound and preparation method and application thereof

Info

Publication number: CN110669044A
Application number: CN201910847790.XA
Authority: CN
Inventors: 孙昊鹏; 林宏智; 乔玉婷; 冯锋; 曲玮; 柳文媛
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2019-09-09
Filing date: 2019-09-09
Publication date: 2020-01-10
Anticipated expiration: 2039-09-09
Also published as: CN110669044B

Abstract

The invention discloses a donepezil-oxadiazole fusion compound, the structural formula of which is shown as formula (I), (II), (III) or (IV), and also discloses a preparation method of the donepezil-oxadiazole fusion compound and application of the donepezil-oxadiazole fusion compound in preparation of a therapeutic drug for treating Alzheimer's disease.

Description

Donepezil-oxadiazole fusion compound and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicaments for treating Alzheimer's disease, and particularly relates to a donepezil-oxadiazole fusion compound and a preparation method and application thereof.

Background

Alzheimer's disease is a neurodegenerative disease with complex etiology, however, most of the drugs on the market for the disease are single-target cholinesterase inhibitors. The mere inhibition of the activity of acetylcholinesterase in the brain to increase the activity of the cholinergic system can temporarily improve the cognitive ability of the patient, but cannot relieve other pathological processes in the brain of the patient. Therefore, cholinesterase inhibitors, such as donepezil, have problems of large individual difference in therapeutic effect and single activity effect when used for treating alzheimer's disease.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a donepezil-oxadiazole fusion compound capable of relieving Alzheimer's disease.

The technical problem to be solved by the present invention is to provide a method for preparing the donepezil-oxadiazole fusion compound.

The invention also aims to solve the technical problem of providing the application of the donepezil-oxadiazole fusion compound in preparing the medicine for treating the Alzheimer disease.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

donepezil-oxadiazole fusion compounds having the structure of formula (i), (II) (iii) or (iv):

wherein R is₁Is halogen, methyl, methoxy, tert-butyl, trifluoromethyl or cyano;

x is methylene or carbonyl;

wherein Ar is a pyridine ring or a 2-substituted furan ring;

wherein R is₂Is hydrogen, fluorine or methyl, X is methylene or carbonyl;

wherein R is₃Is hydrogen, fluorine, or methyl.

A process for the preparation of a compound of the structure of formula (i) comprising the steps of:

(1) using N-boc-4-hydroxymethyl piperidine as a raw material, reacting with methanesulfonyl chloride under the conditions of using TEA as a catalyst and DCM as a solvent, converting alcohol into a mesylate active intermediate, then performing substitution reaction with phenolic hydroxyl on methyl 3-hydroxybenzoate to obtain ether under the condition of using potassium carbonate as a catalyst and DMF as a solvent to obtain an ester intermediate, and then performing substitution reaction on the intermediate in MeOH/H₂In the mixed solvent system, the intermediate 2 is obtained by LiOH catalytic hydrolysis;

(2) condensing the intermediate 2 and 2-methyl-N-hydroxy-5-pyridine formamidine by using CDI as a condensing agent, and performing ring synthesis at 105 ℃ to obtain an intermediate 3;

(3) deprotecting the intermediate 3 in trifluoroacetic acid to obtain an intermediate 4;

(4) using different substituted benzyl bromide or benzyl chloride and the intermediate 4 to react at 105 ℃ by taking DMF as a solvent under the catalysis of potassium carbonate to obtain target products NAD 1-NAD 15;

(5) benzoic acid or 4-fluorobenzoic acid and 4 are dehydrated and condensed into amide under the conditions that T3P is used as a condensing agent and ethyl acetate is used as a solvent, and NAD16 and NAD17 are obtained.

scheme 1 (a), i, MsCl, TEA, DCM; ii, 3-hydroxybenzoic acid methyl ester, K₂CO₃,DMF；ii,LiOH,MeOH/H₂O; (b) 2-methyl-N-hydroxy-5-pyridinecarboxamidine, CDI, DMF; (c) TFA, DCM; (g) 4, K₂CO₃,DMF；(h),4,T3P,TEA,EA.

A process for preparing a compound having the structure of formula (II), comprising the steps of:

(1) using substituted chloromethylpyridine hydrochloride at different positions and an intermediate 4 to react at 105 ℃ by taking DMF as a solvent under the catalysis of potassium carbonate to obtain target products NAD 18-NAD 20;

(2) condensing different substituted 2-furaldehyde and the intermediate 4 by using methanol as a solvent under an acidic condition, and then reducing the condensed product into tertiary amine under the action of sodium cyanoborohydride to obtain target products NAD 21-NAD 23.

scheme 2,(a),4,K₂CO₃,DMF；(b),4,MeOH,then NaBH₃CN,AcOH.

A process for the preparation of a compound of the structure of formula (iii) comprising the steps of:

(1) using N-boc-4-hydroxymethyl piperidine as a raw material, reacting with methanesulfonyl chloride under the conditions of using TEA as a catalyst and DCM as a solvent, converting alcohol into a mesylate active intermediate, then performing substitution reaction with phenolic hydroxyl on methyl 3-hydroxybenzoate to obtain ether under the condition of using potassium carbonate as a catalyst and DMF as a solvent to obtain an ester intermediate, and then performing substitution reaction on the intermediate in MeOH/H₂In the mixed solvent system, the intermediate 6 is obtained by LiOH catalytic hydrolysis;

(2) condensing the intermediate 2 and 2-methyl-N-hydroxy-5-pyridine formamidine by using CDI as a condensing agent, and performing ring synthesis at 105 ℃ to obtain an intermediate 7;

(3) deprotecting the intermediate 7 in trifluoroacetic acid to obtain an intermediate 8;

(4) using different substituted benzyl bromide or benzyl chloride and the intermediate 8 to react at 105 ℃ by taking DMF as a solvent under the catalysis of potassium carbonate to obtain target products NAD 24-NAD 28;

(5) benzoic acid or 4-fluorobenzoic acid and 8 are dehydrated and condensed into amide under the conditions that T3P is used as a condensing agent and ethyl acetate is used as a solvent, and NAD29 and NAD30 are obtained.

scheme 3 (a), i, MsCl, TEA, DCM; ii, 3-hydroxybenzoic acid methyl ester, K₂CO₃,DMF；ii,LiOH,MeOH/H₂O; (b) 2-methyl-N-hydroxy-5-pyridinecarboxamidine, CDI, DMF; (c) TFA, DCM; (g) 8, K₂CO₃,DMF；(h),8,T3P,TEA,EA.

A process for the preparation of a compound of the structure of formula (iv) comprising the steps of:

(1) using 3-bromopropyne and the phenolic hydroxyl of methyl 3-hydroxybenzoate to perform substitution reaction to obtain ether under the condition that potassium carbonate is used as a catalyst and DMF is used as a solvent, and then obtaining an ester intermediate in MeOH/H₂In the mixed solvent system, performing catalytic hydrolysis on the O by NaOH to obtain an intermediate 9;

(2) condensing the intermediate 9 and 2-methyl-N-hydroxy-5-pyridine formamidine by using CDI as a condensing agent, and performing ring synthesis at 105 ℃ to obtain an intermediate 10;

(3) reacting different substituted bromobenzyl or chlorobenzyl with sodium azide under the catalysis of potassium carbonate and with DMF as solvent at 105 ℃ to obtain azide derivative intermediate, and then reacting the intermediate in MeOH/H₂And O is a mixed solvent system, copper sulfate pentahydrate and ascorbic acid are used as catalysts, and target products NAD 31-NAD 35 are obtained through reaction at room temperature.

scheme 3 (a), i, 3-bromopropyne, K₂CO₂DMF,105 ℃; ii, naoh (aq), MeOH; (b) 2-methyl-N-hydroxy-5-pyridinecarboxamidine, CDI, DMF; (c) i, NaN₃,DMF,105℃；ii,10,CuSO₄·5H₂O,ascorbic acid,MeOH,H₂O.

The application of the donepezil-oxadiazole fusion compound in preparing the medicine for treating the Alzheimer disease is within the protection scope of the invention.

Has the advantages that:

the compound has a novel structure, has multiple activities of inhibiting cholinesterase, activating Nrf2 pathway and the like, and can improve the level of acetylcholine in brain and activate the antioxidant stress reaction of cells.

Drawings

FIG. 1 Induction Activity of NAD series compounds on ARE luciferase reporter genes. The fold induction of NAD series compounds at both 5 μ M and 20 μ M concentrations relative to the blank was plotted as the ordinate. Each data is presented as mean ± SD from two independent sets of experiments.

FIG. 2 Effect of NAD series compounds on the cellular activity of PC-12. Effect of NAD series of compounds on cell viability at 20. mu.M and 50. mu.M. Each data is presented as mean ± SEM from three independent experiments.

FIG. 3 Performance in the water maze experiment for control (control), model (model), tacrine and NAD3 mice.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.

Example 1: N-Boc-4-hydroxymethylpiperidine (1)

4-Hydroxymethylpiperidine (5.0g,43.41mmol) is dissolved in 50ml of dichloromethane, 9ml of triethylamine (65.12mmol) are added and the mixture is placed in an ice bath. 10.5g of Boc anhydride was dissolved in 50ml of dichloromethane and added dropwise to the reaction flask in ice bath. After completion, stir at room temperature overnight. Then 50ml of water was added to the reaction solution to quench the reaction, and after that, DCM was washed twice with saturated sodium bicarbonate (50ml), twice with water (50ml) and twice with brine (50 ml). The solvent DCM was removed by rotary evaporation under reduced pressure to give a colorless wax. 50ml of petroleum ether was added to the reaction flask and stirred for 2 hours to obtain a white powder. The powder was filtered off with suction and washed twice with diethyl ether (20 ml). Dried to obtain 8.0g of white powder. The yield thereof was found to be 85.6%. 1H-NMR (300MHz, CDCl3)4.14(br,2H),3.51(br,1H),2.71 (br), 1.74(br,1H),1.71(br,1H),1.69-1.64(m,1H),1.61(d, J ═ 2.3Hz,1H),1.47(s,9H),1.20-1.11(m,2H).

Example 2: 3- (N-Boc-4-piperidinyl) methoxybenzoic acid (2)

N-Boc-4-hydroxymethylpiperidine (5.0g, 23.2mmol) and 5ml triethylamine (36.0mmol) were dissolved in 50ml dichloromethane and placed in an ice bath. Methanesulfonyl chloride (3.2g, 27.87mmol) was dissolved in 50ml of dichloromethane, and dropwise added to the reaction flask, followed by stirring overnight at room temperature. The reaction mixture was quenched with 50ml of water, and then DCM was washed twice with saturated sodium bicarbonate (50ml), twice with water (50ml) and twice with brine (50 ml). The solvent DCM was removed by rotary evaporation under reduced pressure to give a colorless wax. 50ml of petroleum ether was added to the reaction flask and stirred for 2 hours to obtain a white powder. The powder was suction filtered and washed twice with petroleum ether (20 ml). And drying to obtain white powder. The solid was dissolved in 20ml of DMF and methyl 3-hydroxybenzoate (3.5g, 23.2mmol) and potassium carbonate (4.8g, 34.8mmol) were added and the mixture was stirred at 105 ℃ with warming. After 1 hour, a large amount of gum was formed, and 5ml of DMF was added in three portions to keep stirring. After 5 hours the reaction was complete, cooled to room temperature and quenched by the addition of 50ml of water to the reaction flask. After stirring well, the mixture was left standing for 2 hours to give a light brown oil. The solution was decanted and the remaining oil was dissolved in a mixed solution of 30ml methanol and 30ml water. Lithium hydroxide monohydrate (1.17g, 27.87mmol) was added to the reaction flask and the temperature was raised to 80 ℃ and stirred for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature. The reaction solution was washed twice with petroleum ether (40 ml). And placing the aqueous phase in an ice bath, dropwise adding dilute hydrochloric acid to adjust the acid to be weakly acidic, and then generating a large amount of white solid. And (4) carrying out suction filtration on the solid, washing a filter cake for three times, and drying to obtain 4.4g of white powder. The overall yield was 56.5%. 1H-NMR (300MHz, DMSO-d6)7.52(d, J ═ 4.6Hz,2H),7.43(s,1H),7.40(t, J ═ 4.7Hz,1H),7.18(dd, J1 ═ 4.9Hz, J2 ═ 1.1Hz,1H),3.88(d, J ═ 3.8Hz,2H),2.75(br,2H),1.76(d, J ═ 7.0Hz,2H),1.41-1.39(M,11H),1.21-1.13(M,3H), ms (esi): found334.1659, calcd for C18H24NO5[ M-H ] -334.1660.

Example 3: 5- [3- (N-boc-4-piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (3)

Intermediate 2(4.0g, 11.9mmol) and CDI (1.93g, 11.9mmol) were dissolved in 20ml DMF and stirred at room temperature for 45 min. Intermediate 2-methyl-N-hydroxy-5-pyridinecarboxamidine (1.80g, 11.9mmol) was added to the reaction flask, and the mixture was heated to 110 ℃ and stirred for 5 hours. The reaction mixture was cooled to room temperature and poured into 70ml of saturated aqueous sodium bicarbonate solution, and a brown oil precipitated. Standing for 1h, decanting the solution, dissolving the remaining oil in 80ml ethyl acetate, washing twice with water (40ml) and twice with brine (40ml), separating the organic phase, and drying with anhydrous sodium sulfate overnight. The solvent was removed by rotary evaporation under reduced pressure, and column chromatography (DCM: MeOH: 100:1) of the remaining oil gave 78.2% yield. 1H NMR (300MHz, DMSO) δ 9.27(d, J ═ 2.1Hz,1H),8.32(dd, J ═ 8.1,2.2Hz,1H),7.80(d, J ═ 7.7Hz,1H),7.70(dd, J ═ 2.3,1.6Hz,1H),7.46(t, J ═ 8.0Hz,1H),7.31(d, J ═ 8.0Hz,1H),7.14(dd, J ═ 8.2,2.5Hz,1H),3.92(d, J ═ 6.3Hz,2H), 2.83-2.71 (M,3H),2.66(s,3H),1.86(d, J ═ 11.7, 2H),1.70(d, J ═ 4, 2.47H), 1.84 (H, 18H, 3531H), 18H, 1.86(d, 11.7, 2H),1.70(d, J ═ 11H, 18H, 3H), 3H, 1.47 ═ 25H, 18]⁺451.2340.

Example 4: 5- [3- (4-Piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (4)

Intermediate 3(4.0g, 8.9mmol) was dissolved in 30ml dichloromethane and placed in an ice bath. 10ml of trifluoroacetic acid was added dropwise to the reaction flask in an ice bath. Then, the mixture was stirred at room temperature overnight. The solvent was removed by rotary evaporation under reduced pressure and the remaining oil was dissolved in 50ml of ethyl acetate, washed twice with saturated sodium bicarbonate solution (30ml) and twice with halogen (30 ml). The ethyl acetate layer was separated and dried over anhydrous sodium sulfate for 10 hours. The ethyl acetate was evaporated off under reduced pressure and the remaining oil was added to 50ml of petroleum ether and stirred at room temperature overnight to give a pale green solid. And (5) carrying out suction filtration and drying. The next step was carried out without purification. HRMS (ESI) found in found 351.1816, calcd for C20H23N4O2[ M + H ]]⁺351.1816.

Example 5: 5- [3- (N-benzyl-4-piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD1)

Intermediate 4(160mg, 0.46mmol) and benzyl bromide (93mg, 0.55mmol) were dissolved in 3ml DMF and potassium carbonate (95mg, 0.68mmol) was added. Stirring was carried out at 105 ℃ for 2 hours. After cooling to room temperature, 20ml of water was added to the reaction flask, followed by stirring thoroughly and standing for 1 hour. The solvent was decanted to give a black oil which was recrystallized from 5ml of ethanol to give a white powder. Suction filtration, washing with a little ethanol, and drying to obtain white powder 70mg with yield of 34.8%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.35(dd,J＝8.1,2.0Hz,1H),7.82(d,J＝7.7Hz,1H),7.72(s,1H),7.47(t,J＝8.0Hz,1H),7.34(t,J＝6.4Hz,6H),7.16(dd,J＝8.3,2.0Hz,1H),3.93(d,J＝5.8Hz,2H),3.55(s,2H),2.98(d,J＝11.3Hz,2H),2.68(s,3H),2.05(t,J＝11.6Hz,2H),1.88(d,J＝11.7Hz,3H),1.48(q,J＝12.8Hz,2H).HRMS(ESI):found 441.2287,calcd for C27H29N4O2[M+H]⁺441.2285.

Example 6: 5- {3- [ N- (4-Fluorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD2)

In the same manner as in example 5, 4-fluorobenzyl bromide (100mg, 0.55mmol) was used as a starting material to give 60mg of a white powder with a yield of 28.7%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.34(dd,J＝8.0,1.9Hz,1H),7.81(d,J＝7.6Hz,1H),7.72(s,1H),7.47(t,J＝8.0Hz,1H),7.36–7.29(m,3H),7.16(dd,J＝8.2,2.1Hz,1H),7.02(t,J＝8.6Hz,2H),3.93(d,J＝5.8Hz,2H),3.50(s,2H),2.95(d,J＝11.2Hz,2H),2.68(s,3H),2.04(t,J＝11.0Hz,2H),1.87(d,J＝11.2Hz,3H),1.48(q,J＝12.8Hz,2H).HRMS(ESI):found 459.2199,calcd for C27H28FN4O2[M+H]⁺459.2191.

Example 7: 5- {3- [ N- (3-fluorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD3)

In the same manner as in example 5, 3-fluorobenzyl chloride (80mg, 0.55mmol) was used as a starting material to give 110mg of a white powder with a yield of 52.5%.¹H NMR(300MHz,CDCl₃)δ9.29(d,J＝1.8Hz,1H),8.34(dd,J＝8.1,2.2Hz,1H),7.82(d,J＝7.7Hz,1H),7.73(d,J＝1.6Hz,1H),7.47(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.30-7.27(m,1H,solvent peak is involved),7.16(dd,J＝8.0,2.1Hz,1H.),7.16-7.11(m,2H),6.98-6.95(m,1H),3.94(d,J＝5.9Hz,2H),3.53(s,2H),2.96(d,J＝11.3Hz,2H),2.68(s,3H),2.06(t,J＝10.8Hz,2H),1.88(d,J＝11.4Hz,3H),1.49(q,J＝12.0Hz,2H).HRMS(ESI):found 459.2194,calcd for C27H28FN4O2[M+H]⁺459.2191.

Example 8: 5- {3- [ N- (2-fluorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD4)

In the same manner as in example 5, starting from 2-fluorobenzyl bromide (100mg, 0.55mmol), 70mg of a white powder was obtained with a yield of 33.4%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.2Hz,1H),8.34(dd,J＝8.0,1.9Hz,1H),7.81(d,J＝7.6Hz,1H),7.71(s,1H),7.47(t,J＝8.0Hz,1H),7.42(t,J＝7.2Hz,1H),7.33(d,J＝8.0Hz,1H),7.25(t,J＝7.1Hz,1H),7.16–7.12(m,2H),7.05(t,J＝9.1Hz,1H),3.92(d,J＝5.7Hz,2H),3.63(s,2H),3.00(d,J＝11.2Hz,2H),2.68(s,3H),2.12(t,J＝11.3Hz,2H),1,89-1.83(m,3H),1.49(d,J＝9.7Hz,2H).HRMS(ESI):found 459.2195,calcd forC27H28FN4O2[M+H]⁺459.2191.

Example 9: 5- {3- [ N- (4-chlorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD5)

In the same manner as in example 5, 4-chlorobenzyl chloride (88mg, 0.55mmol) was used as a starting material to give 120mg of a white powder with a yield of 57.3%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.9Hz,1H),8.34(dd,J＝8.1,2.2Hz,1H),7.81(d,J＝7.6Hz,1H),7.72(d,J＝1.5Hz,1H),7.47(t,J＝8.0Hz,1H),7.34-7.27(m,5H,solvent peak is involved),7.16(dd,J＝8.3,2.5Hz,1H),3.93(d,J＝5.9Hz,3H),3.50(s,2H),2.94(d,J＝11.3Hz,2H),2.68(s,3H),2.04(t,J＝10.9Hz,2H),1.87(d,J＝11.2Hz,3H),1.47(q,J＝11.9Hz,2H).HRMS(ESI):found 475.1904,calcd for C27H28ClN4O2[M+H]⁺475.1895.

Example 10: 5- {3- [ N- (3-chlorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD6)

In the same manner as in example 5, using 3-chlorobenzyl chloride (88mg, 0.55mmol) as a starting material, 130mg of a white powder was obtained with a yield of 62.1%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.8Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.7Hz,1H),7.73(d,J＝1.7Hz,1H),7.47(t,J＝8.0Hz,1H),7.37(s,1H),7.33(d,J＝8.1Hz,1H),7.27-7.22(m,3H),7.16(dd,J＝8.2,2.1Hz,1H),3.94(d,J＝5.9Hz,2H),3.51(s,2H),2.95(d,J＝11.3Hz,2H),2.68(s,3H),2.06(t,J＝10.8Hz,2H),1.88(d,J＝11.5Hz,3H),1.56-1.33(m,2H).HRMS(ESI):found 475.1895,calcd for C27H28ClN4O2[M+H]⁺475.1895.

Example 11: 5- {3- [ N- (2-chlorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD7)

In the same manner as in example 5, starting from 4-bromobenzyl (88mg, 0.55mmol), 50mg of a white powder was obtained in a yield of 23.9%.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.7Hz,1H),8.34(dd,J＝8.1,2.2Hz,1H),7.82(d,J＝7.7Hz,1H),7.73(dd,J＝2.2,1.6Hz,1H),7.53(dd,J＝7.6,1.4Hz,1H),7.48(t,J＝8.0Hz,1H),7.37(dd,J＝7.9,1.2Hz,1H),7.33(d,J＝8.1Hz,1H),7.26(dd,J＝7.5,1.2Hz,1H),7.21(td,J＝7.7,1.7Hz,1H),7.17(ddd,J＝8.3,2.5,0.7Hz,1H),3.94(d,J＝6.0Hz,2H),3.66(s,2H),3.01(d,J＝11.4Hz,2H),2.68(s,3H),2.18(t,J＝10.9Hz,2H),1.89(d,J＝12.4Hz,3H),1.51(dt,J＝11.5,9.4Hz,2H).HRMS(ESI):found 475.1896,calcdfor C27H28ClN4O2[M+H]⁺475.1895.

Example 12: 5- {3- [ N- (4-bromobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD8)

In the same manner as in example 5, 4-bromochlorobenzyl chloride (137mg, 0.55mmol) was used as a starting material to give 120mg of a white powder with a yield of 50.6%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.6Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.7Hz,1H),7.72(s,1H),7.47(t,J＝8.3Hz,3H),7.33(d,J＝8.1Hz,1H),7.24(d,J＝8.3Hz,2H),7.16(dd,J＝8.3,1.9Hz,1H),3.93(d,J＝5.9Hz,2H),3.49(s,2H),2.94(d,J＝11.3Hz,2H),2.68(s,3H),2.04(t,J＝10.9Hz,2H),1.87(d,J＝9.8Hz,3H),1.47(dd,J＝11.9,2.3Hz,2H).HRMS(ESI):found 519.1398,calcd for C27H28BrN4O2[M+H]⁺519.1390.

Example 13: 5- {3- [ N- (4-methylbenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD9)

In the same manner as in example 5, 4-methylbenzyl bromide (101mg, 0.55mmol) was used as a starting material to give 60mg of a white powder with a yield of 28.9%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.34(dd,J＝8.1,1.9Hz,1H),7.81(d,J＝7.5Hz,1H),7.72(s,1H),7.47(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.24(d,J＝7.8Hz,2H),7.16(d,J＝7.7Hz,3H),3.92(d,J＝5.8Hz,2H),3.51(s,2H),2.98(d,J＝10.9Hz,2H),2.68(s,3H),2.37(s,3H),2.03(t,J＝11.1Hz,2H),1.87(d,J＝10.4Hz,3H),1.47(q,J＝11.9Hz,2H).HRMS(ESI):found 455.2445,calcd for C28H31N4O2[M+H]⁺455.2442.

Example 14: 5- {3- [ N- (4-methoxybenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD10)

In the same manner as in example 5, 4-methylbenzyl chloride (86mg, 0.55mmol) was used as a starting material to give 40mg of a white powder with a yield of 18.6%.¹H NMR(500MHz,CDCl₃)δ9.29(s,1H),8.34(d,J＝7.8Hz,1H),7.81(d,J＝7.5Hz,1H),7.71(s,1H),7.46(t,J＝7.9Hz,1H),7.32(d,J＝8.0Hz,1H),7.29-7.26(m,2H),7.15(d,J＝7.7Hz,1H),6.88(d,J＝8.1Hz,2H),3.92(d,J＝5.5Hz,2H),3.82(s,3H),3.50(s,2H),2.98(d,J＝10.7Hz,2H),2.67(s,3H),2.03(t,J＝11.2Hz,2H),1.87(d,J＝10.6Hz,3H),1.48(d,J＝11.4Hz,2H).HRMS(ESI):found 471.2393,calcd for C28H31N4O2[M+H]⁺471.2391.

Example 15: 5- {3- [ N- (4-tert-butylbenzyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD11)

In the same manner as in example 5, 4-tert-butylbenzyl chloride (100mg, 0.55mmol) was used as a starting material to give 40mg of a white powder with a yield of 39.7%.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.6Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.81(d,J＝7.7Hz,1H),7.72(s,1H),7.47(t,J＝8.0Hz,1H),7.37(d,J＝8.2Hz,2H),7.33(d,J＝8.1Hz,1H),7.28(d,J＝7.4Hz,2H),7.16(dd,J＝8.3,1.8Hz,1H),3.93(d,J＝5.9Hz,2H),3.52(s,2H),2.99(d,J＝11.3Hz,2H),2.68(s,3H),2.04(t,J＝10.9Hz,2H),1.88-1.83(m,3H),1.52-1.45(m,2H),1.35(s,9H).HRMS(ESI):found 497.2909,calcd forC31H37N4O2[M+H]⁺497.2911.

Example 16: 5- {3- [ N- (4-cyanobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD12)

In the same manner as in example 5, 4-cyanobenzyl chloride (83mg, 0.55mmol) was used as a starting material to give 60mg of a white powder with a yield of 28.2%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.5Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.6Hz,1H),7.72(d,J＝1.3Hz,1H),7.63(d,J＝8.2Hz,2H),7.52–7.44(m,3H),7.33(d,J＝8.1Hz,1H),7.16(dd,J＝7.9,2.1Hz,1H),3.94(d,J＝5.8Hz,2H),3.58(s,2H),2.92(d,J＝11.1Hz,2H),2.67(s,3H),2.09(t,J＝11.5Hz,2H),1.89-1.81(m,3H),1.48(d,J＝11.6Hz,2H).HRMS(ESI):found 466.2228,calcd for C28H28N5O2[M+H]⁺466.2238.

Example 17: 5- {3- [ N- (4-trifluoromethylbenzyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD13)

In the same manner as in example 5, 4-trifluoromethylbenzyl chloride (106mg, 0.55mmol) was used as a starting material to obtain 60mg of a white powder with a yield of 25.8%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.35(dd,J＝8.0,1.9Hz,1H),7.82(d,J＝7.6Hz,1H),7.73(s,1H),7.60(d,J＝7.9Hz,2H),7.46(d,J＝7.3Hz,3H),7.33(d,J＝8.2Hz,1H),7.17(dd,J＝8.2,2.4Hz,1H),3.94(d,J＝5.8Hz,2H),3.59(s,2H),2.95(d,J＝11.2Hz,2H),2.68(s,3H),2.08(t,J＝11.1Hz,2H),1.89(d,J＝11.2Hz,3H),1.49(q,J＝12.1Hz,2H).HRMS(ESI):found 509.2159,calcd for C28H28F3N4O2[M+H]⁺509.2159.

Example 18: 5- {3- [ N- (3, 4-difluorobenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD14)

In the same manner as in example 5, 3, 4-difluorobenzyl chloride (89mg, 0.55mmol) was used as a starting material to give 70mg of a white powder in a yield32.2％。¹H NMR(500MHz,CDCl₃)δ9.29(s,1H),8.34(dd,J＝8.0,1.4Hz,1H),7.82(d,J＝7.5Hz,1H),7.72(s,1H),7.47(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.29(s,1H),7.21(t,J＝8.1Hz,,1H),7.16(dd,J＝8.3,2.4Hz,1H),7.11(dd,J＝18.2,8.4Hz,1H),7.06(br,1H),3.94(d,J＝5.8Hz,2H),3.48(s,2H),2.94(dd,J＝23.7,12.6Hz,2H),2.68(s,3H),2.05(t,J＝11.2Hz,2H),1.88(d,J＝11.4Hz,3H),1.48(dd,J＝22.1,12.1Hz,2H).HRMS(ESI):found 477.2101,calcd for C27H27F2N4O2[M+H]⁺477.2097.

Example 19: 5- {3- [ N- (3, 4-dimethylbenzyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD15)

In the same manner as in example 5, 3, 4-dimethylbenzyl chloride (85mg, 0.55mmol) was used as a starting material to give 120mg of a white powder with a yield of 56.1%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.8Hz,1H),8.34(dd,J＝8.1,2.2Hz,1H),7.81(d,J＝7.7Hz,1H),7.72(d,J＝2.0Hz,1H),7.47(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.15(dd,J＝8.3,1.9Hz,1H),7.14–7.05(m,3H),3.93(d,J＝5.9Hz,2H),3.49(s,2H),2.98(d,J＝11.3Hz,2H),2.68(s,3H),2.28(s,3H),2.27(s,3H),2.04(t,J＝11.3Hz,2H),1.87(d,J＝10.8Hz,3H),1.52-1.45m,2H).HRMS(ESI):found 469.2599,calcdfor C29H33N4O2[M+H]⁺469.2589.

Example 20: 5- [3- (N-benzoyl-4-piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD16)

Intermediate 4(160mg, 0.46mmol) and benzoic acid (67mg, 0.55mmol) were dissolved in 5ml DMF and triethylamine (100mg, 1.0mmol) was added. T3P (50% w.t.in DMF,436mg,0.68mmol) was added to the reaction flask and stirred at 105 ℃ for 5 h. After cooling to room temperature, 30ml of ethyl acetate was added to the reaction flask. The ethyl acetate layer was washed three times with 0.5N sodium hydroxide solution (15ml) and waterTwice (15ml) and twice (15ml) brine. The mixture was dried overnight with anhydrous sodium sulfate. Ethyl acetate was removed by rotary evaporation under reduced pressure, and the remaining solid was recrystallized from 10ml of water/methanol (1: 1) to give 40mg of a white powder with a yield of 19.2%.¹H NMR(500MHz,CDCl₃)δ9.29(s,1H),8.34(d,J＝7.6Hz,1H),7.84(d,J＝7.4Hz,1H),7.73(s,1H),7.49(t,J＝7.9Hz,1H),7.44(s,5H),7.33(d,J＝7.9Hz,1H),7.17(d,J＝7.5Hz,1H),3.98(br,2H),2.99(d,J＝108.2Hz,2H),2.68(s,3H),2.18(br,1H),1.96(d,J＝64.1Hz,2H),1.45(d,J＝50.8Hz,4H).HRMS(ESI):found 455.2080,calcd for C27H27N4O3[M+H]⁺455.2078.

Example 21: 5- {3- [ N- (4-fluorobenzoyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD17)

In the same manner as in example 20, starting from 4-fluorobenzoic acid (77mg, 0.55mmol), 30mg of a white powder was obtained in a yield of 13.9%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.7Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.83(d,J＝7.7Hz,1H),7.72(d,J＝2.0Hz,1H),7.49(d,J＝8.0Hz,1H),7.48–7.43(m,2H),7.33(d,J＝8.1Hz,1H),7.16(dd,J＝8.3,1.8Hz,1H),7.12(t,J＝8.6Hz,2H),3.98(br,2H),3.00(d,J＝103.0Hz,2H),2.67(s,3H),2.17(br,1H),1.97(br,2H),1.44(br,2H).HRMS(ESI):found 473.1989,calcd for C27H26FN4O3[M+H]⁺473.1983.

Example 22: 5- {3- [ N- (4-Piperidinylmethyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD18)

In the same manner as in example 5, 4-chloromethylpyridine hydrochloride (90mg, 0.55mmol) and potassium carbonate (190mg, 1,37mmol) were used as acid-binding agents to obtain 50mg of white powder with a yield of 24.8%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.7Hz,1H),8.56(d,J＝5.7Hz,2H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.7Hz,1H),7.73(s,1H),7.47(t,J＝8.0Hz,1H),7.31(dd,J＝9.9,7.0Hz,3H),7.16(dd,J＝8.2,2.1Hz,1H),3.94(d,J＝5.9Hz,2H),3.54(s,2H),2.93(d,J＝11.3Hz,2H),2.67(s,3H),2.09(t,J＝10.9Hz,2H),1.89(d,J＝9.9Hz,3H),1.50(dt,J＝12.2,9.8Hz,2H).HRMS(ESI):found 442.2235,calcd for C26H28N5O2[M+H]⁺442.2238.

Example 23: 5- {3- [ N- (3-Piperidinylmethyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD19)

In the same manner as in example 5, 3-chloromethylpyridine hydrochloride (90mg, 0.55mmol) and potassium carbonate (190mg, 1,37mmol) were used as acid-binding agents to obtain 50mg of white powder with a yield of 24.8%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.6Hz,1H),8.57(s,1H),8.53(d,J＝3.6Hz,1H),8.33(dd,J＝8.1,2.1Hz,1H),7.81(d,J＝7.6Hz,1H),7.71(d,J＝8.5Hz,2H),7.47(t,J＝8.0Hz,1H),7.32(d,J＝8.1Hz,1H),7.28(dd,J＝7.7,4.8Hz,1H),7.15(dd,J＝8.0,2.1Hz,1H),3.93(d,J＝5.7Hz,2H),3.53(d,J＝22.9Hz,2H),2.95(d,J＝11.2Hz,2H),2.67(s,3H),2.08(t,J＝10.9Hz,2H),1.88(d,J＝11.4Hz,3H),1.51-1.44(m,2H).HRMS(ESI):found 442.2234,calcd for C26H28N5O2[M+H]⁺442.2238.

Example 24: 5- {3- [ N- (2-Piperidinylmethyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD20)

In the same manner as in example 5, 2-chloromethylpyridine hydrochloride (90mg, 0.55mmol) and potassium carbonate (190mg, 1,37mmol) were used as acid-binding agents to obtain 90mg of white powder with a yield of 44.6%.¹H NMR(500MHz,CDCl₃)δ9.29(s,1H),8.59(s,1H),8.34-8.33(m,1H),7.80(d,J＝6.6Hz,1H),7.72(s,1H),7.67(d,J＝7.5Hz,1H),7.46(t,J＝7.8Hz,2H),7.32(d,J＝7.9Hz,1H),7.17(dd,J＝15.3,7.6Hz,2H),3.93(d,J＝5.4Hz,2H),3.70(s,2H),3.00(d,J＝9.5Hz,2H),2.67(s,3H),2.18-2.14(m,2H),1.88(d,J＝11.6Hz,3H),1.54(d,J＝11.8Hz,2H).HRMS(ESI):found 442.2236,calcdfor C26H28N5O2[M+H]⁺442.2238.

Example 25: 5- {3- [ N- (4-Furanylmethyl) -4-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD21)

Furfural (53mg,0.55mmol) and intermediate 4(160mg, 0.46mmol) were dissolved in 5ml of methanol, a drop of acetic acid was added, and the mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (56mg, 0.91mmol) was added to the reaction flask and stirred at room temperature overnight. The reaction was quenched by addition of water (20 ml). The aqueous phase was extracted three times with ethyl acetate (20 ml). The ethyl acetate was combined, and dried over anhydrous sodium sulfate for 6 hours. Ethyl acetate was removed by rotary evaporation under reduced pressure and the remaining oil was purified by chromatography (DCM: MeOH ═ 100; 1) to give 45mg of a white powder in 22.9% yield.¹H NMR(300MHz,CDCl₃)δ9.28(d,J＝1.6Hz,1H),8.33(dd,J＝8.1,2.2Hz,1H),7.87–7.77(m,1H),7.70(dd,J＝2.3,1.6Hz,1H),7.46(t,J＝8.0Hz,1H),7.41(dd,J＝1.8,0.8Hz,1H),7.32(d,J＝8.1Hz,1H),7.14(ddd,J＝8.3,2.6,0.9Hz,1H),6.34(dd,J＝3.1,1.9Hz,1H),6.24(d,J＝3.1Hz,1H),3.92(d,J＝5.9Hz,2H),3.60(s,2H),3.01(d,J＝11.5Hz,2H),2.67(s,3H),2.10(t,J＝10.8Hz,2H),1.89(d,J＝10.3Hz,3H),1.59-1.50(m,2H).HRMS(ESI):found 431.2077,calcd for C25H27N4O3[M+H]⁺431.2078.

Example 26: 5- {3- [ N- (5-chloro-2-furylmethyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD22)

In the same manner as in example 25, 5-chlorofurfural (71mg, 0.55mmol) and the starting materials were used to give 40mg of a white powder with a yield of 18.8%.¹H NMR(500MHz,CDCl₃)δ9.28(s,1H),8.33(d,J＝7.9Hz,1H),7.81(d,J＝7.4Hz,1H),7.67(s,1H),7.46(t,J＝8.0Hz,1H),7.33(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.85(s,1H),6.28(d,J＝2.3Hz,1H),4.27(s,2H),4.00(s,1H),3.57(br,2H),2.87(br,2H),2.67(s,3H),2.23-1.96(m,5H).HRMS(ESI):found 465.1693,calcd forC25H26ClN4O3[M+H]⁺465.1688.

Example 27: 5- {3- [ N- (5-bromo-2-furylmethyl) -4-piperidinylmethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD23)

In the same manner as in example 25, starting from 5-bromofurfural (96mg, 0.55mmol) and carbon, 40mg of white powder was obtained with a yield of 17.2%.¹H NMR(500MHz,CDCl₃)δ9.28(s,1H),8.33(d,J＝7.9Hz,1H),7.81(d,J＝7.4Hz,1H),7.67(s,1H),7.46(t,J＝7.9Hz,1H),7.33(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.85(s,1H),6.28(d,J＝2.3Hz,1H),4.27(s,2H),4.00(s,2H),3.57(br,2H),2.87(br,2H),2.67(s,3H),2.16(br,5H).HRMS(ESI):found 509.1170,calcd for C25H26BrN4O3[M+H]⁺509.1183.

Example 28: 5- [3- (3-Piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (8)

The procedure of examples 2,3 and 4 was followed to give intermediate 8 as a brown solid in an amount of 0.6g with a total yield of 21.5%. The next step was carried out without purification.¹HRMS(ESI):found 351.1816,calcd for C20H23N4O2[M+H]⁺351.1816.

Example 29: 5- [3- (N-benzyl-3-piperidinemethoxy) phenyl ] -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD24)

Intermediate 5(160mg, 0.46mmol) and benzyl bromide (93mg, 0.55mmol) were dissolved in 3ml DMF and potassium carbonate was added(95mg, 0.68 mmol). Stirring was carried out at 105 ℃ for 2 hours. After cooling to room temperature, 20ml of water was added to the reaction flask, followed by stirring thoroughly and standing for 1 hour. The solvent was decanted to give a black oil, which was purified over column (DCM: MeOH ═ 200: 1). 120mg of a colorless oil was obtained in 56.7% yield.¹H NMR(300MHz,CDCl₃)δ9.30(d,J＝1.6Hz,1H),8.35(dd,J＝8.1,2.2Hz,1H),7.83-7.80(m,1H),7.71(dd,J＝2.4,1.6Hz,1H),7.47(t,J＝8.0Hz,1H),7.36-7.29(m,7H),7.14(ddd,J＝8.3,2.6,1.0Hz,1H),3.96(d,J＝6.3Hz,2H),3.56(q,J＝13.2Hz,2H),2.89(dd,J＝59.1,10.5Hz,2H),2.68(s,3H),2.27-2.17(m,1H),2.10-2.00(m,2H),1.90-1.84(m,2H),1.75-1.68(m,2H).HRMS(ESI):found 441.2283,calcd for C27H29N4O2[M+H]⁺441.2285.

Example 30: 5- {3- [ N- (4-Fluorobenzyl) -3-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD25)

The procedure is as in example 29, except that 4-fluorobenzyl bromide (100mg, 0.55mmol) is used as the starting material, 110mg of a colorless oil is obtained, and the yield is 52.5%.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.8Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.7Hz,1H),7.71(s,1H),7.47(t,J＝8.0Hz,1H),7.34-7.30(m,3H),7.14(dd,J＝8.2,2.3Hz,1H),7.00(t,J＝8.6Hz,2H),3.96(d,J＝6.3Hz,2H),3.51(dd,J＝34.2,13.2Hz,2H),2.86(dd,J＝98.2,10.2Hz,2H),2.68(s,3H),2.22-2.18(m,1H),2.09-2.00(m,2H),1.88-1.73(m,3H),1.68-1.64(m,1H).HRMS(ESI):found 459.2192,calcd forC27H28FN4O2[M+H]⁺459.2191.

Example 30: 5- {3- [ N- (3-fluorobenzyl) -3-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD26)

In the same manner as in example 29, 3-fluorobenzyl chloride (79mg, 0.55mmol) was used as a starting material to give 110mg of a colorless oil in a yield of 47.8%.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.8Hz,1H),8.35(dd,J＝8.1,2.2Hz,1H),7.82(d,J＝7.7Hz,1H),7.71(s,1H),7.47(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.30-7.24(m,2H),7.15(dd,J＝8.3,2.0Hz,1H),7.10(t,J＝8.1Hz,2H),6.95(td,J＝8.5,2.2Hz,1H),3.97(d,J＝5.9Hz,2H),3.54(dd,J＝33.3,13.5Hz,2H),2.87(dd,J＝100.1,10.3Hz,2H),2.68(s,3H),2.22(ddd,J＝9.9,6.4,3.7Hz,1H),2.07(dt,J＝20.1,9.8Hz,2H),1.94-1.83(m,1H),1.81-1.68(m,3H).HRMS(ESI):found 459.2194,calcd forC27H28FN4O2[M+H]⁺459.2191.

Example 31: 5- {3- [ N- (2-Fluorobenzyl) -3-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD27)

In the same manner as in example 29, starting from 2-fluorobenzyl chloride (79mg, 0.55mmol), 110mg of a colorless oil was obtained in a yield of 23.8%.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.6Hz,1H),8.35(dd,J＝8.1,2.2Hz,1H),7.81(d,J＝7.8Hz,1H),7.71(dd,J＝2.2,1.5Hz,1H),7.47(t,J＝8.0Hz,1H),7.41(td,J＝7.5,1.4Hz,1H),7.33(d,J＝8.1Hz,1H),7.28–7.22(m,1H),7.16-7.13(m,1H),7.11(td,J＝7.5,0.9Hz,1H),7.07-7.02(m,1H),3.96(d,J＝6.4Hz,2H),3.64(d,J＝3.1Hz,2H),2.92(dd,J＝99.0,10.5Hz,2H),2.68(s,3H),2.28-2.18(m,1H),2.14(t,J＝9.7Hz,1H),2.07(t,J＝10.2Hz,1H),1.90-1.61(m,5H).HRMS(ESI):found 459.2193,calcd forC27H28FN4O2[M+H]⁺459.2191.

Example 32: 5- {3- [ N- (4-methylbenzyl) -3-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD28)

In the same manner as in example 29, starting from 4-methylbenzyl chloride (77mg, 0.55mmol), 110mg of a colorless oil was obtained in 23.8% yield.¹H NMR(500MHz,CDCl₃)δ9.30(d,J＝1.6Hz,1H),8.35(dd,J＝8.1,2.2Hz,1H),7.81(d,J＝7.8Hz,1H),7.71(dd,J＝2.2,1.5Hz,1H),7.47(t,J＝8.0Hz,1H),7.41(td,J＝7.5,1.4Hz,1H),7.33(d,J＝8.1Hz,1H),7.28–7.22(m,1H),7.16-7.13(m,1H),7.11(td,J＝7.5,0.9Hz,1H),7.07-7.02(m,1H),3.96(d,J＝6.4Hz,2H),3.64(d,J＝3.1Hz,2H),2.92(dd,J＝99.0,10.5Hz,2H),2.68(s,3H),2.28-2.18(m,1H),2.14(t,J＝9.7Hz,1H),2.07(t,J＝10.2Hz,1H),1.90-1.61(m,5H).HRMS(ESI):found 459.2193,calcd forC27H28FN4O2[M+H]⁺459.2191.

Example 33: 5- [3- (N-benzoyl-3-piperidinylmethoxy) phenyl ] -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD29)

In the same manner as in example 20, starting from benzoic acid (67mg, 0.55mmol), 30mg of a white powder was obtained in a yield of 14.4%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.82(d,J＝7.5Hz,1H),7.66(d,J＝44.2Hz,1H),7.41(s,3H),7.33(d,J＝8.1Hz,1H),7.10(d,J＝52.3Hz,1H),4.60(d,J＝112.2Hz,1H),3.96(d,J＝41.1Hz,3H),3.00(d,J＝69.6Hz,2H),2.67(s,3H),2.21-2.00(m,2H),1.92-1.49(m,5H).HRMS(ESI):found 455.2088,calcd forC28H31N4O2[M+H]⁺455.2088.

Example 34: 5- {3- [ N- (4-fluorobenzoyl) -3-piperidinemethoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD30)

In the same manner as in example 20, starting from 4-fluorobenzoic acid (67mg, 0.55mmol), 30mg of a white powder was obtained in 18.5% yield.¹H NMR(300MHz,CDCl₃)δ9.29(d,J＝1.5Hz,1H),8.35(dd,J＝8.1,2.2Hz,1H),7.84(d,J＝7.6Hz,1H),7.67(s,1H),7.45(dt,J＝12.1,6.1Hz,3H),7.33(d,J＝8.1Hz,1H),7.09(t,J＝8.4Hz,3H),4.57(d,J＝113.2Hz,1H),4.57(d,J＝113.2Hz,1H),3.99(s,3H),3.00(d,J＝76.3Hz,2H),2.68(s,3H),2.19(s,1H),2.06(d,J＝13.8Hz,1H),1.84(s,1H),1.74(s,2H),1.64-1.49(m,2H).HRMS(ESI):found 459.2193,calcd for C27H28FN4O2[M+H]⁺459.2191.

Example 35: 3- [1- (2-Propynyloxy) ] benzoic acid (9)

Methyl 3-hydroxybenzoate (0.5g, 3.3mmol) and 1-bromo-2-propyne (0.47g, 3.94mmol) were dissolved in DMF (5ml), potassium carbonate (0.68g, 0.49mmol) was added, and the mixture was stirred at 105 ℃ for 5 hours. After cooling to room temperature, water (40ml) was added and a light brown oil precipitated, which was left to stand for 1 hour and the solvent was decanted off. The remaining oil was dissolved in methanol (20mL), 50% sodium hydroxide solution (20mL) was added, and the mixture was stirred at 105 ℃ for 2 hours. Cooled to room temperature and acidified with dilute hydrochloric acid. White powder precipitated. Suction filtration and washing twice with water (10 ml). Oven drying to obtain white powder 0.5g, yield 86.4%.¹H NMR(500MHz,CDCl₃)δ7.82–7.77(m,1H),7.74(dd,J＝2.5,1.4Hz,1H),7.44(t,J＝8.0Hz,1H),7.27(ddd,J＝8.2,2.6,0.8Hz,1H),4.79(d,J＝2.4Hz,2H),2.58(t,J＝2.4Hz,1H).

Example 36: 5- {3- [1- (2-propynyloxy) ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (10)

In the same manner as in example 3, 3- [1- (2-propynyloxy)]Benzoic acid (1g, 5.68mmol) was used as the starting material to give compound 6 as a white powder 0.9g, yield 54.4%.¹H NMR(500MHz,CDCl₃)δ9.29(d,J＝1.8Hz,1H),8.34(dd,J＝8.1,2.1Hz,1H),7.89(d,J＝7.7Hz,1H),7.83(d,J＝2.2Hz,1H),7.52(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.26(dd,J＝8.2,2.3Hz,1H),4.83(d,J＝2.3Hz,2H),2.68(s,3H),2.60(t,J＝2.3Hz,1H).

Example 37: 5- {3- [ 1-benzyl-4- (1,2, 3-triazolyl) methoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD31)

Benzyl bromide (112mg, 0.66mmol) was dissolved in 5ml of DMF, and sodium azide (50mg, 0.77mmol) was added and stirred at 110 ℃ for 3 hours. After cooling to room temperature, 20ml of ethyl acetate were added, and the organic phase was washed three times with water (20 ml). Then, ethyl acetate was removed by rotary evaporation under reduced pressure. After the reaction flask was charged with 5ml of methanol and 5ml of water, intermediate 6(160mg, 0.55mmol) was added to the reaction flask, followed by addition of a catalytic amount of copper sulfate pentahydrate and ascorbic acid and stirring at room temperature overnight. Extracted three times with ethyl acetate (20ml), the organic phases were combined and dried for 5 hours with anhydrous sodium sulfate. Ethyl acetate was rotary evaporated under reduced pressure to give a white powder, which was purified by column chromatography (DCM: MeOH: 200:1) to give 50mg of white powder with a yield of 21.45%.¹H NMR(500MHz,CDCl₃)δ9.28(d,J＝1.5Hz,1H),8.33(dd,J＝8.1,2.2Hz,1H),7.85(d,J＝7.8Hz,1H),7.82–7.81(m,1H),7.60(s,1H),7.49(t,J＝8.0Hz,1H),7.40(q,J＝5.4Hz,3H),7.32(dd,J＝9.7,7.4Hz,3H),7.25(dd,J＝8.3,1.9Hz,1H),5.58(s,2H),5.31(s,2H),2.68(s,3H).HRMS(ESI):found 425.1730,calcdfor C24H21N6O2[M+H]⁺425.1721.

Example 38: 5- {3- [1- (4-Fluorobenzyl) -4- (1,2, 3-Triazolyl) methoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD32)

The procedure is as in example 37, except that 4-fluorobenzyl bromide (125mg, 0.66mmol) is used as the starting material to give 70mg of a white powder with a yield of 28.8%.¹H NMR(300MHz,CDCl₃)δ9.28(s,1H),9.28(s,1H),8.33(dd,J＝8.1,2.2Hz,1H),7.91–7.78(m,2H),7.60(s,1H),7.49(t,J＝8.0Hz,1H),7.32(dd,J＝8.9,3.8Hz,3H),7.25(ddd,J＝8.4,2.6,1.0Hz,1H),7.13–7.05(m,2H),5.55(s,2H),5.31(s,2H),2.67(s,3H).HRMS(ESI):found 443.1634,calcd for C24H20FN6O2[M+H]⁺443.1626.

Example 39: 5- {3- [1- (3-Fluorobenzyl) -4- (1,2, 3-Triazolyl) methoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD33)

In the same manner as in example 37, 3-fluorobenzyl chloride (95mg, 0.66mmol) was used as a starting material to obtain 90mg of a white powder with a yield of 37.0%.¹H NMR(500MHz,CDCl₃)δ9.28(d,J＝1.8Hz,1H),8.33(dd,J＝8.1,2.2Hz,1H),7.85(d,J＝7.7Hz,1H),7.83–7.81(m,1H),7.64(s,1H),7.49(t,J＝8.0Hz,1H),7.37(td,J＝8.0,5.9Hz,1H),7.33(d,J＝8.1Hz,1H),7.26(dd,J＝8.3,2.0Hz,1H),7.11–7.07(m,2H),7.01–6.97(m,1H),5.57(s,2H),5.33(s,2H),2.67(s,3H).HRMS(ESI):found 443.1635,calcd for C24H20FN6O2[M+H]⁺443.1626.

Example 40: 5- {3- [1- (2-Fluorobenzyl) -4- (1,2, 3-Triazolyl) methoxy ] phenyl } -3- [3- (6-methylpyridinyl) ] -1,2, 4-oxadiazole (NAD34)

In the same manner as in example 37, 2-fluorobenzyl chloride (95mg, 0.66mmol) was used as a starting material to obtain 90mg of a white powder with a yield of 37.0%.¹H NMR(300MHz,CDCl₃)δ9.29(s,1H),8.38–8.30(m,1H),7.85(d,J＝8.4Hz,2H),7.70(s,1H),7.50(d,J＝8.0Hz,1H),7.36(dd,J＝16.7,7.5Hz,3H),7.27(d,J＝10.1Hz,1H),7.16(dd,J＝16.7,8.6Hz,2H),5.64(s,2H),5.31(s,2H),2.68(s,3H).HRMS(ESI):found 443.1631,calcd for C24H20FN6O2[M+H]⁺443.1626.

Example 41: 5- {3- [1- (4-methylbenzyl) -4- (1,2, 3-triazolyl) methoxy ] phenyl } -3- [3- (6-methylpyridyl) ] -1,2, 4-oxadiazole (NAD35)

In the same manner as in example 37, 4-methylbenzyl chloride (93mg, 0.66mmol) was used as a starting material to give 100mg of a white powder with a yield of 41.5%.¹H NMR(300MHz,CDCl₃)δ9.28(d,J＝1.7Hz,1H),8.33(dd,J＝8.1,2.2Hz,1H),7.89–7.78(m,2H),7.57(s,1H),7.48(t,J＝8.0Hz,1H),7.33(d,J＝8.1Hz,1H),7.25(ddd,J＝8.4,2.6,1.0Hz,1H),7.20(s,3H),5.52(s,2H),5.30(s,2H),2.68(s,3H),2.37(s,3H).HRMS(ESI):found 439.1879,calcd for C25H23N6O2[M+H]⁺439.1877.

Example 41: cholinesterase inhibitory Activity of NAD series Compounds

Medicine and reagent:

NAD series compounds, AChE (e.c.3.1.1.7, Type VI-s, selected from eels), BuChE (e.c.3.1.1.8, selected from horse serum), 5' -dithiobis (2-nitrobenzoic acid) (DTNB), Acetylthiocholine (ATC) iodide, and Butyrylthiocholine (BTC) iodide were all purchased from sigma; tacrine hydrochloride (9-Amin-1,2,3,4-tetrahydroacridine hydrochloride) was purchased from BioTrend corporation.

The instrument comprises the following steps: THERMO Varioskan Flash full-wavelength multifunctional microplate reader.

(II) experimental method:

(1) preparing a buffer solution: 13.6g of potassium dihydrogen phosphate are dissolved in 1L of water and the pH is adjusted to 8. + -. 0.1 with potassium hydroxide. The solution was stored at 4 ℃ until use.

(2) Preparing a 0.01M DTNB solution: 0.396g of DTNB and 0.15g of sodium bicarbonate were dissolved in 100mL of water to prepare a 0.01M DTNB solution, which was stored at-30 ℃ for further use.

(3) Preparing 0.075M ATC and BTC solution: 0.217g ATC is dissolved in 10mL water to prepare 0.075M ATC and BTC solution, and the solution is stored at-30 ℃ for standby; 0.237g BTC was dissolved in 10mL water to make a 0.075M BTC solution, which was stored at-30 ℃ until use.

(4) Preparing AChE and BuChE solutions: dissolving 5000 units of AChE in 1mL of 1% gel solution, diluting with water to 100mL to obtain AChE solution with concentration of 5 units/mL, and storing at-30 deg.C; 5000 units of BuChE was dissolved in 1mL of 1% gel solution, and then diluted to 100mL with water to prepare a BuChE solution having a concentration of 5 units/mL, which was stored at-30 ℃ for use.

(5) Preparing a test solution: test compound was dissolved in ethanol to give a concentration of 10^-3M (ethanol did not affect the test results), and then diluted with water to give concentrations of 10^-4、10^-5、10^-6、10^-7、10^-8、10^-9、10^-10A solution of M.

Before the experiment, all the solutions were warmed to room temperature and the AChE, BuChE solution was diluted one time with water to make an enzyme solution with a concentration of 2.5 units/mL. Background UV absorbance was measured with blank buffer (3 mL). Adding 100 of the tested compound solution, 100 of the DTNB solution and 100 of the enzyme solution into 3mL of buffer solution, immediately timing and simultaneously and rapidly mixing the test solution after 20 of ATC or BTC solution is added to trigger reaction, and measuring the ultraviolet absorbance at 412nM wavelength after 2 min. The blank control was measured using an equal volume of water instead of the test solution. All tests were run in parallel three times. The absorbance of the test compound at each concentration was recorded (value 1) with the UV absorption value of the blank as 100%, and the obtained result was calculated by using GraphPadPrism (GraphPad Software, san Diego, Calif., USA) Software in a nonlinear regression analysis mode (nonlinear regression analysis model) to obtain the corresponding IC50 value, and the result is shown in the following table. 78

The results show that the compounds of series I show the best inhibitory activity against acetylcholinesterase. Wherein NAD3 has inhibitory activity pIC on eeAChE₅₀Values greater than 7 are the most active of all compounds. The analysis of structure-activity relationship shows that the position on benzyl in the series of compounds is very sensitive to steric substitution, and the introduction of substituent with larger steric hindrance can cause the loss of the activity; in addition, when the methylene at the benzyl position is converted into carbonyl to form an amido bond with stronger rigidity, the activity of the compound is greatly reduced. Meanwhile, the series of compounds have weak activity on butyrylcholine esterase and pIC₅₀All values are below 6. The compounds of series II and III both showed moderate cholinesterase inhibitory activity (4. ltoreq. pIC. ltoreq.6). The compounds in series IV almost lost their inhibitory activity against cholinesterase.

Table 1 pIC of a compound of formula (I)₅₀Correspondence table

^aThe inhibitory activity of each compound against the enzyme consisted of three independent experiments, which were expressed as mean ± SEM.^bAChE(EC 3.1.1.7)fromelectric eel。^cBuChE(EC 3.1.1.8)from horse serum.^dAChE(EC3.1.1.7)from human.

TABLE 2 pIC of the Compound of formula (II)₅₀Correspondence table

^aThe inhibitory activity of each compound against the enzyme consisted of three independent experiments, which were expressed as mean ± SEM.^bAChE(EC 3.1.1.7)from electric eel。^cBuChE(EC 3.1.1.8)from horse serum.

TABLE 3 pIC of the Compounds of formula (III) and (IV)₅₀Correspondence table

Example 42: the NAD series compound ARE reporter gene activity.

(1) And (3) in vitro cell culture. HepG2 cells transfected with the ARE luciferase reporter gene-containing plasmid were cultured at 37 ℃ in 5% carbon dioxide using 10% fetal bovine serum in RPMI-1640 medium.

(2) HepG2-ARE-C8 cells in the logarithmic growth phase were treated with 0.25% pancreatin digest to prepare a cell suspension at a concentration of 4X 105. 100uL of the culture solution was added to each 96-well microplate and incubated overnight.

(3) The NAD series compounds were prepared at 2-fold assay concentration in culture medium, and 100uL of tert-butylhydroquinone (tBHQ) was added to the corresponding wells as a positive control and DMSO as a negative control. The 96-well enzyme-linked plate added with the compound is incubated for 12 hours at 37 ℃ under 5% carbon dioxide.

(4) The 5 Xlysate in the luciferase assay kit was prepared as 1 Xlysate for use.

(5) The 96-well plate was removed, the culture medium was aspirated from the well, the cells were washed with 1 × PBS buffer, and after the washing, the PBS buffer was aspirated. 25 or 30uL of 1 × cell lysate was added to each well and lysed on ice for 15 min. After the cracking is finished, standing for 3-5min, and sucking 20uL of supernatant liquid to add into a corresponding 96-hole white enzyme label plate.

(6) The white enzyme-labeled plate is placed into a Thermo Scientific luminoskaasent chemiluminescence micropore reading instrument, 100uL of luciferase detection testing agent (prepared by uniformly mixing 1 bottle of luciferase detection substrate in a luciferase detection kit and 1 bottle of luciferase detection buffer solution) is added into each hole before testing, and reading is carried out within 1min after the detection reagent is added.

The results are shown in FIG. 1. The fold induction of ARE-luciferase by the test compounds NAD1 to NAD35 in the test cells at both concentrations of 5. mu.M and 20. mu.M is plotted on the ordinate, and the results show that all the test compounds exhibit a certain ARE reporter gene-inducing activity at both concentrations tested. For the compounds in the series I, the compounds with hydrophobic group substitution (such as NAD9, NAD11, NAD12 and the like) have stronger activation activity; the compounds with amide structure (NAD16 and NAD17) were most active. For the compounds of series II and III, the ARE reporter gene activity is higher. Similar to the compounds of series I, compounds with amide structures (NAD29 and NAD30) showed the best activity except. The series IV compounds with 1,2, 3-triazole linking structure show the best ARE reporter gene activation activity.

Example 43: and (4) evaluating cytotoxicity.

The cell activity of the compounds was examined at both concentrations of 10. mu.M and 50. mu.M using the MTT method. The results are shown in the following graph, and the cytotoxicity of NAD compounds against PC12 cells was greatly reduced as compared with NAT compounds. The vast majority of compounds in series 1(NAD1 to NAD17) had no significant effect on PC12 cells at a concentration of 50. mu.M. However, the other three groups of compounds show certain cytotoxicity, especially the series 4 compounds with 1,2, 3-triazole structure.

Example 44: the Morris water maze study mouse behavioural studies.

The instrument comprises the following steps: panlab SMART 3.0 behavioural video analyzer

Animals: adult male ICR mice (8-10 weeks, 20-25 grams in weight) were purchased from the university of Yangzhou medical center.

Reagent: scopolamine hydrobromide was purchased from alatin reagent (S107418, shanghai), tacrine (purity > 95%), compound NAD 3.

The experimental method comprises the following steps: the 40 mice were randomly divided into 4 groups (10 mice per group): control, model, tacrine, NAD3 treated. Tacrine, NAD3 were dissolved in CMC-Na solution (0.5g CMC-Na, 100ml distilled water) respectively and administered by intragastric administration (10mg/kg body weight). After 30min, mice in the model group, tacrine group and NAD3 treated group were injected with scopolamine (1mg/kg) intraperitoneally, and mice in the control group were injected with normal saline intraperitoneally. The cognitive function and memory of the mice were tested by the water maze. An escape platform (diameter 10cm) is fixed in a circular water pool (diameter 120cm, height 60cm), a small flag (height 5 cm) is fixed on the platform, water with height 40cm is filled in the water pool, and the water maze is formed by keeping the temperature at 25 ℃. The mice are placed on an escape platform for training on days 1-2 after taking the medicine, the platform is placed 1cm under water on days 3-5, the mice are trained, the platform is moved away on the last day (day 6), the mice are evaluated, and the time, the track and the speed of the mice reaching the position of the platform are recorded. The results of the experiment are shown in fig. 3 and table 4.

TABLE 4 time, trajectory and speed of the mouse to the platform

And (4) analyzing results: as can be seen from fig. 3 and table 4, the average time and distance to the plateau of the model mice were much higher than those of the control group, indicating that scopolamine causes memory deficiency in the mice and that molding was successful. The time and distance spent by tacrine groups was significantly reduced relative to the model group, indicating that tacrine significantly improved memory and cognitive function in mice. While the average time and distance to the plateau for mice in the NAD 3-treated group were slightly higher than in the tacrine group, but significantly lower than in the model group. NAD3 was shown to have an improved effect on mouse memory and cognitive function, but was slightly inferior to tacrine.

Claims

1. A donepezil-oxadiazole fusion compound having the formula (I), (II), (III) or (IV):

wherein R is₁Is halogen, methyl, methoxy, tert-butyl, trifluoromethyl or cyano; x is methylene or carbonyl;

wherein Ar is a pyridine ring or a 2-substituted furan ring;

wherein R is₂Is hydrogen, fluorine or methyl, X is methylene or carbonyl;

wherein R is₃Is hydrogen, fluorine, or methyl.

2. A process for the preparation of donepezil-oxadiazole fusion compound of claim 1, wherein the compound of formula (i) is prepared by the steps of:

(5) benzoic acid or 4-fluorobenzoic acid and the intermediate 4 are dehydrated and condensed into amide under the conditions that T3P is used as a condensing agent and ethyl acetate is used as a solvent, and NAD16 and NAD17 are obtained.

3. A process for the preparation of donepezil-oxadiazole fusion compound of claim 1, wherein the compound of formula (ii) is prepared by the steps of:

4. A method of preparing a donepezil-oxadiazole fusion compound according to claim 1, wherein the compound of formula (iii) is prepared by the steps of:

(5) benzoic acid or 4-fluorobenzoic acid and intermediate 8 are dehydrated and condensed into amide under the condition of T3P as a condensing agent and ethyl acetate as a solvent to obtain NAD29 and NAD 30.

5. A process for the preparation of donepezil-oxadiazole fusion compound of claim 1, wherein the compound of formula (iv) is prepared by the steps of:

(3) reacting different substituted bromobenzyl or chlorobenzyl with sodium azide under the catalysis of potassium carbonate and with DMF as solvent at 105 ℃ to obtain azide derivative intermediate, and then reacting the intermediate in MeOH/H₂Catalyzing by copper sulfate pentahydrate and ascorbic acid in mixed solvent system of OThe reagent is reacted at room temperature to obtain target products NAD31 to NAD 35.

6. Use of donepezil-oxadiazole fusion compound according to claim 1 for the preparation of a therapeutic agent for the treatment of alzheimer's disease.