CN110507650B

CN110507650B - Application of atractylenolide in preparation of antithrombotic drugs

Info

Publication number: CN110507650B
Application number: CN201910970248.3A
Authority: CN
Inventors: 化召辉; 李震; 刘仕睿; 马柯; 张麒
Original assignee: First Affiliated Hospital of Zhengzhou University
Current assignee: First Affiliated Hospital of Zhengzhou University
Priority date: 2019-10-12
Filing date: 2019-10-12
Publication date: 2023-04-07
Anticipated expiration: 2039-10-12
Also published as: CN110507650A

Abstract

The invention discloses application of atractylenolide in preparation of antithrombotic drugs. In recent years, the incidence of cardiovascular diseases is continuously increased, and the death caused by cardiovascular diseases is the first death rate of various diseases. Thrombotic disease is a high proportion of cardiovascular disease, of which thrombotic and embolic complications due to platelet dysfunction are the major causes of cardiovascular disease. The antithrombotic drug is an important means for preventing and treating thrombotic diseases, wherein the traditional Chinese medicine has unique curative effects on antiplatelet and antithrombotic, and has small side effect. The embodiment of the invention shows that the low and high dose of the atractylis ovata lactam can obviously prolong CT and BT of mice and inhibit FeCl ₃ Inducing thrombosis of rat, the atractylenolide has obvious anticoagulation and antithrombotic effects, and can be used for preparing antithrombotic drugs.

Description

Application of atractylenolide in preparation of antithrombotic drugs

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of atractylenolide in preparation of antithrombotic medicines.

Background

In recent years, the incidence of cardiovascular diseases is continuously increased, and the death caused by cardiovascular diseases is the first death rate of various diseases. Thrombotic diseases are a high proportion of cardiovascular diseases, of which thrombotic and embolic complications due to platelet dysfunction are the main causes of cardiovascular disease.

Thrombotic diseases are diseases caused by two pathological processes of thrombosis and thromboembolism, wherein the former refers to that the formed components in blood form emboli in blood vessels to cause partial or complete blockage of the blood vessels; the latter means that the thrombus is detached from the site of formation and partially or completely blocks some blood vessels along with the blood circulation. Thrombotic disease is characterized by high mortality and disability rates, with abnormal platelet activation being an important pathological basis for thrombosis.

The antithrombotic drug is an important means for preventing and treating thrombotic diseases, wherein the traditional Chinese medicine and the preparation thereof have unique curative effect on antithrombotic and small side effect, so that the discovery of natural compounds with antithrombotic activity from the traditional Chinese medicine and the preparation thereof is always a hotspot in the research field of antithrombotic drugs. Song Dynasty and others found that part of veratrum alkaloid has obvious antithrombotic effect, specifically related to the chemical structure of veratrum alkaloid, and the drug effect has obvious structure-activity relationship (research on antithrombotic effect and structure-activity relationship of veratrum alkaloid, chinese medicinal materials, no. 37, no. 11, 2014 11). The wanyangjun et al found that both the high and medium dose of formononetin sodium sulfonate could inhibit thrombosis and platelet aggregation in rats, prolong the Bleeding Time (BT) and the Activated Partial Thrombin Time (APTT) in mice, and have the function of inhibiting thrombosis, and the function mechanism of the formononetin sodium sulfonate could be related to the function of the formononetin sodium sulfonate in inhibiting ADP and thrombin induced platelet aggregation and reducing the endogenous blood coagulation pathway (antithrombotic function and mechanism research, volume 25, 12 of 2016, new Chinese medicine). In the morning and the like, the high, medium and low dose groups of diosmetin in the galium can obviously inhibit thrombosis of the inferior vena cava of rats and protect vascular endothelial cells, and the medium dose group has the effect equivalent to that of aspirin, which indicates that the diosmetin has the effects of anticoagulation and venous thrombosis resistance (experimental study on antithrombotic effect of diosmetin in the galium, traditional Chinese medicine information, vol. 31, no. 6 in 11 months 11 and 2014).

Atractylodes macrocephala lactam (Atractylenylolactam) is an amide natural compound and has the following chemical structure.

The antithrombotic effect of atractylenolide has not been disclosed so far.

Disclosure of Invention

The invention aims to provide application of atractylenolide in preparation of antithrombotic drugs.

The above purpose of the invention is realized by the following technical scheme:

application of atractylenolide in preparing antithrombotic medicine is provided.

An antithrombotic medicine contains Atractylodis rhizoma lactam as active ingredient.

Furthermore, the antithrombotic medicament also contains pharmaceutically acceptable auxiliary materials and is prepared into pharmaceutically acceptable dosage forms.

Still further, the adjuvant comprises a liquid, solid, or semi-solid adjuvant.

Further, the dosage forms include tablets, capsules, and injections.

Has the advantages that:

in recent years, the incidence of cardiovascular diseases is continuously increased, and the death caused by the cardiovascular diseases is the first death rate of all diseases. Thrombotic disease is a high proportion of cardiovascular disease, of which thrombotic and embolic complications due to platelet dysfunction are the major causes of cardiovascular disease. Antithrombotic drugs are important means for preventing and treating thrombotic diseases, wherein traditional Chinese medicines have unique curative effects on antiplatelet and antithrombotic drugs and have small side effects. The embodiment of the invention shows that the low and high dose of the atractylenolide can obviously prolong CT and BT of mice and inhibit FeCl ₃ Inducing thrombosis of rat, the atractylenolide has obvious anticoagulation and antithrombotic effects, and can be used for preparing antithrombotic drugs.

Drawings

FIG. 1 shows CT (unit: s) for each group of mice;

FIG. 2 shows BT (unit: s) of each group of mice;

FIG. 3 shows the thrombus mass (unit: mg) of rats in each group.

Detailed Description

The following examples are given to illustrate the essence of the present invention, but not to limit the scope of the present invention.

Example 1: effect of Atractylodes Macrocephala lactam on clotting time

Healthy Kunming mice with the weight of 18-22 g are randomly divided into 4 groups: a control group, a positive medicine group, a low-dose atractylenolide administration group and a high-dose atractylenolide administration group, wherein each group contains 6 patients. The positive medicine group is administrated with 50mg/kg/d aspirin by gavage, the low-dosage and high-dosage atractylenolide group is administrated with 20mg/kg/d and 50mg/kg/d atractylenolide by gavage respectively, the control group is administrated with equal volume of solvent 0.5 percent of CMC-Na solution by gavage, and the continuous administration is carried out for 7d. After 1h of the last administration, the Clotting Time (CT) was determined by the slide method. The specific method comprises the following steps: blood is collected by eyeballs of mice, 1 drop of blood is respectively dropped at two ends of a glass slide, the diameter of the blood drop is about 5mm, a stopwatch is used for timing immediately, a cleaning pin is used for slightly picking the blood from the edge of the blood inwards for 1 time every 10s, and the condition that blood filaments are picked up is observed. The time from the beginning of blood collection to the picking of blood filaments is CT, the longest observation time of CT is 15min, the non-coagulation time exceeding 15min is calculated according to 30min, and another blood drop is retested. CT was calculated for each group and analyzed using SPSS17.0 software, data were expressed as mean. + -. Standard deviation, comparisons between groups were by One-wayANOVA test, and P <0.05 indicated that the difference was statistically significant.

As shown in table 1 and fig. 1, compared with the control group, the CT of mice in the positive drug group, the low-dose atractylenolide administration group and the high-dose atractylenolide administration group was significantly prolonged, the difference was statistically significant, and the prolonged effect of the low-dose and high-dose atractylenolide administration group on CT showed dose dependence.

TABLE 1 groups of mice CT

Group of	CT (Unit: s)
		Control group	56.7±8.2
Positive drug group	88.3±7.5
		Low-dose administration group of atractylenolide	76.7±8.2
High-dose administration group of atractylenolide	98.3±7.5

Example 2: effect of Atractylodes Macrocephala lactam on bleeding time

Healthy Kunming mice with the weight of 18-22 g are randomly divided into 4 groups: a control group, a positive medicine group, a low-dose atractylenolide administration group and a high-dose atractylenolide administration group, wherein each group contains 6 patients. 50mg/kg/d aspirin is administrated in the gavage of the positive drug group, 20mg/kg/d and 50mg/kg/d atractylenolide are administrated in the gavage of the low-dosage and high-dosage atractylenolide administration groups respectively, the equal volume of the solvent in the gavage of the control group is 0.5 percent of CMC-Na solution, and the continuous administration is 7d. After the last administration for 1h, the Bleeding Time (BT) of the mice was measured by tail-biting method. The specific method comprises the following steps: fixing the mouse, measuring the length of the tail of the mouse by a millimeter ruler, marking, crossing the tail tip of the mouse at a position of 4mm by using a surgical scissors, and sucking blood drops once every 10 seconds when the blood automatically overflows, until the blood flow naturally stops (no blood is sucked by the filter paper), thus obtaining the BT. BT is observed for 30min at the longest time, and bleeding is still occurred after 30min, which is calculated according to 30 min. BT of each group is calculated and analyzed by SPSS17.0 software, data are expressed by mean +/-standard deviation, and comparison among groups is tested by One-wayANOVA, and P <0.05 represents that difference has statistical significance.

As shown in table 2 and fig. 2, compared with the control group, the BT of mice in the positive drug group, the low-dose atractylenolide administration group and the high-dose atractylenolide administration group was significantly prolonged, the difference was statistically significant, and the prolonged action of the low-dose and high-dose atractylenolide administration groups on BT was dose-dependent.

TABLE 2 groups of mice BT

Example 3: effect of Atractylodes Macrocephala lactam on thrombogenesis amount

Clean male SD rats weighing 260-280g were randomly divided into 4 groups: a control group, a positive medicine group, a low-dose atractylenolide administration group and a high-dose atractylenolide administration group, wherein each group contains 6 patients. The low-dosage and high-dosage atractylenolide administration groups are respectively gavaged with 20mg/kg/d and 50mg/kg/d atractylenolide, the control group is gavaged with equal volume of solvent 0.5 percent of CMC-Na solution and is continuously administered for 7d, and the positive medicines are respectively injected with 2000U/kg/d heparin sodium subcutaneously at the back at the 6 th and 7 th days. After the last 2h administration, the rats were anesthetized with 3% pentobarbital sodium, the abdominal cavity was opened by a median abdominal incision, and the abdominal aorta section was isolated. Then 50% FeCl by complete penetration ₃ And (3) encircling a filter paper strip (1.0 cm multiplied by 2.0 cm) of the solution on the separated abdominal aorta section, precisely cutting a 1.0cm long blood vessel section along the edge of the filter paper ring after 50min, sucking residual blood in the blood vessel, precisely weighing the mass, precisely weighing the blood vessel after removing the thrombus, and subtracting the front part from the rear part to obtain the mass of the thrombus in the 1.0cm long blood vessel section. Calculating thrombus mass of each group, analyzing by SPSS17.0 software, expressing data as mean + -standard deviation, comparing between groups by One-wayANOVA test, P<0.05 represents that the difference is statistically significant.

As shown in table 3 and fig. 3, compared with the control group, the thrombus quality of the rats of the positive drug group, the low-dose atractylenolide administration group and the high-dose atractylenolide administration group was significantly reduced, the difference was statistically significant, and the reduction effect of the low-dose and high-dose atractylenolide administration group on thrombus was dose-dependent.

TABLE 3 thrombus quality in groups of rats

Group of	Quality of thrombus (unit: mg)
		Control group	9.5±1.2
Positive drug group	2.3±0.5
		Low-dose administration group of atractylis ovata lactam	5.6±0.7
High-dose administration group of atractylenolide	3.1±0.8

In recent years, the incidence of cardiovascular diseases is continuously increased, and the death caused by the cardiovascular diseases is the first death rate of all diseases. Thrombotic disease is a high proportion of cardiovascular disease, of which thrombotic and embolic complications due to platelet dysfunction are the major causes of cardiovascular disease. The antithrombotic drug is an important means for preventing and treating thrombotic diseases, wherein the traditional Chinese medicine has unique curative effects on antiplatelet and antithrombotic, and has small side effect. The embodiment of the invention shows that the low and high dose of the atractylenolide can obviously prolong CT and BT of mice and inhibit FeCl ₃ Inducing thrombosis of rat, the atractylenolide has obvious anticoagulation and antithrombotic effects, and can be used for preparing antithrombotic drugs.

The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.

Claims

1. Application of atractylenolide in preparing antithrombotic medicine is provided.