A compound for improving vascular endothelial cell function, and a preparation method and application thereof

Abstract

The application relates to a compound for improving vascular endothelial cell function, a preparation method and application thereof. The compound is prepared by taking tyrosol, dopamine, substituted cinnamic acid or 5-carboxyl pentyl-triphenyl phosphonium bromide as a starting material, and then performing amidation or esterification reaction with adamantylamine and adamantane acid. The compound has a significant inhibitory effect on the inflammation reaction of endothelial cells induced by palmitate (PA) as an anti-inflammatory molecule, can be used for preparing drugs for hyperlipidemia and related cardiovascular diseases, treating vascular endothelial injury diseases, treating hyperlipidemia, atherosclerosis and related cardiovascular diseases, and has the potential of being developed into innovative drugs for treating hyperlipidemia, atherosclerosis and related cardiovascular diseases with high efficiency and low toxicity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a compound for improving vascular endothelial cell function, a preparation method and application thereof. BACKGROUND

[0002] Cardiovascular diseases (CVDs) are a class of diseases related to the heart or blood vessels, mainly including coronary heart disease, stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, abnormal heart rhythm, congenital heart disease, valvular heart disease, myocarditis, aortic aneurysm, peripheral arterial disease, thromboembolic disease and venous thrombosis. However, the earliest stage of cardiovascular disease is manifested in vascular endothelial damage, which is caused by multiple pathogenic factors, and endothelial damage and dysfunction are the early signs of arteriosclerosis, an important link in the occurrence and development of various cardiovascular diseases, and an important pathological basis for the occurrence and development of various diseases such as coronary heart disease, hypertension and diabetes. High blood lipids are one of the important risk factors for endothelial damage.

[0003] At present, there are drug and surgical treatment methods for cardiovascular diseases, but the type of disease has high concealment, high mortality and high disability rate, so early prevention is particularly important. Studies have reported that natural active ingredients such as chlorogenic acid, ligusticum and lignan have anti-atherosclerotic function, and functional foods using them as raw materials have been introduced. It is of great significance and prospect to explore more effective new drugs that can improve vascular endothelial damage and treat hyperlipidemia, atherosclerosis and other related cardiovascular diseases. Natural products have become an important source of innovative drugs with novel structures, unique biological activities and specific action targets due to their structural diversity. In fact, 61% of the small molecule new chemical entities of the currently marketed drugs can be traced back to natural products, such as artemisinin, morphine, ephedrine, scopolamine, reserpine, penicillin, camptothecin, paclitaxel and hydroxytyrosol, which are all directly derived from natural products.

[0004] However, there is no report on the design and synthesis of a series of innovative hydroxytyrosol derivatives based on the mother nucleus structure of natural product hydroxytyrosol. SUMMARY

[0005] In order to overcome the defects of the prior art, the purpose of the present application is to provide a compound for improving vascular endothelial cell function, a preparation method and application thereof. The compound can significantly inhibit the inflammatory response of human aortic endothelial cells caused by saturated fatty acids, and can significantly reduce the levels of triglycerides and free fatty acids in the serum of hyperlipidemia, and can be used for preventing and treating hyperlipidemia, atherosclerosis and other related cardiovascular diseases related to vascular endothelial inflammation.

[0006] To achieve the above object, the present application adopts the following technical solutions:

[0007] A compound for improving vascular endothelial cell function, the structure of which is shown in formula I or formula II:

[0008]

[0009] The R group includes a functional structure for introducing a substituent, and the functional structure is any one or more than two optimized structures of a nitro group, fluorine, or chlorine, and X is hydrogen or oxygen.

[0010] The structure can also be any one of a tautomer, a stereoisomer, or a pharmaceutically acceptable salt of the compound shown in formula I or formula II, including the following:

[0011]

[0012] A preparation method of a compound for improving vascular endothelial cell function, comprising the following steps:

[0013] Hydroxytyrosol, dopamine, substituted cinnamic acid, or 5-carboxy-pentyl-triphenylphosphonium bromide is used as a starting material, and is reacted with adamantylamine or adamantane acid through amidation or esterification to obtain the compound.

[0014] Specifically, the method comprises the following steps:

[0015] Hydroxytyrosol A and adamantane acid B, or substituted cinnamic acid C and adamantylamine D, or dopamine E and adamantane acid B, or 5-carboxy-pentyl-triphenylphosphonium bromide F and adamantylamine D are stirred under the condition of a solvent, a condensing agent, and a catalyst to synthesize a series of ester or amide derivatives through intermolecular dehydration, and the compound is obtained through column chromatography for purification:

[0016]

[0017]

[0018] The R group includes a functional structure for introducing a substituent, and the functional structure is any one or more than two optimized structures of a nitro group, fluorine, or chlorine, and X is hydrogen or oxygen.

[0019] The solvent is one or more of tetrahydrofuran, acetonitrile, benzene, dichloromethane, and N,N-dimethylformamide, and the amount is sufficient for complete dissolution; the reaction temperature is 25-30°C, and the reaction time is 15-20 h.

[0020] The condensing agent is one or more of N,N'-dicyclohexyl carbodiimide (DCC), carbonyl diimidazole (CDI), diisopropyl carbodiimide (DIC), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDCI);

[0021] The catalyst is one or more of 4-dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt).

[0022] The molar ratio of the condensing agent to the carboxylic acid in adamantane acid B or substituted cinnamic acid C is 1:1 to 1:2.

[0023] The amount of the catalyst is controlled at 1% to 10% relative to the molar ratio of the carboxylic acid.

[0024] The molar ratio of the amine or alcohol to the carboxylic acid is (1 to 2):1.

[0025] An application for preparing a compound for improving vascular endothelial cell function, which can be used for preparing a drug for hyperlipidemia and related cardiovascular diseases, and treating vascular endothelial injury diseases.

[0026] The compound has anti-inflammatory activity.

[0027] The compound is used as an active ingredient for preparing a drug for treating hyperlipidemia and related cardiovascular diseases.

[0028] The beneficial effects of the present application are embodied in:

[0029] 1. The present application takes natural product hydroxytyrosol, dopamine or cinnamic acid derivative as a lead compound structure, designs and synthesizes derivatives with the two chemical structures as basic skeletons, and obtains anti-inflammatory molecules with more obvious inhibitory effect on inflammatory cytokines produced by Raw264.7 cells under LPS stimulation through toxicity and activity experiments, which helps to prolong the IBD remission period time as an active ingredient of a drug for treating inflammatory diseases.

[0030] 2. The present application acylates or esterifies hydroxytyrosol, dopamine or cinnamic acid derivative through condensation reaction to form a series of ester derivatives or amide derivatives with anti-inflammatory activity, and respectively investigates the influence of introducing different electron-withdrawing groups and electron-donating groups in the benzene ring on the biological activity of the target molecules (i.e. the nitro, fluorine, chlorine substituents in structure C), B. the influence of amide bond or ester bond on the biological activity of the target molecules, C. the influence of the length of the chain on the biological activity of the target molecules. A new way is provided for screening active ingredients of high-efficiency anti-inflammatory drugs.

[0031] 3、The application finds that the amide compound formed by dopamine and adamantane acetic acid has the strongest anti-inflammatory activity, and can be used for preparing therapeutic drugs for vascular endothelial inflammatory diseases, and is beneficial to obtaining innovative drugs for hyperlipidemia, atherosclerosis and other related cardiovascular diseases. BRIEF DESCRIPTION OF DRAWINGS

[0032] Fig. 1(a) is an HRMS spectrum of the compound HTNEADA; Fig. 1(b) is an HRMS spectrum of the compound CPNADA.

[0033] Fig. 2(a) is an HRMS spectrum of the compound HTNTPB; Fig. 2(b) is an HRMS spectrum of the compound HTNADA.

[0034] Fig. 3(a) is an HRMS spectrum of the compound NPNADA; Fig. 3(b) is an HRMS spectrum of the compound FPNADA.

[0035] Figure 4 Fig. 1 is an effect of HT-NEADA produced in the figure on human aortic endothelial cell activity and protective effect, wherein the abscissa is the concentration of the compound, and the ordinate is cell activity.

[0036] Figure 5 Fig. 1 is an inhibitory effect of HT-NEADA produced in the figure on palmitic acid-induced inflammatory reaction of human aortic endothelial cells, wherein the abscissa is the concentration of the compound, and the ordinate is the mRNA level content of interleukin-6 (IL-6).

[0037] Figure 6 Fig. 1 is an inhibitory effect of HT-NEADA produced in the figure on palmitic acid-induced inflammatory reaction of human aortic endothelial cells, wherein the abscissa is the concentration of the compound, and the ordinate is the mRNA level content of metal matrix protease-1 (MMP-1).

[0038] Figures 7A-7B Fig. 1 is that HT-NEADA produced in the figure can reduce the triglyceride and free fatty acid levels in hyperlipidemia mice, wherein: Figure 7A is the triglyceride level, Figure 7B is the free fatty acid level, the abscissa is different drug doses, and the ordinate is the triglyceride and free fatty acid level content. DETAILED DESCRIPTION

[0039] The application will be further described in detail below in combination with the drawings and examples. The examples are only used for explaining the application, and are not used for limiting the protection scope of the application.

[0040] Example One

[0041] Preparation of compound 3 (HTNEADA)

[0042]

[0043] Compound 2 (237.89 mg, 1.2 mmol, 1.2 eq), HOBT (206.83 mg, 1.5 mmol, 1.5 eq), EDCI (234.74 mg, 1.2 mmol, 1.2 eq) were dissolved in 5 ml dry dichloromethane and to this mixture dopamine hydrochloride 1 (189.64 mg, 1 mmol, 1 eq) and triethylamine (0.49 ml, 3.5 mmol, 3.5 eq) dissolved in 2 ml dry dichloromethane was added slowly drop wise at 0°C and stirring was continued for 0.5 h and the reaction mixture was allowed to attain room temperature and stirred overnight. TLC showed the end of the reaction. The solvent was removed and the reaction mixture was diluted with 20 ml water and extracted with ethyl acetate (20 x 3). The organic layers were combined and washed with 1 M HCI solution (10 x 1), saturated sodium chloride solution (20 x 1) and dried over anhydrous sodium sulphate. The organic layer was concentrated under vacuum and column chromatography was performed using P:E = 2:1 as the eluent to get 245.9 mg of compound 3 as a white solid with a yield of 74.64%. The HRMS (ESI) for C 20 H 27 NO3[M+H] + calculated as 329.1991and found to be 330.2058. See Figure 1(a).

[0044] Example Two

[0045] Preparation of compound 6 (HTNTPB)

[0046]

[0047] Compound 4 (933.36 mg, 2 mmol, 1 eq), adamantylamine 5 (370.42 mg, 2.4 mmol, 1.2 eq), DCC (505.31 mg, 2.4 mmol, 1.2 eq), DMAP (124.66 mg, 1 mmol, 0.5 eq) were dissolved in 10 ml dry dichloromethane and stirred at room temperature overnight. TLC showed the end of the reaction. After completion of the reaction, the reaction mixture was filtered under suction and the filtrate was washed with saturated sodium bicarbonate solution (20 x 1) and dried over anhydrous sodium sulphate. The organic layer was concentrated under vacuum and column chromatography was performed using D:M = 10:1 as the eluent to get 316.8 mg of compound 6 as a white solid with a yield of 26.82%. The HRMS (ESI) for C34 H 41 BrNOP[M-Br] + The value was calculated as 510.2920 and found to be 512.2354, as shown in Figure 2(a).

[0048] Example 3

[0049] Preparation of compound 8 (HTNADA)

[0050]

[0051] 3,4-Dihydroxyphenylacetic acid 7 (343.16 mg, 2 mmol, 1 eq) and triethylamine (0.56 mL, 4 mmol, 2 eq) were dissolved in 6.5 mL of DMF and cooled to 0 °C. Amantadine 5 (370.42 mg, 2.4 mmol, 1.2 eq) was added to the mixture, followed by slow dropwise addition of 9.5 mL of anhydrous dichloromethane solution of BOP (1.08 g, 2.4 mmol, 1.2 eq). The mixture was stirred at 0 °C for 30 min, then cooled to room temperature and stirred overnight. DCM was removed by rotary evaporation, and the mixture was diluted with water (20 × 1). Extraction was performed with ethyl acetate (20 × 3). The organic phases were combined and washed with 1 M HCl (10 × 1) and saturated sodium bicarbonate solution (20 × 1). The mixture was dried over anhydrous sodium sulfate, and the organic phase was concentrated under vacuum. Column chromatography was performed at a P:E ratio of 1:1 to give 129.6 mg of compound 8 as a white solid, with a yield of 21.5%. The HRMS (ESI) for C 18 H 23 NO3 [MH] + was calculated as 300.1678 and found to be 300.1595, see Figure 2(b).

[0052] Example 4

[0053] Preparation of compound 10 (NPNADA)

[0054]

[0055] Compound 9 (394.2 mg, 2 mmol, 1 eq), HOBT (330.92 mg, 2.4 mmol, 1.2 eq), and EDCI (469.48 mg, 2.4 mmol, 1.2 eq) were dissolved in 5 mL of anhydrous dichloromethane and stirred at 0 °C for 0.5 h. Amantadine 5 (370.42 mg, 2.4 mmol, 1.2 eq) was added to the mixture, followed by slow dropwise addition of triethylamine (0.97 mL, 7 mmol, 3.5 eq). Stirring continued for another 0.5 h, and the mixture was moved to room temperature and stirred overnight. The reaction endpoint was observed by TLC. The dichloromethane was removed by rotary evaporation, the mixture was diluted with 20 mL of water, and extracted with ethyl acetate (20 × 3). The organic phases were combined and subjected to 1 M... The sample was washed with HCl solution (10 × 1) and saturated sodium bicarbonate solution (20 × 1), dried over anhydrous sodium sulfate, and the organic phase was concentrated under vacuum. Column chromatography was then performed with a P:E ratio of 2:1 to give 467 mg of compound 10 as a white solid, in a yield of 71.54%. The HRMS (ESI) for C 19 H 22 N2O3[MH] + It was calculated as 326.1630 and found to be 327.1679, as shown in Figure 3(a).

[0056] Example 5

[0057] Preparation of compound 12 (FPNADA)

[0058]

[0059] Compound 11 (33.08 mg, 2 mmol, 1 eq), HOBT (330.92 mg, 2.4 mmol, 1.2 eq), and EDCI (469.48 mg, 2.4 mmol, 1.2 eq) were dissolved in 5 mL of anhydrous dichloromethane and stirred at 0 °C for 0.5 h. Amantadine 5 (370.42 mg, 2.4 mmol, 1.2 eq) was added to the mixture, followed by slow dropwise addition of triethylamine (0.97 mL, 7 mmol, 3.5 eq). Stirring continued for another 0.5 h, and the mixture was moved to room temperature and stirred overnight. The reaction endpoint was observed by TLC. The dichloromethane was removed by rotary evaporation, the mixture was diluted with 20 mL of water, and extracted with ethyl acetate (20 × 3). The organic phases were combined and treated with 1 M... The sample was washed with HCl solution (10 × 1) and saturated sodium bicarbonate solution (20 × 1), dried over anhydrous sodium sulfate, and the organic phase was concentrated under vacuum. Column chromatography was then performed with a P:E ratio of 2:1 to give 453.4 mg of compound 12 as a white solid, in a yield of 75.72%. The HRMS (ESI) for C 19 H 22 FNO[M+H]+ was calculated as 299.3894and found to be 300.1757,see Figure 3(b).

[0060] Example Six

[0061] Preparation of Compound 14 (CPNADA)

[0062]

[0063] Compound 13 (372.66 mg, 2 mmol, 1 eq), HOBT (330.92 mg, 2.4 mmol, 1.2 eq), EDCI (469.48 mg, 2.4 mmol, 1.2 eq) were dissolved in dry dichloromethane 5 ml, stirred at 0 °C for 0.5 h, to the mixture was added adamantylamine 5 (370.42 mg, 2.4 mmol, 1.2 eq), triethylamine (0.97 ml, 7 mmol, 3.5 eq) was added dropwise slowly, stirring was continued for 0.5 h, moved to room temperature and stirred overnight, TLC showed the end of the reaction, dichloromethane was distilled off, diluted with 20 ml water and extracted with ethyl acetate (20 x 3), the organic phase was combined and washed with 1 M HC1 solution (10 x 1), saturated sodium bicarbonate solution (20 x 1), dried over anhydrous sodium sulfate, the organic phase was concentrated in vacuum, column chromatography was performed with P:E = 2:1 developing conditions to get 525.9 mg of compound 14 as a white solid with 83.25% yield. The HRMS (ESI) for C 19 H 22 ClNO[M+H] + was calculated as 316.1390and found to be 316.1460,see Figure 1(b).

[0064] Example Seven

[0065] Example of preparation of tablet of drug for improving vascular endothelial cell function

[0066] After mixing 2 g of each of compounds HTNEADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA with 17.5 g of excipient (white lake: lactose = 7:3, mass ratio), granulation was performed with 95% ethanol, dried, and sized (sifted), 0.5 g of sodium stearate was added and mixed uniformly, and then tableting was performed to obtain tablets each weighing 100 mg, the content of compounds HTADA, HTNEADA, HTOADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA being 10 mg per tablet.

[0067] Example 8

[0068] Example of preparation of powder injection of the drug for improving vascular endothelial cell function

[0069] Each 1 g of the compounds HTNEADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA was dissolved in 170 mL of water for injection, and after initial mixing, the volume was made up to 200 mL. The resulting solution was filtered, filled into a vial, 1 mL per vial, lyophilized, sealed and sterilized to obtain a lyophilized powder injection containing 5 mg of the compounds HTADA, HTNEADA, HTOADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA per vial.

[0070] Example 9

[0071] Example of preparation of capsule of the drug for improving vascular endothelial cell function

[0072] Each 3 g of the compounds HTNEADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA was mixed with 27 g of excipients (white lake: lactose = 7:3, mass ratio), and then granulated with 95% ethanol, dried, sized (sifted), and filled into a capsule, about 30 mg per capsule, wherein the content of the compounds HTADA, HTNEADA, HTOADA, HTNTPB, HTNADA, NPNADA, FPNADA and CPNADA was about 3 mg.

[0073] In Examples 7, 8 and 9, the dosage form of the drug is a dosage form that can effectively allow the active ingredient to reach the body (particularly the blood circulation system), and is specifically selected from the group consisting of a tablet, a capsule, a powder, a granule, a syrup, a solution, a suspension, an injection, a tincture, an oral liquid, an aerosol, a buccal agent, a draught, a pill, a powder, and a sustained-release dosage form such as a nano formulation.

[0074] In the drug, in addition to the active ingredient described above, a small amount of a secondary component that does not affect the effectiveness of the active ingredient and / or a pharmaceutically acceptable carrier can be contained, for example, a sweetener to improve the taste, an antioxidant to prevent oxidation, and an excipient necessary for various formulations can be contained.

[0075] Biological activity test

[0076] (1) Experimental materials

[0077] The compound related to the present application is HT-NEADA, the structure of which is as follows:

[0078]

[0079] TRIzol reagent was purchased from Invitrogen; RNA reverse transcription kit, SYBR fluorescent dye were purchased from Dalian Baisheng Company. RNA primer sequences were ordered and synthesized from Xi'an Qikexi Biological Co., Ltd.

[0080] (2) Experimental cell culture and model establishment

[0081] Human aortic endothelial cells (HAEC) were purchased from Shanghai Baili Biological Technology Co., Ltd., and palmitic acid was purchased from Sigma Company. Cells were cultured in a 37°C constant temperature, humidified, sterile incubator containing 95% air and 5% CO2. The experiment was divided into three groups, respectively: (1) control group; (2) 500 μM PA treatment group; (3) HT and 6 HT derivatives pretreatment + 500 μM PA treatment group.

[0082] (3) Specific methods of cell experiment

[0083] ① MTT detection

[0084] Different concentrations of HT and 6 HT derivatives were used to act on HAEC, and the cells were cultured in a cell incubator for 24 h. Then 500 μM palmitic acid was used for 24 h, and then the cells were washed with PBS once, 0.5 mg / ml MTT was added, and the cells were incubated in a 37°C incubator containing 95% air and CO2 for 4 h. Then wash with PBS for three times, add DMSO for dissolution, and detect the absorbance value at 490 nm wavelength.

[0085] ② Detection of IL-6 and MMP-1 mRNA content

[0086] Reverse transcription RNA-real-time fluorescent quantitative PCR method was used for detection. The specific method is as follows:

[0087] A. RNA extraction

[0088] 12-well cell culture plate, add 500 μL of TRIzol reagent, room temperature, shaking bed for 5 min, then collect the cells into 1.5 mL EP tube, then add 200 μl chloroform (1 / 5 total volume) to extract protein, mix vigorously for 15 s, room temperature for 15 min, 12,000 g, 4°C, centrifuge for 10 min. The upper water phase was transferred to another EP tube, and an equal volume of isopropanol was added. Mix well and place at -20°C for 1 h, then centrifuge at 12,000 g, 4°C for 10 min, discard the supernatant. Add 1 mL of pre-cooled 75% ethanol, mix well by inverting, centrifuge at 12,000 g, 4°C for 10 min. Discard the supernatant, and place it on the clean bench for 30 min to completely evaporate the ethanol, and dissolve in 10 μl DEPC water. The concentration was determined by ultraviolet spectrophotometer and used for reverse transcription.

[0089] B. RNA reverse transcription

[0090] The transcription volume was 20 μl. Take 2 μg of RNA and add 0.5 μg of random primers. Add 4 μl of 5X Master Mix, and bring the volume to 20 μl with DEPC water. Incubate at 37°C for 60 min, then at 80°C for 15 s. Store at -20°C for later use.

[0091] C. Real-time quantitative PCR

[0092] The reaction was performed using the SYBR Green method. The reaction mixture consisted of 1 μl cDNA and 5 μl 2× Premix ExTaq TM II. Add 0.5 μl of upstream and downstream primer mixture (10 μM) to a final volume of 10 μl with sterile water. Reaction conditions were performed according to the manufacturer's instructions: PCR was performed at 95°C for 10 min, followed by 40 cycles (each cycle consisting of 95°C for 30 s, 55°C for 30 s, and 72°C for 20 s). The melting curve was observed at the end (95°C for 15 s, 60°C for 15 s, and 95°C for 15 s). β-actin was used as an internal control. The primer sequences used in the experiment were:

[0093] IL-6:

[0094] forward: 5-TTTTGTACTCATCTGCACAGC-3'

[0095] reverse:5-GGATTCAATGAGGAGACTTGC-3'

[0096] MMP-1:

[0097] Forward:5'-ACGCCAGATTTGCCAAGAG-3'

[0098] Reverse:5'-TTGACCCTCAGAGACCTTGGT-3'

[0099] β-actin:

[0100] forward:5'-ATCATGTTTGAGACCTTCAA-3'

[0101] reverse:5'-AGATGGGCACAGTGTGGGT-3'

[0102] (4) Establishment and administration of hyperlipidemia mouse model

[0103] 6-8 weeks old C57BL / 6 male mice were housed in a clean level laboratory animal room, the room was kept at temperature (22-28C), humidity 60%, appropriate ventilation air flow and 12h light and dark alternation. The experimental mice were free to eat and drink water, after adaptive feeding for 1 week, the mice were randomly divided into six groups: control group, hyperlipidemia mouse group, low dose HT (10mg / kg / day, 10 days), high dose HT (50mg / kg / day, 10 days), low dose HT-NEADA (10mg / kg / day, 10 days), high dose HT-NEADA (50mg / kg / day, 10 days), 8 in each group. The control group and the hyperlipidemia mouse group were given the same volume of water by gavage every day, and after 10 days of continuous gavage administration, the hyperlipidemia mouse group, the low dose of punicalagin group and the high dose of punicalagin group were injected intraperitoneally with 0.5g / kg of poloxamer 407 (Polo) to induce hyperlipidemia in mice, while the control group was injected with the same volume of normal saline, and the effect time after intraperitoneal injection was 24h.

[0104] (5) Statistical analysis

[0105] The results were expressed in the form of Mean ± S.E.M, and the data analysis used One Way-ANOVA analysis method, and the significant statistical significance was *p<0.05, **p<0.01, ***p<0.001.

[0106] (6) HT and 6 HT derivatives can increase the viability of human aortic endothelial cells inhibited by palmitic acid

[0107] After HT and 6 HT derivatives (0.01-50μM) acted on human aortic endothelial cells (HAEC) for 24h, MTT was used to detect cell viability, Figure 4 It is shown that HT and 6 HT derivatives have no toxic side effects on human aortic endothelial cells when the concentration is in the range of 0.01-1μM.

[0108] (7) Inhibitory effect of HT and 6 HT derivatives on inflammation of human blood aortic endothelial cells induced by palmitic acid

[0109] HT and 6 HT derivatives (0.1-10μM) were first added to human endothelial cells for 24h, and then 500μM palmitic acid was added for 24h, Figure 5 and Figure 6HT and six HT derivatives can significantly inhibit the inflammatory response of human aortic endothelial cells caused by palmitic acid damage, 500 μM of palmitic acid can cause inflammatory response of human aortic endothelial cells, the inflammatory level of the model group is significantly increased, combined with the levels of IL-6 and MMP-1, among the HT and six HT derivatives, the compounds with anti-inflammatory effect are HT, HT-NEADA, HT-NADA, NP-NADA and HT-NTPB, and the most obvious anti-inflammatory effect is HT-NEADA, which suggests that HT-NEADA has anti-inflammatory effect and potential in the treatment of hyperlipidemia and related cardiovascular diseases.

[0110] (8) HT-NEADA can reduce the levels of triglyceride and free fatty acid in the serum of hyperlipidemia mice

[0111] Triglyceride is mainly involved in energy metabolism in the human body to produce heat energy, and high content of triglyceride in the blood can cause blood to be thick, and lipid to deposit on the blood vessel wall, gradually forming small plaques, i.e. atherosclerosis. The increase of free fatty acid can make blood viscous, thereby blocking blood vessels, in addition, fatty acid oxidation can invade the intima of blood vessels, and blood vessels are prone to atherosclerosis and other diseases. Figure 7A and 7B It is shown that HT-NEADA can effectively reduce the content of triglyceride and free fatty acid in the serum of hyperlipidemia, which indicates that HT-NEADA can improve hyperlipidemia and has potential application value in related cardiovascular diseases. The following applications can be carried out:

[0112] Application of HT-NEADA in preparation of a drug for improving the function of aortic endothelial cells.

[0113] Further, the drug is a drug that can inhibit the inflammatory response of aortic endothelial cells caused by saturated fatty acids.

[0114] The drug is a drug that can reduce the mRNA levels of IL-6 and MMP-1 in aortic endothelial cells.

[0115] The drug is a drug for reducing hyperlipidemia.

[0116] Since endothelial cell function inflammation is the initial manifestation of atherosclerosis and related cardiovascular diseases, the basis and cause of development, hyperlipidemia (high blood triglyceride and free fatty acid levels) is one of the important risk factors of vascular endothelial injury caused by cardiovascular disease. HT-NEADA shows strong anti-inflammatory effect in the model of saturated fatty acid-induced vascular endothelial cell injury, and HT-NEADA can reduce the level of triglyceride and free fatty acid in hyperlipidemia. Therefore, HT-NEADA has good application prospect in preventing and treating endothelial inflammation caused by hyperlipidemia such as atherosclerosis and other cardiovascular diseases, and opens up a new medical approach for preventing and treating endothelial damage caused by unbalanced dietary structure leading to atherosclerosis and other cardiovascular diseases.

[0117] The following applications can be made:

[0118] The application of HT-NEADA in the preparation of drugs for preventing and treating cardiovascular diseases.

[0119] The drug is a drug for treating hyperlipidemia and atherosclerosis.

[0120] The application of HT-NEADA in the preparation of drugs or health products for preventing the occurrence of hyperlipidemia and atherosclerosis.

[0121] The above examples are the preferred examples of the implementation of the present application, and the present application is not limited to the above examples. Any non-essential addition, replacement made by the skilled in the art according to the technical features of the technical scheme of the present application belongs to the protection scope of the present application.

Claims

1. Use of a HT-NEADA compound for the preparation of a medicament for improving vascular endothelial cell function, characterized in that, The compound has the following structure: The compound can be used for preparing drugs for treating hyperlipidemia and atherosclerosis.

2. Use of a HT-NEADA compound for the preparation of a medicament for improving vascular endothelial cell function, characterized in that, The compound has the following structure: The compound is used for preparing drugs for inhibiting inflammation of aortic endothelial cells caused by saturated fatty acids.

3. Use of the HT-NEADA compound according to claim 2 for the preparation of a medicament for improving vascular endothelial cell function, characterized in that, The drugs can reduce the mRNA levels of IL-6 and MMP-1 in aortic endothelial cells.

4. Use of a HT-NEADA compound for the preparation of a medicament for improving vascular endothelial cell function, characterized in that, The compound has the following structure: The compound is used as an active ingredient for preparing drugs for treating hyperlipidemia and related cardiovascular diseases.

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