CN111423407A

CN111423407A - Caffeoylquinic acid derivative and preparation method and application thereof

Info

Publication number: CN111423407A
Application number: CN201910022953.0A
Authority: CN
Inventors: 田瑜; 许旭东; 郭鹏; 吴崇明; 尚海
Original assignee: Institute of Medicinal Plant Development of CAMS and PUMC
Current assignee: Institute of Medicinal Plant Development of CAMS and PUMC
Priority date: 2019-01-10
Filing date: 2019-01-10
Publication date: 2020-07-17
Anticipated expiration: 2039-01-10
Also published as: CN111423407B

Abstract

The invention discloses a caffeoylquinic acid derivative, a preparation method and application thereof, and the caffeoylquinic acid derivative has a structure shown in a general formula (I). The invention takes 3-O-caffeoylquinic acid as a raw material, reserves the structural skeleton of the 3-O-caffeoylquinic acid and is positioned at 1 positionDifferent amino substituent groups are introduced into carboxyl, 4, 5-position hydroxyl groups are substituted or exposed by ketal, and the novel caffeoylquinic acid derivative with lipid-lowering activity is synthesized, the synthesis method is simple and convenient, and the product purity is high. Compared with 3-O-caffeoylquinic acid, the caffeoylquinic acid derivative provided by the invention has better lipid-lowering activity.

Description

Caffeoylquinic acid derivative and preparation method and application thereof

Technical Field

The invention relates to the field of pharmaceutical chemistry and therapeutics, in particular to a caffeoylquinic acid derivative, a preparation method and application thereof.

Background

The hyperlipidemia (hyperlipidemia, H L P) can induce clinical common cardiovascular and cerebrovascular diseases, and is one of the potential factors threatening human health, at present, the dyslipidemia patients in China reach 2.0 hundred million, the morbidity of normal people reaches 35-40%, and the middle-aged and elderly people reach more than 60%.

Currently, the common lipid-lowering drugs in clinic mainly include statin lipid-regulating drugs (atorvastatin and lovastatin), fibrate drugs (gemfibrozil and fenofibrate), nicotinic acid drugs (acipimox) and the like. However, these drugs acting on a single target point have various unsatisfactory side effects in clinical application, such as severe myopathy and rhabdomyolysis, and hepatotoxicity in the current first ten drugs of the world sold atorvastatin (also known as lipitor) as an HMG-CoA reductase inhibitor; even the new drug lomitapide, which has just been approved by the FDA to be marketed in 12 months 2012 for the treatment of familial hypercholesterolemia, is indicated to be at risk for gastrointestinal reactions and hepatotoxicity. Therefore, the search for lipid-lowering drugs with significant efficacy, safety and reliability is a subject worthy of long-term research by medical workers.

However, from the actual situation of our country, we completely imitate the western countries to find the lead compound from the artificially synthesized product, and create a new lipid-lowering drug with high efficiency and low toxicity, which is difficult to get a major breakthrough in a short period, but the natural product rich in the traditional clinically effective drug has the specific structural diversity and multi-target action mechanism, and is more and more favored by researchers at home and abroad, such as berberine which is the main lipid-lowering active ingredient in the Chinese medicine coptis root.

The action mechanism of traditional Chinese medicine lipid regulation is complex compared with western medicines, adverse reactions are less, and lipid reduction is achieved mainly by reducing absorption of exogenous lipid in intestinal tracts, inhibiting synthesis of endogenous lipid and regulating lipid metabolism. For example, a single Chinese medicine (kudzu root, ginseng, salvia miltiorrhiza, ginkgo leaf and the like) with the function of regulating blood fat; the lipid-lowering natural product can be polysaccharides (fructus Lycii polysaccharide, fructus Cucurbitae Moschatae polysaccharide), flavonoids (dihydrotanshinone, folium Ginkgo flavone) and polyphenols (catechin). In recent years, many folk traditional medicines are found to have good lipid regulating effect and are gradually developed into medicines, for example, a Xuezhikang capsule (main component red yeast) developed by limited northern Weixin biotechnology has completed phase II clinical research in the United states at present, and Jianjiandong subject group of institute of biotechnology of Chinese academy of medicine sciences finds that berberine also has good lipid lowering activity, is completely different from the action mechanism of statin lipid lowering medicines, and theoretically provides a new molecular target for searching for novel lipid lowering medicines. Different from the single-target western medicines which are clinically and frequently adopted at the present stage and easily cause adverse reactions, the multi-target characteristics of the natural medicines are probably the reasons for less toxic and side effects, so the method has epoch significance for the deep exploration and research of the natural medicines.

Disclosure of Invention

In view of the above, the present invention aims to provide a caffeoylquinic acid derivative, and a preparation method and a use thereof, wherein the caffeoylquinic acid derivative has a good lipid-lowering effect.

In view of the above object, the present invention provides a caffeoylquinic acid derivative having a structure represented by general formula (i):

in the formula: r₆Is an H atom;

R₄and R₅Is an H atom;

R₂and R₃Is H atom or propylidene;

R₁selected from alkylamines containing 1-18 carbons, arylamines containing 6-10 carbons or saturated or unsaturated cyclic amines containing 3-7 carbons.

In some embodiments of the invention, R₁Selected from alkylamines containing 3-8 carbons, arylamines containing 7-8 carbons or saturated cyclic amines containing 5-6 carbons.

In some embodiments of the invention, R₁Selected from n-butylamino, isobutylamino, n-octylamino, propargylamino, benzylamino, piperidinyl or cyclohexylamino.

The invention also provides a preparation method of the caffeoylquinic acid derivative, which comprises the following steps:

(1) reacting 3-O-caffeoylquinic acid with 2, 2-dimethoxypropane to generate an intermediate;

(2) and carrying out condensation reaction on the intermediate and amine to generate a target compound caffeoylquinic acid derivative.

In some embodiments of the present invention, after step (2), further comprising:

(3): carrying out deprotection reaction on the target compound caffeoylquinic acid derivative.

In some embodiments of the present invention, the method for preparing caffeoylquinic acid derivatives comprises the following steps:

(1) reacting 3-O-caffeoylquinic acid and 2, 2-dimethoxypropane in anhydrous acetone by taking p-toluenesulfonic acid as a catalyst to generate an intermediate shown in a formula (II);

(2) the intermediate shown in the formula (II) is subjected to condensation reaction with amine in a mixed solvent of anhydrous acetonitrile and tetrahydrofuran under the condensation condition of BOP and DIEA to generate the target compound caffeoylquinic acid derivative.

(3): removing acetonylidene protection from a target compound caffeoylquinic acid derivative in a mixed solution of trifluoroacetic acid, dichloromethane and water.

The invention also provides a pharmaceutical composition, which comprises the caffeoylquinic acid derivative and a medicinal carrier.

The invention also provides application of the caffeoylquinic acid derivatives in preparing a medicament for preventing and/or treating hyperlipidemia.

The invention also provides application of the caffeoylquinic acid derivatives in preparing medicaments for preventing and/or treating atherosclerosis.

From the above, the invention can be seen in that 3-O-caffeoylquinic acid is used as a raw material, the structural skeleton of the 3-O-caffeoylquinic acid is kept, different amino substituent groups are introduced to the carboxyl group at the 1 position, and the hydroxyl groups at the 4 and 5 positions are substituted or exposed by ketal, so that the novel caffeoylquinic acid derivative with lipid-lowering activity is synthesized, the synthesis method is simple and convenient, and the product purity is high. Compared with 3-O-caffeoylquinic acid, the caffeoylquinic acid derivative provided by the invention has better lipid-lowering activity. The invention fully utilizes medicinal plant resources found from natural products, deeply researches and develops to find compounds with unique chemical structures, and finds the lipid-lowering medicament for clinical application.

Drawings

FIG. 1 is a graph of the modulating effect of chlorogenic acid and its derivatives on oleic acid-induced lipid accumulation in HepG2 hepatocytes; wherein, A picture shows the regulation effect of

caffeoylquinic acid derivatives

3a, 3b, 3c, 3d, 3e and 3f on lipid accumulation caused by oleic acid in HepG2 liver cells; the B picture shows that the

caffeoylquinic acid derivatives

4a, 4B, 4c, 4d, 4e and 4f have the regulating effect on lipid accumulation initiated by oleic acid in HepG2 liver cells, CA is chlorogenic acid, S is simvastatin, M is oleic acid, K is a control, and # p <0.001 is very obvious compared with the cells (K) treated by the control group; p <0.05 was significantly different relative to cells treated with oleic acid (M); p <0.01 was significant relative to oleic acid (M) -treated cells; the cells differed significantly with p <0.001 versus oleic acid (M) treated cells.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to specific embodiments below.

The invention provides a caffeoylquinic acid derivative, which has a structure shown in a general formula (I):

in the formula: r₆Is an H atom;

R₄and R₅Is an H atom;

R₂and R₃Is H atom or propylidene;

As an embodiment of the present invention, R₁Selected from alkylamines containing 3-8 carbons, arylamines containing 7-8 carbons or saturated cyclic amines containing 5-6 carbons.

As an embodiment of the present invention, R₁Selected from n-butylamino, isobutylamino, n-octylamino, propargylamino, benzylamino, piperidinyl or cyclohexylamino.

As an embodiment of the present invention, when R₆Is an H atom; r₄And R₅Is an H atom; r₂And R₃Is propylidene; the structure shown in the formula (I) is as follows:

when R is₁Is n-butylamino, and is a tertiary amino,

the caffeoylquinic acid derivative is now designated 3 a.

When R is₁Is an isobutylamino group, and the like,

the caffeoylquinic acid derivative is named 3 b.

When R is₁Is a n-octylamino group, and the amino acid is,

the caffeoylquinic acid derivative is named 3 c.

When R is₁Is a propargylamine group, and is a propargylamine group,

the caffeoylquinic acid derivative is named 3 d.

When R is₁Is a benzylamino group, and is a benzyl amine group,

the caffeoylquinic acid derivative is named 3 e.

When R is₁Is a piperidine group, and is a cyclic or cyclic piperidine group,

the caffeoylquinic acid derivative is named 3 f.

When R is₁Is a cyclohexylamino group, and is a cyclohexylamino group,

the caffeoylquinic acid derivative was named 3g at this time.

As an embodiment of the present invention, when R₆Is an H atom; r₄And R₅Is an H atom; r₂And R₃Is an H atom; the structure shown in the formula (I) is as follows:

when R is₁Is n-butylamino, and is a tertiary amino,

the caffeoylquinic acid derivative is now designated 4 a.

When R is₁Is an isobutylamino group, and the like,

the caffeoylquinic acid derivative is named 4 b.

When R is₁Is a n-octylamino group, and the amino acid is,

the caffeoylquinic acid derivative is named 4 c.

When R is₁Is a propargylamine group, and is a propargylamine group,

the caffeoylquinic acid derivative is named 4 d.

When R is₁Is a benzylamino group, and is a benzyl amine group,

the caffeoylquinic acid derivative is named 4 e.

When R is₁Is a piperidine group, and is a cyclic or cyclic piperidine group,

the caffeoylquinic acid derivative is named 4 f.

When R is₁Is a cyclohexylamino group, and is a cyclohexylamino group,

the caffeoylquinic acid derivative was named 4g at this time.

Specifically, the synthetic route of the caffeoylquinic acid derivative is as follows:

(1) synthesis reaction of intermediate 2

Taking a compound 1 (3-O-caffeoylquinic acid) as an initial raw material, and reacting 4, 5-hydroxy of the 3-O-caffeoylquinic acid with 2, 2-Dimethyloxypropane (DMP) in anhydrous acetone under the catalysis of p-toluenesulfonic acid (TsOH) to obtain an intermediate 2;

wherein, the structural formula of the 2, 2-dimethoxypropane is as follows:

(2) synthesis reaction of target Compound

Intermediate 2 was condensed with BOP (benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate) and DIEA (N, N-diisopropylethylamine) in anhydrous acetonitrile (CH)₃CN) and Tetrahydrofuran (THF) to generate a target compound caffeoylquinic acid derivative through a condensation reaction with amine, wherein the target compound caffeoylquinic acid derivative comprises 3a, 3b, 3c, 3d, 3e, 3f and 3 g.

In one embodiment of the present invention, the target compound caffeoylquinic acid derivative of step (2) is subjected to deprotection reaction to obtain the target compounds

caffeoylquinic acid derivatives

4a, 4b, 4c, 4d, 4e, 4f and 4 g.

Specifically, the synthetic route is as follows:

the caffeoylquinic acid derivatives of the target compounds comprise 3a, 3b, 3c, 3d, 3e, 3f and 3g, and the acetonylidene protection is removed from a mixed solution of trifluoroacetic acid (TFA), Dichloromethane (DCM) and water, so that caffeoylquinic acid derivatives of the target compounds, 4a, 4b, 4c, 4d, 4e, 4f and 4g, are respectively obtained.

The general synthetic route of the target compound caffeoylquinic acid derivative is as follows:

wherein, (A) DMP, TsOH, acetone; (B) BOP, DIEA, amines; (C) TFA, DCM, H₂O(9:1:1).

Therefore, according to the structural characteristics of the 3-O-caffeoylquinic acid, the hydroxyl at the 4, 5-position of the 3-O-caffeoylquinic acid is reacted with 2, 2-Dimethoxypropane (DMP), so that the hydroxyl at the 4, 5-position of the 3-O-caffeoylquinic acid is substituted by ketal to obtain an intermediate 2; then reacting the intermediate 2 with various amines, and introducing different amino substituent groups on the 1-position carboxyl group to obtain the caffeoylquinic acid derivatives; or the acetonylidene protection is further removed from the obtained caffeoylquinic acid derivative, so that the hydroxyl groups at the 4 and 5 positions are exposed. The preparation method of the caffeoylquinic acid derivative provided by the invention has the advantages of simple and convenient synthesis method and high product purity.

Example 1 preparation of caffeoylquinic acid derivative 3a

(1) Preparation of intermediate 2

Accurately weighing 7.0g (19.8mmol) of 3-O-caffeoylquinic acid (compound 1) in a round-bottom flask with the thickness of 250m L, adding 60m L anhydrous acetone and 40m L DMP (2, 2-dimethoxypropane), stirring to react to form suspension, adding a catalytic amount of 50mg TsOH (p-toluenesulfonic acid) into the reaction liquid, stirring to react until the reaction liquid is clear, reacting at room temperature, monitoring by T L C, and after the reaction is finished, using anhydrous Na₂CO₃And neutralizing until the pH value is 5-6, performing suction filtration, performing reduced pressure evaporation to remove the solvent, and purifying the residue by silica gel column chromatography to obtain an intermediate 2 with the yield of 87%.

(2) Preparation of caffeoylquinic acid derivative 3a

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred until dissolved, 0.8g (6mmol) of DIEA was added, 3mmol of n-butylamine was added dropwise to the reaction solution until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3a as pale yellow powder with a yield of 81%.

¹H-NMR(600MHz,DMSO):9.58(s,1H,Ph-OH),9.14(s,1H,Ph-OH),7.71 (t,J＝6.0Hz,1H,NH),7.48(d,J＝15.8Hz,1H,H-7′),7.05(d,J＝1.7Hz,1H,H- 2′),7.00(dd,J＝8.3Hz,1.7Hz,1H,H-6’),6.77(d,J＝8.2Hz,1H,H-5’),6.24(d, J＝15.9Hz,1H,H-8′),5.41(s,1H,H-1),5.37-5.32(m,1H,H-3),4.43-4.41(m, 1H,H-4),4.13-4.11(m,1H,H-5),3.11-3.01(m,2H,NH-CH₂),2.26-2.22(m,1H, H-2),1.99-1.94(m,1H,H-2),1.80-1.73(m,2H,H-6),1.42(s,3H,CH₃-C-O), 1.39-1.36(m,2H,NH-CH₂-CH₂),1.26-1.22(m,5H,CH₃-C-O,CH₂-CH₃),0.84(t, J＝7.3Hz,3H,CH₂-CH₃)；¹³C-NMR(150MHz,DMSO):175.0,165.9,148.4,145.6,145.3,125.6,121.4,115.8,114.9,114.1,108.0,76.9,74.1,73.5,70.9,38.2,37.0,34.5,31.2,28.0,25.9,19.4,13.7；HRMS(ESI):Calcd for[M+Na]⁺C₂₃H₃₁NNaO₈:472.1947,found 472.1948。

Example 2 preparation of caffeoylquinic acid derivative 4a

(3) Preparation of caffeoylquinic acid derivative 4a

In a 50m L round-bottomed flask, accurately weighing 2mmol caffeoylquinic acid derivative 3a, respectively, and adding TFA/DCM/H₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4a as a pale yellow powder in 78% yield.

¹H-NMR(600MHz,MeOD):7.58(d,J＝15.9Hz,1H,H-7′),7.05(d,J＝1.9 Hz,1H,H-2′),6.95(dd,J＝8.2Hz,1.9Hz,1H,H-6’),6.78(d,J＝8.1Hz,1H,H- 5’),6.29(d,J＝15.9Hz,1H,H-8′),5.42-5.38(m,1H,H-3),4.24-4.23(m,1H,H- 4),3.72-3.70(m,1H,H-5),3.21-3.19(m,2H,NH-CH₂),2.12-2.07(m,2H,H-2), 2.02-2.00(m,1H,H-6),1.95-1.91(m,1H,H-6),1.52-1.47(m,2H,NH-CH₂-CH₂), 1.36-1.32(m,2H,CH₂-CH₃),0.93(t,J＝7.4Hz,3H,CH₂-CH₃)；¹³C-NMR(150 MHz,MeOD):176.6,169.0,149.6,147.0,146.8,127.8,122.9,116.5,115.4, 115.2,77.8,74.4,72.7,71.9,40.0,39.9,38.7,32.6,21.0,14.1；HRMS(ESI):Calcd for[M+Na]⁺C₂₀H₂₇NNaO₈:432.1634,found 432.1636。

Example 3 preparation of caffeoylquinic acid derivative 3b

(1) Preparation of intermediate 2

(2) Preparation of caffeoylquinic acid derivative 3b

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred until dissolved, 0.8g (6mmol) of DIEA was added, 3mmol of isobutylamine was added dropwise to the reaction solution until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3b as pale yellow powder in 77% yield.

¹H-NMR(600MHz,DMSO):7.70(t,J＝6.1Hz,1H,NH),7.47(d,J＝15.8 Hz,1H,H-7′),7.04(d,J＝2.0Hz,1H,H-2′),7.00(dd,J＝8.2Hz,2.0Hz,1H,H- 6’),6.76(d,J＝8.2Hz,1H,H-5’),6.24(d,J＝15.9Hz,1H,H-8′),5.47(br,1H,H- 1),5.37-5.33(m,1H,H-3),4.43-4.41(m,1H,H-4),4.14-4.11(m,1H,H-5), 2.94-2.86(m,2H,NH-CH₂),2.26-2.22(m,1H,H-2),1.99-1.96(m,1H,H-2), 1.80-1.75(m,2H,H-6),1.74-1.69(m,1H,NH-CH₂-CH),1.42(s,3H,CH₃-C-O), 1.26(s,3H,CH₃-C-O),0.82-0.80(m,6H,2×CH₃)；13C-NMR(150MHz,DMSO) :175.1,165.9,148.5,145.6,145.4,125.6,121.4,115.8,114.9,114.1,108.0,76.9, 74.2,73.6,70.9,45.9,37.1,34.4,28.1,26.0,20.0；HRMS(ESI):Calcd for[M+ Na]+C₂₃H₃₁NNaO₈:472.1947,found 472.1946。

Example 4 preparation of caffeoylquinic acid derivative 4b

(3) Preparation of caffeoylquinic acid derivative 4b

In a 50m L round-bottomed flask, 2mmol of caffeoylquinic acid derivative 3b was accurately weighed out, respectively, and TFA/DCM/H was added₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4b as a pale yellow powder with a yield of 85%.

¹H-NMR(600MHz,MeOD):7.57(d,J＝15.7Hz,1H,H-7′),7.05(s,1H,H- 2′),6.99-6.89(m,1H,H-6’),6.78(d,J＝7.7Hz,1H,H-5’),6.29(d,J＝15.7Hz, 1H,H-8′),5.46-5.37(m,1H,H-3),4.30-4.18(m,1H,H-4),3.78-3.66(m,1H,H- 5),3.09-2.95(m,2H,NH-CH2),2.11-1.94(m,1H,H-2,6),1.85-1.73(m,1H, NH-CH₂-CH),0.94-0.84(m,6H,2×CH₃)；¹³C-NMR(150MHz,MeOD):176.7, 169.0,149.5,147.0,146.7,127.7,122.9,116.5,115.3,115.2,77.8,74.3,72.6,71.9, 47.6,40.0,38.7,29.6,20.3；HRMS(ESI):Calcd for[M+Na]⁺C₂₀H₂₇NNaO₈: 432.1634,found 432.1638.

Example 5 preparation of caffeoylquinic acid derivative 3c

(1) Preparation of intermediate 2

In a 250m L round bottom flask, 7.0g (19.8mmol) of 3-O-caffeoylquinic acid (Compound 1) was accurately weighed and 60 was addedm L anhydrous acetone and 40m L DMP (2, 2-dimethoxypropane), stirring to obtain suspension, adding a catalytic amount of 50mg TsOH (p-toluenesulfonic acid) into the reaction solution, stirring to react until the reaction solution is clear, reacting at room temperature, and monitoring with T L C, and reacting with anhydrous Na₂CO₃And neutralizing until the pH value is 5-6, performing suction filtration, performing reduced pressure evaporation to remove the solvent, and purifying the residue by silica gel column chromatography to obtain an intermediate 2 with the yield of 87%.

(2) Preparation of caffeoylquinic acid derivative 3c

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred until dissolved, 0.8g (6mmol) of DIEA was added, 3mmol of n-octylamine was added dropwise to the reaction solution until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3C as pale yellow powder in 82% yield.

¹H-NMR(600MHz,DMSO):7.74(t,J＝5.9Hz,1H,NH),7.47(d,J＝15.8 Hz,1H,H-7′),7.04(d,J＝2.0Hz,1H,H-2′),7.00(dd,J＝8.2Hz,2.0Hz,1H, H-6’),6.76(d,J＝8.2Hz,1H,H-5’),6.24(d,J＝15.9Hz,1H,H-8′),5.44(br,1H, H-1),5.36-5.31(m,1H,H-3),4.43-4.40(m,1H,H-4),4.13-4.11(m,1H,H-5), 3.10-2.98(m,2H,NH-CH₂),2.25-2.22(m,1H,H-2),1.97-1.94(m,1H,H-2), 1.75-1.74(m,2H,H-6),1.41(s,3H,CH3-C-O),1.40-1.37(m,2H,NH-CH₂-CH₂), 1.25(s,3H,CH₃-C-O),1.25-1.19(m,10H,(CH2)₅-CH₃),0.83(t,J＝7.1Hz,3H, (CH₂)₅-CH₃)；¹³C-NMR(150MHz,DMSO):174.9,165.8,148.4,145.5,145.2,125.5,121.3,115.7,114.8,114.0,107.9,76.8,73.9,73.4,70.8,38.4,37.0,34.5, 31.1,28.9,28.6,28.5,27.9,26.1,25.8,22.0,13.8；HRMS(ESI):Calcd for[M+ Na]⁺C₂₇H₃₉NNaO₈:528.2573,found 528.2578。

EXAMPLE 6 preparation of caffeoylquinic acid derivative 4c

(3) Preparation of caffeoylquinic acid derivative 4c

At 50m L round bottomIn a flask, 2mmol of caffeoylquinic acid derivative 3c is accurately weighed, and TFA/DCM/H is added₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4C as a pale yellow powder with a yield of 76%.

¹H-NMR(400MHz,MeOD):7.58(d,J＝15.9Hz,1H,H-7′),7.05(d,J＝1.9 Hz,1H,H-2′),6.95(dd,J＝8.2Hz,1.9Hz,1H,H-6’),6.78(d,J＝8.2Hz,1H,H- 5’),6.29(d,J＝15.9Hz,1H,H-8′),5.43-5.37(m,1H,H-3),4.24-4.23(m,1H,H- 4),3.72-3.69(m,1H,H-5),3.20-3.17(m,2H,NH-CH₂),2.13-1.90(m,1H,H-2, 6),1.52-1.49(m,2H,NH-CH₂-CH₂),1.34-1.26(m,10H,(CH₂)₅-CH₃),0.87(t,J ＝7.1Hz,3H,-CH₃)；¹³C-NMR(100MHz,MeOD):176.6,169.0,149.6,147.0, 146.8,127.8,123.0,116.5,115.3,115.1,77.7,74.4,72.7,72.0,40.2,40.0,38.7, 33.0,30.5,30.4,27.9,23.7,14.4；HRMS(ESI):Calcd for[M+Na]⁺C₂₄H₃₅NNaO₈:488.2260,found 488.2252。

Example 7 preparation of caffeoylquinic acid derivative 3d

(1) Preparation of intermediate 2

(2) Preparation of caffeoylquinic acid derivative 3d

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred until dissolved. 0.8g (6 m) ofmol) of DIEA, 3mmol of propargylamine was added dropwise to the reaction solution until the reaction was monitored by T L C, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3d as a pale yellow powder in a yield of 75%.

¹H-NMR(600MHz,DMSO):8.16(t,J＝5.8Hz,1H,NH),7.47(d,J＝15.8 Hz,1H,H-7′),7.04(d,J＝2.0Hz,1H,H-2′),7.00(dd,J＝8.2Hz,2.0Hz,1H,H- 6’),6.76(d,J＝8.2Hz,1H,H-5’),6.24(d,J＝15.8Hz,1H,H-8′),5.50(s,1H,H-1), 5.35-5.31(m,1H,H-3),4.43-4.41(m,1H,H-4),4.14-4.12(m,1H,H-5),3.84(dd, J＝5.8Hz,2.5Hz,2H,NH-CH₂),3.03(t,J＝2.5Hz,1H,CCH),2.25-2.22(m,1H, H-2),2.00-1.95(m,1H,H-2),1.79-1.70(m,2H,H-6),1.42(s,3H,CH₃-C-O), 1.26(s,3H,CH₃-C-O)；¹³C-NMR(150MHz,DMSO):175.3,166.0,148.5,145.7, 145.5,125.7,121.5,115.9,114.9,114.2,108.2,81.2,76.9,74.2,73.6,72.6,70.9, 37.0,34.3,28.4,28.1,26.0；HRMS(ESI):Calcd for[M+Na]⁺C₂₂H₂₅NNaO₈:454.1478,found 454.1480。

EXAMPLE 8 preparation of caffeoylquinic acid derivative 4d

(3) Preparation of caffeoylquinic acid derivative 4d

In a 50m L round-bottomed flask, 2mmol of caffeoylquinic acid derivative 3d was accurately weighed out, respectively, and TFA/DCM/H was added₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4d as pale yellow powder in 69% yield.

¹H-NMR(400MHz,MeOD):7.57(d,J＝15.9Hz,1H,H-7′),7.11-7.00(m, 1H,H-2′),6.99-6.89(m,1H,H-6’),6.78(d,J＝8.0Hz,1H,H-5’),6.29(d,J＝15.9 Hz,1H,H-8′),5.43-5.37(m,1H,H-3),4.29-4.19(m,1H,H-4),4.03-4.92(m,2H, NH-CH₂),3.74-3.71(m,1H,H-5),2.61-2.49(m,1H,CCH),2.15-1.95(m,4H,H- 2,6)；¹³C-NMR(100MHz,MeOD):176.5,169.0,149.5,147.0,146.7,127.7, 123.0,116.5,115.3,115.1,80.5,77.8,74.3,72.5,72.1,71.8,39.8,38.5,29.4； HRMS(ESI):Calcd for[M+Na]⁺C₁₉H₂₁NNaO₈:414.1165,found 414.1667。

EXAMPLE 9 preparation of caffeoylquinic acid derivative 3e

(1) Preparation of intermediate 2

(2) Preparation of caffeoylquinic acid derivative 3e

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred to dissolve, 0.8g (6mmol) of DIEA was added, 3mmol of benzylamine was added dropwise to the reaction solution until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3e as pale yellow powder with a yield of 79%.

¹H-NMR(600MHz,DMSO):8.32(t,J＝6.2Hz,1H,NH),7.48(d,J＝15.8 Hz,1H,H-7′),7.31-7.28(m,2H,Ph-H),7.24-7.20(m,3H,Ph-H),7.05(d,J＝2.0 Hz,1H,H-2′),7.01(dd,J＝8.2Hz,2.0Hz,1H,H-6’),6.77(d,J＝8.2Hz,1H,H- 5’),6.25(d,J＝15.9Hz,1H,H-8′),5.55(br,1H,H-1),5.39-5.35(m,1H,H-3), 4.45-4.42(m,1H,H-4),4.32-4.25(m,2H,NH-CH₂),4.15-4.13(m,1H,H-5), 2.29-2.25(m,1H,H-2),2.04-2.01(m,1H,H-2),1.84-1.76(m,2H,H-6),1.43(s, 3H,CH3-C-O),1.26(s,3H,CH3-C-O)；¹³C-NMR(150MHz,DMSO):175.2, 165.8,148.4,145.5,145.3,139.5,128.1,126.9,126.7,125.5,121.3,115.7,114.8, 114.0,108.0,76.8,74.1,73.4,70.8,42.0,37.0,34.5,27.9,25.8；HRMS(ESI):Calcd for[M+Na]⁺C₂₆H₂₉NNaO₈:506.1791,found 506.1790。

EXAMPLE 10 preparation of caffeoylquinic acid derivative 4e

(3) Preparation of caffeoylquinic acid derivative 4e

In a 50m L round-bottomed flask, 2mmol of caffeoylquinic acid derivative 3e was accurately weighed out, respectively, and TFA/DCM/H was added₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4e as a pale yellow powder with a yield of 73%.

¹H-NMR(600MHz,MeOD):7.58(d,J＝15.9Hz,1H,H-7′),7.32-7.22(m, 5H,Ph-H),7.05(s,1H,H-2′),6.96-6.94(m,1H,H-6’),6.78(d,J＝8.1Hz,1H,H- 5’),6.29(d,J＝15.8Hz,1H,H-8′),5.45-5.39(m,1H,H-3),4.39(m,2H,NH-CH₂), 4.26-4.25(m,1H,H-4),3.73-3.69(m,1H,H-5),2.17-1.97(m,4H,H-2,6)；¹³C- NMR(150MHz,MeOD):176.8,169.0,149.5,147.0,146.8,139.9,129.5,128.2, 128.2,128.0,122.9,116.5,115.4,115.2,77.9,74.4,72.7,71.9,43.8,40.0,38.8； HRMS(ESI):Calcd for[M+Na]⁺C₂₃H₂₅NNaO₈:466.1448,found 466.1484。

EXAMPLE 11 preparation of caffeoylquinic acid derivative 3f

(1) Preparation of intermediate 2

(2) Preparation of caffeoylquinic acid derivative 3f

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v＝3:2) stirring to dissolve, after adding 0.8g (6mmol) of DIEA, 3mmol of piperidine was added dropwise to the reaction solution until T L C monitoring the end of the reaction, the solvent was evaporated under reduced pressure and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 3f as a pale yellow powder in 64% yield.

¹H-NMR(600MHz,DMSO):9.59(s,1H,Ph-OH),9.15(s,1H,Ph-OH),7.48 (d,J＝15.8Hz,1H,H-7′),7.04(d,J＝1.5Hz,1H,H-2′),7.00(dd,J＝8.1Hz,1.5 Hz,1H,H-6’),6.77(d,J＝8.0Hz,1H,H-5’),6.22(d,J＝15.9Hz,1H,H-8′),5.62 (s,1H,H-1),5.32-5.29(m,1H,H-3),4.45-4.42(m,1H,H-4),4.08-4.06(m,1H, H-5),3.97-3.59(m,2H,NH-CH₂),3.55-3.43(m,1H,NH-CH),3.35-3.17(m,1H, NH-CH),2.33-2.30(m,1H,H-2),2.01-1.98(m,1H,H-2),1.90-1.86(m,2H,H-6), 1.58-1.50(m,2H,CH₂),1.49-1.37(m,2H,CH3-C-O,(CH₂)₂),1.25(m,3H,CH₃- C-O)；¹³C-NMR(150MHz,DMSO):171.5,165.8,148.5,145.6,145.4,125.5, 121.3,115.8,114.8,114.0,108.0,76.1,74.8,73.2,70.6,37.0,36.8,34.5,29.0,27.9, 25.8,25.5,24.2；HRMS(ESI):Calcd for[M+Na]⁺C₂₄H₃₁NNaO₈:484.1947,found 484.1949。

EXAMPLE 12 preparation of caffeoylquinic acid derivative 4f

(3) Preparation of caffeoylquinic acid derivative 4f

In a 50m L round-bottomed flask, 2mmol of caffeoylquinic acid derivative 3f was accurately weighed out, and TFA/DCM/H was added₂O (v/v ═ 9:1:1)20m L, the reaction was stirred at room temperature for 0.5 h until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give caffeoylquinic acid derivative 4f as a pale yellow powder with a yield of 61%.

¹H-NMR(600MHz,MeOD):7.57(d,J＝15.9Hz,1H,H-7′),7.05(d,J＝1.9 Hz,1H,H-2′),6.95(dd,J＝8.2Hz,2.0Hz,1H,H-6’),6.78(d,J＝8.2Hz,1H,H- 5’),6.26(d,J＝15.9Hz,1H,H-8′),5.36-5.33(m,1H,H-3),4.28-4.27(m,1H,H- 4),4.06-3.79(m,2H,NH-CH₂),3.73-3.71(m,1H,H-5),3.63-3.38(m,2H,NH- CH₂),2.33-2.01(m,4H,H-2,6),1.68-1.61(m,2H,CH₂),1.60-1.48(m,4H, (CH₂)₂),1.90-1.86(m,2H,H-6),1.58-1.50(m,2H,CH₂),1.49-1.37(m,2H, CH3-C-O,(CH₂)₂)；¹³C-NMR(150MHz,MeOD):173.2,168.8,149.6,147.2, 146.8,127.8,123.0,116.5,115.3,115.2,78.6,73.3,72.4,71.6,40.0,39.2,38.7, 30.7,25.6；HRMS(ESI):Calcd for[M+Na]⁺C₂₁H₂₇NNaO₈:444.1634,found 444.1637。

EXAMPLE 13 preparation of caffeoylquinic acid derivatives 3g

(1) Preparation of intermediate 2

(2) Preparation of caffeoylquinic acid derivative 3g

In a 250m L round-bottom flask, exactly 21.2 g (3mmol) of intermediate and 1.5g (3mmol) of BOP are weighed, and 70m L of anhydrous mixed solvent THF/CH is added under the protection of argon₃CN (v/v ═ 3:2) was stirred until dissolved, 0.8g (6mmol) of DIEA was added, 3mmol of cyclohexylamine was added dropwise to the reaction solution until T L C monitored the end of the reaction, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give 3g of caffeoylquinic acid derivative as pale yellow powder with a yield of 71%.

¹H-NMR(400MHz,DMSO):7.47(d,J＝15.8Hz,1H,H-7′),7.38(d,J＝8.2 Hz,1H,NH),7.04-6.99(m,2H,H-2′,6’),6.76(d,J＝8.1Hz,1H,H-5’),6.24(d,J ＝15.8Hz,1H,H-8′),5.44(br,1H,H-1),5.37-5.30(m,1H,H-3),4.42-4.41(m,1H, H-4),4.14-4.11(m,1H,H-5),3.51-3.49(m,1H,NH-CH),2.25-2.20(m,1H,H-2), 1.97-1.91(m,1H,H-2),1.77-1.75(m,2H,H-6),1.67-1.65(m,4H,CH(CH2)2), 1.41(s,3H,CH3-C-O),1.26-1.10(m,9H,CH3-C-O,(CH₂)₃)；¹³C-NMR(100 MHz,DMSO):174.1,165.8,148.4,145.5,145.3,125.5,121.3,115.7,114.8, 114.0,108.0,76.8,74.0,73.5,70.9,47.4,37.0,34.4,32.1,28.0,25.9,25.1,24.4； HRMS(ESI):Calcd for[M+Na]⁺C₂₅H₃₃NNaO₈:498.2104,found 498.2106。

EXAMPLE 14 preparation of caffeoylquinic acid derivative 4g

(3) Preparation of caffeoylquinic acid derivative 4g

In a 50m L round-bottomed flask, 3g of 2mmol caffeoylquinic acid derivative was accurately weighed out, and TFA/DCM/H was added₂O (v/v ═ 9:1:1)20m L, and the reaction was stirred at room temperature for 0.5 h until T L C monitored that the reaction was complete, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to give 4g of caffeoylquinic acid derivative as a pale yellow powder with a yield of 64%.

¹H-NMR(600MHz,MeOD):7.57(d,J＝15.9Hz,1H,H-7′),7.05(d,J＝2.0 Hz,1H,H-2′),6.95(dd,J＝8.2Hz,2.0Hz,1H,H-6’),6.78(d,J＝8.2Hz,1H,H- 5’),6.29(d,J＝16.0Hz,1H,H-8′),5.42-5.36(m,1H,H-3),4.24-4.22(m,1H,H- 4),3.72-3.69(m,1H,H-5),3.65-3.58(m,1H,NH-CH),2.12-1.89(m,4H,H-2,6), 1.84-1.82(m,2H,CH₂),1.77-1.73(m,2H,CH₂),1.65-1.58(m,2H,CH₂), 1.40-1.34(m,2H,CH₂),1.26-1.23(m,2H,CH₂)；¹³C-NMR(150MHz,MeOD): 175.7,169.0,149.6,147.0,146.8,127.8,122.9,116.5,115.4,115.2,77.6,74.4, 72.7,72.0,49.6,40.0,38.7,33.6,30.7,26.5,26.1；HRMS(ESI):Calcd for[M+ Na]+C₂₂H₂₉NNaO₈:458.1791,found 458.1795。

Test example 1 biological Activity test of Caffeoylquinic acid derivatives

1.1 cytology methods

The human liver HepG2 cell line was cultured in DMEM medium containing 10% fetal bovine serum and penicillin/streptomycin (100. mu.g/m L) at 37 ℃ in 5% CO₂When the cells grow to 70-80% of the culture dish, stimulating the cells for 12h by using 100 mu mol/L oleic acid (M) and palmitic acid mixed liquor to prepare a lipid accumulation model, and then giving different caffeoylquinic acid derivativesBiological (10 mu mol/L) and chlorogenic acid (namely 3-O-Caffeoylquinic Acid) (CA) and simvastatin (S) positive control with the same concentration, a blank group (K) control is added, the positive control and the simvastatin (S) positive control are incubated for 24h, oil red O staining is used after the experiment is finished, the absorbance (A) value is measured at 358nm through a spectrophotometer, and effective compounds for inhibiting lipid accumulation in the caffeoylquinic acid compounds are observed, each experiment is repeated for 3 times, and according to three independent experiments, data are expressed as

The value is obtained.

1.2 results of lipid-lowering Activity

The lipid-lowering effect of chlorogenic acid and its derivatives on oleic acid-induced lipid accumulation in HepG2 hepatocytes was examined using an oil red O staining test, the results of oil red O staining indicate that some derivatives have a moderate to good modulating effect on oleic acid-induced lipid accumulation in HepG2 hepatocytes as shown in fig. 1, preliminary testing at 10 μmol/L showed that

caffeoylquinic acid derivatives

3d, 3f, 3g, 4a and 4c more moderate lipid-lowering effect on HepG2 cells, treatment of HepG2 hepatocytes with

caffeoylquinic acid derivatives

3d, 3g, 4a and 4c had better modulating effect than Chlorogenic Acid (CA), after treatment with 10 μmol/L

caffeoylquinic acid derivatives

3d, 3f and 3g in fig. 1A, respectively, the absorbance was reduced from 0.261 (treated with oleic acid alone) to 0.249,0.255 and 0.249, and 0.249 in fig. 1B, after treatment with 10 μmol/L

caffeoylquinic derivatives

3d, 3f and 3g, respectively, to 0.267 c, to 0.271, where the potential modulating effect of the lipid-accumulation of the derivatives was found to 0.271, 3d, 3c, 3d, 3c, and 267 c, 3c, respectively, to show a, as strong modulating effects after treatment with oleic acid derivatives, to 0.271, which were studied.

1.3 Structure-activity relationship analysis

Preliminary structure-activity relationship (SARs) analysis showed that amide derivatives of chlorogenic acid with ketal-substituted 4,5 hydroxyl groups were slightly less active than the amide derivatives with naked 4,5 hydroxyl groups compared to simvastatin (S) and Chlorogenic Acid (CA) positive controls. In addition, the 3d, 4d, 3f, 4f, 3g and 4g derivatives containing propargylamine, piperidyl and cyclohexylamine groups can better regulate lipid accumulation after oleic acid treatment, and show that the introduction of propargylamine, piperidyl and cyclohexylamine groups can improve lipid accumulation. Among them,

derivatives

3d and 4d showed stronger lipid-lowering effect compared to other compounds, which means that propargylamine group is a favorable substituent for regulating lipid accumulation in HepG2 hepatocytes. However, the

derivatives

3b and 4b (including the isobutylamino group) were less active, indicating that the introduction of the isobutylamino group was not favorable for the derivatives to exert lipid-lowering activity. In addition, the above derivatives show lipid-lowering effect in fig. 1, which shows that the 1-position carboxyl group and the 4, 5-position hydroxyl groups of chlorogenic acid are not essential groups for regulating lipid effect, and the influence of different amino substituents on the 1-position carboxyl group on lipid-lowering activity is more obvious.

Those of ordinary skill in the art will understand that: the discussion of any embodiment above is meant to be exemplary only, and is not intended to intimate that the scope of the disclosure, including the claims, is limited to these examples; within the inventive idea, also technical features in the above embodiments or in different embodiments may be combined and there are many other variations of the different aspects of the invention as described above, which are not provided in detail for the sake of brevity. Therefore, any omissions, modifications, substitutions, improvements and the like that may be made without departing from the spirit and principles of the invention are intended to be included within the scope of the invention.

Claims

1. A caffeoylquinic acid derivative, which is characterized by having a structure shown in a general formula (I):

in the formula: r₆Is an H atom;

R₄and R₅Is an H atom;

R₂and R₃Is H atom or propylidene;

R₁selected from 1-18 carbonsAlkyl amines, aryl amines having 6 to 10 carbons or saturated or unsaturated cyclic amines having 3 to 7 carbon atoms.

2. The caffeoylquinic acid derivative of claim 1, wherein R is₁Selected from alkylamines containing 3-8 carbons, arylamines containing 7-8 carbons or saturated cyclic amines containing 5-6 carbons.

3. The caffeoylquinic acid derivative of claim 2, wherein R is₁Selected from n-butylamino, isobutylamino, n-octylamino, propargylamino, benzylamino, piperidinyl or cyclohexylamino.

4. A process for the preparation of caffeoylquinic acid derivatives, as claimed in any one of claims 1-3, comprising the steps of:

5. The process for producing a caffeoylquinic acid derivative according to claim 4, which further comprises, after the step (2):

6. The process for producing a caffeoylquinic acid derivative according to claim 4, which comprises the steps of:

7. The process for producing a caffeoylquinic acid derivative according to claim 6, which further comprises, after the step (2):

8. A pharmaceutical composition comprising the caffeoylquinic acid derivative according to any one of claims 1-3 and a pharmaceutically acceptable carrier.

9. Use of the caffeoylquinic acid derivatives of any one of claims 1-3 in the preparation of a medicament for the prevention and/or treatment of hyperlipidemia.

10. Use of a caffeoylquinic acid derivative as defined in any of claims 1-3 in the preparation of a medicament for the prevention and/or treatment of atherosclerosis.