Astragaloside-salvianolic acid B-paeoniflorin-amygdalin nano-liposome and preparation method thereof

Abstract

The invention discloses an astragaloside-salvianolic acid B-paeoniflorin-amygdalin nano-liposome and a preparation method of the astragaloside-salvianolic acid B-paeoniflorin-amygdalin nano-liposome. The mass ratio of astragaloside to salvianolic acid B to paeoniflorin to amygdalin in the nano-liposome is (1-5): (1-5); (1-10): (1-5). The liposome disclosed by the invention can be used for preparing the medicine for preventing and / or treating cardiovascular diseases, especially for preparing the astragalus-dragon gum, and has a remarkable effect in the aspect of treating cardiovascular diseases.

Description

Technical Field

[0001] The invention relates to the field of medicine, and in particular to astragaloside IV-salvianolic acid B-peonidin-amygdalin nanoliposomes and a preparation method thereof. Background Art

[0002] Qilong Capsule is a medicine, and its main effects include nourishing qi and blood, promoting blood circulation and removing blood stasis, and treating both the symptoms and the root causes. It is widely used in the treatment of cardiovascular diseases. Studies have shown that its active ingredients include salvianolic acid B, paeoniflorin, tanshinone IIA, astragaloside I, amygdalin, and calycosin isoflavone glycosides. Chinese medicine preparations can treat many types of diseases, but they also have low bioavailability and slow effects. In addition, the concentrations of the active ingredients of different batches of Qilong Capsules vary greatly, resulting in large differences in their therapeutic effects. In particular, the main active ingredient, astragaloside I, has a high relative molecular mass and extremely low oral bioavailability, making it difficult to exert its efficacy.

[0003] In 1965, Bangham et al. first proposed the concept of liposomes. Liposomes are ultramicrosphere carrier preparations made by encapsulating drugs in lipid membranes formed by lipid bilayers. They can encapsulate hydrophilic and lipophilic drugs, and the particle size usually ranges from tens of nanometers to thousands of nanometers. They have good biocompatibility, can increase the efficacy of drugs, reduce the therapeutic dose of drugs, and reduce drug toxicity. They can be used as delivery carriers for various therapeutic drugs.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] In view of the problems in the prior art that the utilization rate of traditional Chinese medicine is low and the concentration of active ingredients of different batches of medicines varies greatly, the present invention is proposed. The content of the present invention is as follows:

[0006] In the first aspect of the present invention, a nanoliposome of astragaloside IV-salvianolic acid B-peonidin-amygdalin nanoliposome is provided, wherein the mass ratio of astragaloside IV:salvianolic acid B:peonidin:amygdalin in the nanoliposome is (1-5):(1-5);(1-10):(1-5). Preferably, the mass ratio of astragaloside IV:salvianolic acid B:peonidin:amygdalin in the nanoliposome is 1:3:1:1. In a specific embodiment of the present invention, each 2 mg of nanoliposome comprises astragaloside IV:0.113-0.397 mg, salvianolic acid B:0.164-0.323 mg,peonidin:0.561-0.952 mg,amygdalin:0.014-0.284 mg.

[0007] The second aspect of the present invention provides the use of the nanoliposome in the preparation of a drug for the prevention and / or treatment of cardiovascular diseases. In a specific embodiment of the present invention, the nanoliposome is used to prepare Qilong capsules.

[0008] The third aspect of the present invention provides a method for preparing the nanoliposome, comprising the following steps:

[0009] S1. Mix astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of (1-5):(1-5);(1-10):(1-5) to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture. Preferably, the mass ratio of astragaloside IV:salvianolic acid B:peoniflorin:amygdalin in the nanoliposome is 1:1:3:1.

[0010] S2, dissolving the three substances in anhydrous ethanol at a mass ratio of 10:1-20:1 of lecithin to the mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 2:1-10:1 of lecithin to the lipophilic additive, and mixing them to obtain a liposome suspension;

[0011] S3, after preheating ultrapure water to 40-50°C, the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50°C for hydration for 0.5-1h, and rotary evaporated at -0.01MPa for 5-10min at a temperature of 25°C and a rotation speed of 85-90r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0012] S4, placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0013] S5. Filter the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then store it in a refrigerator at 4° C.

[0014] Furthermore, the lecithin described in step S2 comprises egg yolk lecithin, soybean lecithin, hydrogenated soybean lecithin or hydrogenated egg yolk lecithin.

[0015] Furthermore, the lipophilic additive described in step S2 comprises cholesterol, stearic acid or tocopherol.

[0016] In a specific embodiment of the present invention, each 2 mg of nanoliposomes includes astragaloside IV: 0.113-0.397 mg, salvianolic acid B: 0.164-0.323 mg, paeoniflorin: 0.561-0.952 mg, and amygdalin: 0.014-0.284 mg.

[0017] The beneficial effects of the present invention include but are not limited to:

[0018] 1. The nanoliposomes of the present invention improve the solubility of various components, especially astragaloside IV, solve the problem of low bioavailability, and enhance the pharmacological effects of astragaloside IV, salvianolic acid B, paeoniflorin, and amygdalin;

[0019] 2. Liposome preparations can delay or control the release of drugs, reduce the elimination rate of astragaloside IV, salvianolic acid B, paeoniflorin, and amygdalin, prolong the duration of action of astragaloside IV, and increase the in vivo and in vitro stability of astragaloside IV;

[0020] 3. The liposome preparation process of the present invention is simple, and after being made into liposomes, they can be further processed into various dosage forms such as aerosols and sprays. DETAILED DESCRIPTION

[0021] The present invention is described in detail below with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, the raw materials and catalysts in the examples of the present invention are purchased through commercial channels.

[0022] Example 1

[0023] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 1:1:1:1 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0024] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 10:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 2:1 between egg yolk lecithin and cholesterol, and mixing them to obtain a liposome suspension;

[0025] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at -0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 85-90 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0026] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0027] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0028] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0029] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0030] Through calculation, the encapsulation efficiency of this embodiment is 48.68%.

[0031] Example 2

[0032] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 1:1:3:1 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0033] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 10:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 2:1 between egg yolk lecithin and a lipophilic additive, and mixing them to obtain a liposome suspension;

[0034] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at -0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 85-90 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0035] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0036] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0037] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0038] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0039] Through calculation, the encapsulation efficiency of this embodiment is 49.65%.

[0040] Example 3

[0041] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 2:1:5:2 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0042] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 10:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 2:1 between egg yolk lecithin and a lipophilic additive, and mixing them to obtain a liposome suspension;

[0043] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at -0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 8590 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0044] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0045] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0046] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0047] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0048] Through calculation, the encapsulation efficiency of this embodiment is 48.98%.

[0049] Example 4

[0050] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 1:1:1:1 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0051] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 20:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 5:1 between egg yolk lecithin and cholesterol, and mixing them to obtain a liposome suspension;

[0052] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at -0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 85-90 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0053] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0054] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0055] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0056] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0057] Through calculation, the encapsulation efficiency of this embodiment is 67.62%.

[0058] Example 5

[0059] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 1:1:3:1 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0060] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 20:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 5:1 between egg yolk lecithin and a lipophilic additive, and mixing them to obtain a liposome suspension;

[0061] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at -0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 85-90 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0062] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0063] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0064] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0065] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0066] Through calculation, the encapsulation efficiency of this embodiment is 68.26%.

[0067] Example 6

[0068] (1) mixing astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of 2:1:5:2 to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture;

[0069] (2) dissolving the three substances in anhydrous ethanol at a mass ratio of 20:1 between egg yolk lecithin and a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 5:1 between egg yolk lecithin and a lipophilic additive, and mixing them to obtain a liposome suspension;

[0070] (3) After preheating ultrapure water to 40-50° C., the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50° C. for hydration for 0.5-1 h, and rotary evaporated at 0.01 MPa for 5-10 min at a temperature of 25° C. and a rotation speed of 85-90 r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin;

[0071] (4) placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution;

[0072] (5) filtering the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain the filtrate, and then storing it in a refrigerator at 4° C.;

[0073] (6) Determination of encapsulation efficiency: 2 mL of the prepared nanoliposomes were placed in a Millipore ultrafiltration centrifuge tube (molecular weight cutoff 10,000), centrifuged at 3000 r / min for 30 min, the filtrate was collected, diluted to an appropriate concentration with the mobile phase, filtered through a 0.22 μm filter membrane, and the content of free drug after separation and elution was determined by high performance liquid chromatography to calculate the encapsulation efficiency;

[0074] Encapsulation efficiency determination formula: Encapsulation efficiency = [encapsulated drug concentration / (free drug concentration + encapsulated drug concentration)] × 100%.

[0075] Through calculation, the encapsulation efficiency of this embodiment is 71.28%.

[0076] Example 7

[0077] The liposomes prepared in Examples 1-6 were used to replace the astragalus, angelica, salvia miltiorrhiza, red peony root, ligusticum chuanxiong, safflower and peach kernel in Qilong Capsules and mixed with the earthworm extract of Eisenia fetida to prepare Qilong Capsules (for specific steps, see CN1541683A), and the prepared Qilong Capsules were verified for their anti-experimental thrombosis and thrombolytic effects in rats. Qilong Capsules were used as the control group (Qilong Capsules were produced by Jining Huaneng Pharmaceutical Factory Co., Ltd., batch number: 240943).

[0078] 7-1 Rat arteriovenous bypass thrombosis experiment

[0079] Rats (Wistar rats, weighing 230-260g) were randomly divided into 8 groups, namely, a blank control group, a control group Qilong capsule 0.3g / kg group, and a group Qilong capsule 0.3g / kg prepared by Example 1-6 (numbered 1-6 groups). The daily dose was given twice for a total of 3 days. Surgery was performed 1 hour after the last administration. The rats were anesthetized with 25% urethane 1.5g / kg ip, and arteriovenous bypass was established. After 30 minutes, the cannula was removed, the thrombus was removed, the floating blood was removed from the filter paper, and the wet weight of the thrombus was weighed; then it was placed in a 60℃ oven for drying for 1 hour to constant weight, and the dry weight of the thrombus was weighed after cooling. The experimental results are shown in Table 1.

[0080] Table 1

[0081] serial number Group dose Thrombus wet weight (mg) Thrombus dry weight (mg) 1 Example 1 Liposome-prepared Qilong capsules 0.3g / kg 72.14±11.26 16.73±3.36 2 Example 2 Qilong capsule prepared by liposome 0.3g / kg 64.28±10.42 12.64±4.51 3 Example 3 Liposome-prepared Qilong capsules 0.3g / kg 71.32±12.85 17.84±3.63 4 Example 4 Liposome-prepared Qilong capsules 0.3g / kg 71.78±13.06 17.61±4.75 5 Example 5: Qilong capsules prepared by liposomes 0.3g / kg 65.51±11.25 13.58±4.48 6 Example 6 Liposome-prepared Qilong capsules 0.3g / kg 72.48±12.69 16.23±2.27 7 Qilong Capsules (Huaneng Pharmaceutical) 0.3g / kg 74.62±13.35 18.28±4.85 8 Blank control group — 120.64±48.73 33.21±12.94

[0082] As can be seen from Table 1, the Qilong capsules prepared with liposomes of Examples 1-6 are more effective in reducing the wet weight and dry weight of thrombus than traditional Qilong capsules, and when the mass ratio of astragaloside IV: salvianolic acid B: paeoniflorin: amygdalin in the nanoliposomes is 1:1:3:1 (Groups 2 and 5), the effect is better than other ratios.

[0083] 7-2 Experiment on the dissolution of freshly formed blood clots in vitro by serum containing Qilong capsule

[0084] The grouping and administration were the same as in 7-1. Male rats were taken and bled through the common carotid artery into a plastic tube with a diameter of 3 mm. Both ends were clamped and placed vertically at room temperature for 24 hours before the blood clots were removed. The rats were anesthetized by ip 10% chloral hydrate solution (350 mg / kg). The common carotid artery thrombosis model of rats was established according to the literature method (Chen Qi, ed. Methodology of Chinese Medicine Pharmacology Research [M]. People's Medical Publishing House, September 1993: 572.), the current was set to 4 mA, the stimulation time was 7 minutes, and then the skin was sutured and gentamicin sulfate was injected to prevent infection. The rats were immediately administered, and the control group was given an equal volume of distilled water. The rats were returned to the cage for breeding. The administration was continued for 3 days. On the morning of the third day, 1 hour after the last administration, the rats were anesthetized by ip 10% chloral hydrate solution (350 mg / kg), fixed in the supine position, bled through the common carotid artery, and anticoagulated with 3.8% sodium citrate (V / V 1:9). Centrifuge at 3000r / ·min for 6min, take out 1.5mL of upper plasma and add it to a vial, then inactivate it in a 56℃ water bath for 30min to obtain the drug-containing serum. Weigh 50mg of freshly prepared blood clots, add serum, incubate at 37℃ for 24h, take out the blood clots, blot dry and weigh accurately, and calculate the thrombolysis rate (relative thrombolysis rate (%) = (wet weight of blood clots before lysis - wet weight of blood clots after lysis) / wet weight of blood clots before lysis × 100%). The results are shown in Table 2.

[0085] Table 2

[0086] serial number Group dose Blood clot weight (mg) Dissolution rate (%) 1 Example 1 Liposome-prepared Qilong capsules 0.3g / kg 26.41±3.62 38.37±3.63 2 Example 2 Qilong capsule prepared by liposome 0.3g / kg 24.82±3.24 42.46±4.15 3 Example 3 Liposome-prepared Qilong capsules 0.3g / kg 27.23±3.58 37.42±3.38 4 Example 4 Liposome-prepared Qilong capsules 0.3g / kg 27.85±3.63 38.16±4.57 5 Example 5: Qilong capsules prepared by liposomes 0.3g / kg 25.18±3.52 43.83±4.84 6 Example 6 Liposome-prepared Qilong capsules 0.3g / kg 28.84±3.86 39.32±2.74 7 Qilong Capsules (Huaneng Pharmaceutical) 0.3g / kg 31.37±3.31 36.09±3.58 8 Blank control group — 36.38±2.73 27.53±4.64

[0087] As can be seen from Table 2, the Qilong capsules prepared with liposomes of Examples 1-6 have a better effect on dissolving fresh blood clots in vitro than the traditional Qilong capsules after oral administration of the drug-containing serum, and when the mass ratio of astragaloside IV: salvianolic acid B: paeoniflorin: amygdalin in the nanoliposomes is 1:1:3:1 (Groups 2 and 5), the effect is better than other ratios.

[0088] The above description is only an embodiment of the present invention. The protection scope of the present invention is not limited by these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent substitution, improvement, etc. made within the technical ideas and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A nanoliposome of astragaloside IV-salvianolic acid B-peonidin-amygdalin nanoliposome, characterized in that: The mass ratio of astragaloside IV: salvianolic acid B: paeoniflorin: amygdalin in the nanoliposome is (1-5): (1～5)；(1～10)：(1～5)。 2. The nanoliposome according to claim 1, characterized in that The mass ratio of astragaloside IV:salvianolic acid B:peonidin:amygdalin in the nanoliposome is 1:1:3:

1.

3. The nanoliposome according to claim 1, characterized in that Each 2 mg of nanoliposomes includes astragaloside IV: 0.113-0.397 mg, salvianolic acid B: 0.164-0.323 mg, paeoniflorin: 0.561-0.952 mg, and amygdalin: 0.014-0.284 mg.

4. Use of the nanoliposomes according to any one of claims 1 to 3 in the preparation of drugs for preventing and / or treating cardiovascular diseases.

5. Use of the nanoliposomes as described in any one of claims 1 to 3 in the preparation of Qilong Capsules.

6. A method for preparing the nanoliposome according to any one of claims 1 to 3, characterized in that: The method comprises the following steps: S1. Mix astragaloside IV, salvianolic acid B, paeoniflorin and amygdalin in a mass ratio of (1-5):(1-5);(1-10):(1-5) to obtain an astragaloside IV-salvianolic acid B-peoniflorin-amygdalin mixture; S2, dissolving the three substances in anhydrous ethanol at a mass ratio of 10:1-20:1 of lecithin to the mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin and a mass ratio of 2:1-10:1 of lecithin to the lipophilic additive, and mixing them to obtain a liposome suspension; S3, after preheating ultrapure water to 40-50°C, the liposome suspension obtained in step S2 is introduced into ultrapure water through a peristaltic pump, and then placed at 50°C for hydration for 0.5-1h, and rotary evaporated at -0.01MPa for 5-10min at a temperature of 25°C and a rotation speed of 85-90r / min to remove anhydrous ethanol to obtain a liposome solution of a mixture of astragaloside IV-salvianolic acid B-peonidin-amygdalin; S4, placing the astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture liposome solution obtained in step S3 in an ice water bath and ultrasonicating with a 100W probe for 5-10 minutes, wherein the probe works for 10 seconds and the interval is 5 seconds, to obtain an astragaloside IV-salvianolic acid B-peonidin-amygdalin mixture nanoliposome solution; S5. Filter the astragaloside nanoliposome solution obtained in step S4 through a microporous membrane filter with a pore size of 0.22 μm to obtain a filtrate.

7. The method for preparing nanoliposomes according to claim 6, characterized in that: The lecithin described in step S2 includes egg yolk lecithin, soybean lecithin, hydrogenated soybean lecithin or hydrogenated egg yolk lecithin.

8. The method for preparing nanoliposomes according to claim 6, characterized in that: The lipophilic additive described in step S2 comprises cholesterol, stearic acid or tocopherol.

9. The method for preparing nanoliposomes according to claim 6, characterized in that: Each 2 mg of nanoliposomes includes astragaloside IV: 0.113-0.397 mg, salvianolic acid B: 0.164-0.323 mg, paeoniflorin: 0.561-0.952 mg, and amygdalin: 0.014-0.284 mg.

10. The method for preparing nanoliposomes according to claim 6, characterized in that: The mass ratio of astragaloside IV:salvianolic acid B:peonidin:amygdalin in the nanoliposomes described in step S1 is 1:1:3:1.