CN1539429A - Application of 3'-deoxyadenosine in the preparation of blood lipid-lowering drugs - Google Patents

Abstract

The invention discloses the use of 3'-deoxyadenosine shown in the general formula (I) for lowering blood fat, especially the use of 3'-deoxyadenosine for lowering serum triglyceride.

Description

Application of 3'-deoxyadenosine in the preparation of blood lipid-lowering drugs

technical field

The invention relates to the application of 3'-deoxyadenosine in the preparation of blood lipid-lowering drugs, in particular to the application of 3'-deoxyadenosine in the preparation of serum triglyceride-lowering drugs.

Background technique

Blood lipid is the general term for the lipids contained in the blood. Lipids in blood mainly include triglycerides, phospholipids, cholesterol and free fatty acids. The lipid content in the blood accounts for only a very small part compared with the total body lipid, but it is transported between various tissues and can often reflect the lipid metabolism in the body. Normal adult plasma lipid content is relatively stable, with a certain fluctuation range. Lipid levels are also susceptible to non-disease factors, including those caused by diet or short-term starvation. But this effect is only temporary and can return to normal conditions. However, if one or more lipids in the plasma are continuously higher than the normal value due to abnormal fat metabolism or operation, it is called hyperlipidemia (Hyperlipemia, HP).

Hyperlipidemia can be divided into the following three types according to the abnormally changed blood lipid components:

1. Hypercholesterolemia: The total blood cholesterol of normal people should be lower than 5.2 mmol/L (mmol/L). If it exceeds 5.7 mmol/L, it can be diagnosed as hypercholesterolemia. The total blood cholesterol level is between the two Those in between are borderline or critically elevated, which is also abnormal.

2. Hypertriglyceridemia: The ideal value of blood triglycerides should be less than 1.70 mmol/L; any blood triglycerides exceeding 1.7 mmol/L is the disease.

3. Compound hyperlipidemia: The disease can be diagnosed when the total cholesterol and triglycerides in the blood increase at the same time.

A large number of research data show that hyperlipidemia, including hypercholesterolemia (Hypercholesterolemia), hypertriglyceridemia (Hypertriglyceridemia) and complex hyperlipidemia is an important risk of stroke, coronary heart disease, myocardial infarction, and sudden cardiac death factor. 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, cholelithiasis, pancreatitis, fundus hemorrhage, blindness, peripheral vascular disease, claudication, and hyperuricemia. It has long been known that eating a high-fat and high-cholesterol diet can induce atherosclerosis, and it is more obvious in some people with genetic defects in lipid metabolism. These people have difficulty clearing excessive neutral fat and cholesterol in blood circulation. , causing elevated blood cholesterol and triglyceride concentrations.

Hyperlipidemia is a common disease, especially for the elderly. Hyperlipidemia is one of the diseases of modern civilization. In China, the number of people with hyperlipidemia increases with the development of the economy, and they are getting younger and younger, which is worrying. It is of great significance to find drugs or health foods that can effectively lower blood lipids, and will bring great social and economic benefits.

At present, there are many kinds of blood-lipid-lowering drugs, which can be divided into lowering total cholesterol (TC), mainly lowering total cholesterol and lowering triglycerides, lowering triglycerides (TG), and mainly lowering triglycerides and lowering triglycerides. The four categories of total cholesterol.

Bile acid integrators such as: resinous cholestyramine (ChoIestyramine) (also known as cholestyramine) and colestipol (CoIestipol) (also known as colestipol) have been clinically studied and applied abroad, and have been As the first-line drug for lowering TC. But these drugs taste bad and can easily cause constipation.

HMG-CoA reductase inhibitors are a novel class of lipid-lowering drugs, which can inhibit the biosynthesis of cholesterol in the body. In most patients with hypercholesterolemia, TC in the blood mainly comes from the synthesis in the body. These drugs have a stronger TC-lowering effect than bile acid integrators and have fewer side effects. Lovastatin (formerly known as Mevinolin) (also known as Levatin, Metrol) belongs to this category of blood lipid-lowering drugs. Lovastatins can not only significantly reduce serum TC and LDL-C levels, but also moderately reduce serum TG levels. But it also has many toxic and side effects, for example: about 2%-3% of patients after taking the medicine have gastrointestinal dysfunction, nausea, insomnia, muscle tenderness and skin rash. About 2% of patients treated with lovastatin can see elevated liver-derived transaminases, which can return to normal after drug withdrawal. Long-term administration of lovastatin to dogs with large doses (60-180mg/kg) per day can show cataracts, and human lovastatin with therapeutic doses. After 2 to 3 years of detailed ophthalmological examination, no crystal turbidity occurred. Its toxic and side effects can be divided into two categories in theory: one is caused by the reduction of metabolites of mevalonate; the other is caused by its own toxic effects.

Niacin (Nicotinic Acid, also known as Niacin) (belonging to B vitamins) and its derivatives such as Acipimox (also known as Olbetam) have the effect of anti-fat decomposition, among them, Asi The effects of Limus last longer and are more potent.

Contents of the invention

In order to overcome the deficiencies of anti-hyperlipidemia drugs in the prior art, the present invention provides the application of 3'-deoxyadenosine as shown in general formula (I) in the preparation of blood lipid-lowering drugs.

Another object of the present invention is to provide a pharmaceutical composition in which 3'-deoxyadenosine represented by general formula (I) is an active ingredient.

Specifically, the blood lipid-lowering effect of 3'-deoxyadenosine of the present invention is preferably lowering serum triglyceride in blood fat.

Hyperlipidemia in this application includes hypercholesterolemia (Hypercholesterolemia), hypertriglyceridemia (Hypertriglyceridemia) and complex hyperlipidemia.

Hyperlipidemia can also cause stroke, coronary heart disease, myocardial infarction, and sudden cardiac death; it can also promote high blood pressure, impaired glucose tolerance, and diabetes. Hyperlipidemia can also lead to fatty liver, liver cirrhosis, cholelithiasis, pancreatitis, fundus hemorrhage, blindness, peripheral vascular disease, claudication, and hyperuricemia.

In addition, high-fat and high-cholesterol diets can induce atherosclerosis, which is more obvious in some people with inherited lipid metabolism defects.

Therefore, the 3'-deoxyadenosine of the present invention can be prepared to prevent and/or treat hyperlipidemia-related cardiovascular and cerebrovascular diseases. The cardiovascular and cerebrovascular diseases are preferably stroke, coronary heart disease, myocardial infarction, sudden cardiac death, hypertension, atherosclerosis, and peripheral vascular disease.

The 3'-deoxyadenosine of the present invention can also be prepared to prevent and/or treat hyperlipidemia-related abnormal glucose tolerance, diabetes, fatty liver, liver cirrhosis, cholelithiasis, pancreatitis, fundus hemorrhage, blindness, lameness, hyperuricemia Hyperemia drugs.

The 3'-deoxyadenosine of the present invention can also be used in medicines for treating hyperlipidemia in patients with hereditary lipid metabolism defects.

The pharmaceutical composition of the present invention contains an effective dose of 3'-deoxyadenosine represented by general formula (I), and a pharmacodynamically acceptable carrier.

According to the present invention, the pharmaceutical composition can be tablets, capsules, pills, injections, sustained-release preparations, controlled-release preparations and various particle delivery systems.

In the safety experiment, 3'-deoxyadenosine was intragastrically administered to mice at 5.0 g/kg. After administration, observe continuously for two weeks, record the behavior, activity, body weight, food intake, feces and death of the tested mice, and execute them after two weeks for autopsy. After administration, the mice had smooth hair, normal behavioral activities, weight gain, food intake, and feces, and there was no significant difference compared with the solvent control group; and no death or other abnormalities occurred within two weeks. Autopsy was carried out after execution, and no abnormal changes were found in the major organs (heart, liver, spleen, lung, kidney) by naked eye, and there was no significant difference compared with the solvent control group. It shows that 3'-deoxyadenosine has no obvious toxic reaction, and the maximum tolerated dose in mice is 5.0 g/kg, that is, the LD ₅₀ of 3'-deoxyadenosine administered orally is greater than 5.0 g/kg.

Compared with the model group, the TG level of the hyperlipidemia model mice taking 3'-deoxyadenosine can significantly reduce the serum TG level in the 3'-deoxyadenosine 20mg/kg and 10mg/kg dose groups. Statistics show that there are highly significant differences between the 3'-deoxyadenosine 10mg/kg dose group and the hyperlipidemia model group. 3'-deoxyadenosine 20mg/kg, 10mg/kg dose group compared with fenofibrate group TG level has no significant difference, indicating that 3'-deoxyadenosine at 10mg/kg, 20mg/kg dose and fenofibrate Fibrate was equally effective in lowering serum triglycerides at a dose of 25 mg/kg.

CHO in hyperlipidemia model mice was significantly increased. There was no significant difference in CHO levels between each administration group and the model group, indicating that 3'-deoxyadenosine had no effect on lowering total cholesterol. 3'-deoxyadenosine has a strong effect on hyperlipidemia model mice mainly by reducing serum triglycerides.

Compared with the TG level of the model group, 3'-deoxyadenosine 100mg/kg, 50mg/kg, 25mg/kg, the three dosage groups can significantly reduce the serum TG level, and statistics show that there are highly significant differences, It shows that 3'-deoxyadenosine has a better effect of lowering triglycerides, but there is no significant difference in the intensity of lowering triglycerides among the three dose groups of 3'-deoxyadenosine. It shows that the administration of 3'-deoxyadenosine has a better effect of reducing serum triglycerides in the hyperlipidemia model rats fed with high-fat diet.

The pharmaceutical compositions of the present invention can be prepared according to methods known in the art. When used for this purpose, the active ingredient can be combined with one or more solid or liquid pharmaceutical excipients and/or adjuvants, if necessary, to make an appropriate administration form or dosage form for human medicine.

The pharmaceutical composition of the present invention can be administered in the form of a unit dose, and the route of administration can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneal or rectal, etc.

The administration route of the pharmaceutical composition of the present invention can be injection administration. Injection includes intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and acupoint injection.

The dosage forms for administration may be liquid dosage forms or solid dosage forms. For example, the liquid dosage forms can be true solutions, colloids, particulate dosage forms, emulsion dosage forms, and suspension dosage forms. Other dosage forms such as tablets, capsules, drop pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injections, etc.

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

Various carriers known in the art can be widely used for tableting unit dosage forms. Examples of carriers are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid Aluminum, etc.; wetting agents and binders, such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, acacia mucilage, gelatin paste, sodium carboxymethylcellulose , shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants, such as dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbate Sugar alcohol fatty acid esters, sodium lauryl sulfonate, methylcellulose, ethylcellulose, etc.; disintegration inhibitors, such as sucrose, tristearin, cocoa butter, hydrogenated oil, etc.; absorption enhancers , such as quaternary ammonium salts, sodium lauryl sulfate, etc.; lubricants, such as talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc. 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 formulate a dosage unit into a pellet, various carriers known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talc, etc.; binders such as acacia, tragacanth, Gelatin, ethanol, honey, liquid sugar, rice paste or batter, etc.; disintegrants, such as agar powder, dry starch, alginate, sodium dodecylsulfonate, methylcellulose, ethylcellulose, etc.

In order to formulate the administration unit into a suppository, various carriers known in the art can be widely used. Examples of carriers are, for example, polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides and the like.

To form a dosage unit into a capsule, the active ingredient is mixed with the various carriers mentioned above, and the mixture thus obtained is placed in a hard gelatin capsule or a soft capsule. The active ingredients can also be made into microcapsules, suspended in an aqueous medium to form a suspension, and can also be packed into hard capsules or made into injections for application.

For example, the composition of the present invention is made into injection preparations, such as solutions, suspension solutions, emulsions, and freeze-dried powder injections. This preparation can be aqueous or non-aqueous, and can contain one and/or more A pharmaceutically acceptable carrier, diluent, binder, lubricant, preservative, surface active agent or dispersant. For example, the diluent may be selected from water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid ester, and the like. In addition, in order to prepare isotonic injection, an appropriate amount of sodium chloride, glucose or glycerin can be added to the preparation for injection, and in addition, conventional solubilizers, buffers, pH regulators, etc. can also be added. These excipients are commonly used in the field

In addition, colorants, preservatives, fragrances, correctives, sweeteners or other materials can also be added to the pharmaceutical preparations, if necessary.

The dosage of the pharmaceutical composition of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, body weight, personality and individual response of the patient or animal, the route of administration, the frequency of administration, etc. , so the therapeutic dosage of the present invention can vary widely. Generally speaking, the dosages of the pharmaceutical ingredients in the present invention are well known to those skilled in the art. According to the actual amount of effective drug contained in the final preparation of the pharmaceutical composition of the present invention, it can be adjusted appropriately to meet the requirement of its therapeutically effective dose and complete the therapeutic purpose of the present invention. Usually, for patients with a body weight of about 75 kg, the daily dose of the compound of the present invention is 0.5 mg/kg body weight to 20 mg/kg body weight, preferably 4 mg/kg body weight to 16 mg/kg body weight. The above-mentioned dosage may be administered in a single dosage form or divided into several, for example, two, three or four dosage forms, which are limited by the clinical experience of the administering physician and the dosage regimen.

the term:

TG: Triglycerides

CHO: total cholesterol

LDL-C: low-density lipoprotein

HDL-C: high-density lipoprotein

VLDL-C: very low-density lipoprotein

Fibrates drugs such as Clofibrate mainly reduce the content of cAMP in adipocytes by inhibiting adenylate cyclase, inhibit the hydrolysis of adipose tissue, and reduce the content of non-esterified fatty acids in blood. Lead to decreased synthesis and secretion of hepatic VLDL. At the same time, it can enhance the activity of lipoprotein lipase and accelerate the catabolism of VLDL and TG. These eventually reduce the content of VLDL, TG, LDL-C and TC in the blood. In addition, it can also reduce the content of TC in the blood by inhibiting the synthesis of cholesterol by liver cells and increasing the excretion of cholesterol from the intestine.

A new generation of lipid-lowering drugs, such as fenofibrate, Lezhiping, gemfibrozil, etc. They all have a strong effect on lowering cholesterol and triglycerides, and have less toxic side effects. However, there are still some contraindications to be aware of. Such as pregnant women and breastfeeding women are contraindicated; patients with active hepatitis and impaired renal function are prohibited; patients who take the drug for a long time must regularly check the liver, kidney function and blood lipid levels. Adverse reactions include stomach discomfort, loss of appetite, diarrhea, headache, dizziness, paresthesia, leukopenia, elevated alanine aminotransferase, rash and other symptoms.

To sum up, at present, although many lipid-lowering drugs have been used clinically, they have played an important role in the treatment of hyperlipidemia. However, they more or less have certain toxic and side effects, and lipid-lowering drugs need to be continuously optimized to improve efficacy and reduce toxic and side effects.

3'-deoxyadenosine (3-deoxyadenosine, also known as cordycepin: Cordycepin), molecular formula C ₁₀ H ₁₃ N ₅ O ₃ , MW 251, is a nucleoside substance isolated from Cordyceps militaris, nest building Aspergillus nidulans can also produce this substance. Its structural formula is as follows:

There is no report on the blood lipid-lowering function of 3'-deoxyadenosine in the prior art. According to the retrieval of the Patent Retrieval and Consultation Center of the State Intellectual Property Office of China [entrustment number (G) 030358], there is no report on the blood lipid lowering of 3'-deoxyadenosine.

Detailed ways

The following examples are used to further illustrate the present invention, but this does not imply any limitation to the present invention.

1. Efficacy test

Example 1: Effect of 3'-deoxyadenosine prophylactic administration on blood lipid content of hyperlipidemia model mice

Materials and Methods

[Experimental animals] Kunming mice, male, weighing 18-20 g, were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences of the Chinese People's Liberation Army, license number: SCXK (Army) 2002-001.

[Test product] 3'-deoxyadenosine (isolated from the fruiting body of Cordyceps militaris, the analyzed purity is above 99.9%).

[Feed] Basic feed and high-fat feed were provided by the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: Jingdong Xuzi 003, certificate number: 0015790. 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% basic feed, 10% lard, 10% egg yolk powder, 1% cholesterol.

[Positive drug] Fenofibrate (Lipingzhi), French Liboforni Pharmaceutical Company. Specification: 100mg/capsule, batch number: 64801, production date: 2000.11

[Kit] Triglyceride (TG) kit, total cholesterol (CHO) kit, Beijing Beihua Kangtai Clinical Reagent Co., Ltd., batch number: 030320.

[Instrument] Seperate ^TM Max 190 microplate reader, Molecular Devices Corporation Sunnyvale, CA.

method

[Grouping and Administration] Mice were randomly divided into six groups according to body weight: normal control group, high-fat model group, 3'-deoxyadenosine high-dose group (20mg/kg), 3'-deoxyadenosine medium-dose group (10mg /kg), 3'-deoxyadenosine low-dose group (5mg/kg), fenofibrate group (25mg/kg), 10 rats in each group. The normal control group was given ordinary feed, and the other groups were given high-fat feed for 14 consecutive days, all of which were given unlimited supply. At the same time, each administration group was intragastrically administered once a day, and the administration volume of each group was 0.1ml/10g, and the administration was continued for 14 days. The high-fat model group was given an equal volume of normal saline every day.

[Efficacy evaluation] After 14 days, the mice were taken from eyeballs to take blood to measure the content of serum TG and CHO. The determination method was carried out according to the instructions of the TG and CHO kit, and the absorbance was measured at a wavelength of 500nm with a microplate reader. Calculate triglyceride content according to the following formula: TG (mg/dl) = absorbance of measuring tube/absorbance of standard tube ^* 200 (mg/dl); calculate total cholesterol content: CHO (mg/dl) = absorbance of measuring tube/absorbance of standard tube ^* 193 (mg/dl).

[Data analysis] The data are expressed as mean ± standard deviation, and the data analysis adopts t test.

result

1. Compared with the TG level of the normal control group (64.1±8.1mg/dl), the TG level of the model group (134.20±31.30mg/dl) has a very significant difference, indicating that the establishment of the high-fat mouse model in this experiment is successful . (See Table 1)

2. Compared with the TG level of the model group, both the 3'-deoxyadenosine high and medium dose groups can significantly reduce the serum TG level, and statistics show that there is a highly significant difference in the 3'-deoxyadenosine medium dose group ; Compared with the TG level of the fenofibrate group, there was no significant difference in the TG level of the 3'-deoxyadenosine and high-dose groups, indicating that the 3'-deoxyadenosine had the same effect as fenofibrate at 10mg/kg and 20mg/kg doses. Fibrate was equally effective in lowering serum triglycerides at a dose of 25 mg/kg (see Table 1).

3. Compared with the CHO level of the normal group, the CHO level of the model group has a very significant difference, indicating that the CHO level of the high-fat model mice is significantly increased. There was no significant difference in CHO levels between each administration group and the model group, indicating that 3'-deoxyadenosine had no effect on lowering total cholesterol (see Table 2).

in conclusion

3'-deoxyadenosine has a better effect on reducing serum triglycerides in hyperlipidemia model mice fed high-fat diet.

The influence of table 1.3'-deoxyadenosine on serum TG content of hyperlipidemia model mice (mean ± SD).

Group Dose (mg/kg) TG content (mg/dl)

Normal control group -- 64.1±8.1

Model group -- 134.20±31.30##

20 101.7±14.64 ^*

3'-Deoxyadenosine 10 93.73±17.19 ^**

5 128.16±22.54 ^ΔΔ

Fenofibrate group 25 97.77±12.19 ^*

^## p<0.01 compared with normal control group; ^* p<0.05, ^** p<0.01 compared with model group; ^ΔΔ p<0.01 compared with fenofibrate group.

The influence of table 2.3'-deoxyadenosine on serum CHO content of hyperlipidemia model mice (mean ± SD).

Group Dose (mg/kg) CHO content (mg/dl)

Normal control group -- 136.5±3.5

Model group -- 274.5±7.1 ^**

20 279.9±7.2

3’-deoxyadenosine 10 264.3±6.8

5 289.5±7.5

^** p<0.01 vs normal control group.

Example 2: Effect of 3'-deoxyadenosine preventive administration on blood triglyceride content in hyperlipidemia model rats

Materials and methods

[Experimental animals] Wistar rats, male, weighing about 220±20 g, were purchased from the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: SCXK-11-00-0006.

[Test product] 3'-deoxyadenosine (isolated from the fruiting body of Cordyceps militaris, the analyzed purity is above 99.9%).

[Feed] Basic feed and high-fat feed were provided by the Institute of Experimental Animals, Chinese Academy of Medical Sciences, license number: Jingdong Xuzi 003, certificate number: 0015790. 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% basic feed, 10% lard, 10% egg yolk powder, 1% cholesterol.

[Positive drug] Fenofibrate (Lipingzhi), French Liboforni Pharmaceutical Company. Specification: 100mg/capsule, batch number: 64801, production date: 2000.11

[Kit] Triglyceride (TG) kit, total cholesterol (CHO) kit, low-density lipoprotein-cholesterol (LDL-C) kit, high-density lipoprotein-cholesterol (HDL-C) kit, all Purchased from Zhongsheng Beikong Biotechnology Co., Ltd., batch number: 030313.

[Instrument] Seperate ^TM Max 190 microplate reader, Molecular Devices Corporation Sunnyvale, CA.

[Grouping and administration] Rats were fed with ordinary feed to adapt to the experimental environment for 5 days, then blood was collected from the fundus venous plexus of the capillary, centrifuged at 300G for 10 minutes to obtain serum, and the blood lipid index values before modeling were measured. Stratified randomization was performed according to TG level. Rats were divided into six groups: normal control group, high-fat model group, 3'-deoxyadenosine high-dose group (100mg/kg), 3'-deoxyadenosine medium-dose group (50mg/kg), 3'-deoxyadenosine Glycoside low-dose group (25mg/kg), fenofibrate group (36mg/kg), 10 rats in each group. The normal control group was given ordinary feed, and the other groups were given high-fat feed for 8 consecutive days, all of which were given unlimited supply. At the same time, each administration group was intragastrically administered once a day, and the administration volume of each group was 1.0ml/100g body weight, and the administration was continued for 8 days. The high-fat model group was given an equal volume of normal saline every day.

[Drug efficacy evaluation] After 8 days, blood was collected from the fundus venous plexus of rats to measure serum total cholesterol (CHO), triglyceride (TG), low-density lipoprotein (LDL-C) and high-density lipoprotein (HDL-C) content. The assay method was carried out according to the instructions of the kit, and the absorbance was measured with a microplate reader (wavelength: 500 nm). Very low density lipoprotein (VLDL-C) was calculated from CHO-LDL-C.

[Data analysis] The data are expressed as mean ± standard deviation, and the data analysis adopts t test. The blood lipid levels before and after administration were analyzed by paired t test.

result

1. Compared with the TG level of the normal control group (125.5±30.8mg/dl), the TG level of the model group (216.0±83.2mg/dl) has a very significant difference, indicating that the high-fat rat model in this experiment is successful. . (See Table 3)

2. Compared with the TG level of the model group, the 3'-deoxyadenosine high, medium and low dose groups can significantly reduce the serum TG level, and statistics show that there are highly significant differences, indicating that 3'-deoxyadenosine Adenosine has a good effect on lowering triglycerides, but there is no significant difference in the strength of the three doses of 3'-deoxyadenosine in lowering triglycerides. (See Table 3)

in conclusion

Preventive administration of 3'-deoxyadenosine has a better effect on reducing serum triglycerides in hyperlipidemia model rats fed high-fat diet.

Table 3 Effect of 3'-deoxyadenosine preventive administration on serum triglyceride content in hyperlipidemia model rats (mean±SD)

Lipids

Before administration After administration

Dosage Very Low Density Very Low Density

Groups Low Density Lipids Low Density Lipids

(mg/kg) Triglycerides Total Cholesterol Lipoproteins High Density Lipoproteins Triglycerides Total Cholesterol Lipoproteins High Density Lipoproteins

Protein

(TG) (CHO) (VLDL-C White (HDL-C) (TG) (CHO) (VLDL-C White (HDL-C)

(LDL-C) (LDL-C) (LDL-C)

) ) )

Normal pair 125.5±30.

Normal saline 102.9±24.1 126.3±5.8 27.3±12.4 66.9±11.6 40.3±10.7 125.6±7.0 19.7±9.0 76.6±10.6 35.1±21.3

Group Photo 8

High fat mold 216.0±83.

Normal saline 104.3±30.2 132.9±37.6 20.5±10.2 79.4±6.6 31.5±6.3 135.5±35.3 21.9±13.5 80.3±10.7 21.3±13.6 ^Δ

Type group 2 ^##ΔΔ

132.9±37.88.3± ^15.8Δ

100 101.5±29.2 131.1±36.0 29.7±9.5 75.5±10.5 31.7±7.1 135.1±24.4 23.1±8.6 39.7±35.6 ⁺

6 ^{**△△ ++++}

3'-Deoxy 121.1±36. 69.3±8.4 ^*Δ

Adenosine High 50 102.1±25.7 118.0±42.0 12.2±8.1 81.9±9.9 31.3±4.8 99.5±27.8 ^* 17.5±9.7 18.4±5.2 ^ΔΔ

0 ^{**++ Δ++}

Dose group 138.0±42. 68.7±7.6 ^*Δ

25 101.1±29.4 144.0±12.5 33.3±11.5 85.7±9.4 29.9±6.3 112.5±11.8 46.0±20.4 26.2±7.7 ⁺⁺

0 ^{**Δ++ Δ++}

Fenofib 82.0±12.5 102.9±14.7 42.9±12.6 ^*

36 102.6±26.0 138.1±21.3 35.9±15.7 79.9±13.1 34.2±6.1 49.8±22.5 14.0±5.0 ^ΔΔ

Special group ^{**Δ * *ΔΔ}

^** P<0.01, ^* P<0.05, compared with the high-fat model group; ^## P<0.01, ^# P<0.05, compared with the normal control group;

⁺⁺ P<0.01, ⁺ P<0.05, compared with fenofibrate group; ^ΔΔ P<0.01, ^Δ P<0.05, paired t test, compared with blood lipid levels before administration and after administration.

2. Safety experiment

Example 3 3'-deoxyadenosine acute toxicity test

Materials and methods

[Experimental animals] Kunming mice, weighing 18-22g, half male and half male, provided by the Department of Experimental Animal Science, Peking University Health Science Center, certificate number 01-3049

[Test product] 3'-deoxyadenosine (is isolated from the fruiting body of Cordyceps militaris for this laboratory, see embodiment 1).

[Test method] Take 40 Kunming mice, weighing 18-22g, half male and half male. They were randomly divided into two groups, 20 in each group, namely the 3'-deoxyadenosine group and the solvent control group, and administered after 12 hours of fasting (without water). The 3'-deoxyadenosine group used 12.5% 3-deoxyadenosine to give the mice intragastric administration of 0.4ml/10g once, and then the dosage was 5.0g/kg; the solvent control group was given the same amount of distilled water. After administration, observe continuously for two weeks, record the behavior, activity, body weight, food intake, feces and death of the tested mice, and execute them after two weeks for autopsy.

result

After administration, the mice in the 3'-deoxyadenosine group had smooth hair, normal behavioral activities, weight gain, food intake, and feces, which were not significantly different from those in the solvent control group; and no death or other abnormalities occurred within two weeks. The two groups were sacrificed at the same time, and autopsy was performed. The major organs (heart, liver, spleen, lung, and kidney) had no abnormal changes when compared with the solvent control group. The results are shown in Table 4.

in conclusion

The results of the maximum tolerated dose test of 3'-deoxyadenosine once administered orally showed that there was no obvious toxic reaction after administration to 20 mice, and no death occurred within two weeks. Therefore, the maximum tolerated dose of 3'-deoxyadenosine oral administration in mice is 5.0g/kg, that is, the LD50 of 3'-deoxyadenosine oral administration is greater than 5.0g/kg.

                                                                                                

Group Concentration Volume Volume Dose Number of Animals Number of Dead Animals Maximum Tolerable Dose

(%) (ml/10g) (g/kg) (only) (only) (g/kg)

3’-deoxyadenosine 12.5 0.4 5.0 20 0 5.0

Group

Solvent Control Group - 0.4 - 20 0 0 -

Claims (8)

1. the application of 3 '-deoxyadenosine shown in general formula (I) in the preparation blood lipid-lowering medicine.

2. application according to claim 1 is characterized in that described blood fat is a serum triglycerides.

3. 3 '-deoxyadenosine shown in general formula (I) prevents and/or treats application in the medicine of the relevant cardiac and cerebral vascular diseases of hyperlipemia in preparation.

4. application according to claim 3 is characterized in that, described cardiac and cerebral vascular diseases is:

Apoplexy, coronary heart disease, myocardial infarction, cardiac sudden death, hypertension, atherosclerosis, peripheral vascular disease.

5. 3 '-deoxyadenosine shown in general formula (I) prevents and/or treats the application of the medicine of the relevant impaired glucose tolerance of hyperlipemia, diabetes, fatty liver, liver cirrhosis, cholelithiasis, pancreatitis, retinal hemorrhage, blind, limping, hyperuricemia in preparation.

6. 3 '-deoxyadenosine shown in general formula (I) prevents and/or treats application in heritability lipid metabolism disorder patient's the medicine of hyperlipemia in preparation.

7. pharmaceutical composition is characterized in that: contain the 3 '-deoxyadenosine shown in general formula (I) of effective dose, and acceptable carrier on the pharmacodynamics.

8. according to the pharmaceutical composition of claim 7, it is characterized in that described pharmaceutical composition can be tablet, capsule, pill, injection, slow releasing preparation, controlled release preparation and various particulate delivery system.