Composition for relieving cerebral ischemia-reperfusion injury as well as preparation method and application thereof

Abstract

The invention provides a composition for relieving cerebral ischemia reperfusion injury as well as a preparation method and application thereof, and relates to the technical field of biological pharmacy. The composition comprises icariside I, squalene, an oil phase, an emulsifier and a co-emulsifier, the emulsifier is selected from one or a mixture of more of a nonionic surfactant, an anionic surfactant and an ampholytic surfactant; the particle size of the composition ranges from 50 nm to 500 nm. The icariside I and the squalene are mixed according to a proper proportion, and the oil phase, the emulsifying agent and the co-emulsifying agent are introduced, so that the solubility and the bioavailability of the icariside I are remarkably improved, and the drug effect is remarkably improved; meanwhile, the preparation method of the composition provided by the invention is simple in process and mild in condition, provides a universal method for preparing the hydrophobic drug microemulsion, is beneficial to large-scale production, and has wide application and market prospects.

Description

Technical Field

[0001] The invention relates to the technical field of biopharmaceuticals, and in particular to a composition for alleviating cerebral ischemia-reperfusion injury, and a preparation method and application thereof. Background Art

[0002] At present, stroke is the most common sudden cardiovascular and cerebrovascular disease, with high incidence, high recurrence rate, high disability rate, high mortality rate and other harmful characteristics. The prevalence of stroke in my country is increasing, and the incidence trend is younger. About 87% of strokes are ischemic strokes, and the main strategy for dealing with ischemic strokes is thrombolysis. However, since blood flow reperfusion after thrombolysis is prone to cause cerebral ischemia-reperfusion injury, namely CI RI, its application is limited.

[0003] Epimedium belongs to the genus Epimedium of the Berberidaceae family. Icariin II is one of the main flavonoid active ingredients of Epimedium. The latest studies have shown that Icariin II can significantly improve brain tissue infarction and brain function scores caused by cerebral ischemia-reperfusion injury through various pathways, and has a good neuroprotective effect. However, the solubility of Icariin II in water is 15.04 mg / L at 25°C. The poor solubility of Icariin II in water leads to its low bioavailability of only 4.4%, which greatly limits its efficacy. Squalene is a bioactive substance with an all-trans isoprene structure. It has the effects of promoting blood circulation, activating the body's functional cells, anti-tumor, and increasing immunity. It has good absorbability and stability. Studies have shown that squalene can reduce the occurrence of cell ferroptosis and has a protective effect on brain tissue after stroke.

[0004] In view of the above defects, developing a new composition to improve the poor solubility and low bioavailability of icariin II is of great significance for improving cerebral ischemia-reperfusion injury. Summary of the invention

[0005] The purpose of the present invention is to provide a composition for alleviating cerebral ischemia-reperfusion injury, and a preparation method and application thereof, so as to solve the technical problems of poor solubility and low bioavailability of icariin II existing in the prior art.

[0006] In order to solve the above technical problems, the present invention provides a composition for reducing cerebral ischemia-reperfusion injury, the composition comprising icariin II, squalene, an oil phase, an emulsifier, and an emulsifier assistant;

[0007] The icariin II is preferably icariin II extracted from natural plants; the oil phase is the oil phase in an oil-in-water microemulsion, preferably animal oil, vegetable oil, natural oil, synthetic oil, semi-synthetic derivatives thereof and a combination thereof; the viscosity and interfacial tension of different oil phases have a certain influence on the particle size of the microemulsion composition.

[0008] The emulsifier is selected from one or more mixtures of nonionic surfactants, anionic surfactants, and amphoteric surfactants;

[0009] The main function of emulsifiers is to improve surface tension. With appropriate emulsifiers, oily substances and aqueous substances can be mixed to form a stable composition;

[0010] The particle size of the composition is between 50nm and 500nm;

[0011] Further, the particle size of the composition is 80nm-200nm;

[0012] Preferably, the particle size of the composition is 100 nm to 160 nm.

[0013] Furthermore, the components in the composition are calculated by mass percentage as follows:

[0014] Icariside II: 0.09%-4.5%, preferably 0.4%-4%, more preferably 1%-4%;

[0015] Squalene: 1%-10%, preferably 2.5%-9.5%;

[0016] Oil phase: 0.9%-10%, preferably 1.5%-9.5%, more preferably 2.2%-9%;

[0017] Emulsifier: 0.45%-4.5%, preferably 1.3%;

[0018] Emulsifier: 0.09%-1%, preferably 0.5%;

[0019] The balance was the aqueous phase.

[0020] Furthermore, the surface negative charge of the composition is -50mV to -5mV, preferably -40mV to -30mV.

[0021] Further, the oil phase is a mixture of one or more of polyoxyethylene hydrogenated castor oil, palm oil, olive oil, soybean oil, triolein, ethyl oleate, and isopropyl myristate;

[0022] From the perspective of the particle size of the final microemulsion composition, the oil phase is preferably polyoxyethylene hydrogenated castor oil.

[0023] Further, the emulsifier is a mixture of one or more of C6-C30 fatty acids, C6-C30 aromatic acids, polyethylene glycol-distearoylethanolamine, Tween 80, Tween 40, Span 85, phospholipids, soybean lecithin, egg yolk lecithin, and polyoxyethylene ether;

[0024] Preferably, the emulsifier is soy lecithin.

[0025] The auxiliary emulsifier is a mixture of one or more of polyethylene glycol-15 hydroxystearate, polyethylene glycol, sodium alginate and glycerol; preferably polyethylene glycol-15 hydroxystearate.

[0026] Furthermore, the aqueous phase is sterile and pyrogen-free water, preferably ultrapure water or water for injection.

[0027] The present invention also provides a method for preparing the above composition, the specific steps are as follows:

[0028] Step 1, taking 0.09-4.5 parts of icariin II, mixing it with an organic solvent, and fully dissolving it to obtain an icariin II solution;

[0029] Step 2, mixing 1-10 parts of squalene with 0.45-4.5 parts of an emulsifier and 0.9-10 parts of an oil phase, and after fully dissolving, adding the icariin II solution, and stirring to obtain a first mixture;

[0030] Step 3, adding 0.09-1 part of the auxiliary emulsifier into water to obtain an aqueous solution of the auxiliary emulsifier; adding the aqueous solution of the auxiliary emulsifier dropwise into the first mixture under continuous stirring at 2000 r / min, and stirring for 10 minutes to obtain colostrum;

[0031] Step 4, subjecting the colostrum to ultrasonic treatment for 5-15 minutes, and then removing the organic solvent by rotary evaporation to obtain an icariin II-squalene composition.

[0032] The above components are all calculated by mass.

[0033] Furthermore, in step 1, the organic solvent is selected from dichloromethane and anhydrous ethanol.

[0034] Preferably, the mass fraction of icariin II used in step 1 is 0.4-4 parts;

[0035] In step 2, the mass fraction of the oil phase used is 1.5-9.5 parts.

[0036] On the other hand, the present invention also provides a method for alleviating cerebral ischemia-reperfusion injury, using the above-mentioned composition and the preparation method of the present invention to obtain an icariin II-squalene composition.

[0037] Furthermore, the method uses the icariin II-squalene composition in a manner selected from oral administration and intravenous injection.

[0038] By adopting the above technical solution, the present invention has the following beneficial effects:

[0039] The present invention provides a medicine for alleviating cerebral ischemia-reperfusion injury, a preparation method thereof and an application thereof. The effective ingredients include icariin II and squalene. By appropriately adding an oil phase, an emulsifier, an emulsifier aid and an aqueous phase substance, the solubility and bioavailability of icariin II are significantly improved, and the drug efficacy is significantly improved. At the same time, the preparation method of the composition provided by the present invention has a simple process and mild conditions, and provides a universal method for preparing a hydrophobic drug microemulsion, which is conducive to large-scale production and has broad application and market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0041] Figure 1 This is a transmission electron microscope scanning image of the icariin II-squalene composition provided by the present invention;

[0042] Figure 2 A schematic diagram of the icariin II-squalene composition provided by the present invention;

[0043] Figure 3 This is a particle size distribution diagram of the icariin II-squalene composition prepared in Example 2;

[0044] Figure 4 This is a surface charge distribution diagram of the icariin II-squalene composition prepared in Example 2;

[0045] Figure 5 This is a bar graph comparing the neurological function scores of the five groups of SD rats in Example 3 after intragastric administration of the icariin II-squalene composition;

[0046] Figure 6 This is a schematic diagram comparing the brain tissue damage volumes of rats in each group in Example 3;

[0047] Figure 7 This is a comparison chart of the volume percentage of cerebral infarction in the brain tissue of each group of rats in Example 3;

[0048] Figure 8 A comparison chart showing the effects of the icariin II-squalene composition provided by the present invention on the level of inflammatory factor IL-1β;

[0049] Fig. 9 A comparison chart showing the effects of the icariin II-squalene composition provided by the present invention on the level of inflammatory factor TNF-α;

[0050] Fig.10 This is a comparison chart of the effects of the icariin II-squalene composition provided by the present invention on the level of inflammatory factor IL-6. DETAILED DESCRIPTION

[0051] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0052] The present invention is further explained below in conjunction with specific implementation modes.

[0053] The method for preparing the icariin II-squalene composition provided by the present invention comprises the following specific steps:

[0054] Step 1, taking 0.09-4.5 parts of icariin II, mixing it with an organic solvent, and fully dissolving it to obtain an icariin II solution;

[0055] Step 2, mixing 1-10 parts of squalene with 0.45-4.5 parts of an emulsifier and 0.9-10 parts of an oil phase, and after fully dissolving, adding the icariin II solution, and stirring to obtain a first mixture;

[0056] Step 3, adding 0.09-1 part of the auxiliary emulsifier into water to obtain an aqueous solution of the auxiliary emulsifier; adding the aqueous solution of the auxiliary emulsifier dropwise into the first mixture under continuous stirring at 2000 r / min, and stirring for 10 minutes to obtain colostrum;

[0057] Step 4, subjecting the colostrum to ultrasonic treatment for 5 minutes, and then removing the organic solvent by rotary evaporation to obtain an icariin II-squalene composition.

[0058] The above components are all calculated by mass.

[0059] Example 1

[0060] like Figure 1 As shown, in this example, an icariin II-squalene composition was prepared according to the preparation method provided by the present invention.

[0061] Step 1, take 10.15 mg of icariin II, put it in a centrifuge tube, add 2 mL of dichloromethane and 2 mL of anhydrous ethanol, and after sufficient dissolution, obtain an icariin II solution for standby use;

[0062] Step 2, mixing 107.2 mg of squalene with 25 mg of Tween-40 and 60.4 mg of triolein, and adding the icariin II solution after the mixture is fully dissolved, and stirring to obtain a first mixture;

[0063] Step 3, taking 2 mL of 5% polyethylene glycol aqueous solution, adding it dropwise to the first mixture under continuous stirring at 2000 r / min, and stirring for 10 minutes to obtain colostrum;

[0064] Step 4, subjecting the colostrum to ultrasonic treatment for 5 minutes using an ultrasonic cell disruptor at an ultrasonic power of 260 W, and then removing dichloromethane and anhydrous ethanol by rotary evaporation to obtain an icariin II-squalene composition.

[0065] Take 20 μL of the icariin II-squalene composition microemulsion prepared in this example and drop it on the sample plate; put the front side of the copper mesh on the surface of the droplet, remove it after 3 minutes and dry it at room temperature; put it on a 2% phosphotungstic acid dye solution for dyeing for 1-2 minutes, remove the copper mesh, absorb the excess dye with filter paper, and dry the sample at room temperature; use a JEM-1400FLASH transmission electron microscope under 200 kV conditions to observe the copper mesh by transmission electron microscopy, and observe the following Figure 1 The microstructure of the icariin II-squalene composition is shown.

[0066] Example 2

[0067] like Figure 2-Figure 4 As shown, this embodiment adopts the optimal conditions provided by the present invention to prepare the icariin II-squalene composition.

[0068] Step 1, take 10.15 mg of icariin II, put it in a centrifuge tube, add 2 mL of dichloromethane and 2 mL of anhydrous ethanol, and after sufficient dissolution, obtain an icariin II solution for standby use;

[0069] Step 2, taking 107.2 mg of squalene, 230.8 mg of soybean lecithin, and 56.4 mg of polyoxyethylene hydrogenated castor oil / RH-40, after fully dissolving, then adding the icariin II solution, stirring to obtain a first mixture;

[0070] Step 3, taking 2 mL of an aqueous solution containing 0.5% polyethylene glycol-15-hydroxystearate, adding the solution dropwise to the first mixture under continuous stirring at 2000 r / min, and stirring for 10 minutes after the addition is complete to obtain colostrum;

[0071] Step 4, the colostrum is subjected to ultrasonic treatment for 5 minutes using an ultrasonic cell crusher at an ultrasonic power of 260 W, and then dichloromethane and anhydrous ethanol are removed by rotary evaporation to obtain Figure 2 The white microemulsion shown is the icariin II-squalene composition.

[0072] The particle size and surface charge of the composition were measured using a Zetasizer Nano ZS90 laser particle size analyzer:

[0073] Take 10 μL of the icariin II-squalene composition prepared in this example, dilute it to 3 mL with ultrapure water, and put the above samples into the sample pool for detection, and the measurement temperature is set to 25°C. Figure 3 As shown, the particle size of the composition sample prepared in this example was measured to be 147 nm; Figure 4 As shown, the potential of the composition sample prepared in this example was measured to be -33 mV, and its particle polydispersity coefficient was less than 0.3, indicating that the particles were uniformly distributed and in a stable state.

[0074] The encapsulation efficiency of icariin II in the icariin II-squalene composition prepared in this example was measured:

[0075] The icariin II-squalene composition prepared in this embodiment was taken, and the unencapsulated icariin II was separated by a glucose gel G-50 column; the filtrate was collected, 100 μL of the filtrate was taken, 0.1 mL of a surfactant model Triton-X, 0.3 mL of tetrahydrofuran, and 0.3 mL of anhydrous ethanol were added to break the emulsion, and the supernatant was obtained by centrifugation after vortexing. The content of icariin II was obtained by high performance liquid chromatography, and the encapsulation efficiency was calculated to be 84.85±5.47%, and the drug loading could reach 4.60±0.65%.

[0076] Example 3

[0077] like Figure 5-Figure 7 As shown, this example verifies the beneficial effect of the icariin II-squalene composition provided by the present invention on alleviating cerebral ischemia-reperfusion injury.

[0078] Step A1, 30 SPF SD male rats fed for 7 days, with a body weight of 200-250 g, were divided into groups using a simple random method: control group, model group, icariin II group, squalene group, and icariin II-squalene combination group;

[0079] Step A2, establish a rat focal CIRI model by using the suture embolization method to establish a rat cerebral ischemia model. During the operation, the cerebral blood flow of SD rats was detected by a laser Doppler blood flow monitor to confirm that the CIRI model was successfully established;

[0080] Step A3, each experimental group was gavaged and administered 4h, 16h, 28h, and 40h after modeling, and the animals could eat freely during the administration period. The administration conditions of each experimental group were as follows:

[0081] Control group: Administered normal saline at a dose of 5 mg / kg;

[0082] Model group: administrated normal saline at a dose of 5 mg / kg;

[0083] Icariin II group: administered with icariin II at a dose of 5 mg / kg;

[0084] Squalene group: 100 mg / kg of squalene was administered;

[0085] Icariside II-squalene group: the icariside II-squalene composition provided by the present invention was administered at a dose of 5 mg / kg;

[0086] Step A4, after stopping the administration, continue to feed normally for 3 days, and use the Longa 5-point scoring standard to score the neurological function damage, the higher the score, the greater the damage. The Longa 5-point scoring standard is shown in Table 1 below:

[0087] Table 1 Scoring criteria for neurological deficits

[0088] score Neurobehavioral changes 0 points No neurological impairment, walking in a nearly straight line 1 point Mild neurological impairment, incomplete extension of the contralateral forepaw, and walking in a tilted manner 2 points Moderate neurological impairment, walking with a leaning posture toward the opposite side (turning in a large circle) 3 points Severe neurological impairment, walking almost centered on the hind limbs (turning in small circles) 4 points Unable to walk spontaneously, or even lose consciousness

[0089] The test results are as follows Figure 5 As shown, the neurological function scores of CIRI model rats treated with the icariin II-squalene combination by oral administration were significantly lower than those of the model group.

[0090] Step A5, taking the SD rats that have completed the neurological function scoring experiment in step A4, collecting blood from the orbital vein, and anesthetizing the rats with 2% sodium pentobarbital anesthetic solution by intraperitoneal injection. After the rats are anesthetized, the rats are dislocated, and the brain tissue of the rats is quickly peeled out, washed with PBS solution, and quickly placed in a -20°C refrigerator for 20 minutes;

[0091] Step A6, remove the brain tissue and cut it into 5 equal sections, put it into a light-proof bottle of 2,3,5-triphenyltetrazolium chloride dye solution, place it horizontally in a 37°C oven for staining, turn it over at a frequency of 10 minutes per time, and stain for 1 hour;

[0092] Step A7: After ensuring that the staining is sufficient, the brain tissue sections of each group are placed in a 4% paraformaldehyde solution, fixed in a 4°C refrigerator for 48 hours, and then photographed. The results are as follows: Figure 6 As shown, the cerebral infarction volume in the brain tissue of rats in the icariin I I-squalene group was significantly smaller than that in the model group;

[0093] Step A8, the brain tissues of each group after photographing are cut and weighed according to the normal non-infarct area and the infarct area, and the cerebral infarction volume is calculated according to the following calculation formula:

[0094] Percentage of cerebral infarction volume = (cerebral infarction volume / whole brain tissue) × 100%.

[0095] The calculation results of the percentage of brain tissue infarction volume of rats in each experimental group are as follows Figure 7 As shown, the percentage of cerebral infarction volume in rats treated with the icariin II-squalene combination by gavage was significantly lower than that in other model groups.

[0096] This example verified through multiple groups of experiments that the icariin II-squalene composition provided by the present invention has a significant improvement effect on cerebral ischemia-reperfusion injury.

[0097] Example 4

[0098] like Figure 8-Figure 10 As shown, this example verifies that the icariin II-squalene composition provided by the present invention has a significant inhibitory effect on inflammatory factors.

[0099] Step B1, 30 SPF SD male rats fed for 7 days, with a body weight of 200-250 g, were divided into groups using a simple random method: a control group, a model group, an icariin II group, a squalene group, and an icariin II-squalene combination group;

[0100] Step B2, establish a rat focal CIRI model by using the suture embolization method to establish a rat cerebral ischemia model. During the operation, the cerebral blood flow of SD rats was detected by a laser Doppler blood flow monitor to confirm that the CIRI model was successfully established;

[0101] Step B3, each experimental group was gavaged and administered 4h, 16h, 28h, and 40h after modeling, and the animals could eat freely during the administration period. The administration conditions of each experimental group were as follows:

[0102] Control group: Administered normal saline at a dose of 5 mg / kg;

[0103] Model group: administrated normal saline at a dose of 5 mg / kg;

[0104] Icariin II group: administered with icariin II at a dose of 5 mg / kg;

[0105] Squalene group: 100 mg / kg of squalene was administered;

[0106] Icariside II-squalene group: the icariside II-squalene composition provided by the present invention was administered at a dose of 5 mg / kg;

[0107] Step B4, after stopping the administration, the rats were kept normally for 3 days, the blood of the rats in each group was collected, and then centrifuged at a speed of 3000 r / min and a temperature of 4°C for 20 min, and the upper serum was collected;

[0108] Step B5, the serum taken from each experimental group is used to determine the inflammatory factors by enzyme-linked immunosorbent assay. The specific determination steps are as follows:

[0109] First, take the protein-coated ELISA plate, set up standard wells, sample wells and blank wells, among which the blank control wells do not add samples and enzyme-labeled reagents, and add 50μL of standards of different concentrations to each standard well; first add 40μL of sample diluent to the sample to be tested on the coated ELISA plate, and then add 10μL of the sample to be tested, add the sample to the bottom of the ELISA plate well, try not to touch the well wall, and gently shake to mix. Add 100μL of enzyme-labeled reagent to each well, except for the blank well. Seal the plate with a sealing film and incubate at 37℃ for 60min. Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with washing solution, let it stand for 30s and then discard it, repeat this 5 times, and pat dry. Then add 50μL of color developer A to each well, then add 50μL of color developer B, gently shake and mix, and color at 37℃ in the dark for 15min. Add 50 μL of stop solution to each well to terminate the reaction, adjust the blank well to zero, and measure the absorbance of each well in sequence at a wavelength of 450 nm to obtain the OD value.

[0110] The OD value measured by the above method is as follows: Figure 8-Figure 10 The effects of different groups on the level of inflammatory factor IL-1β in rats are shown in Figure 8 As shown, the normal group was compared with the model group: ****p<0.0001; the icariin II I-squalene group was compared with the model group: ****p<0.0001; the icariin II I-squalene group was compared with the squalene group: ****p<0.0001; the icariin II I-squalene group was compared with the icariin II group: ****p<0.0001. Figure 8 It can be seen that after oral administration of the icariin II-squalene composition, the level of the inflammatory factor IL-1β in rats was significantly reduced, and could even reach a level lower than that of the normal group.

[0111] Effects of different groups on the level of inflammatory factor TNF-α in rats Fig. 9 As shown, the comparison between the icariin II-squalene group and the squalene group: *p<0.1; the comparison between the icariin II group and the model group: **p<0.01; the comparison between the normal group and the model group: ****p<0.0001; the comparison between the microemulsion group and the model group: ****p<0.0001. Fig. 9 It can be seen that after oral administration of the icariin II-squalene composition, the level of inflammatory factor TNF-α in rats was significantly reduced.

[0112] Effects of different groups on the level of inflammatory factor IL-6 in rats Fig.10 As shown, the comparison between the icariin II-squalene group and the model group: *p<0.1; the comparison between the icariin II-squalene group and the icariin II group: *p<0.1; the comparison between the icariin II-squalene group and the squalene group: ***p<0.001; the comparison between the normal group and the model group: ****p<0.1. Fig.10 It can also be seen that after oral administration of the icariin II-squalene composition, the level of inflammatory factor IL-6 in rats was significantly reduced.

[0113] After comparing the test data in this example, it can be seen that after intragastric administration of the icariin II-squalene composition, the inflammatory factors IL-1β, IL-6 and TNF-α in the CIRI model rats were significantly reduced. This proves that the use of the icariin II-squalene composition can effectively reduce reperfusion injury after cerebral ischemia.

[0114] In summary, the icariin II-squalene composition provided by the present invention effectively improves the solubility and bioavailability of icariin II, has a significant positive effect on improving neurological deficits, reducing cerebral infarction area, reducing the degree of brain tissue edema, and combating inflammatory damage, and can significantly reduce cerebral ischemia-reperfusion injury; at the same time, the preparation method of the icariin II-squalene composition provided by the present invention has a simple and rapid process and mild conditions, is a universal method for preparing hydrophobic drug microemulsions, is conducive to large-scale production, and has broad application and market prospects.

[0115] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition for alleviating cerebral ischemia-reperfusion injury, characterized in that: The composition comprises icariin II, squalene, an oil phase, an emulsifier, and an auxiliary emulsifier; The emulsifier is selected from a mixture of one or more nonionic surfactants, anionic surfactants, and amphoteric surfactants; The particle size of the composition is between 50nm and 500nm.

2. The composition according to claim 1, characterized in that The components in the composition are calculated by mass percentage as follows: Icariin II: 0.09%-4.5%; Squalene: 1%-10%; Oil phase: 0.9%-10%; Emulsifier: 0.45%-4.5%; Emulsifier: 0.09%-1%; The balance was the aqueous phase.

3. The composition according to claim 1, characterized in that The surface of the composition has a negative charge of -50 mV to -5 mV.

4. The composition according to claim 1, characterized in that The oil phase is a mixture of one or more of polyoxyethylene hydrogenated castor oil, palm oil, olive oil, soybean oil, triolein, ethyl oleate, and isopropyl myristate.

5. The composition according to claim 1, characterized in that The emulsifier is a mixture of one or more of C6-C30 fatty acids, C6-C30 aromatic acids, polyethylene glycol-distearoylethanolamine, Tween 80, Tween 40, Span 85, phospholipids, soybean lecithin, egg yolk lecithin, and polyoxyethylene ether; The auxiliary emulsifier is a mixture of one or more of polyethylene glycol-15 hydroxystearate, polyethylene glycol, sodium alginate and glycerol.

6. A method for preparing the composition according to claims 1-5, characterized in that: The specific steps are as follows: Step 1, taking 0.09-4.5 parts of icariin II, mixing it with an organic solvent, and fully dissolving it to obtain an icariin II solution; Step 2, mixing 1-10 parts of squalene with 0.45-4.5 parts of an emulsifier and 0.9-10 parts of an oil phase, and after fully dissolving, adding the icariin II solution, and stirring to obtain a first mixture; Step 3, adding 0.09-1 part of the co-emulsifier into water to obtain an aqueous solution of the co-emulsifier; adding the aqueous solution of the co-emulsifier dropwise into the first mixture under continuous stirring, and stirring to obtain colostrum; Step 4, subjecting the colostrum to ultrasonic treatment for 5-15 minutes, and then removing the organic solvent by rotary evaporation to obtain an icariin II-squalene composition; The above components are all calculated by mass.

7. The preparation method according to claim 6, characterized in that: The organic solvent in step 1 is selected from dichloromethane and anhydrous ethanol.

8. The preparation method according to claim 6, characterized in that: The mass fraction of icariin II in step 1 is 0.4-4 parts; In step 2, the mass fraction of the oil phase is 1.5-9.5 parts.

9. A method for alleviating cerebral ischemia-reperfusion injury, characterized in that: Use the composition according to claims 1-5.

10. The method according to claim 9, characterized in that The method of using the composition is selected from oral administration and intravenous injection.

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