Compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponins and preparation method thereof

Abstract

The application discloses a compound nanomicelle co-loading icariin and glycyrrhiza triterpene saponin and a preparation method thereof. The compound nanomicelle comprises icariin, glycyrrhiza triterpene saponin and an amphiphilic block copolymer carrier material, and through a self-assembly process in an aqueous medium, the compound nanomicelle is formed, in which icariin and glycyrrhiza triterpene saponin are simultaneously and stably wrapped by the block copolymer carrier material. The prepared compound nanomicelle has uniform particle size, high encapsulation rate and good stability, can effectively overcome the medicine-making defects of icariin and glycyrrhiza triterpene saponin, and obtain a synergistic pharmacological effect of combination.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of compound nanomicelles based on amphiphilic block copolymer material simultaneously carrying icariin and glycyrrhiza triterpene saponins ingredients, and provide the preparation method of the compound nanomicelles, and its use, belong to biological medicine technical field. BACKGROUND

[0002] Epimedium is also known as Xianlingpi, first recorded in Shennong Bencao Jing, bitter, sweet, warm, liver and kidney, is commonly used to Yang medicine. Modern pharmacological studies show that Epimedium can enhance human sexual function, improve immune function, anti-aging, antioxidant, anti-inflammatory, anti-tumor, hypoglycemic, antidepressant and other pharmacological effects. Epimedium contains a variety of flavonoids, which are important active ingredients of Epimedium, including icariin (ICA), icariin I (ICAI), icariin II (ICAII), epimedium (ICT), epimedin A (epimedin A), epimedin B (epimedin B), epimedin C (epimedin C) and so on. Studies have found that icariin can regulate cellular immune function and inhibit cancer cell metastasis. Icariin, epimedin B and baohuoside I have inhibitory effects on the proliferation of human breast cancer cells (MCF-7) and human hepatoma cells (HepG2). However, due to the low bioavailability, the drug has poor quality. Moreover, how to further improve the effect of icariin on inhibiting tumor proliferation has become one of the current research difficulties. SUMMARY

[0003] To solve the above technical problems, the present application aims to provide a compound nanomicelle co-loaded with icariin and glycyrrhiza triterpene saponins and a preparation method thereof. The compound micelle drug delivery system co-loaded with icariin and glycyrrhiza triterpene saponins provided by the present application comprises icariin, glycyrrhiza triterpene saponins and amphiphilic block copolymer.

[0004] The technical solution of the present application is as follows:

[0005] The compound nanomicelle co-loaded with icariin and glycyrrhiza triterpene saponins comprises at least one icariin, at least one glycyrrhiza triterpene saponin and at least one amphiphilic block copolymer, and the mass ratio of icariin, glycyrrhiza triterpene saponin and amphiphilic block copolymer is 1: (0.2-10): (5-100).

[0006] The icariin includes but is not limited to Icaritin (ICA), Icaritin (ICA I), Icaritin (ICA II), Epimedium (ICT), Epimedin A (epimedin A), Epimedin B (epimedin B), Epimedin C (epimedin C);

[0007] The glycyrrhiza triterpene saponin includes but is not limited to glycyrrhizic acid (GL), glycyrrhizic acid (GA);

[0008] The amphiphilic block copolymer is a two-block copolymer of A-B type, wherein the A block is a hydrophilic segment, including polyethylene glycol, polyethylene glycol monomethyl ether, and the number average molecular weight is 400-20000; the B block is a hydrophobic polyester segment, including a hydrophobic group modified or unmodified poly-lactide, poly-glycolide, poly-lactide-co-glycolide, polycaprolactone polymer, and the number average molecular weight is 500-100000, wherein the hydrophobic group is selected from one of acetyl, tert-butyryl, tert-butylacetyl, benzoyl, amino acid residue or amino acid derivative residue;

[0009] And the preparation method comprises the following steps:

[0010] (1) the icariin, glycyrrhiza triterpene saponin and amphiphilic block copolymer are added into an excess of organic solvent according to the mass ratio, and then the organic solvent is removed slowly to prepare a drug-loaded polymer film with uniform dispersion,

[0011] The organic solvent is selected from any one or several of acetone, methanol, ethanol, chloroform, dichloromethane, tetrahydrofuran and acetonitrile;

[0012] (2) the drug-loaded polymer film obtained in step (1) is added into an aqueous medium, and then the aqueous medium is hydrated at 25-70 DEG C for 1-60 min to obtain a self-assembled micelle,

[0013] The aqueous medium is any one or several of pure water, physiological saline, glucose injection, buffer solution, tissue culture medium or body fluid.

[0014] Furthermore, the compound nanomicelles co-loaded with icariin and glycyrrhiza triterpene saponin can also be prepared by a film dispersion method, a solvent evaporation method or a dialysis method.

[0015] Preferably, the icariin is Icaritin II; the glycyrrhiza triterpene saponin is glycyrrhizic acid; and the amphiphilic block copolymer is tert-butoxycarbonyl phenylalanine modified polyethylene glycol monomethyl ether-poly-lactide (mPEG-PLA-BP).

[0016] Preferably, the mass ratio of the icariin, glycyrrhiza triterpene saponin and the amphiphilic block copolymer is 1: (0.25-1): (7-15).

[0017] The compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin prepared in the application are prepared into lyophilized powder by adding pharmaceutically acceptable excipients using the preparation technology known in the art of pharmacy, for injection, oral administration, mucosal or external use.

[0018] Another purpose of the application is to provide the use of the aforementioned compound nanomicelles in the preparation of antitumor drugs. Beneficial effects

[0019] The compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin prepared in the application have uniform particle size, high encapsulation efficiency and good stability. The compound nanomicelles prepared in the application can effectively overcome the defects of icariin and glycyrrhiza triterpene saponin in terms of poor water solubility and low bioavailability. The antitumor effect of the compound nanomicelles prepared in the application is significantly improved, and the synergistic effect of combination is obtained. The antitumor effect of icariin is significantly improved through the branch joint co-loading of glycyrrhiza triterpene saponin. Compared with the antitumor activity of single icariin, the antiproliferative effect of the compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin on hepatocarcinoma and breast cancer is significantly improved. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 Figure 1 is a transmission electron microscope image of the compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin loaded by mPEG-PLA-BP. (A) ICAII and GA are loaded, (B) ICAII and GL are loaded, (C) ICAI and GA are loaded, and (D) ICAI and GL are loaded.

[0021] Figure 2 Figure 2 is a transmission electron microscope image of the compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin loaded by mPEG-PCL. (E) ICAII and GA are loaded, (F) ICAII and GL are loaded, (G) ICAI and GA are loaded, and (K) ICAI and GL are loaded.

[0022] Figure 3 Figure 3 is a transmission electron microscope image of the compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin loaded by mPEG-PLA. (L) ICAII and GA are loaded, (M) ICAII and GL are loaded, (N) ICAI and GA are loaded, and (O) ICAI and GL are loaded.

[0023] Figure 4 Figure 4 is a comparison of the in vitro proliferation inhibition effect of free drugs and the compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponin loaded by mPEG-PLA-BP on MCF-7 cells.

[0024] Figure 5To compare the in vitro proliferation inhibition effect of free drug and mPEG-PLA-BP loaded complex nanomicelles on HepG2 cells. Embodiments

[0025] In order to facilitate the understanding of the present application, examples and test examples are listed to further illustrate the present application, but not to limit the present application in any way.

[0026] Example 1. mPEG 2000- PLA 2000 Preparation of mPEG-PLA-BP loaded complex nanomicelles and freeze-dried powder

[0027] (1) Complex nanomicelles and freeze-dried powder co-loaded with ICAII and GA

[0028] Take the high molecular excipient mPEG 2000- PLA 2000 -BP 45 mg, ICAII 4 mg, GA 1 mg, in a 500 mL round-bottom flask, add 5 mL of acetone to completely dissolve the material, slowly rotary evaporate (50-100 rpm) under water bath (~ 45℃) to remove organic solvents, and obtain a uniform dry drug-loaded film, then add preheated normal saline (4 mL) to the film to completely hydrate, pass through a 0.45 μm polyether sulfone membrane, and obtain a complex nanomicelle solution; after freeze-drying, a freeze-dried powder is obtained.

[0029] The drug loading and encapsulation efficiency were determined by high performance liquid chromatography, and the micelle particle size and its distribution (polydispersity coefficient, PDI) were determined by laser particle size analyzer. The results showed that the average particle size of the prepared complex nanomicelles was 22.3 nm, and the PDI was 0.29; the drug loading of ICAII and GA was 9.62% and 1.81%, respectively; the encapsulation efficiency of ICAII and GA was 96.3% and 90.5%, respectively. It is shown that the complex nanomicelles have high drug encapsulation efficiency, small and uniform micelle particle size, and the transmission electron microscope image is shown in the attached Figure 1 .

[0030] The freeze-dried powder of the micelles can maintain stability after being reconstituted with normal saline and placed at room temperature for 7 days, and the particle size and PDI have no obvious change, and the contents of ICAII and GA in the dissolved state are greater than 95%.

[0031] (2) Complex nanomicelles co-loaded with ICAII and GL

[0032] Take the high molecular excipient mPEG 2000- PLA 2000- BP 45 mg, ICAII 4 mg, GL 1 mg, placed in a 500 mL round bottom flask, add 5 mL of acetone to completely dissolve the material, remove the organic solvent slowly under water bath (~ 45℃) (50-100 rpm) and obtain a uniform dry drug-loaded film, then add preheated normal saline (4 mL) to the film until it is completely hydrated, pass through a 0.45 μm polyether sulfone membrane to obtain a compound nanomicelle solution; freeze-dried to obtain a freeze-dried powder.

[0033] The drug loading and encapsulation efficiency were determined by high performance liquid chromatography, and the micelle particle size and its distribution (polydispersity coefficient, PDI) were determined by laser particle size analyzer. The results showed that the average particle size of the prepared compound nanomicelles was 29.5 nm, and the PDI was 0.33. The transmission electron microscope image is shown in Figure 1. Figure 1 The drug loading of ICAII and GL was 8.64% and 1.12%, respectively; the encapsulation efficiency of ICAII and GL was 86.3% and 84.6%, respectively.

[0034] The micelle freeze-dried powder can maintain stability after being reconstituted with normal saline and placed at room temperature for 7 days, and the particle size and PDI do not change significantly. The contents of ICAII and GL in the dissolved state are greater than 95%.

[0035] Compound nanomicelles co-loading ICAI and GA

[0036] Take high molecular auxiliary material mPEG 2000- PLA 2000 - BP 45 mg, ICAI 4 mg, GA 1 mg, placed in a 500 mL round bottom flask, add 5 mL of acetone to completely dissolve the material, remove the organic solvent slowly under water bath (~ 45℃) (50-100 rpm) and obtain a uniform dry drug-loaded film, then add preheated normal saline (4 mL) to the film until it is completely hydrated, pass through a 0.45 μm polyether sulfone membrane to obtain a compound nanomicelle solution; freeze-dried to obtain a freeze-dried powder.

[0037] The drug loading and encapsulation efficiency were determined by high performance liquid chromatography, and the micelle particle size and its distribution (polydispersity coefficient, PDI) were determined by laser particle size analyzer. The results showed that the average particle size of the prepared compound nanomicelles was 27.7 nm, and the PDI was 0.31. The transmission electron microscope image is shown in Figure 1. Figure 1 The drug loading of ICAII and GL was 8.64% and 1.12%, respectively; the encapsulation efficiency of ICAII and GL was 86.3% and 84.6%, respectively.

[0038] The micelle freeze-dried powder can maintain stability after being reconstituted with normal saline and placed at room temperature for 7 days, and the particle size and PDI do not change significantly. The contents of ICAII and GL in the dissolved state are greater than 95%.

[0039] Compound nanomicelles co-loading ICAI and GL

[0040] Take polymer adjuvant mPEG 2000- PLA 2000 - BP 45 mg, ICAI 4 mg, GL 1 mg, placed in a 500 mL round bottom flask, add acetone 5 mL to make the material completely dissolved, water bath (~ 45℃) slowly rotary evaporation (50-100 rpm) to remove organic solvents, and obtain a uniform dry drug-loaded film, then add preheated normal saline (4 mL) to the film to completely hydrate, pass through a 0.45 μm polyether sulfone membrane to obtain a compound nanomicelle solution; freeze-dried to obtain a freeze-dried powder.

[0041] The drug loading and encapsulation efficiency were determined by high performance liquid chromatography, and the micelle particle size and its distribution (polydispersity coefficient, PDI) were determined by laser particle size analyzer. The results showed that the average particle size of the prepared compound nanomicelles was 31.7 nm, and the PDI was 0.38. The transmission electron microscopy image is shown in Figure Figure 1 The drug loading of ICAI and GL was 6.84% and 0.92% respectively; the encapsulation efficiency of ICAI and GL was 68.9% and 82.4% respectively.

[0042] The micelle freeze-dried powder can maintain stability after being reconstituted with normal saline and placed at room temperature for 7 days, and the particle size and PDI do not change significantly. The contents of ICAI and GA in the dissolved state are greater than 95%.

[0043] Example 2. mPEG 2000 -PCL 2000 Preparation of compound nanomicelles co-loading ICAII and GA

[0044] (1) Compound nanomicelles co-loading ICAII and GA

[0045] Take mPEG 2000 -PCL 2000 Polymer adjuvant 55 mg, ICAII 4 mg, GA 1 mg, placed in a 500 mL round bottom flask, and other operation steps are the same as in Example 1. The average particle size of the prepared compound nanomicelles is 38.7 nm, and the PDI is 0.39. The transmission electron microscopy image is shown in Figure Figure 2 The drug loading of ICAII and GA was 7.17% and 1.75% respectively; the encapsulation efficiency of ICAII and GA was 71.7% and 87.7% respectively.

[0046] Compound nanomicelles co-loading ICAII and GL

[0047] Take mPEG 2000 -PCL 2000Polymer excipient 55 mg, ICAII 4 mg, GL 1 mg, placed in a 500 mL round bottom flask, other operation steps same as example 1. The prepared compound nano-micelles average particle size is 20.6 nm, PDI is 0.33, its transmission electron micrograph is attached Figure 2 . The drug loading of ICAII and GL is 7.05%, 1.62% respectively; the encapsulation efficiency of ICAII and GL is 70.6%, 83.5% respectively.

[0048] Compound nano-micelles co-loading ICAI and GA

[0049] Take mPEG 2000 -PCL 2000 Polymer excipient 55 mg, ICAI 4 mg, GA 1 mg, placed in a 500 mL round bottom flask, other operation steps same as example 1. The prepared compound nano-micelles average particle size is 32.8 nm, PDI is 0.36, its transmission electron micrograph is attached Figure 2 . The drug loading of ICAI and GA is 6.65%, 1.32% respectively; the encapsulation efficiency of ICAI and GA is 66.8%, 79.3% respectively.

[0050] Compound nano-micelles co-loading ICAI and GL

[0051] Take mPEG 2000 -PCL 2000 Polymer excipient 55 mg, ICAI 4 mg, GL 1 mg, placed in a 500 mL round bottom flask, other operation steps same as example 1. The prepared compound nano-micelles average particle size is 37.8 nm, PDI is 0.34, its transmission electron micrograph is attached Figure 2 . The drug loading of ICAI and GL is 6.05%, 0.96% respectively; the encapsulation efficiency of ICAI and GL is 60.3%, 70.6% respectively.

[0052] Example 3. mPEG 2000 -PLA 2000 Preparation of compound nano-micelles loaded

[0053] (1) Compound nano-micelles co-loading ICAII and GA

[0054] Take mPEG 2000 -PLA 2000 Polymer excipient 45 mg, ICAII 4 mg, GA 1 mg, placed in a 500 mL round bottom flask, other operation steps same as example 1. The prepared compound nano-micelles average particle size is 20.1 nm, PDI is 0.26, transmission electron micrograph is attached Figure 3The drug loading of ICAII and GA were 6.66%, 1.82% respectively, and the encapsulation efficiency of ICAII and GA were 66.6%, 91.1% respectively.

[0055] Compound nanomicelles co-loading ICAII and GL

[0056] Take mPEG 2000 -PLA 2000 The polymer excipient 45 mg, ICAII 4 mg, GL 1 mg were placed in a 500 mL round-bottom flask, and other operation steps were the same as in Example 1. The prepared compound nanomicelles had an average particle size of 24.2 nm, a PDI of 0.25, and a transmission electron micrograph as shown in FIG. 2. The drug loading of ICAII and GL were 6.53%, 1.62% respectively, and the encapsulation efficiency of ICAII and GL were 65.6%, 89.7% respectively. Figure 3

[0057] (3) Compound nanomicelles co-loading ICAI and GA

[0058] Take mPEG 2000 -PLA 2000 The polymer excipient 45 mg, ICAI 4 mg, GA 1 mg were placed in a 500 mL round-bottom flask, and other operation steps were the same as in Example 1. The prepared compound nanomicelles had an average particle size of 25.5 nm, a PDI of 0.24, and a transmission electron micrograph as shown in FIG. 3. The drug loading of ICAI and GA were 6.16%, 1.44% respectively, and the encapsulation efficiency of ICAI and GA were 61.8%, 87.9% respectively. Figure 3

[0059] (4) Compound nanomicelles co-loading ICAI and GL

[0060] Take mPEG 2000 -PLA 2000 The polymer excipient 45 mg, ICAI 4 mg, GL 1 mg were placed in a 500 mL round-bottom flask, and other operation steps were the same as in Example 1. The prepared compound nanomicelles had an average particle size of 26.3 nm, a PDI of 0.29, and a transmission electron micrograph as shown in FIG. 4. The drug loading of ICAI and GL were 6.26%, 1.67% respectively, and the encapsulation efficiency of ICAI and GL were 62.8%, 89.6% respectively. Figure 3

[0061] Example 4. Determination of the inhibitory activity of compound nanomicelles on the in vitro proliferation of tumor cells

[0062] ​​​The in vitro tumor cell toxicity activity of the compound nanomicelles prepared in Example 1 was investigated with free compounds of icariin II (ICA II), glycyrrhizic acid (GA), icariin I (ICAI) and glycyrrhizic acid (GL) as controls.

[0063] (1) Inhibition activity determination on the in vitro proliferation of breast cancer MCF-7 cells

[0064] The prepared human breast cancer MCF-7 cell suspension was inoculated in a 96-well plate at a density of 4×10 3 / well and placed in a 5% CO2, 37 ℃ cell incubator for 24 h, and different concentrations of test drugs (0.5-10 μmol / L) were added, including DMSO solutions of ICA II, ICAI, GA, GL, mPEG-PLA-BP @ ICAII / GA compound micelles, mPEG-PLA-BP @ ICAII / GL compound micelles, mPEG-PLA-BP @ ICAI / GA compound micelles and mPEG-PLA-BP @ ICAI / GL compound micelles.

[0065] After 24 h of continuous culture, 10 μl of MTT test solution (final concentration 500 μg / ml) was added to each well, and incubation was continued for 4 h. The culture solution was discarded, 150 μL of DMSO was added to each well, and shaking was performed until the blue-purple formazan crystals were completely dissolved. The absorbance A570 at 570 nm was measured with 630 nm as the reference wavelength. Three parallel duplicate wells were set for each concentration of test drug, and a blank group was determined simultaneously. The cell growth inhibition rate (%) was calculated according to the absorbance determination results.

[0066] The test results are shown in Table 1: Figure 4 As shown in Table 1, the tumor cell growth inhibition rate of glycyrrhizic acid triterpenoid saponins was less than 5% at the experimental concentration, and the glycyrrhizic acid triterpenoid saponins basically had no anti-tumor activity. The in vitro proliferation inhibition effect of icariin flavonoids on MCF-7 cells showed a concentration-dependent manner within the test concentration range, and the inhibition effect gradually increased with the increase of drug concentration. At a concentration of 10 umol / L, the cell proliferation inhibition rate of free ICA II on MCF-7 was 29%, and that of ICAI was slightly lower, at about 23%. However, the compound nanomicelles co-loaded with icariin flavonoids and glycyrrhizic acid triterpenoid saponins had significant synergistic anti-breast cancer cell proliferation activity at the same concentration, and the anti-cell proliferation activity was increased by more than 2 times compared with that of icariin flavonoids alone. At a concentration of 10 umol / L, the cell proliferation inhibition rate of ICA II / GA compound micelles on MCF-7 was 70%, and that of ICA II / GL compound micelles was 64%. The activity of compound micelles loaded with ICAI was slightly lower than that of ICA II, and the inhibition rate of ICAI / GA compound micelles was 59%, and that of ICAI / GL was 56%.

[0067] (2) Inhibition activity of the proliferation of liver cancer HepG2 cells in vitro

[0068] The prepared human liver cancer HepG2 cell suspension was inoculated in a 96-well plate at a density of 4x10 3 / well, and was placed in a 5% CO2, 37°C cell incubator for 24 h. Different concentrations of test drugs (0.5-10 μmol / L) were added, including DMSO solution of ICAII, DMSO solution of ICAI, DMSO solution of GA, DMSO solution of GL, mPEG-PLA-BP@ICAII / GA compound micelle solution, mPEG-PLA-BP@ICAII / GL compound micelle solution, mPEG-PLA-BP@ICAI / GA compound micelle solution, and mPEG-PLA-BP@ICAI / GL compound micelle solution.

[0069] After 24 h of continuous culture, 10 μl of MTT test solution (final concentration 500 μg / ml) was added to each well, and incubation was continued for 4 h. The culture solution was discarded, 150 μL of DMSO was added to each well, and shaking was performed until the blue-purple formazan crystals were completely dissolved. The absorbance A570 at 570 nm was measured with 630 nm as the reference wavelength. Three parallel duplicate wells were set for each concentration of test drug, and a blank group was also determined. The cell growth inhibition rate (%) was calculated according to the absorbance determination results.

[0070] The test results are shown in Table 1: Figure 5 As shown in Table 1, under the experimental concentrations, licorice triterpenoid saponins had a concentration-dependent growth inhibitory effect on liver cancer cells, but the activity was still low, less than 10%. The in vitro proliferation inhibitory effect of icariin on HepG2 cells also showed a concentration-dependent effect, and the inhibitory effect gradually increased with increasing drug concentration. At a concentration of 10 umol / L, the cell proliferation inhibition rate of free ICAII on HepG2 was 40%, and that of ICAI was slightly lower, at about 33%. However, the compound nanomicelles co-loaded with icariin and licorice triterpenoid saponins had a significantly synergistic anti-proliferation activity on liver cancer cells at the same concentration, and the anti-proliferation activity was also increased by about 2 times compared with that of icariin alone. At a concentration of 10 umol / L, the cell proliferation inhibition rate of ICAII / GA compound micelles on HepG2 was as high as 85%, and that of ICAII / GL compound micelles was 78%. The activity of compound micelles loaded with ICAI was slightly lower than that of ICAII, and the inhibition rate of ICAI / GA compound micelles was 69%, and that of ICAI / GL was 65%.

[0071] The prepared compound nanomicelles have the synergistic pharmacodynamic effect of the combination of icariin and glycyrrhiza triterpene saponins, significantly improve the anti-tumor effect of icariin, and compared with the anti-tumor activity of single icariin, the anti-proliferation effect on liver cancer and breast cancer is significantly improved. The innovation of the present application is not in the drug targeting and characteristics of glycyrrhiza triterpene saponins itself, which has no significant anti-proliferation activity on liver cancer and breast cancer, but in the structural characteristics of the amphiphilic substance of glycyrrhiza triterpene saponins, and in the interaction with cholesterol or phospholipid of cell membrane, the pores are induced in the membrane by combining with cholesterol in the lipid bilayer membrane, the membrane permeability is enhanced, the drug transmembrane absorption is promoted, the in vivo bioavailability is improved, and the purpose of significantly improving the anti-tumor proliferation is achieved. The compound preparation of co-loading glycyrrhiza triterpene saponins and icariin has stronger pharmacodynamic effect and lower toxic and side effects.

[0072] The above only describes the preferred embodiments of the present application and is not intended to limit the present application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application is included in the protection scope of the present application.

Claims

1. A compound nanomicelle co-loading icariin and glycyrrhiza triterpene saponins, characterized in that, The compound nanomicelles comprise at least one icariin, at least one glycyrrhizic acid saponin and at least one amphiphilic block copolymer, and the mass ratio of icariin, glycyrrhizic acid saponin and amphiphilic block copolymer is 1:0.2-10:5-100. The icariin is selected from icariin I and icariin II. The glycyrrhizic acid saponin is selected from glycyrrhizic acid and glycyrrhetic acid. The amphiphilic block copolymer is an A-B type two-block copolymer, wherein the A block is a hydrophilic segment selected from polyethylene glycol, polyethylene glycol monomethyl ether with a number average molecular weight of 400-20,000; the B block is a hydrophobic polyester segment selected from hydrophobic group-modified or unmodified polylactide, polyglycolide, poly(lactide-co-glycolide) and polycaprolactone polymer with a number average molecular weight of 500-100,000, wherein the hydrophobic group is selected from acetyl, tert-butyryl, tert-butylacetyl, benzoyl, amino acid residue or amino acid derivative residue. The icariin, glycyrrhizic acid saponin and amphiphilic block copolymer are added into an excess of organic solvent in the mass ratio, dissolved thoroughly, and then the organic solvent is removed slowly to obtain a drug-loaded polymer film with uniform dispersion. The organic solvent is selected from acetone, methanol, ethanol, chloroform, dichloromethane, tetrahydrofuran and acetonitrile. The drug-loaded polymer film obtained in step (1) is added into an aqueous medium, hydrated at 25-70°C for 1-60 min, and then self-assembled to obtain the compound nanomicelles. The aqueous medium is pure water, physiological saline, glucose injection, buffer solution, tissue culture medium or body fluid. The icariin is icariin II; the glycyrrhizic acid saponin is glycyrrhetic acid; and the amphiphilic block copolymer is tert-butyloxycarbonyl phenylalanine-modified polyethylene glycol monomethyl ether-polylactide.

2. The compound nanomicelles co-loading icariin and glycyrrhiza triterpene saponins according to claim 1, characterized in that The mass ratio of icariin, glycyrrhizic acid saponin and amphiphilic block copolymer is 1:0.25-1:7-15.

3. The compound nanomicelle of co-loading icariin and glycyrrhiza triterpene saponins according to claim 1, characterized in that, ​

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