Ginsenoside coenzyme Q10 liposome as well as preparation method and application thereof

Abstract

The invention discloses a ginsenoside coenzyme Q10 liposome as well as a preparation method and application thereof. The invention provides a ginsenoside coenzyme Q10 liposome which comprises the following components in parts by mass: 1 part of coenzyme Q10, 0.3-0.5 part of ginsenoside Rx, 1-3 parts of sunflower lecithin and 1.5-3 parts of maltodextrin, and the mass ratio of sunflower lecithin to maltodextrin is 1: (1-3). Wherein the ginsenoside Rx is a ginsenoside mixture, and the ginsenoside Rx comprises 20 (S)-ginsenoside Rg3, ginsenoside Rg5 and ginsenoside Rk1; the ginsenoside coenzyme Q10 liposome disclosed by the invention does not contain cholesterol. The ginsenoside coenzyme Q10 liposome disclosed by the invention has better bioavailability; compared with the conventional cholesterol coenzyme Q10, the gastrointestinal absorption of the coenzyme Q10 is improved by more than 1.9 times.

Description

TECHNICAL FIELD

[0001] The present application relates to a ginsenoside coenzyme Q10 liposome, a preparation method and application thereof. BACKGROUND

[0002] Liposomes are a kind of targeted drug delivery system, which is a special dosage form of targeted drug delivery system. It can embed drugs in microparticles with nanometer diameter. The microparticles are similar to the double-molecular layer microvesicles in biological membrane structure, and are mainly phagocytosed by the reticuloendothelial system in the human body, and change the in vivo distribution of the encapsulated drugs, so that the drugs are mainly accumulated in the target tissues, thereby improving the therapeutic index of the drugs, reducing the therapeutic dose of the drugs and reducing the toxicity of the drugs.

[0003] Cholesterol is contained in traditional liposomes, and cholesterol plays an important role in stabilizing and increasing the fluidity of the phospholipid bilayer. However, excessive absorption of cholesterol can cause hyperlipidemia and allergic reactions, which has attracted attention in clinical applications.

[0004] In the food and health food industry, the scientific research and industrial circles are faced with several problems: 1) the stability of active ingredients during processing and storage is poor; 2) the bioavailability of these active ingredients is usually low and difficult to be effectively absorbed; 3) it is a major challenge to achieve targeted delivery to specific tissues or cells; 4) it is also a major issue to enhance the nutritional value and sensory properties of food without affecting the quality of food.

[0005] Therefore, by means of liposomes, the stability, bioavailability and targeted delivery of active ingredients in food and health food can be significantly improved, so as to enhance the nutritional value, improve the taste and prolong the shelf life. Liposome technology is a very effective means. In dairy products, liposomes can reduce the cheese ripening time and improve the efficiency; in nutritional supplements, liposomes enhance the protection and bioavailability of compounds, making the product more suitable for commercial application, and providing consumers with better food and health benefits.

[0006] Especially in the processing of food containing polyunsaturated fatty acids, liposome technology is used to improve the sensory properties of marine source food such as fish oil, and to protect these easily oxidized fatty acids, prolong the shelf life of the product, while maintaining its health benefits for protecting cardiovascular diseases, cancer and the like.

[0007] Coenzyme Q10 has certain antioxidant, immune enhancement, and cardiovascular protection effects, and is widely used. The recommended dosage of Coenzyme Q10 soft capsules (health products) is 100-300 mg / day / person (Xiamen Jindawei), 200-400 mg / day / person (Canada GNC), and 45 mg / day / person (Beijing Tongrentang). The daily dosage of Coenzyme Q10 soft capsules on Amazon is basically 100-300 mg / day / person. Therefore, the oral dosage of the designed Coenzyme Q10 liposome should be 100-320 mg / day / person.

[0008] CN202210672651.X discloses a preparation method of phytosteryl laurate Coenzyme Q10 composite liposome. The patent dissolves Tween-80, soybean lecithin, laurate, Coenzyme Q10, etc. in an organic solvent, concentrates into a film, hydrates with a phosphate buffer solution, controls the particle size by ultrasonic head, and prepares laurate Coenzyme Q10 liposome. The patent does not show that the liposome has a significant difference or advantage in in vivo efficacy compared with ordinary cholesterol liposome.

[0009] CN201910876853.4 discloses a preparation method of Coenzyme Q10 liposome against ultraviolet damage. The patent dissolves Coenzyme Q10, thioctic acid, soybean lecithin, and cholesterol in a mixed solvent of anhydrous ethanol and Tween-80 to prepare an oil phase; uses a triethanolamine aqueous solution as an aqueous phase. Then, the oil phase is mixed with the aqueous phase to prepare a Coenzyme Q10 liposome against ultraviolet damage. The patent is mainly used for external skin care and is not suitable for oral use. According to the prescription content, the Coenzyme Q10 content is only 0.9%, and it cannot complete the experiment at a dosage of 100-320 mg / day.

[0010] CN201811598383.1 discloses a preparation method of a health food of Coenzyme Q10 liposome. The patent dissolves Coenzyme Q10, lecithin, cholesterol, Tween-80, and VE acetate in ethanol, concentrates into a film under reduced pressure, dissolves in polyvinylpyrrolidone and glycerol again, adds an aqueous phase, hydrates to obtain an emulsion, controls the particle size by ultrasonic, and prepares a Coenzyme Q10 liposome. According to the patent example, the prescription is: Q10 800 mg, cholesterol 360 mg, soybean lecithin 1.5 g, Tween 80 400 mg, and VE acetate 80 mg, and the proportion of Coenzyme Q10 is 25.48%, and the proportion of cholesterol is 11.46%. The particle size is 115.1 nm. The liposome does not add a protective agent such as glucose, and cannot be spray dried. The excipients are all inert excipients, and the efficacy does not show a significant difference or advantage compared with conventional cholesterol liposome.

[0011] CN200510137887.X discloses a kind of coenzyme Q10 liposome and its preparation process, which dissolves coenzyme Q10, soy lecithin and cholesterol in organic solvent, concentrates into film under reduced pressure, hydrates with phosphate buffer solution, and pushes through the film to prepare oral coenzyme Q10 liposome oral solution. According to the embodiment of the invention, the prescription is: 50mg coenzyme Q10, 300mg soy lecithin, 300mg cholesterol, coenzyme Q10 accounts for 7.69%, and cholesterol accounts for 46.15%. The liposome of the invention is a conventional coenzyme Q10 liposome.

[0012] CN03115914.1 discloses a kind of coenzyme Q10 precursor liposome and its preparation method, which directly heats and melts coenzyme Q10, ceramide, egg yolk lecithin and cholesterol, then hydrates with phosphate buffer solution, controls particle size by high-pressure homogenization, and then spray-dries to prepare coenzyme Q10 precursor liposome. According to the embodiment of the invention, the prescription is: 30g coenzyme Q10, 50g ceramide, 30g soy lecithin, 100g cholesterol, 40g poloxamer F68, and 200g glucose. Coenzyme Q10 accounts for 6.67%, and cholesterol accounts for 22.22%. The liposome of the invention is a conventional coenzyme Q10 liposome.

[0013] CN201610693884.2 discloses a kind of blank liposome with ginsenoside Rg5 and its derivatives as membrane material, its preparation method and application. The prescription ratio of the patent is: drug: ginsenoside: phospholipid: protective agent = 1:2-6:0.5-4:3-10 (total mass of liposome 20-70%). The drug such as paclitaxel is loaded in the membrane composed of Rg5 and lecithin, thereby improving the anti-tumor efficacy and having active targeting to cancer cells. In the patent, the content of ginsenoside is 2-6 times the amount of drug, and the recommended daily dose of ginsenoside Rg3 drug instruction (Shen Yi capsule) is 40mg / day. The designed dosage of coenzyme Q10 liposome is 100-320mg / day, therefore, the prescription ratio of CN201610693884.2 is not suitable for liposome oral preparation with high coenzyme Q10 content and low ginsenoside content.

[0014] CN201811447245.3 discloses a blank liposome with ginsenoside derivatives as membrane material, its preparation method and application. The prescription ratio of the patent is drug: ginsenoside: phospholipid: protective agent = 1:2-4:3-10:3-10 (total mass of liposome 30-60%). The drugs such as paclitaxel are loaded in the membrane composed of Rg5H and lecithin, thereby improving the anti-tumor efficacy and having significant advantages such as active targeting to cancer cells. In this patent, the content of ginsenoside is 2-4 times the amount of drug, so this prescription ratio is not suitable for liposome oral preparations with high coenzyme Q10 content and low ginsenoside content.

[0015] CN201811447243.4 discloses a blank liposome with ginsenoside Rg3 and its derivatives as membrane material and application. The prescription ratio of the patent is drug: ginsenoside: phospholipid: protective agent = 1:0.5-2:3-10:3-10 (total mass of liposome 30-60%). The drugs such as paclitaxel are loaded in the membrane composed of Rg3 and lecithin, thereby improving the anti-tumor efficacy and having significant advantages such as active targeting to cancer cells. The active substance described in this patent is a cytotoxic anti-tumor drug, and the liposome prepared in this patent is mainly used for injection and mainly uses egg yolk lecithin and soybean lecithin, and mainly uses the freeze-drying process. Taking the prescription of the patent as an example: drug: saponin: phospholipid: protective agent = 1:0.5:3:3, the minimum oral dose of ginsenoside is 50-150 mg / day, which is not suitable for liposome oral preparations with high coenzyme Q10 content and low ginsenoside content (Shenyi Capsule is the only ginsenoside Rg3 product on the market, with a maximum oral dose of 40 mg / day).

[0016] CN202011229476.4 discloses a compound ginsenoside paclitaxel liposome, its preparation method and application. The prescription ratio of the patent is drug: ginsenoside: phospholipid: protective agent = 1:1-1.5:8-12:25-35. The drugs such as paclitaxel are loaded in the membrane composed of Rg3 and lecithin, thereby improving the anti-tumor efficacy and having significant advantages such as active targeting to cancer cells. In this patent, the content of ginsenoside is 1-1.5 times the amount of drug, so this prescription ratio is also not suitable for liposome oral preparations with high coenzyme Q10 content and low ginsenoside content.

[0017] The recommended daily oral dose of ginsenoside Rg3 is ≤40mg, and the recommended daily oral dose of total ginsenosides (or secondary mixed saponins) is ≤300mg. The recommended daily oral dose of coenzyme Q10 is 100-300mg. Based on a daily oral liposome dose of 1200mg, the reasonable mass percentage of coenzyme Q10 in the total oral liposome (including the protectant) should be 10-30%, ginsenoside Rx 5-15%, the lyophilized protectant 35-50%, and lecithin 20-40%. Therefore, monomers such as ginsenoside Rg3 and Rh2 are not suitable for use in oral coenzyme Q10 liposomes.

[0018] The core of compound preparations lies in the synergistic interaction of drugs in vivo, which can significantly improve the clinical therapeutic effect. The reasonable ratio range of each functional component in a compound preparation is the core of its composition. In particular, the changes in drug synergy, pharmacokinetics, tissue distribution, and efficacy caused by changes in the functional components of compound liposomes have rarely been addressed.

[0019] Therefore, a great deal of research and technological breakthroughs are needed to select the best compound drug combination and to develop the best preparation process in order to produce an oral ginsenoside coenzyme Q10 liposome with better efficacy, stability and bioavailability to meet the market demand for food and health food. Summary of the Invention

[0020] The technical problems to be solved by this invention are: 1) overcoming the instability of coenzyme Q10 to light and heat; 2) the disadvantages of slow gastrointestinal absorption and low bioavailability after oral administration; and 3) improving the various technical indicators of liposomes while increasing the oral dosage of coenzyme Q10. This invention provides a ginsenoside coenzyme Q10 liposome, which has one or more advantages, including stable properties, reasonable particle size, high drug encapsulation rate, good drug stability, higher gastrointestinal absorption rate, and reasonable compatibility. Furthermore, this invention also provides a method for preparing ginsenoside coenzyme Q10 liposomes and their application; it has a good preparation process, the preparation conditions are easy to achieve, which is conducive to industrialization; and it achieves optimization of the combination of preparation process and product performance.

[0021] This invention provides a ginsenoside coenzyme Q10 liposome, which comprises the following components in parts by mass: 1 part coenzyme Q10, 0.3-0.5 parts ginsenoside Rx, 1-3 parts sunflower lecithin and 1.5-3 parts maltodextrin, wherein the mass ratio of sunflower lecithin to maltodextrin is 1:(1-3).

[0022] The ginsenoside Rx is a mixture of ginsenosides, which includes 20(S)-ginsenoside Rg3, ginsenoside Rg5, and ginsenoside Rk1.

[0023] The ginsenoside coenzyme Q10 liposomes described herein do not contain cholesterol.

[0024] In one embodiment of the present invention, the ginsenoside Rx is 0.3 parts, 0.4 parts, or 0.5 parts.

[0025] In one embodiment of the present invention, the sunflower lecithin is 1 part, 2 parts, or 3 parts.

[0026] In one embodiment of the present invention, the maltodextrin is 1.5 parts, 2 parts, or 3 parts.

[0027] In one embodiment of the present invention, the mass ratio of coenzyme Q10 to sunflower lecithin may be 1:(1-3), for example 1:1, 1:2 or 1:3.

[0028] In one embodiment of the present invention, the mass ratio of coenzyme Q10 to ginsenoside Rx can be 1:(0.3-0.5); for example, 1:0.3, 1:0.4 or 1:0.5.

[0029] In one embodiment of the present invention, the mass ratio of the maltodextrin to the sunflower lecithin may be (1-1.5):1; for example, 1:1 or 1.5:1.

[0030] In one aspect of the present invention, the sunflower lecithin may be sunflower lecithin with a content of 60% or more, for example, composed of 60% sunflower lecithin and 40% fatty acids.

[0031] In one embodiment of the present invention, the ginsenoside Rx is derived from the stems and leaves of American ginseng, preferably an extract of the stems and leaves of American ginseng.

[0032] In one embodiment of the present invention, the ginsenoside Rx may be an alcohol extract of the stem and leaf extract of American ginseng; for example, a n-butanol extract.

[0033] In one embodiment of the present invention, the ginsenoside Rx is prepared by the following steps:

[0034] Step 1: Mix the American ginseng stem and leaf extract with a citric acid aqueous solution and heat to obtain mixed solution 1;

[0035] Step 2: Filter, extract, wash, and concentrate the mixed solution 1 to obtain solid 2;

[0036] Step 3: Redissolve solid 2, filter at low temperature, and concentrate to obtain the product ginsenoside mixture Rx;

[0037] The definition of American ginseng stem and leaf extract is as described in the above-mentioned ginsenoside coenzyme Q10 liposomes.

[0038] In one embodiment of the present invention, in step 1, the citric acid aqueous solution is preferably a 20% citric acid aqueous solution.

[0039] In one embodiment of the present invention, in step 1, the heating temperature is a conventional heating temperature in the art, such as heating to reflux. In another embodiment of the present invention, in step 2, the extraction is performed using an organic solvent, preferably an alcohol solvent, more preferably n-butanol; the number of extractions is adjusted reasonably according to experimental needs, preferably 3-4 times, for example 3 times.

[0040] In one embodiment of the present invention, the cleaning in step 2 is a conventional cleaning method in the art, preferably using an inorganic solution, such as one or more of a 10% NaOH aqueous solution, saturated saline solution, or purified water; the number of cleaning cycles can be reasonably adjusted according to experimental needs, preferably 3 times, for example, washing once with a 10% NaOH aqueous solution, washing once with 200L of saturated saline solution, and washing once with 200L of purified water.

[0041] In one embodiment of the present invention, in step 2, the concentration is a conventional concentration in the art, and the concentration can be vacuum concentration; the concentration is carried out until dry.

[0042] In one embodiment of the present invention, in step 3, the resolution is performed using an organic solution, wherein the organic solution is an alcohol solution, preferably anhydrous ethanol.

[0043] In one embodiment of the present invention, in step 3, the temperature of the low-temperature filtration is 0-15°C, for example 2-8°C.

[0044] In one embodiment of the present invention, in step 3, the concentration is a conventional concentration in the art, and the concentration can be vacuum concentration; the concentration is carried out until all the ethanol has completely evaporated.

[0045] Preferably, the preparation method described herein can be based on Example 1 of this application.

[0046] In one embodiment of the present invention, the ginsenoside Rx may be a conventional mixture of ginsenosides in the art, for example, the total content of each ginsenoside in the ginsenoside Rx is about 90-95%; wherein, the content of 20(S)-ginsenoside Rg3 is 20-25%; the content of ginsenoside Rg5 is 45-50%; the content of ginsenoside Rk1 is 20-25%, and the content of 20(R)-ginsenoside Rg3 in the ginsenoside Rx is less than 2%.

[0047] In one embodiment of the present invention, the mass ratio of 20(S)-ginsenoside Rg3 to ginsenoside Rg5 can be 1:(1.5 to 2.5), for example 1:1.93 or 1:2.

[0048] In one embodiment of the present invention, the mass ratio of 20(S)-ginsenoside Rg3 to ginsenoside Rk1 can be 1:(0.5 to 1.5), for example 1:0.86 or 1:1.

[0049] In one embodiment of the present invention, the ginsenoside coenzyme Q10 liposomes contain 10-30% coenzyme Q10 by mass, 5-15% ginsenoside Rx by mass, 20-40% sunflower lecithin by mass, and 35-50% maltodextrin by mass, wherein the mass percentages are the proportions of each component to the total mass of the raw materials.

[0050] In one embodiment of the present invention, the ginsenoside Rx has an HPLC purity of (Rg3+Rg5+Rk1) ≥ 90% (e.g., 90%-95%).

[0051] In one embodiment of the present invention, the particle size D50 of the ginsenoside coenzyme Q10 liposomes can be adjusted according to production requirements, for example, D50≤500nm, or D50≤400nm, or D50≤300nm, or even 100nm, 200nm, 300nm or 400nm.

[0052] In one embodiment of the present invention, the ginsenoside coenzyme Q10 liposome encapsulation rate is ≥80%, preferably ≥90%, and more preferably ≥95%.

[0053] In one embodiment of the present invention, the ginsenoside coenzyme Q10 liposome may include any of the following components by mass fraction:

[0054] 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0055] Alternatively, 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0056] Alternatively, 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0057] Alternatively, 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0058] Alternatively, 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0059] Alternatively, 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0060] Alternatively, 1 part Q10, 0.3 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0061] Alternatively, 1 part Q10, 0.4 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0062] Alternatively, 1 part Q10, 0.5 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0063] Alternatively, 1 part Q10, 0.3 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin;

[0064] Alternatively, 1 part Q10, 0.4 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin;

[0065] Alternatively, 1 part Q10, 0.5 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin.

[0066] In one embodiment of the present invention, the formulation of the ginsenoside coenzyme Q10 liposomes comprises the following components by mass fraction:

[0067] 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0068] Alternatively, 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0069] Alternatively, 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin;

[0070] Alternatively, 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0071] Alternatively, 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0072] Alternatively, 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin;

[0073] Alternatively, 1 part Q10, 0.3 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0074] Alternatively, 1 part Q10, 0.4 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0075] Alternatively, 1 part Q10, 0.5 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin;

[0076] Alternatively, 1 part Q10, 0.3 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin;

[0077] Alternatively, 1 part Q10, 0.4 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin;

[0078] Alternatively, 1 part Q10, 0.5 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin.

[0079] In one embodiment of the present invention, the formulation of the ginsenoside coenzyme Q10 liposomes comprises any of the following components by mass fraction:

[0080] 26.32% Q10, 7.89% Rx, 26.32% sunflower lecithin, 39.47% maltodextrin;

[0081] Alternatively, 25.64% Q10, 10.26% Rx, 25.64% sunflower lecithin, and 38.46% maltodextrin;

[0082] Alternatively, 25.0% Q10, 12.5% ​​Rx, 25.0% sunflower lecithin, and 37.5% maltodextrin;

[0083] Alternatively, 23.28% Q10, 6.98% Rx, 23.28% sunflower lecithin, and 46.46% maltodextrin;

[0084] Alternatively, 22.73% Q10, 9.09% Rx, 22.73% sunflower lecithin, and 45.45% maltodextrin;

[0085] Alternatively, 22.25% Q10, 11.11% Rx, 22.25% sunflower lecithin, and 44.39% maltodextrin;

[0086] Alternatively, 18.86% Q10, 5.66% Rx, 37.74% sunflower lecithin, and 37.74% maltodextrin;

[0087] Alternatively, 18.52% Q10, 7.40% Rx, 37.04% sunflower lecithin, and 37.04% maltodextrin;

[0088] Alternatively, 18.19% Q10, 9.09% Rx, 36.36% sunflower lecithin, and 36.36% maltodextrin;

[0089] Alternatively, 13.70% Q10, 4.10% Rx, 41.10% sunflower lecithin, and 41.10% maltodextrin;

[0090] Alternatively, 13.51% Q10, 5.41% Rx, 40.54% sunflower lecithin, and 40.54% maltodextrin;

[0091] Alternatively, 13.33% Q10, 6.67% Rx, 40.0% sunflower lecithin, and 40.0% maltodextrin.

[0092] In one embodiment of the present invention, the formulation of the ginsenoside coenzyme Q10 liposome is the composition of the following components in the indicated mass fractions:

[0093]

[0094] This invention also provides a method for preparing ginsenoside coenzyme Q10 liposomes, which includes the following steps:

[0095] Step 1: Dissolve coenzyme Q10, ginsenoside Rx and sunflower lecithin in ethanol to obtain solution A1, and concentrate it under reduced pressure to form a film;

[0096] Step 2: The membrane obtained in Step 1 is hydrated by keeping it warm in an aqueous solution containing maltodextrin to obtain liposome solution A2.

[0097] Step 3: The liposome solution A2 obtained in Step 2 is subjected to high-pressure homogenization to control the particle size D50 to below 300 nm, thereby obtaining a solution A3 containing the liposomes;

[0098] The definitions of coenzyme Q10, ginsenoside Rx, sunflower lecithin, maltodextrin, and their proportions are the same as those described in the ginsenoside coenzyme Q10 liposomes above.

[0099] In one embodiment of the present invention, the ethanol has a mass content of ≥70%, where the mass content is the percentage of the mass of the component relative to the total mass of the raw materials.

[0100] In one embodiment of the present invention, the amount of ethanol used is not specifically limited, as long as it can dissolve coenzyme Q10 ginsenosides and sunflower lecithin; for example, the mass-volume ratio of coenzyme Q10 to ethanol is 1g / 3-15mL, such as 1g / 5mL.

[0101] In one embodiment of the present invention, in step 1, solution A1 is obtained by heating and dissolving coenzyme Q10, ginsenosides, sunflower lecithin, etc. in ethanol.

[0102] In one embodiment of the present invention, the heating may be water bath heating.

[0103] In one embodiment of the present invention, the water bath temperature is 35-65°C, for example 40-45°C or 55-60°C; preferably 55°C.

[0104] In one embodiment of the present invention, the concentration in step 1 may be vacuum concentration.

[0105] In one aspect of the present invention, in step 1, the concentration can be performed under a vacuum of -0.04 MPa to -0.1 MPa; preferably -0.04 to -0.05 MPa.

[0106] In one embodiment of the present invention, in step 1, the concentration is carried out until all the ethanol has completely evaporated.

[0107] In one embodiment of the present invention, the total concentration time in step 1 is less than 4 hours.

[0108] In one embodiment of the present invention, in step 1, the concentration can be carried out in a rotary evaporator at a rotation speed of 40-60 rpm, for example, 50 rpm.

[0109] In one embodiment of the present invention, in step 2, the concentration of the aqueous solution containing maltodextrin can be 100-300 mg / mL, for example, 100 mg / mL.

[0110] In one embodiment of the present invention, the hydration temperature in step 2 can be 35-65°C, for example 45-50°C.

[0111] In one embodiment of the present invention, in step 2, the hydration is carried out in a rotary evaporator at a rotation speed of 40 to 70 rpm, for example, 50 rpm or 65 rpm.

[0112] In one embodiment of the present invention, in step 2, the hydration is sufficient to achieve a homogeneous solution, for example, 2-4 hours.

[0113] In one embodiment of the present invention, in step 2, the coenzyme Q10: maltodextrin solution = 1g: 10-50mL, for example 1g / 15mL or 1g / 30mL.

[0114] In one embodiment of the present invention, in step 3, the high-pressure homogenization is performed by using a 0-10°C chilled water cooling cycle in a homogenizer; preferably, the temperature of the liposome solution is ensured to be 5-10°C.

[0115] In one embodiment of the present invention, in step 3, the pressure of the high-pressure homogenization is between 800-1600 bar, for example 1200 bar.

[0116] In one embodiment of the present invention, in step 3, the high-pressure homogenization may be performed 3-4 times, for example, 4 times.

[0117] In one embodiment of the present invention, the method for preparing ginsenoside coenzyme Q10 liposomes further includes the following steps:

[0118] Step 4: Spray dry the liposome solution A3 to obtain ginsenoside coenzyme Q10 liposomes.

[0119] In the preparation method described above, the conditions and operations for spray drying can be conventional conditions and operations in this type of process in the art; preferably, the following are preferred in this invention:

[0120] In one embodiment of the present invention, in step 4, the spray drying process involves an inlet air temperature of 165-185°C, such as 165°C, 170°C, 175°C, or 180°C, or 185°C; an outlet air temperature of 70-90°C, such as 70°C, 75°C, 80°C, or 85°C; and a spray flow rate that can be adjusted according to the model of the spray drying equipment, based on the requirement that powder solids can be obtained after spraying. For example, when the machine model is PLG-30, the flow rate is 5-30 L / h, such as 5 L / h, 10 L / h, 15 L / h, 20 L / h, 25 L / h, or 30 L / h; and a spray atomizer rotation speed of 6000-18000 rp / min, such as 6000 rp / min, 12000 rp / min, 15000 rp / min, or 18000 rp / min.

[0121] Preferably, the flow rate of the spray increases as the inlet air temperature rises.

[0122] In one aspect of the present invention, in step 4, the spray drying satisfies any of the following conditions:

[0123] Option 1:

[0124] The air inlet temperature is 165℃, the flow rate is 5L / h, and the atomizer speed is 18000rp / min;

[0125] Option 2:

[0126] The inlet air temperature is 165℃, the flow rate is 10L / h, and the atomizer speed is 18000rp / min;

[0127] Option 3:

[0128] The air inlet temperature is 170℃, the flow rate is 15L / h, and the atomizer speed is 18000rp / min;

[0129] Option 4:

[0130] The inlet air temperature is 175℃, the flow rate is 20L / h, and the atomizer speed is 18000rp / min;

[0131] Option 5:

[0132] The air inlet temperature is 180℃, the flow rate is 25L / h, and the atomizer speed is 18000rp / min;

[0133] Option Six:

[0134] The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 18000rp / min;

[0135] Option Seven:

[0136] The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 12000rp / min;

[0137] Option 8:

[0138] The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 6000rp / min;

[0139] Option Nine:

[0140] The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 15000rp / min;

[0141] Option 10:

[0142] The inlet air temperature is 185℃, the flow rate is 30L / h, and the atomizer speed is 15000rp / min.

[0143] The present invention also provides a ginsenoside coenzyme Q10 liposome, which is prepared by the ginsenoside coenzyme Q10 liposome preparation method described above.

[0144] This invention also provides the application of ginsenoside coenzyme Q10 liposomes in the preparation of drugs or health foods for the treatment and / or prevention of cardiovascular and cerebrovascular diseases; the ginsenoside coenzyme Q10 liposomes are the ginsenoside coenzyme Q10 liposomes described above.

[0145] The term "particle size D50" refers to the average particle size at which the cumulative particle size distribution percentage of a sample reaches 50%. Physically, it means that 50% of the particles are smaller than this value.

[0146] The abbreviations for the prescriptions of this invention are explained below:

[0147]

[0148] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0149] The reagents and raw materials used in this invention are all commercially available.

[0150] The positive and progressive effects of this invention are as follows: the ginsenoside coenzyme Q10 liposomes provided by this invention have high bioavailability. In the examples, the ginsenoside coenzyme Q10 liposomes exhibit significantly better efficacy than ordinary cholesterol liposomes; this demonstrates that Rx plays a better role as a "drug, excipient, and membrane material" in the ginsenoside coenzyme Q10 liposomes, resulting in a good synergistic effect. Specifically:

[0151] 1. With a significant reduction in ginsenoside dosage, how can we ensure the stability, encapsulation efficiency, and drug loading of liposomes, and more importantly, how can we ensure improved bioavailability?

[0152] 2. Spray drying technology is used, which involves high temperatures. Phospholipids are prone to denaturation at high temperatures. How can we ensure that the particle size of the liposomes remains within a reasonable range and that the phospholipids do not denature?

[0153] 3. By administering the drug orally to animals, the optimal prescription can be screened to achieve higher blood drug concentrations and thus improve bioavailability;

[0154] 4. Through prescription screening, Q10 can prolong its circulation time in vivo, thereby improving its bioavailability. Attached Figure Description

[0155] Figure 1 Particle size diagram of ginsenoside coenzyme Q10 liposomes.

[0156] Figure 2 This is a particle size diagram of ordinary cholesterol coenzyme Q10 liposomes.

[0157] Figure 3 This is a particle size diagram of ordinary cholesterol coenzyme Q10 liposomes.

[0158] Figure 4 This is an image of the isolated small intestine of a mouse. Detailed Implementation

[0159] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0160] Experimental drugs and equipment

[0161] Experimental drugs: Ginsenoside mixture Rx (abbreviated as Rx) is derived from American ginseng stems and leaves (Jilin Hongjiu Biotechnology Co., Ltd.). The ginsenoside mixture Rx, coenzyme Q10 (Xi'an Tianfeng Biotechnology Co., Ltd.), sunflower lecithin (PC with a content of 60% or higher) (Hebei Meiyesiwei Biotechnology Co., Ltd.), and maltodextrin (Xi'an Tianfeng Biotechnology Co., Ltd.) of this invention are all commercially available in the field.

[0162] The molecular structure of ginsenosides described in this invention is as follows:

[0163]

[0164]

[0165] Experimental Instruments: The instruments used in the following examples are from Shanghai Bensu Pharmaceutical Technology Co., Ltd. and the School of Pharmacy of Fudan University. The equipment models and sources are as follows:

[0166] Agilent liquid chromatography: one Agilent 1100 system, an Altech 3300 ELSD system, Agilent Technologies (China) Co., Ltd.;

[0167] Rotary evaporator: ZX98-1 5L, Shanghai Luyi Industry & Trade Co., Ltd.;

[0168] Ultrasonic cleaning machine (SB3200DT, Ningbo Xinzhi Biotechnology Co., Ltd.);

[0169] Nitrogen blowing device (HGC-12A, Tianjin Hengao Technology Development Co., Ltd.);

[0170] Probe ultrasonic transducer (JYD-650, Shanghai Zhixin Instrument Co., Ltd., China);

[0171] High-pressure homogenizer (B15, AVESTIN, Canada);

[0172] Spray dryers (LPG-5, PLG-30, Changzhou Erle Drying Equipment Co., Ltd.)

[0173] Laser particle size analyzer (Nano ZS, Malvern Ltd., UK);

[0174] Malvern Nanosizer ZS90 particle size analyzer (Malvern, UK);

[0175] Microplate reader (Infinitie 200, Tecan Trading Co., Ltd., Switzerland);

[0176] Flow cytometer (CytoFlex S, Beckman Coulter, Inc., USA);

[0177] Fluorescence microscopy observation (Zeiss LSM 710, Oberkochen, Germany);

[0178] Laser confocal microscope (Leica, DMI 4000D, Germany);

[0179] Upright two-photon microscope (DM5500 Q; Nikon);

[0180] Clean bench (SW-CJ-1FD, Suzhou Antai Air Technology Co., Ltd.);

[0181] 20L rotary evaporator: R5002K, Shanghai Xiafeng Industrial Co., Ltd.;

[0182] Electronic balance: CPA2250 (accuracy 0.00001g), Sartorius (Shanghai) Trading Co., Ltd.;

[0183] Electronic balance: JY3003 (accuracy 0.001g), Shanghai Sunny Hengping Scientific Instruments Co., Ltd.;

[0184] Photoelectric microscope (XDS-1B, Chongqing Optoelectronic Instrument Co., Ltd.);

[0185] Cell incubator (CCL-170B-8, Singapore ESCO).

[0186] High Performance Liquid Chromatography (LC-20AD)

[0187] Animals and cell lines

[0188] Animals: SD rats, 3-4 weeks old, purchased from Shanghai Slack Laboratory Animal Co., Ltd.

[0189] Particle size determination: The liposomes were first diluted in pure water at a volume ratio of 1:100, and the particle size of the liposomes was determined using a Malvern particle size analyzer.

[0190] Encapsulation efficiency (EE%) detection: Take 40 mg of liposomes, add purified water to a final volume of 10 mL, and shake to disperse, forming an emulsion. 1) Add sample: Measure 0.2 mL of the test solution and add it to the gel column. Then, gently stir the sample layer with a glass rod and rinse the glass rod with 1 mL of purified water. 2) Begin elution with purified water, discarding 2 mL of the initial filtrate; collect 15 mL in a graduated cylinder, transfer all of it to a 25 mL volumetric flask, rinse the graduated cylinder with ethanol, dissolve the ethanol, and dilute to the mark. Mix well to obtain solution M1 for determining the encapsulated content. Elute with 4 mL of 50% ethanol, then rinse with anhydrous ethanol. Collect the elution with a graduated cylinder to a 25 mL volumetric flask to obtain solution M2 for determining the free coenzyme Q10 content. Transfer 0.2 mL of the test solution to a 25 mL volumetric flask, dissolve in ethanol, dilute to the mark with purified water, and mix well. This is solution M3 for recovery determination. 3) Following the chromatographic method under the Assay section, inject solutions M1, M2, and M3 into the liquid chromatograph separately and record the chromatograms. Calculate the encapsulation efficiency based on the peak area using the following formula:

[0191]

[0192]

[0193] In the formula: A1 is the peak area of ​​the drug encapsulated in liposomes in solution M1.

[0194] A2 represents the peak area of ​​the unencapsulated free drug in solution M2.

[0195] A3 represents the peak area of ​​the total drug recovered from solution M3.

[0196] Note: Chromatography column brand: Lianhua, specifications: ground glass column with sand core, PTFE gate, outer diameter × length: 17 × 300 mm, 19# G3 sand core.

[0197] Drug loading capacity (DL%) detection: The amount of coenzyme Q10 (WE) in the liposome powder was measured by high performance liquid chromatography (HPLC). The formula for calculating DL% is as follows: DL% = WE / WT * 100%.

[0198] Example 1: Preparation of Ginsenoside Mixture (abbreviated as: Ginsenoside Rx)

[0199] 100g of American ginseng stem and leaf extract (purchased from Jilin Hongjiu) was dissolved in 500mL of citric acid aqueous solution containing 20% ​​citric acid and heated under reflux for 2 hours. After the reaction was completed, the solution was cooled to room temperature and filtered to remove insoluble matter. The aqueous phase was extracted with 200L of n-butanol each time, for a total of 3 extractions, and the organic phases were combined. The organic phase was washed once with 200L of 10% NaOH aqueous solution, once with 200L of saturated brine, and once with 200L of purified water. The solution was concentrated to dryness under reduced pressure, and then redissolved in 250mL of anhydrous ethanol in a 60℃ water bath. The solution was placed in a refrigerator at 2-8℃ for 12 hours, and 20(R)-ginsenoside Rg3 was removed by suction filtration. The ethanol solution was concentrated to dryness under reduced pressure, and the resulting solid was dried to obtain 56g of ginsenoside mixture (abbreviated as: ginsenoside Rx). HPLC analysis revealed that the total ginsenoside content was approximately 90-95%, including 20(S)-ginsenoside Rg3 (24.15%), ginsenoside Rg5 (46.71%), ginsenoside Rk1 (20.81%), and 20(R)-ginsenoside Rg3 (1.63%).

[0200] Example 2: Preparation of Ginsenoside Coenzyme Q10 Liposomes

[0201] 1. Prescription: Coenzyme Q10 100g, Ginsenoside Rx 50g, Sunflower Lecithin 100g, Maltodextrin 150g, Anhydrous Ethanol 500mL, Purified Water 1500mL.

[0202] 2. Film formation: Dissolve the prescribed amount of coenzyme Q10 in anhydrous ethanol and set aside. Then add the prescribed amounts of ginsenoside Rx and sunflower lecithin to the organic solvent, heat to dissolve, transfer to a 1L rotary evaporator, concentrate under reduced pressure, water bath temperature 55℃, rotation speed 50rp / min, vacuum degree -0.04~-0.05MPa, and rotary evaporate until all solvent has evaporated.

[0203] 3. Hydration: Prepare maltodextrin aqueous solution: Add 30g of maltodextrin to 1500mL of purified water, stir to dissolve, and prepare maltodextrin aqueous solution. Heat in a 40℃ water bath for later use.

[0204] Add 1500 mL of maltodextrin aqueous solution to the rotary evaporator after film formation. Hydrate and completely dissolve the liposomes in a water bath at 40-45℃ and a rotation speed of 50 rpm for about 2 hours to obtain a liposome aqueous solution with a solid content of 26.67%.

[0205] 4. High-pressure homogenization: The hydrated solution is transferred to a homogenizer. The homogenizer uses 0-10℃ chilled water for cooling and circulation. The homogenization pressure is set to 1200 bar, and the homogenization is repeated 3-4 times.

[0206] 5. Spray drying: The spray drying equipment is model PLG-30, with an inlet air temperature of 180℃, an outlet air temperature of 70-90℃, and a flow rate of 30L / h. The above liposome solution is spray dried to obtain 338.4g of ginsenoside coenzyme Q10 liposomes.

[0207] 6. Packaging and Testing: The above liposomes were sampled and tested, then packaged in PE bags to obtain ginsenoside coenzyme Q10 liposomes. The average particle size (D50) was measured to be 191 nm, and the encapsulation efficiency was 98.9% (see...). Figure 1 ).

[0208] Example 3: Preparation of a series of ginsenoside coenzyme Q10 liposomes

[0209] Following the same method as in Example 2, a series of ginsenoside coenzyme Q10 liposomes were prepared according to the prescription ratios in Table 1.

[0210] Coenzyme Q10 (Q10), Ginsenoside Rx (Rx), Sunflower Lecithin (PC), Maltodextrin (MD)

[0211] Table 1. Formulation ratio of each component in liposomes

[0212]

[0213] Example 4: Preparation of ordinary cholesterol coenzyme Q10 liposomes

[0214] 1. Prescription: Coenzyme Q10 100g, cholesterol 50g, sunflower lecithin 100g, maltodextrin 150g, anhydrous ethanol 500mL, purified water 1500mL.

[0215] Other liposomes were prepared using the same method as in Example 2 above, resulting in ordinary cholesterol-coenzyme Q10 liposomes. The average particle size (D50) was measured to be 219 nm, and the encapsulation efficiency was 90.8% (see [link to example 2]). Figure 2 ).

[0216] Example 5: Preparation of ordinary cholesterol-coenzyme Q10 liposomes (according to the method of Example 1, CN201811598383.1)

[0217] 1. Prescription: Coenzyme Q10 800mg, cholesterol 360mg, 90% soy lecithin 1.5g, Tween 80 400mg, vitamin E acetate 80mg, anhydrous ethanol 4mL, purified water 20mL.

[0218] 2. Preparation method: Dissolve the prescribed amount of Q10, soybean lecithin, cholesterol, Tween 80, and VE acetate in ethanol, heat to dissolve according to the method in Example 2, then concentrate under reduced pressure to dryness, hydrate, and sonicate with an ultrasonic head to obtain ordinary cholesterol Q10 liposomes for later use.

[0219] The average particle size (D50) was measured to be 274 nm, and the encapsulation efficiency was 91.1% (see [link to data]). Figure 3 ).

[0220] Example 1

[0221] (a) Based on the formulations in Table 2 below, and using the same preparation method, spray drying conditions, and spray drying equipment as in Example 2, the formulation screening experiment for the spray drying protectant was conducted as follows:

[0222] Table 2. Formulation ratio of each component in liposomes

[0223]

[0224] Through the above experiments, the ratio of spray drying protectant to sunflower lecithin was found to be: sunflower lecithin / protectant ≤ 1 (mass ratio). Based on a comprehensive evaluation of the oral dosage of coenzyme Q10, this invention selected sunflower lecithin: protectant = 1:1-3.

[0225] (b) Based on the formulation in Table 3 below and using the same preparation method as in Example 2, the following screening experiments were conducted on the spray drying conditions:

[0226] Spray drying equipment model: PLG-30, flow rate range: 5-30L / h, inlet air temperature range: 120-250℃; outlet air temperature range: 65-150℃ (related to flow rate adjustment, not a key technical indicator); atomizer speed: 6000-18000rp / min.

[0227] Table 3 Screening of spray drying conditions

[0228]

[0229]

[0230] Note: Sunflower lecithin = SPC, maltodextrin = MD

[0231] Based on the above experiments, the spray drying conditions of the present invention are as follows:

[0232] 1) Inlet air temperature: 165-185℃;

[0233] 2) Flow rate: Adjust according to the equipment model. Taking PLG-30 as an example, the flow rate range is 5-30L / h;

[0234] 3) Atomizer speed: 6000-18000 rp / min;

[0235] 4) Outlet air temperature: It needs to be controlled according to the equipment model and the feed flow rate. Taking PLG-30 as an example, the optimal control range for outlet air temperature is 70-90℃.

[0236] (c) Comparison of stability test results

[0237] Liposome preparation method: the same method as in Example 2.

[0238] According to the formulation in Table 4 below, the accelerated stability test conditions were verified as follows: 25℃, 60% humidity;

[0239] Table 4 Accelerated stability test conditions

[0240]

[0241]

[0242] Example 2: Effect of 20(R)-ginsenoside Rg3 content on liposomes

[0243] 1. Prescription: Coenzyme Q10 1g, Ginsenoside Rx (20(R)-Ginsenoside Rg3 with different contents) 0.5g, Sunflower lecithin 1g, Maltodextrin 1.5g, Anhydrous ethanol 5mL, Purified water 15mL.

[0244] 2. Take the ginsenoside Rx obtained in Example 1, wherein the content of 20(R)-ginsenoside Rg3 is 1.63%, and add 99% 20(R)-ginsenoside Rg3 according to the table below:

[0245]

[0246] 3. Hydration: The same method as in Example 2.

[0247] 4. High-pressure homogenization: Following the same method as in Example 2, liposome control solutions 1 (corresponding to sample 1), 2 (corresponding to sample 2), 3 (corresponding to sample 3), and 4 (corresponding to sample 4) were prepared respectively.

[0248] 5. Post-experimental processing: After high-pressure homogenization, the state of the liposome solution was visually inspected. Then, the solution was filtered through filter paper, the filter paper was washed with purified water, and the white solids remaining on the filter paper were detected by HPLC.

[0249] The implementation results are shown in the table below:

[0250] Sample name R-Rg3 content Liposome appearance White residue (confirmed by HPLC detection) Example 2 liposome sample 1.63％ Clear and transparent No residue Comparative solution 1 2.02％ Clear and transparent No residue Comparative solution 2 2.57％ Slightly turbid 20(R)-ginsenoside Rg3 Comparative solution 3 3.52％ Slightly turbid 20(R)-ginsenoside Rg3 Comparative solution 4 5.36％ More turbid 20(R)-ginsenoside Rg3

[0251] Conclusion: According to the formulation and preparation method of the present invention, when the content of 20(R)-ginsenoside Rg3 exceeds 2.57%, 20(R)-ginsenoside Rg3 cannot be encapsulated in liposomes. The present invention determines that the content of 20(R)-ginsenoside Rg3 should not exceed 2%.

[0252] Application Example 1: In vivo imaging to examine the gastrointestinal absorption of ginsenoside-modified liposomes

[0253] 1. Prepare cholesterol liposomes (C-Lip / Dir) and ginsenoside liposomes (G-Lip / Dir) loaded with Dir, and prepare free Dir solutions. Dilute with PBS to obtain three formulations with a final Dir concentration of 20 μg / mL.

[0254] 2. Mice were fasted overnight and randomly divided into three groups (n=6). Each group of mice was orally administered 200 μL of Dir-Free, C-Lip / Dir, or G-Lip / Dir, respectively. One hour later, the mice were dissected, and the distribution of ginsenoside liposomes (G-Lip / Dir) in the small intestine was observed using a small animal in vivo imaging system (see...). Figure 4 ).

[0255] The experimental results showed that, compared with free Dir and ordinary cholesterol liposomes (C-Lip), ginsenoside-modified liposomes (G-Lip) had a higher signal intensity in the small intestine and a lower signal intensity in the large intestine, indicating that ginsenosides can promote the absorption of liposomes in the proximal small intestine.

[0256] Application Example 2: Comparison of the pharmacokinetics of three forms of coenzyme Q10 liposomes

[0257] Experimental animals: 9 healthy male and 9 healthy female Sprague-Dawley (SD) rats, totaling 18 (weighing 180-200g), were randomly divided into 3 groups.

[0258] Experimental drug: Q10-Free. Take 100mg of coenzyme Q10, dissolve it in 10mL of vegetable oil, stir well, and you will get a 10mg / mL coenzyme Q10 solution.

[0259] Q10-Lip-Rx was prepared according to the method in Example 2. 1g of liposome powder (Q10 content = 13.70%) was dispersed in 50mL of purified water to obtain a 2.74mg / mL coenzyme Q10 liposome solution.

[0260] Q10-Lip-Cho was prepared according to the method in Example 2, wherein Rx was replaced with an equal amount of cholesterol. 1 g of liposome powder was dispersed in 50 mL of purified water to obtain a 2.74 mg / mL coenzyme Q10 liposome solution.

[0261] Administration and Sampling: Each rat was orally administered 100 mg / kg of coenzyme Q10, available in three forms: Q10-Free, Q10-Lipo-Cho, and Q10-Lipo-Rx. At 0, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, 24, 48, and 72 hours post-administration, approximately 200 μL of blood was collected via tail vein into heparinized centrifuge tubes. Each blood sample was immediately centrifuged at 3000 rpm for 10 minutes to separate the plasma, which was then stored at -30°C until subsequent sample preparation and analysis.

[0262] Plasma sample preparation: Take 50 μL of plasma sample and add 400 μL of acetonitrile containing 1% formic acid for protein precipitation. After centrifugation at 14000 rpm for 10 minutes at 4 °C, transfer the supernatant to a new tube and dry it under a nitrogen atmosphere. The dried sample is redissolved in 50 μL of internal standard working solution and then analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC).

[0263] The HPLC analysis conditions were as follows: LabSolutions CS system was used; the column was a Waters C18 (4.6×150mm, 5μm); the flow rate was 1mL / min; the column temperature was 35℃; the circulating phase was methanol (A) and anhydrous ethanol (B), with isogradient elution of A:B at 50:50; and the detection wavelength was 275nm.

[0264] Table 5. Plasma Concentrations of Different Forms of Q10 - Timeline

[0265]

[0266] Table 6. Plasma pharmacokinetic parameters of male and female SD rats after oral administration of 15 mg / kg Q10 (free Q10, Q10-Lipo-Cho, and Q10-Lipo-Rx, respectively): expressed as mean ± standard deviation (n=6).

[0267]

[0268] C max Maximum plasma concentration;

[0269] T max Time to reach maximum plasma concentration;

[0270] t 1 / 2 :half life;

[0271] AUC 0-t The area under the curve from time 0 to the best time point;

[0272] AUC 0-∞ The area under the curve from time 0 to time infinity;

[0273] CL / F: Clearance rate, the amount of drug cleared from the body per unit time;

[0274] V / F: Apparent volume of distribution, a constant ratio of the amount of drug in the body to the blood concentration when the drug reaches dynamic equilibrium in the body;

[0275] MRT 0-t : Indicates the dwell time from time point 0 to the last time point;

[0276] MRT 0-∞ The average dwell time from time 0 to time infinity;

[0277] There was no significant difference in ns between the free Q10 group and the Q10-Lipo-Cho group;

[0278] The significance level between the free Q10 group and the Q10-Lipo-Rx group was less than 0.001.

[0279] The significance level between the Q10-Lipo-Cho and Q10-Lipo-Rx groups was less than 0.01.

[0280] * The significance level between the free Q10 group and the Q10-Lipo-Rx group was less than 0.0001;

[0281] #, the significance level between the Q10-Lipo-Cho and Q10-Lipo-Rx groups was less than 0.001;

[0282] The significance level between the free Q10 group and the Q10-Lipo-Cho group was less than 0.05.

[0283] The in vivo metabolism of Q10-Lipo-Rx was investigated by oral gavage of free Q10, Q10-Lipo-Cho, and Q10-Lipo-Rx at a dose of 15 mg / kg. As shown in Table 5, the free drug reached peak plasma concentration at 1 hour after oral administration, while Q10-Lipo-Cho and Q10-Lipo-Rx reached peak plasma concentration at 2 hours. Liposomes prolong the drug's residence time in the small intestine, promoting drug absorption and resulting in a delayed peak concentration for Q10, significantly increasing the peak plasma concentration. The peak plasma concentration in the Q10-Lipo-Rx group was 2.3 times that of free Q10 and 1.9 times that of Q10-Lipo-Cho, significantly improving intestinal absorption. The in vivo half-life of the Q10-Lipo-Rx group was 2.3 times that of the Q10 group and 1.8 times that of the Q10-Lipo-Cho group, showing a significant difference. The area under the pharmacokinetic curve (AUC0-∞) of Q10-Lipo-Rx was 3.3 times that of the free Q10 group and 2.6 times that of the Q10-Lipo-Cho group. This may be because conventional nano-formulations, after entering the body, adsorbed plasma proteins on their surface are recognized and taken up by the reticuloendothelial system, leading to faster clearance from the circulatory system. Compared to free Q10 and Q10-Lipo-Cho, the hydrophilic glycosyl coating on the Q10-Lipo-Rx lipid membrane surface reduces protein adsorption, thus prolonging its residence time in the bloodstream. These results indicate that Rx-containing Q10 liposomes significantly improve the absorption rate of coenzyme Q10 compared to traditional cholesterol liposomes, further prolonging Q10 circulation time and improving bioavailability.

[0284] As shown in Table 6, the PK parameters in female rats followed the same trend as in male rats. These results indicate that Rx-containing liposomes exhibit a lower sex-related effect compared to conventional cholesterol liposomes, suggesting that Rx-encapsulated Q10 liposomes have greater potential for clinical applications.

Claims

1. A ginsenoside coenzyme Q10 liposome, comprising the following components in parts by mass: 1 part coenzyme Q10, 0.3-0.5 parts ginsenoside Rx, 1-3 parts sunflower lecithin and 1.5-3 parts maltodextrin, wherein the mass ratio of sunflower lecithin to maltodextrin is 1:(1-3). in, The ginsenoside Rx is a mixture of ginsenosides, which includes 20(S)-ginsenoside Rg3, ginsenoside Rg5, and ginsenoside Rk1; The ginsenoside coenzyme Q10 liposomes described herein do not contain cholesterol.

2. The liposomes as described in claim 1, characterized in that, It satisfies one or more of the following conditions: (1) The ginsenoside Rx mentioned above is 0.3 parts, 0.4 parts or 0.5 parts; (2) The sunflower lecithin mentioned above is 1 part, 2 parts or 3 parts; (3) The maltodextrin is 1.5 parts, 2 parts or 3 parts; (4) The mass ratio of the coenzyme Q10 to the sunflower lecithin is 1:(1-3), for example 1:1, 1:2 or 1:3; (5) The mass ratio of coenzyme Q10 to ginsenoside Rx is 1:(0.3-0.5); for example, 1:0.3, 1:0.4 or 1:0.5; (6) The mass ratio of the maltodextrin to the sunflower lecithin is (1-1.5):1; for example, 1:1 or 1.5:1; (7) The sunflower lecithin mentioned above is sunflower lecithin with a content of 60% or more, for example, it is composed of 60% sunflower lecithin and 40% fatty acids.

3. The liposomes as described in claim 1, characterized in that, It satisfies one or more of the following conditions: (1) The ginsenoside Rx is derived from the stems and leaves of American ginseng, preferably an extract of the stems and leaves of American ginseng; the extract of the stems and leaves of American ginseng is preferably an alcohol extract of the extract of the stems and leaves of American ginseng, such as n-butanol extract; (2) The total content of each ginsenoside in the ginsenoside mixture is about 90-95%; wherein, the content of 20(S)-ginsenoside Rg3 is 20-25%; the content of ginsenoside Rg5 is 45-50%; the content of ginsenoside Rk1 is 20-25%, and the content of 20(R)-ginsenoside Rg3 in the ginsenoside mixture is less than 2%; (3) The mass ratio of 20(S)-ginsenoside Rg3 to ginsenoside Rg5 can be 1:(1.5 to 2.5), for example 1:1.93 or 1:2; (4) The mass ratio of 20(S)-ginsenoside Rg3 to ginsenoside Rk1 can be 1:(0.5 to 1.5), for example 1:0.86 or 1:1; (5) In the ginsenoside coenzyme Q10 liposomes, the mass percentage of coenzyme Q10 is 10-30%, the mass percentage of ginsenoside Rx is 5-15%, the mass percentage of sunflower lecithin is 20-40%, and the mass percentage of maltodextrin is 35-50%. The mass percentages are the proportions of the mass of each component to the total mass of the raw materials. (6) The purity of (Rg3+Rg5+Rk1) in the ginsenoside Rx is ≥90% by HPLC; for example, 90%-95%.

4. The liposomes as described in claim 1, characterized in that, The ginsenoside Rx was prepared by the following steps: Step 1: Mix the American ginseng stem and leaf extract with a citric acid aqueous solution and heat to obtain mixed solution 1; Step 2: Filter, extract, wash, and concentrate the mixed solution 1 to obtain solid 2; Step 3: Redissolve solid 2, filter at low temperature, and concentrate to obtain the product ginsenoside mixture Rx; The definition of American ginseng stem and leaf extract is as described in the above-mentioned ginsenoside coenzyme Q10 liposomes; Preferably, in step 1, the citric acid aqueous solution is a 20% citric acid aqueous solution; In step 1, the heating temperature is the temperature at which the water is heated to reflux. In step 2, the extraction is performed using an organic solvent, preferably an alcohol solvent, and more preferably n-butanol; the number of extractions is adjusted according to experimental needs, preferably 3-4 times, for example 3 times. In step 2, the cleaning is performed using an inorganic solution, such as one or more of a 10% NaOH aqueous solution, saturated saline solution, or purified water; the number of cleaning cycles can be 3, for example, one alkali wash with a 10% NaOH aqueous solution, one salt wash with 200L of saturated saline solution, and one water wash with 200L of purified water. In step 2, the concentration can be vacuum concentration; the concentration can be carried out until dry. In step 3, the redissolution is performed using an organic solution, which is an alcohol solution, preferably anhydrous ethanol. In step 3, the temperature of the low-temperature filtration is 0-15℃, for example, 2-8℃; In step 3, the concentration can be carried out under reduced pressure; the concentration is carried out until all the ethanol has completely evaporated.

5. The liposomes as described in claim 1, characterized in that, The ginsenoside coenzyme Q10 liposomes comprise any of the following components by mass fraction: (1) 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (2) 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (3) 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (4) 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (5) 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (6) 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (7) 1 part Q10, 0.3 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (8) 1 part Q10, 0.4 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (9) 1 part Q10, 0.5 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (10) 1 part Q10, 0.3 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin; (11) 1 part Q10, 0.4 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin; (12) 1 part Q10, 0.5 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin; Alternatively, the formulation of the ginsenoside coenzyme Q10 liposomes may consist of the following components in mass fractions: (1) 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (2) 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (3) 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 1.5 parts maltodextrin; (4) 1 part Q10, 0.3 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (5) 1 part Q10, 0.4 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (6) 1 part Q10, 0.5 parts Rx, 1 part sunflower lecithin and 2 parts maltodextrin; (7) 1 part Q10, 0.3 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (8) 1 part Q10, 0.4 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (9) 1 part Q10, 0.5 parts Rx, 2 parts sunflower lecithin and 2 parts maltodextrin; (10) 1 part Q10, 0.3 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin; (11) 1 part Q10, 0.4 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin; (12) 1 part Q10, 0.5 parts Rx, 3 parts sunflower lecithin and 3 parts maltodextrin.

6. The liposomes according to claim 1, characterized in that, The formulation of the ginsenoside coenzyme Q10 liposomes includes any of the following components by mass fraction: (1) 26.32% Q10, 7.89% Rx, 26.32% sunflower lecithin, 39.47% maltodextrin; (2) 25.64% Q10, 10.26% Rx, 25.64% sunflower lecithin, 38.46% maltodextrin; (3) 25.0% Q10, 12.5% ​​Rx, 25.0% sunflower lecithin, 37.5% maltodextrin; (4) 23.28% Q10, 6.98% Rx, 23.28% sunflower lecithin, 46.46% maltodextrin; (5) 22.73% Q10, 9.09% Rx, 22.73% sunflower lecithin, 45.45% maltodextrin; (6) 22.25% Q10, 11.11% Rx, 22.25% sunflower lecithin, 44.39% maltodextrin; (7) 18.86% Q10, 5.66% Rx, 37.74% sunflower lecithin, 37.74% maltodextrin; (8) 18.52% Q10, 7.40% Rx, 37.04% sunflower lecithin, 37.04% maltodextrin; (9) 18.19% Q10, 9.09% Rx, 36.36% sunflower lecithin, 36.36% maltodextrin; (10) 13.70% Q10, 4.10% Rx, 41.10% sunflower lecithin, 41.10% maltodextrin; (11) 13.51% Q10, 5.41% Rx, 40.54% sunflower lecithin, 40.54% maltodextrin; (12) 13.33% Q10, 6.67% Rx, 40.0% sunflower lecithin, 40.0% maltodextrin; Alternatively, the formulation of the ginsenoside coenzyme Q10 liposomes may be the components in the following mass fractions as described in any of the following schemes: (1) 26.32% Q10, 7.89% Rx, 26.32% sunflower lecithin, 39.47% maltodextrin; (2) 25.64% Q10, 10.26% Rx, 25.64% sunflower lecithin, 38.46% maltodextrin; (3) 25.0% Q10, 12.5% ​​Rx, 25.0% sunflower lecithin, 37.5% maltodextrin; (4) 23.28% Q10, 6.98% Rx, 23.28% sunflower lecithin, 46.46% maltodextrin; (5) 22.73% Q10, 9.09% Rx, 22.73% sunflower lecithin, 45.45% maltodextrin; (6) 22.25% Q10, 11.11% Rx, 22.25% sunflower lecithin, 44.39% maltodextrin; (7) 18.86% Q10, 5.66% Rx, 37.74% sunflower lecithin, 37.74% maltodextrin; (8) 18.52% Q10, 7.40% Rx, 37.04% sunflower lecithin, 37.04% maltodextrin; (9) 18.19% Q10, 9.09% Rx, 36.36% sunflower lecithin, 36.36% maltodextrin; (10) 13.70% Q10, 4.10% Rx, 41.10% sunflower lecithin, 41.10% maltodextrin; (11) 13.51% Q10, 5.41% Rx, 40.54% sunflower lecithin, 40.54% maltodextrin; (12) 13.33% Q10, 6.67% Rx, 40.0% sunflower lecithin, 40.0% maltodextrin.

7. A method for preparing ginsenoside coenzyme Q10 liposomes, comprising the following steps: Step 1: Dissolve coenzyme Q10, ginsenoside Rx and sunflower lecithin in ethanol to obtain solution A1, and concentrate it under reduced pressure to form a film; Step 2: The membrane obtained in Step 1 is hydrated by keeping it warm in an aqueous solution containing maltodextrin to obtain liposome solution A2. in, The definitions of coenzyme Q10, ginsenoside Rx, sunflower lecithin, maltodextrin, and their proportions are the same as those in the ginsenoside coenzyme Q10 liposomes as described in any one of claims 1-6; Preferably, it satisfies one or more of the following conditions: (1) The ethanol content is ≥70%, where the mass content is the percentage of the component's mass in the total mass of the raw material; (2) The amount of ethanol used is sufficient to dissolve coenzyme Q10 ginsenosides and sunflower lecithin; for example, the mass-volume ratio of coenzyme Q10 to ethanol is 1g / 3-15mL, such as 1g / 5mL. (3) In step 1, the solution A1 is obtained by heating and dissolving coenzyme Q10, ginsenosides, sunflower lecithin, etc. in ethanol; (4) The heating temperature is 35-65℃, for example 40-45℃ or 55-60℃; preferably 55℃; (5) In step 1, the concentration is vacuum concentration; (6) In step 1, the concentration is carried out under vacuum of -0.04 MPa to -0.1 MPa; preferably -0.04 to -0.05 MPa. (7) In step 1, the concentration is complete until all the ethanol has evaporated. (8) In step 1, the concentration is carried out in a rotary evaporator flask, and the rotation speed can be 40-60 rp / min, for example 50 rp / min; (9) In step 2, the concentration of the aqueous solution containing maltodextrin is 100-300 mg / mL, for example 100 mg / mL; (10) In step 2, the hydration temperature is 35-65℃, for example 45-50℃; (11) In step 2, the hydration is carried out in a rotary evaporator at a speed of 40 to 70 rp / min, for example 50 rp / min or 65 rp / min. (12) In step 2, the hydration is sufficient to achieve a homogeneous solution; (13) In step 2, the coenzyme Q10: maltodextrin solution = 1g: 10-50mL, for example 1g / 15mL or 1g / 30mL.

8. The method for preparing ginsenoside coenzyme Q10 liposomes as described in claim 7, characterized in that, It also includes the following steps: Step 3: The liposome solution A2 obtained in Step 2 is subjected to high-pressure homogenization to obtain a solution A3 containing the liposomes; And / or, in step 4, spray-dry the liposome solution A3 to obtain ginsenoside coenzyme Q10 liposomes; In step 3, the high-pressure homogenization is performed using a 0-10°C chilled water cooling cycle in a homogenizer; preferably, the temperature of the liposome solution is maintained at 5-10°C. In step 3, the pressure of the high-pressure homogenization is between 800-1600 bar, for example, 1200 bar; In step 3, the high-pressure homogenization is performed 3-4 times, for example, 4 times; In the preparation method described above, the conditions and operations for spray drying can be conventional conditions and operations used in this type of process in the art; preferably as follows: In step 4, the spray drying process has an inlet air temperature of 165-185℃, for example, 165℃, 170℃, 175℃ or 180℃, 185℃; and an outlet air temperature of 70-90℃, for example, 70℃, 75℃, 80℃ or 85℃. The spray flow rate should be determined based on the amount of powder solid obtained after spraying. For example, when the machine model is PLG-30, the flow rate is 5-30L / h, such as 5L / h, 10L / h, 15L / h, 20L / h, 25L / h, and 30L / h. The atomizer speed for the spray is 6000-18000 rpm, for example, 6000 rpm, 12000 rpm, 15000 rpm or 18000 rpm; Preferably, the flow rate of the spray increases as the inlet air temperature increases; In step 4, the spray drying satisfies any of the following conditions: Option 1: The air inlet temperature is 165℃, the flow rate is 5L / h, and the atomizer speed is 18000rp / min; Option 2: The inlet air temperature is 165℃, the flow rate is 10L / h, and the atomizer speed is 18000rp / min; Option 3: The air inlet temperature is 170℃, the flow rate is 15L / h, and the atomizer speed is 18000rp / min; Option 4: The inlet air temperature is 175℃, the flow rate is 20L / h, and the atomizer speed is 18000rp / min; Option 5: The air inlet temperature is 180℃, the flow rate is 25L / h, and the atomizer speed is 18000rp / min; Option Six: The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 18000rp / min; Option Seven: The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 12000rp / min; Option 8: The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 6000rp / min; Option Nine: The air inlet temperature is 180℃, the flow rate is 30L / h, and the atomizer speed is 15000rp / min; Option 10: The inlet air temperature is 185℃, the flow rate is 30L / h, and the atomizer speed is 15000rp / min.

9. A ginsenoside coenzyme Q10 liposome as described in any one of claims 1-6, which is prepared by the method for preparing ginsenoside coenzyme Q10 liposomes as described in claim 7 or 8.

10. The use of a ginsenoside coenzyme Q10 liposome in the preparation of drugs or health foods for the treatment and / or prevention of cardiovascular and cerebrovascular diseases; wherein the ginsenoside coenzyme Q10 liposome is the ginsenoside coenzyme Q10 as described in any one of claims 1-6.

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