CN102040603B - The purposes of the adjacent methoxycarbonyl benzyl tetrahydro Berberine of bromination N-and treatment hyperlipidemia thereof - Google Patents

Abstract

The invention discloses N-o-methoxycarbonylbenzyl tetrahydroberberine bromide and its application for treating hyperlipidemia. The structure of the brominated N-o-methoxycarbonyl benzyl tetrahydroberberine is shown in formula (I), also discloses the synthetic preparation method of the compound, and the pharmaceutical composition containing the compound, the compound for use in It is used to treat hyperlipidemia, or to treat hyperlipidemia in combination with other blood lipid-regulating drugs.

Description

N-o-methoxycarbonylbenzyltetrahydroberberine bromide and its use in treating hyperlipidemia

technical field

The present invention relates to the compound N-o-methoxycarbonylbenzyltetrahydroberberine bromide, the synthesis and preparation method of the compound, the pharmaceutical composition containing the compound, the use of the compound for treating hyperlipidemia, or the combination with other modulators Combination of blood lipid drugs.

Background technique

A large number of basic research data and clinical practice have proved that hyperlipidemia, including hypercholesterolemia, hypertriglyceridemia and compound hyperlipidemia, is an important risk factor for stroke, coronary heart disease, myocardial infarction, and sudden cardiac death . In addition, hyperlipidemia is also an important risk factor for hypertension, impaired glucose tolerance, and diabetes. Hyperlipidemia can also lead to fatty liver, liver cirrhosis, pancreatitis, fundus hemorrhage, blindness, peripheral vascular disease, and hyperuricemia.

Currently clinically used blood lipid-lowering drugs include cholesterol biosynthesis enzyme inhibitors represented by statins, which can reduce the synthesis of endogenous cholesterol in the body and reduce low-density lipoprotein cholesterol (LDL) by inhibiting HMG-CoA reductase. source of LDL-C). At the same time, this type of drug increases or activates the level of LDL receptor expression on the surface of liver cells to reduce the content of LDL in the blood. The other type is fibrate drugs that activate peroxisome proliferator-activated receptors (PPARs) transcription factors to reduce triglyceride levels. The dosage is small and the bioavailability is good, but serious adverse reactions such as liver injury and rhabdomyolysis have become the most worrying and concerned safety issues of statins and fibrates today.

Compared with the mode of developing new drugs by western pharmaceutical companies, finding new leading compounds for regulating blood lipids from traditional Chinese herbal medicines with definite curative effects in my country is a new drug research mode in line with my country's national conditions. Many Chinese herbal medicine ingredients have good blood lipid regulating effect and less adverse reactions, and have broad application prospects. For example, the triterpenoids contained in hawthorn can inhibit the synthesis of cholesterol in the body and accelerate the clearance of cholesterol; the active ingredients of anthraquinones contained in rhubarb, cassia and Polygonum multiflorum can promote intestinal peristalsis and increase the excretion of cholesterol: the lipid-lowering effect of Alisma and its relationship with The triterpenoids contained in it are related to the interference with the metabolism of endogenous cholesterol; the in vitro model of Danshen A in Danshen shows that it has the effect of inhibiting cholesterol synthesis; red peony can increase the excretion of bile acids; the total glycosides of Jiaogulan can make LDL-C in blood , Total cholesterol and triglyceride levels are reduced, and HDL-C levels are increased.

Berberine (berberine) is a quaternary ammonium alkaloid extracted from Coptis chinensis and other plants. Early researches were mainly on its antibacterial, antifungal and antiviral effects. But later, with the continuous in-depth research of clinical pharmacology, it was found that the drug also has hypoglycemic, hypolipidemic, anti-tumor and vascular system effects. Its mechanism of hypolipidemic action may have the following aspects:

(1) Regulatory effect on blood lipids Animal experiments have shown that berberine has a good regulatory effect on abnormal lipid metabolism in high-fat rats, and can reduce serum triacylglycerol (TG) in high-fat rats by 57%. The deposition of TG in the liver and skeletal muscle of high-fat rats was reduced by 36% and 23% respectively. Berberine can enhance the activity of lipoprotein lipase, which mainly decomposes TG in chylomicrons and very low-density lipoproteins, thereby achieving The effect of lowering blood fat. Recent studies have found that berberine can effectively inhibit the synthesis of fat in liver cells by activating AMP-active protein kinase, so as to achieve the purpose of reducing TG.

(2) The effect on adipocytes Berberine can inhibit the differentiation of adipocytes. Adipocyte differentiation will directly affect glucose and lipid metabolism, and then affect the occurrence and development of metabolic diseases; promote the secretion of adiponectin in adipocytes. Adiponectin[11] is an adipocyte factor discovered in recent years. It is a kind of collagen, which is specifically expressed in adipocytes and exists in a relatively high concentration in serum. Blood glucose concentration, fasting insulin concentration, and insulin resistance were negatively correlated, and positively correlated with insulin sensitivity. Studies have shown that BBR can directly increase the gene expression of adiponectin in isolated adipocytes, but the specific mechanism is still unclear.

(3) Enhancing the expression of low-density lipoprotein receptors Berberine has a good lipid-lowering effect, and its mechanism is different from that of statins, and even has advantages in some aspects[. Berberine promotes the transcription of cells, enhances the expression of low-density lipoprotein receptor (LDLR) protein, binds lipoproteins with higher affinity, mediates the uptake and metabolism of lipoproteins by cells, and maintains plasma LDL at a relatively stable Levels, thereby regulating the homeostasis of extracellular lipoproteins.

Contents of the invention

The technical problem to be solved by the present invention is to provide a new compound, i.e. N-o-methoxycarbonylbenzyltetrahydroberberine bromide.

Another technical problem to be solved by the present invention is to provide a preparation method of this compound.

Another technical problem to be solved by the present invention is to provide a pharmaceutical composition containing this compound.

Another technical problem to be solved by the present invention is to provide the use of this compound in the preparation of medicines, especially the use of treating hyperlipidemia, and also including treating hyperlipidemia in combination with other blood lipid-regulating drugs.

In order to solve the technical problem of the present invention, the present invention adopts the following technical solutions.

The N-o-methoxycarbonylbenzyltetrahydroberberine bromide involved in the present invention is shown in formula I

The present invention also provides a preparation method for the compound of formula I, which uses berberine hydrochloride as a raw material and is synthesized according to the following synthetic route:

1, the synthesis of tetrahydroberberine

The raw material used in the reaction is berberine hydrochloride, and the reduction reaction is carried out to obtain tetrahydroberberine.

A preferred reducing agent is borohydride; a more preferred reducing agent is NaBH4 _.

The molar ratio of borohydride and berberine hydrochloride is 1-4:1, more preferably 2-3:1, most preferably 2.4:1.

Solvent: The reaction is preferably carried out in the presence of lower alcohols, including methanol, ethanol, propanol, etc.; methanol is more preferred, and anhydrous methanol is most preferred.

The reaction time is 0.5-4 hours; the reaction time is 0.5-2 hours; the reaction time is 1 hour;

The temperature of the reaction is 20-100°C, preferably 50-80°C, more preferably 60-70°C, more preferably the temperature of solvent reflux;

The reaction product can be purified by recrystallization or silica gel column, preferably recrystallization purification, the solvent in the recrystallization purification is preferably chloroform, C ₁ -C ₄ alkyl alcohol, ethyl acetate, acetone, n-hexane, A mixture of one or more of petroleum ether, cyclohexane, dichloromethane, and water; preferably a mixture of ethanol and water.

2. Synthesis of N-o-methoxycarbonylbenzyltetrahydroberberine bromide

Tetrahydroberberine and methyl 2-bromomethylbenzoate are reacted to obtain N-o-methoxycarbonylbenzyltetrahydroberberine bromide.

The molar ratio of methyl 2-bromomethylbenzoate and tetrahydroberberine is 1.0-2.0:1, more preferably 1.0-1.5:1, most preferably 1.1:1.

Solvent: the reaction is preferably carried out in the presence of an aprotic solvent, preferably an aprotic polar solvent, and the preferred aprotic polar solvent is nitriles; the preferred nitriles are acetonitrile.

The reaction time is 4-12 hours; the reaction time is 6-10 hours; the reaction time is 8 hours;

The temperature of the reaction is 30-100°C, preferably 70-90°C, more preferably 80-85°C, most preferably the temperature at which the solvent refluxes;

The progress of the reaction can be monitored by TLC or HPLC; preferably by TLC.

The reaction product can be purified by recrystallization or silica gel column, preferably by silica gel column. The eluent is preferably halogenated lower alkanes, C ₁ -C ₄ alkyl alcohols, ethyl acetate, acetone, n-hexane, petroleum ether , a mixture of one or more of cyclohexane; preferably a mixture of dichloromethane and methanol, the preferred volume ratio of the mixture of dichloromethane and methanol is CH ₂ Cl ₂ :CH ₃ OH=98:2.

Wherein, the key intermediate 2-bromomethylbenzoic acid methyl ester is synthesized by the following synthetic route:

1. Synthesis of methyl o-toluate

In the presence of concentrated sulfuric acid, 2-methylbenzoic acid and anhydrous methanol were heated to reflux, and the reaction was completed in about 5 hours. After the reaction, the methanol was removed by steaming, then the pH was adjusted to neutral with 2N sodium hydroxide, and finally extracted with ethyl acetate, the extract was dried and evaporated to dryness to obtain methyl o-toluate.

2, Synthesis of methyl 2-bromomethylbenzoate

Take 2-methylbenzoic acid methyl ester and N-bromosuccinimide (NBS) and heat to reflux in the presence of carbon tetrachloride, and the reaction is completed in about 3 hours. After the reaction, the solid was filtered off and the solvent was evaporated, and finally water was added and extracted with ethyl acetate, and the extract was dried and passed through a decompression column (petroleum ether: ethyl acetate = 8:1) to obtain methyl 2-bromomethylbenzoate .

Still another aspect of the present invention relates to a pharmaceutical composition comprising the compound of the present invention as an active ingredient. The pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be prepared by combining the compounds of the present invention with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The content of the compound of the present invention in its pharmaceutical composition is usually 0.1-95% by weight.

The compound of the present invention or the pharmaceutical composition containing it can be administered in the form of a unit dosage, and the route of administration can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and Respiratory tract, skin, vagina, rectum, etc.

The dosage form for administration may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. Liquid dosage forms can be solutions (including true solutions and colloid solutions), emulsions (including o/w type, w/o type and double emulsion), suspensions, injections (including aqueous injections, powder injections and infusion solutions), eye drops Agents, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules ( Including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, gas (powder) aerosols, sprays, etc.; semi-solid dosage forms can be ointments, Gels, pastes, etc.

The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

To form the compound of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, iso Propanol, etc.; binders can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, arabic mucilage, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hypromellose Base cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked poly Vinylpyrrolidone, croscarmellose sodium, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecylsulfonate, etc.; lubricant and flow aid The agent can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol and the like.

Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

In order to make the administration unit into a capsule, the active ingredient compound of the present invention can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient compound of the present invention can also be made into granules or pellets with diluents, binders, and disintegrants, and then placed in hard capsules or soft capsules. Various diluents, binders, wetting agents, disintegrants, and glidants used in the preparation of tablets of the compound of the present invention can also be used in the preparation of capsules of the compound of the present invention.

In order to make the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixtures can be used as solvent and an appropriate amount of commonly used solubilizers, cosolvents, pH regulators and osmotic pressure regulators in this field can be added. The solubilizer or co-solvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be Sodium chloride, mannitol, glucose, phosphate, acetate, etc. For preparation of freeze-dried powder injection, mannitol, glucose, etc. can also be added as proppants.

In addition, coloring agents, preservatives, fragrances, flavoring agents or other additives can also be added to the pharmaceutical preparations, if necessary.

The inventors found that N-o-methoxycarbonylbenzyltetrahydroberberine bromide has obvious blood lipid regulating activity by modifying the structure of berberine.

The present invention observes the effect of the compound of the present invention on the blood lipid level of mice with hypercholesterolemia. The experimental method is as follows: the hypercholesterolemia model of mice is established by feeding high-fat feed, and the test product is administered by intragastric administration at a dose of 12.5 mg/kg for 7 days, once a day, and the total cholesterol level in the serum of the mice is measured. The experimental results show that the total cholesterol (TC) level in the serum of hypercholesterolemia mice is significantly reduced after 7 days of administration of the compound of the present invention, which is significantly different from that of the model control group. The compound of formula I of the present invention has a similar effect on reducing the total cholesterol level of mice at a dose of 12.5 mg/kg as that of simvastatin at a dose of 2.5 mg/kg.

The present invention also observes the influence of the compound of the present invention on the blood lipid and blood sugar levels of golden hamsters with hyperlipidemia. The experimental method is as follows: establish a hyperlipidemia model of golden hamster by feeding high-fat feed, and administer the test product by intragastric administration at a dose of 20 mg/kg for 14 days, once a day, and measure the serum total cholesterol of the model animals after administration. (TC), triglyceride (TG), high/low density lipoprotein-cholesterol (HDL-C/LDL-C), free fatty acid (FFA) and blood glucose (GLU) levels. Experimental result shows: after compound of the present invention is administered 14 days, TC, TG, LDL-C, FFA level significantly reduce in hyperlipidemia golden hamster serum, high-density lipoprotein cholesterol/low-density lipoprotein cholesterol (HDL-C/LDL -C) The ratio is significantly increased; at the same time, the compound of the present invention can inhibit the increase of blood sugar level in golden hamsters with hyperlipidemia.

The compound of formula I of the present invention can also be used in combination with other lipid-regulating drugs for the treatment of hyperlipidemia. Preferred other lipid-lowering drugs are selected from statins or fibrates.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, individual conditions of the patient or animal, administration route and dosage form, etc. Generally speaking, the suitable daily dosage range of the compound of the present invention is 0.001-150 mg/Kg body weight, preferably 0.1-100 mg/Kg body weight, more preferably 1-60 mg/Kg body weight, most preferably 2-30 mg/Kg body weight. The above-mentioned dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other therapeutic means.

The compound or composition of the present invention can be taken alone, or used in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dose should be adjusted according to the actual situation.

detailed description

The following examples illustrate the preparation of compounds of formula I. The examples are provided merely to illustrate how to prepare and use the compounds of the invention and are not to be construed as limiting the scope of the invention in any way. ¹ H-NMR was in all cases consistent with the given structures. Characteristic chemical shifts (δ) are expressed in parts per million below the magnetic field of tetramethylsilane, where conventional abbreviations are used to designate major peaks: e.g., s, singlet; d, doublet; t, triplet; q , quartet; m, multiplet; br, broad; dd, doublet of doublets. Mass spectra (m/z) were recorded by electrospray ionization. The following abbreviations are used for common solvents: DMSO, deuterated dimethylsulfoxide.

Example 1:

1. Synthesis of methyl o-toluate

Add 13.6g (100mmol) of 2-methylbenzoic acid and 100ml of anhydrous methanol into a 250ml single-necked bottle, then add 5ml of concentrated sulfuric acid, heat to reflux, and the reaction is completed in about 5 hours. After the reaction, the methanol was removed by steaming, then the pH was adjusted to neutral with 2N sodium hydroxide, and finally extracted with ethyl acetate (100ml×3), the extract was dried and evaporated to dryness to obtain 14.5g of the product with a yield of 96.7%.

2, Synthesis of methyl 2-bromomethylbenzoate

Add 7.5g (50mmol) of methyl 2-methylbenzoate, 9.79g (55mmol) of NBS, and 50ml of carbon tetrachloride into a 100ml single-necked bottle, heat to reflux, and the reaction is completed in about 3 hours. After the reaction, the solid was filtered off, and then the solvent was evaporated. Finally, 50ml of water was added and extracted with ethyl acetate (100ml×2). , yield 61.1%.

3. Synthesis of Tetrahydroberberine

Add 18.55g (50mmol) of berberine hydrochloride and 200ml of anhydrous methanol into a 500ml single-necked bottle, add 4.56g (120mmol) of NaBH ₄ in portions at 40°C, and then reflux for 1 hour after adding. After the reaction was completed, a light yellow solid was obtained by filtration, and then recrystallized with ethanol/water to obtain 16.5 g of a light yellow solid, with a yield of 88.9%.

¹ H-NMR (CDCl ₃ ): δ6.868(d, 1H), 6.788(d, 1H), 6.729(s, 1H), 6.594(s, 1H), 5.942(s, -O-CH2-O- ), 4.246(dd, 1H), 3.850(s, 6H), 3.607-3.523(m, 2H), 3.261-3.097(m, 3H), 2.871-2.781(m, 1H), 2.684-2.639(m, 2H ).

ESI-MS m/z: 488.2472 (M ⁺ ).

4. Synthesis of N-o-methoxycarbonylbenzyltetrahydroberberine bromide

Tetrahydroberberine (3.39g, 10mmol), 100ml of acetonitrile, and methyl 2-bromomethylbenzoate (2.52g, 11mmol) were sequentially added into the reaction flask, and the mixture was stirred and refluxed for 8h. TLC monitored the completion of the reaction. The solvent was recovered by concentration under reduced pressure to obtain a crude product, which was subjected to silica gel column chromatography (CH ₂ Cl ₂ :CH ₃ OH=98:2) to obtain the target compound, 3.9 g of a yellow solid, with a yield of 68.7%.

¹ H-NMR (CDCl ₃ ): δ8.105 (d, 1H), 8.024 (d, 1H), 7.701 (t, 1H), 7.659 (p, 1H), 7615-7.504 (m, 2H), 7.004 ( p, 1H), 6.700(s, 1H), 5.977(s, -O-CH2-O-), 5.643(d, 1H), 5.436(d, 1H), 4.794(dd, 1H), 4.779(d, 1H), 4.418(d, 1H), 3.849(s, 3H), 3.808(s, 3H), 3.791(s, 3H), 4.426-4.205(m, 2H), 3.278-3.138(m, 4H).

ESI-MS m/z: 340.1563 (M+H ⁺ ).

Pharmacological experiment

Experimental example 1: Effect of N-o-methoxycarbonylbenzyltetrahydroberberine bromide (code name WS0704035, the same below) on serum total cholesterol levels in hypercholesterolemia mice

Materials and methods

Experimental animals: Kunming mice, male, weighing 18-22 g, purchased from the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: SCXK (Beijing)-2007-0001.

Test product: WS0704035, white powder, provided by Wu Song's laboratory. Prepare the required concentration with 0.25% CMC before use, and store at 4°C for later use.

Positive control drug: Simvastatin, produced by Hangzhou Merck Pharmaceutical Co., Ltd., batch number: 07313. Prepare the required concentration with 0.25% CMC before use, and store at 4°C for later use.

Kit: Total Cholesterol Kit, produced by Zhongsheng Beikong Biotechnology Co., Ltd. Specifications: general purpose 10ml×10; batch number: 080611.200804; expiry date: 201103; store at 2-8°C.

Instrument: SeperateTMMax190 microplate reader, Molecular Devices Corporation Sunnyvale, CA.

Feed: Basic feed and high-fat feed were provided by the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: SCXK Beijing 2006-0003. Basic feed formula: 20% flour, 10% rice flour, 20% corn, 20% soybean flour, 25% bran, 2% bone meal, 2% fish meal. High-fat feed formula: 78.6% basic feed, 10% lard, 10% egg yolk powder, 1% cholesterol, 0.4% bile salt.

Modeling and grouping: mice were adaptively fed for 1 week and randomly divided into 5 groups, which were normal control group, model control group, positive control drug simvastatin group (2.5mg/kg), test product WS0704035 (12.5mg /kg), 10 animals in each group, wherein the animals in the normal control group were fed with common feed; the animals in the model control group and each administration group were fed with high-fat feed.

Administration and measurement of total cholesterol level: each administration group was intragastrically administered once a day in the morning, and the administration volume was 0.1ml/10g, and the administration was continued for 7 days. The normal control group and the high-fat model group were given an equal volume of 0.25% CMC. After 1 week of administration and fasting for 12 hours, blood was collected from orbital veins in each group of animals, and serum was routinely prepared, and serum TC content was measured with a kit. The assay method was carried out according to the instructions of the kit, and the absorbance was measured at a wavelength of 500 nm with a microplate reader. The total cholesterol content was calculated according to the following formula: CHO (mg/dl)=(OD value of serum to be tested-OD value of blank)/(OD value of standard serum-OD value of blank)*208.

Data processing and analysis: All data were statistically processed by analysis of variance, and the results were expressed as mean ± standard deviation.

result

Effects of WS0704035 on Serum Total Cholesterol Content in Hypercholesterolemic Mice

The results showed that the serum cholesterol level was significantly increased after the mice were given high-fat diet continuously for 1 week, and the test product WS0704035 was administered intragastrically at a dose of 12.5 mg/kg for 7 days, and the serum total cholesterol level was significantly reduced, compared with the positive control drug simvastatin The 2.5mg/kg dose was of equivalent strength (see Table 1.).

Table 1. Effect of WS0704035 on serum total cholesterol levels in hypercholesterolemic mice

Compared with the normal group ^# P<0.05, ^## P<0.01; compared with the model group ^* P<0.05.

Experimental example 2: Effect of WS0704035 on blood lipid and blood sugar levels in golden hamsters with hyperlipidemia

Materials and Methods

Experimental animals: golden hamster, male, 90-110 g, provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., certificate number: SCXK (Beijing) 2007-0001.

Test product: WS0704035, white powder, provided by Wu Song's laboratory. Prepare the required concentration with 0.25% CMC before use, and store at 4°C for later use.

Positive control drugs: Fenofibrate Tablets, produced by Liberfone Pharmaceuticals, France. Solvay Pharmaceuticals, batch number: 90894; Simvastatin Tablets, produced by Hangzhou Merck Pharmaceutical Co., Ltd., batch number: 07313.

Kits: total cholesterol (TC), lot number: 090751.200902; triglyceride (TG), lot number: 082861.200812; high-density lipoprotein-cholesterol (HDL-C/LDL-C), lot number: 080221.200906; low-density lipoprotein-cholesterol (HDL-C/LDL-C), batch number: 080211.200904, blood glucose (GLU), batch number: 081651.200812, all provided by Zhongsheng Beikong Biotechnology Co., Ltd. Free fatty acid (FFA) detection kit, produced by Beijing Pulilai Gene Technology Co., Ltd., batch number: 090201. Instrument: SeperateTMMax190 microplate reader, Molecular Devices Corporation Sunnyvale, CA.

Basic feed and high-fat feed: both were provided by the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: SCXK Beijing 2006-0003. Basic feed formula: 20% flour, 10% rice flour, 20% corn, 20% soybean flour, 25% bran, 2% bone meal, 2% fish meal. High-fat feed formula: 79.8% basic feed, 20% lard, 0.2% cholesterol.

Animal model establishment and grouping: Golden hamsters were randomly divided into 5 groups after adaptive feeding for 1 week, which were normal control group, model control group, fenofibrate group (40mg/kg), simvastatin group (2mg/kg ), WS0704035 (20mg/kg) four groups, each group of 8 animals, wherein the animals in the normal control group were fed with common feed; the animals in the model control group and each administration group were fed with high-fat feed.

Administration: The hamsters in each group were intragastrically administered once a day from the day of modeling to 2 weeks after modeling, and the normal control group and model control group were given the same volume of 0.25% CMC.

Determination of blood lipid and blood sugar: After 2 weeks of administration, animals in each group were fasted for 12 hours, anesthetized by intraperitoneal injection of 300 mg/kg chloral hydrate, blood was collected from the abdominal aorta, serum was routinely prepared, and serum TC, TG, HDL-C, LDL-C, TG, FFA, GLU content.

Data processing and analysis: All data were statistically processed by analysis of variance, and the results were expressed as (x±S).

result

Effect of WS0704035 on Blood Lipid Level in Hyperlipidemic Golden Hamsters

The results showed that after the hamsters were given high-fat feed continuously for 2 weeks, the content of serum cholesterol, triglyceride, low-density lipoprotein, and free fatty acid increased significantly, and the ratio of high-density lipoprotein/low-density lipoprotein decreased significantly, indicating that this model animal has obvious Hyperlipidemia characteristics; two positive control drugs - fenofibrate and simvastatin can significantly inhibit the rise of various blood lipid indicators in hyperlipidemic hamsters, indicating that the hyperlipidemia hamster model in this test system has been replicated successfully .

After the test article WS0704035 was administered orally for 14 days at a dose of 20 mg/kg, the contents of TC, TG and LDL-C in the serum of hyperlipidemia hamsters were significantly reduced, and the high-density lipoprotein cholesterol/low-density lipoprotein cholesterol (HDL- C/LDL-C) ratio significantly increased. Statistical analysis results show that WS0704035 reduces hyperlipidemia hamster serum total cholesterol and low-density lipoprotein cholesterol at a dose of 20 mg/kg significantly higher than 40 mg/kg fenofibrate (Table 1); The effect of lowering serum free fatty acids at a dose of 2 mg/kg was significantly stronger than that of 2 mg/kg simvastatin (Table 2).

Table 1. Effect of WS0704035 on blood lipid levels in hyperlipidemic hamsters

Compared with the normal control group: ^## P<0.01, ^### P<0.001; compared with the model control group: ^* P<0.05.

Table 2. Effect of WS0704035 on blood glucose and free fatty acid content in hyperlipidemic hamsters

Compared with the normal control group: ^# P<0.05, ^## P<0.01; compared with the model control group: ^* P<0.05.

In summary, pharmacodynamic experiments have confirmed that the compound of formula I of the present invention has the effect of regulating the blood lipid level of golden hamster hyperlipidemia model animals.

Claims (8)

1. such as formula the adjacent methoxycarbonyl benzyl tetrahydro of the bromination N-shown in (I) Berberine

2. prepare the method for claim 1 Chinese style (I) compound, it is characterized in that, comprise the steps:

The synthesis of (a) N-1

Take berberine hydrochloride as raw material, carry out reduction reaction and obtain N-1;

The synthesis of (b) bromination N-adjacent methoxycarbonyl benzyl tetrahydro Berberine

N-1 and 2-bromomethyl-benzoic acid methyl ester are reacted, obtains bromination N-adjacent methoxycarbonyl benzyl tetrahydro Berberine.

3. a pharmaceutical composition, is characterized in that, containing acceptable carrier in the adjacent methoxycarbonyl benzyl tetrahydro Berberine of the bromination N-shown in claim 1 Chinese style (I) and pharmacodynamics.

4. pharmaceutical composition according to claim 3, is characterized in that, also containing the second lipid regulating agent.

5. pharmaceutical composition according to claim 4, is characterized in that, described the second lipid regulating agent is selected from statins or fibrate.

6., according to the pharmaceutical composition of any one of claim 3-5, described pharmaceutical composition is selected from tablet, capsule, pill or injection.

7., according to the pharmaceutical composition of any one of claim 3-5, described pharmaceutical composition is selected from sustained release preparation or controlled release preparation.

8., according to the pharmaceutical composition of any one of claim 3-5, described pharmaceutical composition is selected from particulate delivery system.