CN114591202A - Ferulic acid derivative, preparation method and application - Google Patents

Abstract

The invention discloses a ferulic acid derivative as well as a preparation method and use thereof. Specifically, the following steps are: using ferulic acid as a raw material, first refluxing methyl ferulic acid with methanol under the condition of concentrated sulfuric acid to obtain methyl ferulic acid; The acyl chloride reacts to generate ferulic acid carbamate substituted derivatives, and the methyl protecting group is further removed under the action of lithium hydroxide to expose the carboxyl end, and finally, under the action of HATU and DIPEA, it reacts with different substituted anilines to generate the target A Wei acid derivatives. Through in vitro cholinesterase activity tests on synthetic ferulic acid derivatives, most of the compounds have inhibitory activity on cholinesterase, which is an ideal anti-cholinesterase chemical entity, which is useful for the treatment of Alzheimer's disease. Haimer's disease is important.

Description

A kind of ferulic acid derivative and preparation method and use

technical field

The invention belongs to the technical field of medicinal chemical synthesis, in particular to a ferulic acid derivative, and also to a preparation method and application of the ferulic acid derivative.

Background technique

my country has entered an aging society, and the health of the elderly has received much attention. Alzheimer's disease (AD), as the most common type of Alzheimer's disease, has no specific drug at present, so the research on AD has become a hot issue of many scientific researches. Although there are various theories about its pathogenesis, it may be related to gene mutation, β-amyloid (Aβ) deposition, damage to the central cholinergic system, toxicity of excitatory amino acids, inflammatory response and immune system impairment, Oxidative stress and calcium homeostasis disorders are related, but its therapeutic drugs are still mainly acetylcholinesterase inhibitors (ACEI). Therefore, finding new anti-AD drugs from natural products is one of the hotspots of pharmaceutical research in recent years.

Due to the complex pathogenesis of AD, it is difficult to effectively cure AD with a single target drug. Multi-target directed ligands (MTDLs) can modulate multiple targets and are an ideal strategy for the synthesis of AD small molecule drugs. MTDLs have become one of the hot spots in the field of anti-AD drug research. Ferulic acid (FA) is a phenolic hydroxyl-containing phenylpropanoid natural product with a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, inhibition of Aβ aggregation and other functions.

Ristigmine, an FDA-approved drug for the treatment of Alzheimer's disease, is an N-methyl-D-aspartate receptor antagonist with N-ethyl-N- The methyl carbamate group is the reactive functional group region. The clinical application of FA in the treatment of AD is limited due to its poor blood-brain barrier (BBB) permeability and low bioavailability. Puris et al. applied an L-type amino acid transporter (LAT1)-mediated prodrug approach to deliver FA into the mouse brain and synthesized three novel LAT1-utilizing FA prodrugs. The results demonstrate that amide-based prodrugs with aromatic rings can efficiently bind to LAT1, utilize the transporter for cellular uptake in vitro, and cross the BBB after in situ perfusion in mice.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a ferulic acid derivative which has inhibitory effect on acetylcholinesterase and butyrylcholinesterase.

The second object of the present invention is to provide a preparation method of a ferulic acid derivative.

The third object of the present invention is to provide the use of a ferulic acid derivative in the preparation of a drug for Alzheimer's disease.

The fourth object of the present invention is to provide a drug for treating Alzheimer's disease, comprising ferulic acid derivatives or pharmaceutically acceptable salts and hydrates thereof synthesized with acids.

The technical scheme adopted in the present invention is, a kind of ferulic acid derivative, and its general structural formula is shown in formula (I):

Wherein: R ₁ is any one of -N(CH ₃ ) ₂ , -N(CH ₃ )CH ₂ CH ₃ , -N(CH ₂ CH ₃ ) ₂ ; R ₂ is H or NO ₂ substituted at different positions , halogen, hydroxyl, methoxy.

The present invention is also characterized in that,

Preferably, the ferulic acid derivative is (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenyldimethylcarbamate, (E)-4-(3-(((4-Chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate, (E) -2-Methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenyldimethylcarbamate, (E)-4 -(3-(((2-Bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate, (E)-4-(3 -(((2-Fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate, (E)-2-methoxy-4 -(3-oxo-3-(phenylamino)-1-propenyl)phenylethyl(methyl)carbamate, (E)-4-(3-((((2-fluorophenyl) )amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate, (E)-4-(3-((((2-bromophenyl) ) amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl (methyl)carbamate, (E)-4-(3-((((2-chlorophenyl) ) amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate, (E)-2-methoxy-4-(3-(( (4-Nitrophenyl)amino)-3-oxo-1-propenyl)phenylethyl(methyl)carbamate, (E)-2-methoxy-4-(3-oxo Substituted-3-(phenylamino)-1-propenyl)phenyldiethylcarbamate, (E)-4-(3-(((2-chlorophenyl)amino)-3-oxo -1-Propenyl)-2-methoxyphenyldiethylcarbamate, (E)-2-methoxy-4-(3-((4-nitrophenyl)amino)-3 -Oxo-1-propenyl)phenyldiethylcarbamate, (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl) -2-Methoxyphenyldiethylcarbamate, (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2- any one in methoxyphenyl diethyl carbamate;

Most preferably, the ferulic acid derivative is (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldi Methylcarbamate, (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylamino Formate, (E)-2-Methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenylethyl(methyl)carbamate, (E )-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate, (E )-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate, (E )-4-(3-(((2-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate, (E )-2-methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenylethyl(methyl)carbamate, ( E) any one of-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldiethylcarbamate .

The beneficial effect of the present invention is that the present invention firstly synthesizes the ferulic acid derivative through four-step reaction, the reaction conditions are mild, the repetition is easy, and the total yield is high. For the first time, the anticholinesterase activity of ferulic acid carbamate aniline substituted derivatives has been investigated, all of which have different anti-acetylcholine and butyrylcholine activities and are potential new chemical entities for the treatment of Alzheimer's disease. It is of great significance for the research and development of Alzheimer's disease drugs.

Detailed ways

The present invention will be described in detail below with reference to specific embodiments.

The preparation method of a kind of ferulic acid derivative of the present invention is specifically implemented according to the following steps:

Step 1, using ferulic acid as a raw material, under the condition of concentrated sulfuric acid and methanol reflux reaction to prepare methyl ferulate;

The mass ratio of ferulic acid to methanol was 1:8; the reflux reaction temperature was 70°C, and the reflux reaction time was 3h;

In step 2, methyl ferulate and carbamoyl chlorides with different substitutions are mixed, and acetone K ₂ CO ₃ is added to carry out a reflux reaction. The reaction temperature is 60° C. and the reaction time is 7h; Derivatives;

Different substituted carbamoyl chlorides are N-methyl-N-ethylformyl chloride, N,N-dimethylformyl chloride or N,N-diethylformyl chloride;

The mass ratio of methyl ferulate to carbamoyl chloride is 1:1.1;

Step 3, the ferulic acid carbamate substituted derivative is reacted with lithium hydroxide in a mixed solution of tetrahydrofuran (THF), methanol (MeOH) and water (H ₂ O) for 24 h, and the methyl protecting group is removed to expose the The carboxyl terminal is obtained to obtain ferulic acid carbamate;

The mass ratio of ferulic acid carbamate substituted derivatives to lithium hydroxide is 1:3;

Step 4 in 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and N,N-diisopropylethylamine Under the action of (DIPEA), the ferulic acid carbamate was reacted with anilines for 2 h to generate the target ferulic acid derivative:

The mass ratio of ferulic acid carbamate and aniline substances is 1:1, and the reaction is carried out at room temperature;

The aniline substance is any one of aniline, 4-chloroaniline, p-nitroaniline, 2-bromoaniline, o-chloroaniline and o-fluoroaniline;

The general chemical reaction formula is:

Wherein: R ₁ is -N(CH ₃ ) ₂ , -N(CH ₃ )CH ₂ CH ₃ , -N(CH ₂ CH ₃ ) ₂ ; R ₂ is H or NO ₂ substituted at different positions, halogen, hydroxyl, Methoxy.

In the above-mentioned ferulic acid derivatives, when R ₁ is -N(CH ₃ ) ₂ , R ₂ is H or NO ₂ , halogen, hydroxyl, methoxy substituted at different positions;

When R ₁ is -N(CH ₃ )CH ₂ CH ₃ , R ₂ is H or NO ₂ substituted at different positions, halogen, hydroxyl, methoxy;

When R ₁ is -N(CH ₂ CH ₃ ) ₂ , R ₂ is H or NO ₂ substituted at different positions, halogen, hydroxyl or methoxy.

Through in vitro cholinesterase activity tests on synthetic ferulic acid derivatives, most of the compounds have inhibitory activity on cholinesterase, which is an ideal anti-cholinesterase chemical entity, which is useful for the treatment of Alzheimer's disease. Haimer's disease is important.

Synthesis of Example 1 Intermediate

a, (E)-3-(4-hydroxy-3-ethoxyphenyl) methyl acrylate (2)

Add 3 g of ferulic acid (FA) (15.5 mmol) to a 100 mL eggplant-shaped flask, dissolve it in 30 mL of methanol, slowly add 1.2 mL (22.5 mmol) of concentrated sulfuric acid, and heat the system to 70 °C for 3 h. After the reaction was completed, the pH was adjusted to 7 with 10% NaOH after concentration under reduced pressure, ethyl acetate was added for extraction, washed with water, the organic phase was Na ₂ SO ₄ and concentrated under reduced pressure to obtain 3.1 g of a yellow oil with a yield of 97%; It is (E)-3-(4-hydroxy-3-ethoxyphenyl) methyl acrylate.

Among them, the product is characterized as follows:

(E)-Methyl 3-(4-hydroxy-3-ethoxyphenyl)acrylate: ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.64 (1H, d, J=16.0 Hz, H-7) ,7.08(1H,d,J=8.2Hz,H-5),7.04(1H,s,H-3),6.93(1H,d,J=8.0Hz,H-6),6.31(1H,d, J=16.0 Hz, , H-8), 6.07 (1H, brs, OH), 3.93 (3H, s, OCH ₃ ), 3.82 (3H, s, OCH ₃ ); ESI-MS m/z: 208.21 [M +H] ⁺ .

b. Synthesis of methyl ferulate carbamate (3a-3c)

Compound 2 (1 g, 4.8 mmol) was dissolved in an appropriate amount of acetone, and then the corresponding acid chloride and K ₂ CO ₃ were added. The reaction system was heated to 60° C., stirred and refluxed for 7 h, cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate. The filtrate was reduced Concentrate to dryness under pressure, add ethyl acetate for extraction, wash with water, combine the organic phases, dry over Na ₂ SO ₄ and concentrate under reduced pressure to obtain the compound, namely (E)-3-(4-((dimethylcarbamoyl) Methyl oxy)-3-methoxyphenyl)acrylate (3a)

Yellow crystals; yield 64.9%; mp 127-129°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.65 (1H, d, J=16.0 Hz, H-7), 7.26 (1H, s, H- 3), 7.10(1H,d,J=8Hz,H-5),7.08(1H,d,J=8Hz,H-6,6.37(1H,d,J=16.0Hz,H-8),3.86( 3H,s, _OCH3 ), 3.81(3H,s, _OCH3 ), 3.12(3H,s, _-CH3 ), 3.01(3H,s, _-CH3 ); ESI-MS m/z: 280.1 [M +H] ⁺ .

(E)-Methyl 3-(4-((ethyl(methyl)carbamoyl)oxy)-3-methoxyphenyl)acrylate (3b)

White solid; yield 62.3%; mp 83-85°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.65 (1H, d, J=16.0 Hz, H-7), 7.11, 7.08 (3H, m, H -3,5,6), 6.38(1H,d,J=16.0Hz,H-8), 3.86(3H,s, _OCH3 ), 3.81(3H,s, _OCH3 ), 3.05(5H,m, -CH2CH3), 1.20 (3H, t, _{-CH3 )} ; ESI _- MS m/z: 294.1 [M+H] ⁺ .

(E)-Methyl 3-(4-((diethylcarbamoyl)oxy)-3-methoxyphenyl)acrylate (3c)

Yellow crystals; yield 63.4%; mp 59-61°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.65 (1H, d, J=16.0 Hz, H-7), 7.11, 7.08 (3H, d, J=12Hz,H-3,5,6), 6.37(1H,d,J=16.0Hz,,H-8), 3.86(3H,s, _OCH3 ), 3.81(3H,s, _OCH3 ), 3.45 (4H, q, _-CH2 ), 1.24 (6H, tt, _-CH3 ); ESI-MS m/z: 308.1 [M+H] ⁺ .

c. Synthesis of ferulic acid carbamate (4a～4c)

Compound (3a-3c) and 3 times the amount of LiOH·H ₂ O were added to the mixed solution of THF/MeOH/H ₂ O (15/5/5) mL, stirred at room temperature for 24 h, and the solvent was removed under reduced pressure, and 30 mL of The residue was diluted with water, adjusted to pH=4 with 10% HCl, extracted with ethyl acetate (3×15 mL), the organic layers were combined, washed with water, dried over anhydrous Na ₂ SO ₄ , and concentrated under reduced pressure to obtain the corresponding product ( 4a to 4c).

(E)-3-(4-((dimethylcarbamoyl)oxy)-3-methoxyphenyl)acrylic acid (4a)

White powder; yield 75.5%; mp 201-202°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 12.39 (1H, s, COOH), 7.60 (1H, d, J=16.0 Hz, H-7) ,7.46(1H,s,H-3),7.25(1H,d,J=8.0Hz,H-5),7.11(1H,d,J=8.0Hz,H-6),6.58(1H,d, J=16.0Hz, H-8), 3.83 (3H,s, _OCH3 ), 3.05 (3H,s, _-CH3 ), 2.91 (3H,s, _-CH3 ); ESI-MS m/z: 266.1 [M+H] ⁺ .

(E)-3-(4-((ethyl(methyl)carbamoyl)oxy)-3-methoxyphenyl)acrylic acid (4b)

White solid; yield 78%; mp 126-128°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 12.41 (1H, s, COOH), 7.59 (1H, d, J=16.0 Hz, H-7) ,7.47(1H,s,H-3),7.25(1H,d,J=8.0Hz,H-5),7.11(1H,d,J=8.0Hz,H-6),6.59(1H,d, J = 16.0 Hz, H-8), 3.83 (3H, s, OCH ₃ ), 3.01 (5H, d, -CH ₂ , -CH ₃ ), 1.19 (3H, t, -CH ₃ ); ESI-MS m /z:280.1[M+H] ⁺ .

(E)-3-(4-((diethylcarbamoyl)oxy)-3-methoxyphenyl)acrylic acid (4c)

White solid; yield 76%; mp 126-127°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.72 (1H, d, J=16.0 Hz, H-7), 7.13, 7.10 (3H, d, J=12Hz,H-3,5,6), 6.36(1H,d,J=16.0Hz,,H-8), 3.87(3H,s, _OCH3 ), 3.40(4H,dd, _-CH2 ) , 1.24 (6H, tt, -CH ₃ ); ESI-MS m/z: 294.1 [M+H] ⁺ .

Example 2 Preparation of (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenyldimethylcarbamate (5a)

First add compound (4a) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add aniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105: 30) Purify by column chromatography; white solid; yield 46.2%; identified as (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl) Phenyldimethylcarbamate.

mp 178-180°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.34 (1H, s, H-9), 7.65 (2H, d, J=8.0 Hz, H-11, 15), 7.47 (1H ,d,J＝16Hz,H-7),7.32(2H,t,H-12,14),7.09(1H,t,H-13),7.04(1H,d,J＝8.0Hz,H-4 ),6.96(1H,d,J=8.0Hz,H-3),6.89(1H,s,H-6),6.00(1H,d,J=16Hz,H-8),3.66(3H,s, _OCH3 ), 3.19 (3H,s, _-CH3 ), 3.10 (3H,s, _-CH3 ); ESI-MS m/z: 341.1 [M+H] ⁺ .

Example 3 (E)-4-(3-(((4-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate ( 5b) Preparation

First add compound (4a) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 4-chloroaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White solid; 60.7% yield; identified as (E)-4-(3-(((4-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Dimethyl carbamate.

mp 164-166°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.52 (1H, s, H-9), 7.62 (2H, d, J=8.0 Hz, H-11, 15), 7.42 (1H, d,J＝16Hz,H-7),7.28(2H,d,H-12,14),7.04(1H,d,J＝8.0Hz,H-4),6.93(1H,d,J＝8.0Hz ,H-3),6.84(1H,s,H-6),5.82(1H,d,J=16Hz,H-8),3.62(3H,s,OCH ₃ ),3.20(3H,s,-CH ₃ ), 3.11 (3H, s, -CH ₃ ); ESI-MS m/z: 375.0 [M+H] ⁺ .

Example 4 (E)-2-methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenyldimethylcarbamate Preparation of (5c)

First add compound (4a) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 4-nitroaniline (0.2mol), react at room temperature for 2h, spot plate detection After the reaction is completed, use petroleum ether: Acetone (105:30) was purified by column chromatography. Yellow powder; yield 44.8%; identified as (E)-2-methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)benzene dimethylcarbamate.

mp 168-169°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.74 (1H, dd, J=8 Hz, H-12), 8.45 (1 H, dd, J=8.0 Hz, H-14), 8.22 (1H,d,J＝8.0Hz,H-11),8.04(1H,d,J＝16Hz,H-7),7.83(2H,d,J＝8.0Hz,H-15),7.45(1H, dd,J＝8.0Hz,H-11),7.24(1H,dd,J＝8.0Hz,H-4)7.19(1H,s,H-6),7.18(2H,d,J＝8.0Hz,H -3), 6.72 (1H, d, J=16Hz, H-8), 3.91 (3H, s, OCH ₃ ), 3.15 (3H, s, -CH ₃ ), 3.04 (3H, s, -CH ₃ ) ; ESI-MS m/z: 386.0 [M+H] ⁺ .

Example 5 (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate ( 5d) Preparation

First add compound (4a) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 2-bromoaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White powder; yield 37.5%; identified as (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Dimethyl carbamate.

mp 175-176°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.75 (1H, dd, J=8Hz, H-11), 8.45 (1H, dd, J=8.0 Hz, H-14), 8.04 (1H,d,J＝16Hz,H-7),7.45(1H,dd,J＝8.0Hz,H-12),7.25(1H,dd,J＝8.0Hz,H-4)7.19(1H,s , H-6), 7.18 (2H, d, J=8.0Hz, H-3), 6.72 (1H, d, J=16Hz, H-8), 3.91 (3H, s, OCH ₃ ), 3.14 (3H , s, -CH ₃ ), 3.03 (3H, s, -CH ₃ ); ESI-MS m/z: 420.9 [M+H] ⁺ .

Example 6 (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldimethylcarbamate ( 5e) Preparation

First add compound (4a) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 4-fluoroaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White powder; yield 74.1%; identified as (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Dimethyl carbamate.

mp 174-175°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.45 (1H, t, H-9), 7.97 (1H, s, H-11), 7.59 (1H, d, J=16 Hz, H-7), 7.16 (1H, d, J=8.0Hz, H-12), 7.12 (1H, d, J=8.0Hz, H-14), 7.07 (1H, d, J=8.0Hz, H- 13), 7.05 (2H, s, H-3, 6), 6.99 (1H, s, H-4), 6.40 (1H, d, J=16Hz, H-8), 3.80 (3H, s, OCH ₃ ), 3.15 (3H,s, _-CH3 ), 3.04 (3H,s, _-CH3 ); ESI-MS m/z: 359.1 [M+H] ⁺ .

Example 7 (E) of 2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenylethyl(methyl)carbamate (5f) preparation

First add compound (4b) 0.2mol and HATU (1.2eq), DIEPA (2eq), add aniline (0.2mol) after stirring for 15min, react at room temperature for 2h, after the reaction is detected by spot plate, use petroleum ether:acetone (105: 30) Carry out column chromatography separation and purification. White oil; 64% yield; identified as (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenylethyl(methyl) carbamate.

¹ H-NMR (400MHz, CDCl ₃ ), δ 8.48 (1H, s, H-9), 7.65 (2H, d, J=8.0 Hz, H-11, 15), 7.44 (1H, d, J= 16Hz, H-7), 7.32 (2H, t, H-12, 14), 7.08 (1H, d, J=8.0Hz, H-13), 7.04 (1H, d, J=8.0Hz, H-4 ),6.94(1H,d,J=8.0Hz,H-3),6.85(1H,s,H-6),5.93(1H,d,J=16Hz,H-8),3.62(3H,s, OCH ₃ ), 3.55 (2H, q, -CH ₂ ), 3.12 (3H, d, -CH ₃ ), 1.29 (3H, t, -CH ₃ ); ESI-MS m/z: 355.1 [M+H] ⁺ .

Example 8 (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamic acid Preparation of (5g)

First add compound (4b) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 2-fluoroaniline (0.2mol), react at room temperature for 2h, after the reaction is detected by spot plate, use petroleum ether:acetone (105:30) for separation and purification by column chromatography. White oil; yield 55.6%; identified as (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Ethyl (methyl) carbamic acid.

¹ H-NMR (400MHz, CDCl ₃ ) δ 8.43 (1H, s, H-9), 7.92 (1H, d, H-11), 7.60 (1H, d, J=16Hz, H-7), 7.16 (1H,d,J＝8.0Hz,H-4),7.12(1H,d,J＝8.0Hz,H-12),7.07(3H,d,J＝8.0Hz,H-3,6,14) ,7.00(1H,d,H-13),6.40(1H,d,J=16Hz,H-8),3.80(3H,s, _OCH3 ),3.55(2H,q, _-CH2 ),3.12( 3H,d, _-CH3 ), 1.25 (3H,t, _-CH3 ); ESI-MS m/z: 373.1 [MH] ⁺ .

Example 9 (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)carbamate Preparation of acid ester (5h)

First add compound (4b) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 2-bromoaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White powder; yield 77.4%; identified as (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Ethyl (methyl) carbamate.

mp 141-142°C; 1H-NMR (400MHz, CDCl ₃ ) δ 8.75 (1H, dd, J=8Hz, H-11), 8.45 (1H, dd, J=8.0Hz, H-14), 8.05 ( 1H,d,J=16Hz,H-7),7.45(1H,dd,J=8.0Hz,H-12),7.26(1H,s,H-6,),7.19(2H,s,H-3 ), 6.72 (1H, d, J=16Hz, H-8), 3.91 (3H, s, OCH ₃ ), 3.46 (2H, q, -CH ₂ ), 3.15 (3H, d, -CH ₃ ), 1.25 (3H,t, _-CH3 ); ESI-MS m/z: 434.1 [M+H] ⁺ .

Example 10(E)-4-(3-(((2-Chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenylethyl(methyl)aminomethyl Preparation of acid ester (5i)

First add compound (4b) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 2-chloroaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White oil; yield 64.2%; identified as (E)-4-(3-(((2-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Ethyl (methyl) carbamate.

¹ H-NMR (400MHz, CDCl ₃ ) δ 8.75 (1H, dd, J=8Hz, H-11), 8.45 (1H, dd, J=8.0Hz, H-14), 7.72 (1H, d, J =16Hz,H-7),7.45(1H,dd,J=8.0Hz,H-12),7.26(1H,s,H-4,),7.19(1H,s,H-6),6.53(1H , d, J=16Hz, H-8), 3.89 (3H, s, OCH ₃ ), 3.45 (2H, dd, -CH ₂ ), 3.05 (3H, d, -CH ₃ ), 1.24 (3H, t, -CH ₃ ); ESI-MS m/z: 389.1 [M+H] ⁺ .

Example 11 (E)-2-methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenylethyl(methyl)amino Preparation of formic acid (5j)

First add compound (4b) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 4-nitroaniline (0.2mol), react at room temperature for 2h, after the reaction is detected by spot plate, use petroleum ether: Acetone (105:30) was purified by column chromatography. Yellow powder; yield 57.1%; identified as (E)-2-methoxy-4-(3-(((4-nitrophenyl)amino)-3-oxo-1-propenyl)benzene ethyl (methyl) carbamic acid.

mp 77-79°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.90 (1H, s, H-9), 8.22 (2H, d, J=9.2 Hz, H-12, 14), 8.04 (1H ,d,J＝16Hz,H-7),7.84(2H,d,J＝9.2Hz,H-11,15),7.19(1H,d,J＝8.0Hz,H-4),7.09(1H, dd,J＝8.0Hz,H-6),6.97(1H,dd,J＝8.0Hz,H-3),6.73(1H,d,J＝16Hz,H-8),3.90(3H,s,OCH ₃ ), 3.45 (2H, q, -CH ₂ ), 3.12 (3H, t, -CH ₃ ), 1.28 (3H, t, -CH ₃ ); ESI-MS m/z: 398.1 [MH] ⁺ .

Example 12 Preparation of (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenyldiethylcarbamate (5k)

First add compound (4c) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add aniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105: 30) Carry out column chromatography separation and purification. Light yellow solid; 88% yield; identified as (E)-2-methoxy-4-(3-oxo-3-(phenylamino)-1-propenyl)phenyldiethylcarbamide acid ester.

mp 140-142°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.41 (1H, s, H-9), 7.65 (2H, d, J=8.0 Hz, H-11, 15), 7.45 (1H ,d,J＝16Hz,H-7),7.32(2H,t,H-12,14),7.08(1H,d,J＝8.0Hz,H-13),7.05(1H,d,J＝8.0 Hz,H-4),6.97(1H,d,J=8.0Hz,H-3),6.86(1H,s,H-6),5.92(1H,d,J=16Hz,H-8),3.63 (3H, s, OCH ₃ ), 3.50 (4H, q, 2×-CH ₂ ), 1.30 (6H, tt, —CH ₃ ); ESI-MS m/z: 369.1 [M+H] ⁺ .

Example 13 (E)-4-(3-(((2-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldiethylcarbamate ( 5l) Preparation

First add compound (4c) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, then add 2-chloroaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White powder; yield 51.9%; identified as (E)-4-(3-(((2-chlorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Diethylcarbamate.

mp 124-125°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.74 (1H, dd, J=8.0 Hz, H-11), 8.45 (1H, dd, J=8.0 Hz, H-14), 8.05(1H,d,J＝16.0Hz,H-7),7.45(1H,dd,J＝8.0Hz,H-12),7.23(1H,d,J＝12.0Hz,H-4),7.19( 1H, s, H-6), 6.72 (1H, d, J=16Hz, H-8), 3.91 (3H, s, OCH ₃ ), 3.43 (4H, q, 2×-CH ₂ ), 1.24 (6H , tt, -CH ₃ ); ESI-MS m/z: 403.1 [M+H] ⁺ .

Example 14 (E)-2-methoxy-4-(3-((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenyldiethylcarbamate ( 5m) preparation

First add compound (4c) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 4-nitroaniline (0.2mol), react at room temperature for 2h, spot plate detection After the reaction is completed, use petroleum ether: Acetone (105:30) was purified by column chromatography. Yellow powder; 42.9% yield; identified as (E)-2-methoxy-4-(3-((4-nitrophenyl)amino)-3-oxo-1-propenyl)phenyl Diethylcarbamate.

mp 125-126°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.74 (1H, dd, J=8Hz, H-11), 8.45 (1H, dd, J=8.0 Hz, H-14), 8.05 (1H,d,J＝16Hz,H-7),7.46(1H,dd,J＝8.0Hz,H-12),7.22(1H,d,J＝12Hz,H-3),7.19(1H,s ,H-6)6.72(1H,d,J=16Hz,H-8),3.91(3H,s,OCH ₃ ),3.43(4H,q,2×-CH ₂ ),1.24(6H,tt,- CH ₃ ); ESI-MS m/z: 412.1 [MH] ⁺ .

Example 15 (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldiethylcarbamate ( 5n) Preparation

First add compound (4c) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 2-bromoaniline (0.2mol), react at room temperature for 2h, after the reaction is detected by spot plate, use petroleum ether:acetone (105:30) for separation and purification by column chromatography. White solid; 58.1% yield; identified as (E)-4-(3-(((2-bromophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Diethylcarbamate.

mp 100-102°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.75 (1H, dd, J=8Hz, H-11), 8.45 (1H, dd, J=8.0 Hz, H-14), 8.04 (1H,d,J=16Hz,H-7),7.45(1H,dd,J=8.0Hz,H-12),7.25(1H,d,H-6,),7.20(2H,d,H- 4, 13), 7.09 (1H, d, H-3), 6.72 (1H, d, J=16Hz, H-8), 3.90 (3H, s, OCH ₃ ), 3.43 (4H, q, 2×- _CH2 ), 1.25 (6H, tt, _-CH3 ); ESI-MS m/z: 448.1 [M+H] ⁺ .

Example 16 (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyldiethylcarbamate ( 5o) Preparation

First add compound (4c) 0.2mol, HATU (1.2eq), DIEPA (2eq), stir for 15min, add 2-fluoroaniline (0.2mol), react at room temperature for 2h, spot plate detection after the reaction is completed, use petroleum ether: acetone (105:30) for separation and purification by column chromatography. White oil; 50% yield; identified as (E)-4-(3-(((2-fluorophenyl)amino)-3-oxo-1-propenyl)-2-methoxyphenyl Diethylcarbamate.

¹ H-NMR (400MHz, CDCl ₃ ) δ 8.43 (1H, s, H-9), 7.93 (1H, s, H-11), 7.60 (1H, d, J=16Hz, H-7), 7.15 (1H,d,J＝8.0Hz,H-6),7.09(1H,d,J＝8.0Hz,H-12),7.07(2H,d,J＝8.0Hz,H-3,14),7.01 (1H, s, H-13), 6.40 (1H, d, J=16Hz, H-8), 3.81 (3H, s, OCH ₃ ), 3.45 (4H, q, 2×-CH ₂ ), 1.23 ( 6H,tt, _-CH3 ); ESI-MS m/z: 387.1 [M+H] ⁺ .

Table 1 Ferulic acid derivatives prepared by the present invention

Evaluation of Anticholinesterase Activity of Ferulic Acid Derivatives

(1) Experimental principle

The basic principle of the Ellman method is as follows: AChE catalyzes the hydrolysis of the synthetic substrate thioacetylcholine iodide to generate sulfhydryl-containing compounds, and the sulfhydryl-containing compounds can react with 5-5'-dithio(2-nitrobenzoic acid) (DTNB), This leads to the cleavage of its disulfide bond and converts the colorless DTNB into yellow 5-thio-2-nitrobenzoic acid (TNB). The maximum absorption of TNB is around 412nm, but DTNB has no absorption here, so the amount of thiol compounds can be calculated by measuring the absorbance of TNB at 412nm, and its content is proportional to the activity of AchE within a certain range. Therefore, the activity of the enzyme can be determined by measuring the absorbance at 412 nm after the enzyme reaction. The principle of butyrylcholinesterase is the same as that of acetylcholinesterase, and the substrate is replaced by thioacetylcholine iodide with thiobutyrylcholine iodide.

(2) Experimental method

The inhibitory activities of AChE and BuChE were determined by Ellman method.

Determination of AChE inhibitory activity: The compound was dissolved in DMSO, prepared into a stock solution with an initial concentration of 10 mM, diluted with buffer B to the desired concentration, and the DMSO content in the prepared solution was controlled to be less than 1%. Add: 120 μL PBS, 20 μL ChE (0.1 U/mL, prepared in buffer A) and 20 μL of compounds of different concentrations in sequence in a 96-well plate; incubate at 37°C for 6 min, quickly add 20 μL DTNB and 20 μL ATCI (prepared in buffer B), 37 Incubate at ℃ for 30min. Absorbance values were determined at 412 nm.

Determination of BuChE inhibitory activity: The compound was dissolved in DMSO, prepared into a stock solution with an initial concentration of 10 mM, diluted with buffer B to the desired concentration, and the DMSO content in the prepared solution was controlled to be less than 1%. Add: 120 μL PBS, 20 μL BuChE (0.1 U/mL, prepared in buffer A) and 20 μL compounds of different concentrations in sequence in a 96-well plate; incubate at 37°C for 6 min, quickly add 20 μL DTNB and 20 μL BTCI (prepared in buffer B), Incubate at 37°C for 30min. Absorbance values were determined at 412 nm.

The formula for calculating the inhibition rate is:

Inhibition(%)=(control-sample)/control*100%

(3) Experimental results

Table 2 acetylcholinesterase inhibitory activity

Table 3 Butyrylcholinesterase inhibitory activity

From Table 2, it can be seen that the five compounds of compounds 5c, 5f, 5j, 5g, and 5m all showed inhibitory effects on acetylcholinesterase. Table 3 shows that except for 4a, 4b, 4c, 5l, and 5m, almost all compounds The compounds all showed a good inhibitory effect on butyrylcholinesterase, and 5c, 5d, 5e, and 5f showed much higher inhibition rates than the positive drugs, and the inhibition rates were all greater than 90% at 100 μM concentration. As mentioned above, the ferulic acid carbamate aniline derivative is an ideal anticholinesterase chemical entity with important implications for the treatment of Alzheimer's disease.

Claims (6)

1. A ferulic acid derivative is characterized in that the structural general formula is shown as the formula (I):

wherein: r₁is-N (CH)₃)₂、-N(CH₃)CH₂CH₃、-N(CH₂CH₃)₂Any one of the above; r₂Being H or NO substituted in different positions₂Halogen, hydroxy, methoxy.

2. A ferulic acid derivative according to claim 1, wherein the ferulic acid derivative is (E) -2-methoxy-4- (3-oxo-3- (phenylamino) -1-propenyl) phenyldimethylcarbamate, (E) -4- (3- (((4-chlorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl dimethylcarbamate, (E) -2-methoxy-4- (3- (((4-nitrophenyl) amino) -3-oxo-1-propenyl) phenyldimethylcarbamate, (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl dimethylcarbamate, (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl dimethylcarbamate, (E) -2-methoxy-4- (3-oxo-3- (phenylamino) -1-propenyl) phenylethyl (methyl) carbamate, (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenylethyl (methyl) carbamate, (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl phenyl ethyl (methyl) carbamate, (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl Ethyl (methyl) carbamate, (E) -4- (3- (((2-chlorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenylethyl (methyl) carbamate, (E) -2-methoxy-4- (3- (((4-nitrophenyl) amino) -3-oxo-1-propenyl) phenylethyl (methyl) carbamate, (E) -2-methoxy-4- (3-oxo-3- (phenylamino) -1-propenyl) phenyldiethyl carbamate, (E) -4- (3- (((2-chlorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyldiethyl carbamate An ester, (E) -2-methoxy-4- (3- ((4-nitrophenyl) amino) -3-oxo-1-propenyl) phenyldiethyl carbamate, or (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl diethyl carbamate, or (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl diethyl carbamate.

3. A ferulic acid derivative according to claim 2, wherein the ferulic acid derivative is (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl dimethylcarbamate, (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenyl dimethylcarbamate, (E) -2-methoxy-4- (3-oxo-3- (phenylamino) -1-propenyl) phenylethyl (methyl) carbamate, (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) phenylethyl (methyl) carbamate ) -2-methoxyphenylethyl (methyl) carbamate, (E) -4- (3- (((2-bromophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenylethyl (methyl) carbamate, (E) -4- (3- (((2-chlorophenyl) amino) -3-oxo-1-propenyl) -2-methoxyphenylethyl (methyl) carbamate, (E) -2-methoxy-4- (3- (((4-nitrophenyl) amino) -3-oxo-1-propenyl) phenylethyl (methyl) carbamate, (E) -4- (3- (((2-fluorophenyl) amino) -3-oxo-1-propenyl) phenylethyl (methyl) carbamate, and mixtures thereof Any one of propenyl) -2-methoxyphenyl diethyl carbamate.

4. The process for the preparation of ferulic acid derivative according to any one of claims 1 to 3, which is carried out in particular according to the following steps:

step 1, taking ferulic acid as a raw material, and carrying out reflux reaction on the ferulic acid and methanol under the condition of concentrated sulfuric acid to prepare ferulic acid methyl ester;

step 2, performing reflux reaction on the ferulic acid methyl ester and different substituted carbamoyl chlorides respectively to generate a ferulic acid carbamate substituted derivative;

step 3, placing the derivative substituted by the ferulic acid carbamate and lithium hydroxide in a mixed solution of tetrahydrofuran, methanol and water for reaction for 24 hours to obtain the ferulic acid carbamate;

and 4, reacting the ferulic acid carbamate with aniline substances for 2 hours under the action of 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate HATU and DIPEA to generate the ferulic acid derivative.

5. Use of a ferulic acid derivative according to any of claims 1-3 for the preparation of a medicament for the treatment of alzheimer's disease.

6. A medicament for treating alzheimer's disease, comprising the ferulic acid derivative of claims 1 to 3 or a pharmaceutically acceptable salt or hydrate thereof with an acid.