A Ginsenoside Rh2 Composition, Its Preparation Method and Application

Abstract

The present invention relates to the field of nano-drug technology, and specifically discloses a ginsenoside Rh2 composition, a preparation method thereof and an application. The composition of the present invention comprises ginsenoside Rh2, oleic acid / linoleic acid, and an amphiphilic auxiliary material, and is prepared into nanoparticles with a particle size of 50-1000 nm by an anti-solvent precipitation method, which can effectively solve the problem of poor solubility and difficult administration of ginsenoside Rh2, and at the same time significantly improve the bioavailability of ginsenoside Rh2.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, relates to a nano-drug, and particularly relates to a nano-form of ginsenoside Rh2 composition with a high drug loading amount, a preparation method thereof, and an application thereof. Background Art

[0002] Ginseng is a perennial herbaceous plant, sweet and slightly bitter in taste, warm and flat in nature. It has the effects of greatly tonifying primordial qi, restoring pulse and arresting collapse, invigorating the spleen and benefiting the lung, promoting fluid production, calming the nerves, etc. Among the various chemical components included in ginseng, such as saponins, amino acids and polypeptides, volatile oils, sugars, trace elements and other components, ginsenosides are one of the most important active components.

[0003] Among them, ginsenoside Rh2 (G-Rh2) has been widely studied due to its outstanding anti-tumor activity. (20S)G-Rh2 has a more excellent anti-tumor effect compared with (20R)G-Rh2. It can directly kill tumor cells, inhibit the proliferation of tumor cells, induce apoptosis of tumor cells, and has obvious effects on the metastasis and invasion of tumor cells. Research reports show that G-Rh2 inhibits cancer cells by inducing apoptosis of various types of cancer cells through multiple pathways; Chen et al. obtained through microarray result analysis that ginsenoside Rh2 inhibits liver cancer by regulating the differential expression of IncRNAs (long non-coding RNAs) and mRNAs. In addition, ginsenoside Rh2 has a protective effect on myocardial damage, can enhance the immune function of the body, and also has pharmacological effects such as anti-allergy and anti-asthma. Its structure is as follows:

[0004]

[0005] Ginsenoside Rh2 belongs to the secondary glycoside of ginseng and has a very low content. At present, it is mostly obtained by methods such as degradation and transformation of ginsenosides with a relatively high content. Ginsenoside Rh2 mainly exerts its anti-tumor effect by enhancing the immune function, inhibiting the occurrence, development and metastasis of tumors, inducing apoptosis of tumor cells and the differentiation of cancer cells, changing the drug resistance of tumor cells to anti-cancer drugs and enhancing the efficacy of anti-cancer drugs, and can also prevent or reduce certain toxic and side effects caused by chemotherapy in common cancer treatment methods, etc.

[0006] A large number of basic and clinical studies have shown that ginsenoside Rh2-containing has broad application prospects in tumor treatment, but traditional preparations still have many problems at present, such as relatively large particle size, insoluble in water, low bioavailability, etc., which limit its clinical application. If new ginsenoside-containing traditional Chinese medicines can be prepared by improving traditional preparation processes and adopting modern technologies, so as to improve their bioavailability, reduce the drug dosage, reduce toxic and side effects and improve the curative effect, it will have important clinical significance. Summary of the Invention

[0007] In view of the above, the object of the present invention is to provide a ginsenoside Rh2 composition, which is a composition that can improve the oral bioavailability of ginsenoside Rh2.

[0008] It should be noted that ginsenoside Rh2 is a poorly soluble component and an active ingredient that has received much attention. However, different from conventional poorly soluble drugs, the preparation of ginsenoside nanoparticles is extremely difficult. So far, the research on the nano-delivery system of ginsenoside Rh2 is extremely limited. There are only sub-microemulsions, self-microemulsions, lipid nanoparticles (using EPC, PLGA, DSPE-PEG as excipients, drug loading of 7.2%), nanoparticles prepared with chitosan and sodium tripolyphosphate as excipients with a drug loading of 7.89%; Rh2 micelles with PLGA-PEG as excipients, and the drug loading is <4% under the optimized conditions.

[0009] The research on ginsenoside Rh2 liposomes is relatively more. So far, the liposome has the highest drug loading, and its maximum drug loading is only 23%. The preparation of liposomes requires the use of a relatively large amount of phospholipids, and the cost of phospholipids is high; at the same time, the chemical stability of phospholipids is poor, and they are prone to oxidation during transportation and storage, which also has an adverse impact on the stability of the product. A series of measures taken to avoid or delay the oxidation of phospholipids will further increase the cost of the product. Therefore, in order to truly achieve clinical application, it is necessary to prepare Rh2 nano-drugs with higher drug loading using more stable excipients.

[0010] Among all nano-delivery systems, the drug loadings of nano-suspensions and nanocrystals are the highest. It seems logical to prepare nanocrystals or nano-suspensions of Rh2. However, ginsenoside Rh2 is rather special. The present invention has tried various excipients as stabilizers to prepare nano-suspensions of Rh2 by the most economical grinding method (top-down method), and set the drug-to-carrier ratio at 1:2 and above, but all ended in failure without exception. Either the particle size was extremely large or the stability was poor (irreversible precipitation aggregation occurred during storage); later, the anti-solvent precipitation method was tried to screen nano-suspensions that could prepare Rh2 nanoparticles with good stability at a drug-to-carrier ratio of 1:2, but the results were also unsuccessful. It shows that it is impossible to successfully prepare nano-suspensions of Rh2 with conventional methods and conventional excipients, and a new approach is needed to obtain a nano-composition of Rh2 with a high drug loading.

[0011] During the preliminary exploration of the present invention, it was unexpectedly found that after mixing oleic acid into the ethanol solution of Rh2 and using TPGS as a stabilizer, nanoparticles similar to the nano-suspension of ginsenoside Rh2 could be successfully prepared, which were stable when placed and stable in artificial gastrointestinal fluids, and had a drug loading as high as 40%. When oleic acid was replaced with linoleic acid, and mixtures of different ratios of oleic acid and linoleic acid, they still had excellent stability and high drug loading. However, when oleic acid was replaced with soybean oil, corn oil, or olive oil, stable Rh2 nanoparticles could not be successfully prepared, suggesting that there may be a certain specific interaction between oleic acid / linoleic acid and Rh2, which can help Rh2 complete assembly under the stabilizing effect of TPGS to form high-drug-loading nanoparticles similar to the nano-suspension; and when TPGS was replaced with other amphiphilic excipients, it was found that Rh2 nanoparticles could basically be prepared.

[0012] In order to achieve the object of the present invention, based on a large amount of research, the present invention adopts the following technical solutions:

[0013] The first technical object of the present invention is to provide a ginsenoside Rh2 composition, which includes ginsenoside Rh2, oleic acid / linoleic acid, and an amphiphilic excipient; wherein,

[0014] The weight ratio of ginsenoside Rh2, oleic acid / linoleic acid, and the amphiphilic excipient is 4:2 - 8:1 - 8.

[0015] Optionally, the composition includes pharmaceutically acceptable excipients, and the excipients include excipients for adjusting osmotic pressure and pH, specifically including glucose, sodium chloride, phosphate buffer solution, and glycerol.

[0016] Furthermore, the oleic acid / linoleic acid is a mixed solution of oleic acid and linoleic acid in any ratio, and the amphiphilic excipient includes one or more combinations of TPGS, PLA-PEG, PLGA-PEG, PCL-PEG, DSPE-PEG, phospholipids, and sodium oleate.

[0017] It should be noted that the average particle size of the ginsenoside Rh2 composition is 50 - 1000 nm. The presence of oleic acid / linoleic acid is essential for the formation of the ginsenoside Rh2 nano-composition. The ginsenoside Rh2 composition can effectively solve the problem of poor solubility and difficult administration of ginsenoside Rh2, and can significantly improve the bioavailability of ginsenoside Rh2 by oral administration. Incorporating borneol into the composition can further improve the oral bioavailability.

[0018] Moreover, the ginsenoside Rh2 composition can basically maintain its original particle size (particle size change < 100 nm) in gastrointestinal fluids and can be orally administered in the form of nanoparticles; in physiological saline, PBS, and 5% glucose, the particle size is basically stable (particle size change < 100 nm) and can be intravenously injected in the form of nanoparticles.

[0019] The second technical object of the present invention is to provide a preparation method of the ginsenoside Rh2 composition as described above, and the method comprises the following steps:

[0020] Dissolve ginsenoside Rh2, amphiphilic excipient, oleic acid / linoleic acid in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution into water, and evaporate the organic solvent under reduced pressure to finally obtain the ginsenoside Rh2 composition.

[0021] Or, the preparation method of the ginsenoside Rh2 composition, and the method comprises the following steps:

[0022] Dissolve ginsenoside Rh2, oleic acid / linoleic acid, amphiphilic excipient in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution into an aqueous solution containing sodium oleate, and evaporate the organic solvent under reduced pressure to finally obtain the ginsenoside Rh2 composition.

[0023] Optionally, the organic solvent includes one or more combinations of ethanol, methanol, and acetone, the volume of water is 3-50 times that of the organic solvent, the stirring speed is 300-2000 rpm, and the water bath ultrasonic power is 100-500 W.

[0024] Optionally, for the aqueous dispersion system prepared by the method, after adding 1% mannitol and mixing evenly, freeze-dry to obtain the solid powder of the ginsenoside Rh2 composition, which is convenient for long-term storage.

[0025] After adding mannitol to the aqueous nano-dispersion system and then freeze-drying, the solid powder of the composition is further obtained, which can be filled into capsules for oral administration or packed as a granule. Before use, add water and stir evenly before taking.

[0026] The third technical object of the present invention is to provide an application of the ginsenoside Rh2 composition as described above in pharmaceutical preparations.

[0027] The ginsenoside Rh2 composition disclosed by the present invention can be used as an active ingredient together with pharmaceutically acceptable excipients to prepare a pharmaceutical composition, and the pharmaceutical composition can be prepared into various dosage forms by conventional methods in the pharmaceutical field, such as tablets, powders, granules, pills, capsules, tinctures, oral liquids, ointments, creams, emulsions or patches, etc. According to the different dosage forms, the excipients used in the pharmaceutical composition are also different, and the common excipients include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, emulsifiers, and osmotic pressure regulators.

[0028] Moreover, the ginsenoside Rh2 nano-composition disclosed by the present invention is used for the treatment and prevention of tumors.

[0029] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0030] In the present invention, ginsenoside Rh2, oleic acid / linoleic acid, and amphiphilic excipients are dissolved in an appropriate amount of water-miscible organic solvents such as ethanol, and then added to the aqueous phase under stirring or ultrasonic action. Then, the organic solvent is removed by reduced pressure evaporation to obtain a nano-composition with an average particle size of 50-1000 nm. The drug loading can easily reach 30%, and the optimized prescription drug loading is 39.6%. It can be directly administered orally or by intravenous injection. The preparation process of this composition is simple. Compared with the physical suspension of Rh2, the oral bioavailability in rats is increased by more than 2.5 times. After adding a small amount of borneol to the prescription, the bioavailability is increased to 5.1 times. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained according to the provided drawings without creative efforts.

[0032] Figure 1 It is the particle size diagram of the Rh2 composition prepared in Example 2.

[0033] Figure 2 It is the change diagram of the particle size and particle size distribution of the Rh2 composition prepared in Example 2 during storage at room temperature for one week.

[0034] Figure 3 It is the particle size diagram of the Rh2 composition prepared in Example 3.

[0035] Figure 4 It is the change diagram of the particle size and particle size distribution of the Rh2 composition prepared in Example 3 during storage at room temperature for one week.

[0036] Figure 5 It is the change diagram of the particle size of the Rh2 composition prepared in Example 3 during incubation in different physiological media at 37°C.

[0037] Figure 6 It is the morphological photograph of the Rh2 composition prepared in Example 3 under transmission electron microscopy.

[0038] Figure 7 It is the particle size diagram of the Rh2 composition prepared in Example 4.

[0039] Figure 8 It is the change diagram of the particle size and particle size distribution of the Rh2 composition prepared in Example 5 during storage at room temperature for one week.

[0040] Figure 9 It is the particle size diagram of the Rh2 composition prepared in Example 8.

[0041] Figure 10 It is the particle size change diagram of the Rh2 composition prepared in Example 47 when incubated in different physiological media at 37 °C.

[0042] Figure 11 It is the particle size and particle size distribution change diagram of the Rh2 composition prepared in Example 47 during the two-week storage at room temperature. Detailed implementation manners

[0043] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present invention.

[0044] The special term "embodiment" used here does not necessarily mean that any embodiment described as "exemplary" is better than or superior to other embodiments. For the performance index tests in the embodiments of the present application, unless otherwise specified, the conventional test methods in the art are adopted. It should be understood that the terms described in the present application are only used to describe specific embodiments and are not intended to limit the content disclosed in the present application.

[0045] Unless otherwise specified, the technical and scientific terms used herein have the same meanings as those commonly understood by those of ordinary skill in the technical field to which this application belongs; other test methods and technical means not specifically noted in this application refer to the experimental methods and technical means commonly adopted by those of ordinary skill in the art.

[0046] To better illustrate the content of this application, numerous specific details are given in the following specific embodiments. Those skilled in the art should understand that this application can still be implemented without certain specific details. In the embodiments, some methods, means, instruments, equipment, etc. well-known to those skilled in the art are not described in detail in order to highlight the gist of this application.

[0047] On the premise of no conflict, the technical features disclosed in the embodiments of this application can be combined arbitrarily, and the obtained technical solutions belong to the content disclosed in the embodiments of this application.

[0048] To better understand the present invention, the present invention will be further specifically described below through the following embodiments, but it should not be understood as a limitation of the present invention. For some non-essential improvements and adjustments made by those skilled in the art according to the above-mentioned invention content, they are also considered to fall within the protection scope of the present invention.

[0049] Example 1: Rh2:OA:TPGS = 2:1:2, Rh2 = 2 mg / mL, theoretical drug loading 40%

[0050] Method: Weigh 8 mg of Rh2, 4 mg of oleic acid OA, and 8 mg of TPGS and dissolve them in 0.6 mL of ethanol. Under a 100 W water bath ultrasonic bath, it is dropped into 4 mL of water, and the ethanol is removed by evaporation under reduced pressure at 45 °C.

[0051] Result: The average particle size is (209.0 ± 11.9) nm, and the PDI value is 0.210 ± 0.029

[0052] Example 2: Rh2:OA:TPGS = 2:2:1, Rh2 = 2.5 mg / mL, theoretical drug loading 40%

[0053] Method: Weigh 10 mg of Rh2, 10 mg of oleic acid OA, and 5 mg of TPGS and dissolve them in 0.5 mL of ethanol. Under a 250 W water bath ultrasonic bath, it is dropped into 4 mL of water, and the ethanol is removed by evaporation under reduced pressure at 45 °C.

[0054] Result: The particle size is (143.5 ± 1.709) nm, and the PDI value is 0.199 ± 0.012

[0055] Moreover, after standing at room temperature for one week, there is no obvious change in the particle size and particle size distribution, indicating good storage stability.

[0056] Example 3: Rh2:OA:TPGS = 2:2:1, Rh2 = 5 mg / mL, theoretical drug loading 40%

[0057] Method: Weigh 15 mg of Rh2, 15 mg of oleic acid, and 7.5 mg of TPGS and dissolve them in 0.6 mL of ethanol. Under a 250 W water bath ultrasonic bath, it is dropped into 3 mL of water, and the ethanol is removed by evaporation under reduced pressure at 45 °C.

[0058] Result: The average particle size is (164.3 ± 2.203) nm, the PDI value is 0.177 ± 0.015, and the measured drug loading is 39.7%.

[0059] Moreover, after standing at room temperature for one week, there is no obvious change in the particle size and particle size distribution, indicating good storage stability.

[0060] Example 4: Rh2:OA:TPGS = 2:2:1, Rh2 = 8 mg / mL, theoretical drug loading 40%

[0061] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid, and 8 mg of TPGS and dissolve them in 0.6 mL of ethanol. Under a 250 W water bath ultrasonic bath, it is dropped into 2 mL of water, and the ethanol is removed by evaporation under reduced pressure at 45 °C.

[0062] Results: The average particle size was (244.3 ± 5.776) nm, the PDI value was 0.148 ± 0.024, the measured drug loading was 39.8%, and there was no obvious change in the particle size and particle size distribution after storing at room temperature for one week, indicating good storage stability.

[0063] Example 5 Rh2:OA:TPGS = 2:2:1, Rh2 = 12 mg / mL, theoretical drug loading 40%

[0064] Method: Weigh 24 mg of Rh2, 24 mg of oleic acid, and 12 mg of TPGS, dissolve them in 0.6 ml of ethanol, add dropwise to 2 ml of water under 300 W ultrasonic treatment, and evaporate ethanol under reduced pressure at 45 °C.

[0065] Results: The average particle size was (238.6 ± 6.950) nm, the PDI value was 0.208 ± 0.021, it was stable when stored at room temperature, and the measured drug loading was 39.5%.

[0066] Investigation on physiological medium stability: Mix the prepared ginsenoside Rh2 nanoparticles with 2×PBS (PBS with twice the concentration), 1.8% NaCl solution, 10% glucose solution in equal volumes, or mix with 4 times the volume of artificial gastric juice and artificial intestinal juice, incubate at 37 °C, and measure the particle size and particle size distribution at specific time points to observe whether there is turbidity, aggregation or precipitation.

[0067] The results showed that it was stable in normal saline, PBS, 5% glucose, artificial gastric juice and artificial intestinal juice except that the particle size increased in fetal bovine serum FBS, and it was suitable for both intravenous injection and oral administration.

[0068] Transmission electron microscopy showed that the nano - composition presented spherical or less regular circular shapes.

[0069] Example 6 Rh2:OA:TPGS = 2:2:2, Rh2 = 15 mg / mL

[0070] Method: Weigh 30 mg of Rh2, 30 mg of oleic acid, and 30 mg of TPGS, dissolve them in 0.6 ml of ethanol, add dropwise to 2 ml of water under 300 W ultrasonic treatment, and evaporate ethanol under reduced pressure at 45 °C.

[0071] Results: The average particle size was (471.8 ± 13.13) nm, the PDI value was 0.423 ± 0.057.

[0072] After homogenization at 1500 bar for 10 times, the average particle size was (93.51 ± 4.576) nm, the PDI value was 0.169 ± 0.062.

[0073] Example 7 Rh2:OA:TPGS = 2:2:2, Rh2 = 20 mg / mL

[0074] Method: Weigh 40 mg of Rh2, 40 mg of oleic acid, and 40 mg of TPGS, dissolve them in 0.6 ml of ethanol, add the solution dropwise to 2 ml of water under 300 W ultrasonic treatment, and remove ethanol by reduced pressure evaporation at 45 °C.

[0075] Result: The average particle size is (536.1 ± 10.15) nm, and the PDI value is 0.399 ± 0.038.

[0076] After homogenization at 1500 bar for 15 times, the average particle size is (123.03 ± 4.354) nm, and the PDI value is 0.192 ± 0.078.

[0077] Example 8 Rh2:OA:TPGS = 2:2:4, Rh2 = 10 mg / mL

[0078] Method: Weigh 40 mg of Rh2, 40 mg of oleic acid, and 80 mg of TPGS, dissolve them in 0.8 ml of ethanol, add the solution dropwise to 4 ml of water under 250 W ultrasonic treatment, and remove ethanol by reduced pressure evaporation at 45 °C.

[0079] Result: The average particle size is (238.6 ± 8.5) nm, the PDI value is 0.179 ± 0.038, and no visible change is observed after standing at room temperature for one week.

[0080] Example 9 Rh2:OA:TPGS = 2:4:2, Rh2 = 5 mg / mL

[0081] Method: Weigh 40 mg of Rh2, 80 mg of oleic acid, and 40 mg of TPGS, dissolve them in 0.8 ml of ethanol, add the solution dropwise to 8 ml of water under 250 W ultrasonic treatment, and remove ethanol by reduced pressure evaporation at 45 °C.

[0082] Result: The average particle size is (247.7 ± 7.2) nm, the PDI value is 0.192 ± 0.057, and no visible change is observed after standing at room temperature for one week.

[0083] Example 10 Rh2:OA:TPGS = 4:4:1, Rh2 = 5 mg / mL, methanol

[0084] Method: Weigh 40 mg of Rh2, 40 mg of oleic acid, and 10 mg of TPGS, dissolve them in 0.8 ml of methanol, add the solution dropwise to 8 ml of water under 250 W ultrasonic treatment, and remove methanol by reduced pressure evaporation at 45 °C.

[0085] Result: The average particle size is (286.8 ± 9.5) nm, the PDI value is 0.242 ± 0.096, and no visible change is observed after standing at room temperature for one week.

[0086] Example 11 Rh2:OA:TPGS = 4:4:2, Rh2 = 5 mg / mL, acetone

[0087] Method: Weigh 20 mg of Rh2, 20 mg of oleic acid, and 10 mg of TPGS, dissolve them in 0.6 ml of acetone, and add the solution dropwise to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate acetone under reduced pressure at 45 °C.

[0088] Results: The average particle size is (226.7 ± 5.332) nm, the PDI value is 0.158 ± 0.035, and there is no visible change after standing at room temperature for one week.

[0089] Example 12 Rh2:LA:TPGS = 4:4:2, Rh2 = 5 mg / mL, acetone, linoleic acid

[0090] Method: Weigh 20 mg of Rh2, 20 mg of linoleic acid (LA), and 10 mg of TPGS, dissolve them in 0.6 ml of acetone, and add the solution dropwise to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate acetone under reduced pressure at 45 °C.

[0091] Results: The average particle size is (203.4 ± 4.897) nm, the PDI value is 0.166 ± 0.045, and there is no visible change after standing at room temperature for one week.

[0092] Example 13 Rh2:LA:TPGS = 4:4:2, Rh2 = 5 mg / mL, methanol, linoleic acid

[0093] Method: Weigh 20 mg of Rh2, 20 mg of linoleic acid (LA), and 10 mg of TPGS, dissolve them in 0.6 ml of methanol, and add the solution dropwise to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate methanol under reduced pressure at 45 °C.

[0094] Results: The average particle size is (235.8 ± 5.921) nm, the PDI value is 0.178 ± 0.063, and there is no visible change after standing at room temperature for one week.

[0095] Example 14 Rh2:oleic acid:TPGS:sodium oleate = 4:4:2:1, drug loading ratio 4:7, theoretical drug loading = 36.36% (Rh2 concentration is 5 mg / mL)

[0096] Method: Weigh 20 mg of ginsenoside Rh2 and 20 mg of oleic acid, dissolve them in 0.5 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate, dissolve them in 4 mL of water as the aqueous phase. Drop the organic phase into the 4 ml aqueous phase under ultrasonic treatment, and evaporate ethanol by rotary evaporation at 45 °C.

[0097] Results: The average particle size is (155.8 ± 1.343) nm, the PDI value is 0.205 ± 0.006, and the surface potential is -(52.3 ± 2.11) mV.

[0098] Particle size stability during storage at room temperature: The newly prepared ginsenoside Rh2 nanoparticles were stored at room temperature for 30 days. Samples were taken on days 0, 7, 14, and 30 to measure the particle size and particle size distribution. At the same time, careful observation was made for any turbidity, aggregation, or precipitation. As a result, after one month of storage at room temperature, the average particle size remained almost unchanged, and the PDI showed slight fluctuations within a very small range, indicating good stability of the prepared ginsenoside Rh2 nanoparticles during storage at room temperature.

[0099] Example 15 Rh2:OA:TPGS = 2:1:2, Rh2 = 2 mg / mL, stirring

[0100] Method: Weigh 8 mg of Rh2, 4 mg of oleic acid OA, and 8 mg of TPGS and dissolve them in 0.6 mL of ethanol. While stirring at 300 rpm, the solution was dropped into 4 mL of water. Ethanol was removed by evaporation under reduced pressure at 45 °C, and then high-pressure homogenization was carried out 5 times at 1500 bar.

[0101] Result: The average particle size was (226.0 ± 8.9) nm, and the PDI value was 0.221 ± 0.027.

[0102] Example 16 Rh2:OA:TPGS = 2:2:1, Rh2 = 2.5 mg / mL, stirring

[0103] Method: Weigh 10 mg of Rh2, 10 mg of oleic acid OA, and 5 mg of TPGS and dissolve them in 0.5 mL of ethanol. While stirring at 500 rpm, the solution was dropped into 4 mL of water. Ethanol was removed by evaporation under reduced pressure at 45 °C.

[0104] Result: The particle size was (190.1 ± 1.002) nm, and the PDI value was 0.178 ± 0.011.

[0105] Example 17 Rh2:OA:TPGS = 2:2:1, Rh2 = 5 mg / mL, stirring

[0106] Method: Weigh 15 mg of Rh2, 15 mg of oleic acid, and 7.5 mg of TPGS and dissolve them in 0.6 mL of ethanol. While stirring at 1000 rpm, the solution was dropped into 3 mL of water. Ethanol was removed by evaporation under reduced pressure at 45 °C.

[0107] Result: The average particle size was (171.2 ± 1.891) nm, the PDI value was 0.186 ± 0.018, and the measured drug loading was 39.3%.

[0108] Example 18 Rh2:OA:TPGS = 2:2:1, Rh2 = 8 mg / mL, stirring

[0109] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid, and 8 mg of TPGS and dissolve them in 0.6 mL of ethanol. While stirring at 1500 rpm, the solution was dropped into 2 mL of water. Ethanol was removed by evaporation under reduced pressure at 45 °C.

[0110] Results: The average particle size was (229.1 ± 4.582) nm, the PDI value was 0.153 ± 0.023, and the measured drug loading was 39.6%.

[0111] Example 19 Rh2:OA:TPGS = 2:2:1, Rh2 = 12 mg / mL, stirring

[0112] Method: Weigh 24 mg of Rh2, 24 mg of oleic acid, and 12 mg of TPGS and dissolve them in 0.6 ml of ethanol. While stirring at 1800 rpm, add the solution dropwise to 2 ml of water, and evaporate ethanol under reduced pressure at 45°C.

[0113] Results: The average particle size was (241.7 ± 6.214) nm, the PDI value was 0.213 ± 0.019, and the measured drug loading was 39.2%.

[0114] Example 20 Rh2:OA:TPGS = 2:2:2, Rh2 = 15 mg / mL, stirring

[0115] Method: Weigh 30 mg of Rh2, 30 mg of oleic acid, and 30 mg of TPGS and dissolve them in 0.6 ml of ethanol. While stirring at 2000 rpm, add the solution dropwise to 2 ml of water, evaporate ethanol under reduced pressure at 45°C, and measure the particle size.

[0116] Results: The average particle size was (538.8 ± 12.49) nm, the PDI value was 0.505 ± 0.061. After homogenization at 1500 bar for 10 times, the average particle size was (106.43 ± 3.869) nm, and the PDI value was 0.157 ± 0.052.

[0117] Example 21 Rh2:OA:TPGS = 2:2:2, Rh2 = 20 mg / mL, stirring

[0118] Method: Weigh 40 mg of Rh2, 40 mg of oleic acid, and 40 mg of TPGS and dissolve them in 0.6 ml of ethanol. While stirring at 2000 rpm, add the solution dropwise to 2 ml of water, and evaporate ethanol under reduced pressure at 45°C.

[0119] Results: The average particle size was (672.1 ± 15.82) nm, the PDI value was 0.471 ± 0.074. After homogenization at 1500 bar for 15 times, the average particle size was (158.93 ± 4.132) nm, and the PDI value was 0.201 ± 0.067.

[0120] Example 22 Rh2:OA:TPGS = 2:2:4, Rh2 = 10 mg / mL, stirring

[0121] Method: Weigh 40 mg of Rh2, 40 mg of oleic acid, and 80 mg of TPGS, dissolve them in 0.8 ml of ethanol, and dropwise add the solution to 4 ml of water under stirring at 1600 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0122] Result: The average particle size is (241.6 ± 6.4) nm, and the PDI value is 0.183 ± 0.041.

[0123] Example 23 Rh2: oleic acid: TPGS: sodium oleate = 4:4:2:1, stirring

[0124] Method: Weigh 20 mg of ginsenoside Rh2 and 20 mg of oleic acid, dissolve them in 0.5 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate, dissolve them in 4 mL of water as the aqueous phase. Dropwise add the organic phase to the 4 ml of aqueous phase under stirring at 1600 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0125] Result: The average particle size is (151.6 ± 3.4) nm, and the PDI value is 0.161 ± 0.025.

[0126] Example 24 Rh2: LA: TPGS = 2:1:2, Rh2 = 2 mg / mL, linoleic acid

[0127] Method: Weigh 8 mg of Rh2, 4 mg of linoleic acid LA, and 8 mg of TPGS, dissolve them in 0.6 mL of ethanol, and dropwise add the solution to 4 ml of water under stirring at 300 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0128] Result: The average particle size is (203.1 ± 5.4) nm, and the PDI value is 0.197 ± 0.023

[0129] Example 25 Rh2: LA: TPGS = 2:2:1, Rh2 = 2.5 mg / mL, linoleic acid

[0130] Method: Weigh 10 mg of Rh2, 10 mg of linoleic acid LA, and 5 mg of TPGS, dissolve them in 0.5 mL of ethanol, and dropwise add the solution to 4 ml of water under stirring at 500 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0131] Result: The particle size is (187.2 ± 1.023) nm, and the PDI value is 0.171 ± 0.012

[0132] Example 26 Rh2: LA: TPGS = 2:2:1, Rh2 = 5 mg / mL, linoleic acid

[0133] Method: Weigh 15 mg of Rh2, 15 mg of linoleic acid LA, and 7.5 mg of TPGS, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 3 ml of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0134] Results: The average particle size was (167.5 ± 2.673) nm, the PDI value was 0.175 ± 0.015, the measured drug loading was 39.5%, and no visible change was observed after storing at room temperature for one week.

[0135] Example 27 Rh2:LA:TPGS = 2:2:1, Rh2 = 8 mg / mL, linoleic acid

[0136] Method: Weigh 16 mg of Rh2, 16 mg of linoleic acid LA, and 8 mg of TPGS and dissolve them in 0.6 ml of ethanol. While stirring at 1500 rpm, add the solution dropwise to 2 ml of water, and evaporate the ethanol under reduced pressure at 45°C.

[0137] Results: The average particle size was (207.6 ± 3.893) nm, the PDI value was 0.161 ± 0.031, the measured drug loading was 39.2%, and no visible change was observed after storing at room temperature for one week.

[0138] Example 28 Rh2:LA:TPGS = 2:2:1, Rh2 = 12 mg / mL, linoleic acid

[0139] Method: Weigh 24 mg of Rh2, 24 mg of linoleic acid LA, and 12 mg of TPGS and dissolve them in 0.6 ml of ethanol. While stirring at 1800 rpm, add the solution dropwise to 2 ml of water, and evaporate the ethanol under reduced pressure at 45°C.

[0140] Results: The average particle size was (237.6 ± 5.860) nm, the PDI value was 0.201 ± 0.017, it was stable after storing at room temperature, and the measured drug loading was 39.6%.

[0141] Example 29 Rh2:LA:TPGS = 2:2:4, Rh2 = 10 mg / mL, linoleic acid

[0142] Method: Weigh 40 mg of Rh2, 40 mg of linoleic acid LA, and 80 mg of TPGS and dissolve them in 0.8 ml of ethanol. While under 250 W ultrasonic treatment, add the solution dropwise to 4 ml of water, and evaporate the ethanol under reduced pressure at 45°C.

[0143] Results: The average particle size was (252.1 ± 7.9) nm, the PDI value was 0.203 ± 0.043.

[0144] Example 30 Rh2:LA:TPGS:sodium oleate = 4:4:2:1, linoleic acid

[0145] Method: Weigh 20 mg of ginsenoside Rh2, 20 mg of linoleic acid LA and dissolve them in 0.8 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate and dissolve them in 4 mL of water as the aqueous phase. While under 250 W ultrasonic treatment, add the organic phase dropwise to the 4 ml of aqueous phase, and evaporate the ethanol by rotary evaporation at 45°C.

[0146] Results: The average particle size was (142.1 ± 3.0) nm, the PDI value was 0.157 ± 0.021, and no visible change was observed after storing at room temperature for one week.

[0147] Example 31 Rh2:LA:OA:TPGS = 2:1:1:1, Rh2 = 8 mg / mL, oleic acid + linoleic acid

[0148] Method: Weigh 16 mg of Rh2, 8 mg of oleic acid OA, 8 mg of linoleic acid LA, and 8 mg of TPGS, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 2 ml of water under stirring at 1500 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0149] Results: The average particle size was (198.5 ± 3.562) nm, the PDI value was 0.160 ± 0.029, the measured drug loading was 39.7%, and no visible change was observed after storing at room temperature for one week.

[0150] Example 32 Rh2:OA:LA:TPGS = 2:2:1, Rh2 = 12 mg / mL, oleic acid + linoleic acid

[0151] Method: Weigh 24 mg of Rh2, 12 mg of oleic acid OA, 12 mg of linoleic acid LA, and 12 mg of TPGS, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 2 ml of water under ultrasonic treatment at 250 W. Evaporate ethanol under reduced pressure at 45 °C.

[0152] Results: The average particle size was (218.9 ± 5.769) nm, the PDI value was 0.196 ± 0.015, the measured drug loading was 39.3%, and no visible change was observed after storing at room temperature for one week.

[0153] Example 33 Rh2:OA:LA:TPGS = 2:2:4, Rh2 = 10 mg / mL, oleic acid + linoleic acid

[0154] Method: Weigh 40 mg of Rh2, 20 mg of oleic acid OA, 20 mg of linoleic acid LA, and 80 mg of TPGS, dissolve them in 0.8 ml of ethanol, and dropwise add the solution to 4 ml of water under ultrasonic treatment at 250 W. Evaporate ethanol under reduced pressure at 45 °C.

[0155] Results: The average particle size was (235.9 ± 6.1) nm, the PDI value was 0.178 ± 0.038, and no visible change was observed after storing at room temperature for one week.

[0156] Example 34 Rh2:OA:LA:TPGS:sodium oleate = 4:2:2:2:1, oleic acid + linoleic acid

[0157] Method: Weigh 20 mg of ginsenoside Rh2, 10 mg of oleic acid (OA), and 10 mg of linoleic acid (LA), dissolve them in 0.6 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate, dissolve them in 4 mL of water as the aqueous phase. While stirring at 1600 rpm, add the organic phase dropwise to the 4 ml of aqueous phase, and remove ethanol by rotary evaporation at 45 °C.

[0158] Result: The average particle size is (160.8 ± 3.6) nm, the PDI value is 0.170 ± 0.029, and no visible change is observed after standing at room temperature for one week.

[0159] Example 35 Rh2:LA:OA:PLA2000 - PEG2000 = 2:1:1:1, Rh2 = 8 mg / mL, oleic acid + linoleic acid, PLA - PEG

[0160] Method: Weigh 16 mg of Rh2, 8 mg of oleic acid (OA), 8 mg of linoleic acid (LA), and 8 mg of PLA2000 - PEG2000, dissolve them in 0.6 ml of ethanol, and add the solution dropwise to 2 ml of water while stirring at 1500 rpm. Remove ethanol by evaporation under reduced pressure at 45 °C.

[0161] Result: The average particle size is (213.4 ± 4.572) nm, the PDI value is 0.176 ± 0.035, and no visible change is observed after standing at room temperature for one week.

[0162] Example 36 Rh2:OA:LA:PLA2000 - PEG2000 = 2:2:1, Rh2 = 12 mg / mL, oleic acid + linoleic acid, PLA - PEG

[0163] Method: Weigh 24 mg of Rh2, 12 mg of oleic acid (OA), 12 mg of linoleic acid (LA), and 12 mg of PLA2000 - PEG2000, dissolve them in 0.6 ml of ethanol, and add the solution dropwise to 2 ml of water under 250W ultrasonic treatment. Remove ethanol by evaporation under reduced pressure at 45 °C.

[0164] Result: The average particle size is (226.9 ± 6.231) nm, the PDI value is 0.208 ± 0.018, the measured drug loading is 39.2%, and no visible change is observed after standing at room temperature for one week.

[0165] Example 37 Rh2:OA:PLGA2000 - PEG2000 = 2:2:1, Rh2 = 8 mg / mL, oleic acid + linoleic acid, PLGA - PEG

[0166] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid (OA), and 8 mg of PLGA2000 - PEG2000, dissolve them in 0.6 ml of ethanol, and add the solution dropwise to 2 ml of water while stirring at 1500 rpm. Remove ethanol by evaporation under reduced pressure at 45 °C.

[0167] Results: The average particle size was (205.4 ± 4.218) nm, the PDI value was 0.171 ± 0.030, and there was no visible change after being placed at room temperature for one week.

[0168] Example 38 Rh2:LA:PLGA2000-PEG2000 = 2:2:1, Rh2 = 12 mg / mL, oleic acid + linoleic acid, PLGA-PEG

[0169] Method: Weigh 24 mg of Rh2, 24 mg of linoleic acid LA, and 12 mg of PLGA2000-PEG2000, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 2 ml of water under 250 W ultrasonic treatment. Then evaporate the ethanol under reduced pressure at 45°C.

[0170] Results: The average particle size was (216.7 ± 5.8652) nm, the PDI value was 0.198 ± 0.014, and there was no visible change after being placed for one week.

[0171] Example 39 Rh2:OA:DSPE-PEG2000 = 2:2:1, Rh2 = 8 mg / mL, oleic acid + linoleic acid, DSPE-PEG

[0172] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid OA, and 8 mg of DSPE-PEG2000, dissolve them in 0.6 ml of ethanol, and drop the solution into 2 ml of water under stirring at 1500 rpm. Then evaporate the ethanol under reduced pressure at 45°C.

[0173] Results: The average particle size was (193.6 ± 4.003) nm, the PDI value was 0.168 ± 0.026, and there was no visible change after being placed at room temperature for one week.

[0174] Example 40 Rh2:LA:DSPE-PEG2000 = 2:2:1, Rh2 = 12 mg / mL, oleic acid + linoleic acid, PLGA-PEG

[0175] Method: Weigh 24 mg of Rh2, 24 mg of linoleic acid LA, and 12 mg of DSPE-PEG2000, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 2 ml of water under 250 W ultrasonic treatment. Then evaporate the ethanol under reduced pressure at 45°C.

[0176] Results: The average particle size was (203.5 ± 5.003) nm, the PDI value was 0.187 ± 0.019, and there was no visible change after being placed at room temperature for one week.

[0177] Example 41 Rh2:OA:TPGS:DSPE-PEG2000 = 4:4:1:1, Rh2 = 8 mg / mL, TPGS + DSPE-PEG

[0178] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid OA, 4 mg of TPGS, and 4 mg of DSPE-PEG2000, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 2 ml of water under stirring at 1500 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0179] Result: The average particle size is (211.6 ± 6.329) nm, the PDI value is 0.175 ± 0.032, and there is no visible change after standing at room temperature for one week.

[0180] Example 42 Rh2:LA:TPGS:PLA2000-PEG2000 = 4:4:1:1, TPGS + PLA-PEG

[0181] Method: Weigh 16 mg of Rh2, 16 mg of linoleic acid LA, 4 mg of TPGS, and 4 mg of PLA2000-PEG2000, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate ethanol under reduced pressure at 45 °C.

[0182] Result: The average particle size is (198.2 ± 5.294) nm, the PDI value is 0.173 ± 0.033, and there is no visible change after standing at room temperature for one week.

[0183] Example 43 Rh2:LA:OA:PCL2000-PEG2000:PLA1000-PEG2000 = 4:2:2:1:1, PCL-PEG + PLA-PEG

[0184] Method: Weigh 16 mg of Rh2, 8 mg of oleic acid OA, 8 mg of linoleic acid LA, 4 mg of PCL2000-PEG2000, and 4 mg of PLA1000-PEG2000, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate ethanol under reduced pressure at 45 °C.

[0185] Result: The average particle size is (214.8 ± 6.025) nm, the PDI value is 0.183 ± 0.040, and there is no visible change after standing at room temperature for one week.

[0186] Example 44 Rh2:OA:SPC, SPC

[0187] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid OA, and 8 mg of soy lecithin SPC, dissolve them in 0.6 ml of ethanol, and dropwise add the solution to 4 ml of water under ultrasonic treatment in a 250 W water bath. Evaporate ethanol under reduced pressure at 45 °C.

[0188] Result: The average particle size is (205.7 ± 8.425) nm, the PDI value is 0.191 ± 0.051.

[0189] Example 45 Rh2: Linoleic Acid: EPC, EPC

[0190] Method: Weigh 16 mg of Rh2, 16 mg of linoleic acid LA, and 8 mg of egg yolk lecithin EPC and dissolve them in 0.6 ml of ethanol. Dropwise add the solution to 4 ml of water under ultrasonic bath at 250 W, and evaporate ethanol under reduced pressure at 45 °C.

[0191] Result: The average particle size is (183.7 ± 6.429) nm, and the PDI value is 0.182 ± 0.043.

[0192] Example 46 Rh2: Linoleic Acid: DSPC, DSPC

[0193] Method: Weigh 16 mg of Rh2, 16 mg of linoleic acid LA, and 8 mg of DSPC and dissolve them in 0.6 ml of ethanol. Dropwise add the solution to 4 ml of water under ultrasonic bath at 250 W, and evaporate ethanol under reduced pressure at 45 °C.

[0194] Result: The average particle size is (196.1 ± 6.032) nm, and the PDI value is 0.171 ± 0.032.

[0195] Example 47 Rh2: LA: Borneol: TPGS

[0196] Method: Weigh 12 mg of Rh2, 13 mg of linoleic acid LA, 5 mg of borneol, and 7.5 mg of TPGS and dissolve them in 0.6 ml of ethanol. Dropwise add the solution to 3 ml of water under ultrasonic bath at 250 W, and evaporate ethanol under reduced pressure at 45 °C.

[0197] Result: The average particle size is (164.3 ± 2.203) nm, and the PDI value is 0.177 ± 0.015. No visible change was observed after storing at room temperature for two weeks, and no significant change in particle size and particle size distribution was found (see Figure 11 ).

[0198] Particle size stability in physiological media: As Figure 10 shown, during the 12-hour incubation period, no visible turbidity, aggregation, or precipitation occurred in the Rh2 composition at all time points. The particle size in glucose, artificial gastrointestinal tract, artificial intestinal fluid, PBS, and FBS hardly changed, indicating stability in these physiological media. Therefore, the prepared ginsenoside Rh2 nanoparticles can be directly administered orally and can be used for injection after being dispersed in 5% glucose or PBS; during incubation in physiological saline, the particle size increased to more than 2000 nm, suggesting poor stability in physiological saline. Avoid using physiological saline as the dispersion medium during injection.

[0199] Example 48 Rh2: OA: TPGS = 2:2:1, ethanol-acetone (volume ratio 1:1) as the organic phase

[0200] Method: Weigh 16 mg of Rh2, 16 mg of oleic acid, and 8 mg of TPGS, dissolve them in 0.6 ml of ethanol-acetone mixture (volume ratio 1:1), and dropwise add the solution to 2 ml of water under ultrasonic bath at 250 W. Evaporate ethanol under reduced pressure at 45 °C.

[0201] Result: The average particle size is (223.5 ± 5.801) nm, the PDI value is 0.153 ± 0.031. After standing at room temperature for one week, there is no obvious change in the particle size and particle size distribution, indicating good storage stability.

[0202] Example 49 Rh2:LA:TPGS = 2:2:1, methanol-acetone (volume ratio 1:1) as the organic phase

[0203] Method: Weigh 16 mg of Rh2, 16 mg of linoleic acid, and 8 mg of TPGS, dissolve them in 0.6 ml of methanol-acetone mixture (volume ratio 1:1), and dropwise add the solution to 2 ml of water under ultrasonic bath at 250 W. Evaporate ethanol under reduced pressure at 45 °C.

[0204] Result: The average particle size is (253.8 ± 6.013) nm, the PDI value is 0.163 ± 0.033. After standing at room temperature for one week, there is no obvious change in the particle size and particle size distribution, indicating good storage stability.

[0205] To further prove the beneficial effects of the present invention for better understanding of the present invention, the following comparative examples are used to further clarify the technical features disclosed in the present invention, but it should not be construed as a limitation to the present invention. For other improvements made by those skilled in the art based on the above-mentioned inventive content without creative work, they are also considered to fall within the protection scope of the present invention.

[0206] Comparative Example 1 Rh2:OA = 1:1, Rh2 = 2.5 mg / mL

[0207] Method: Weigh 10 mg of Rh2 and 10 mg of oleic acid OA, dissolve them in 0.5 mL of ethanol, and dropwise add the solution to 4 ml of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0208] Result: A large amount of turbidity appears immediately after injection, and a large amount of precipitation appears after removing ethanol.

[0209] Comparative Example 2 Rh2:TPGS = 1:1, Rh2 = 2.5 mg / mL

[0210] Method: Weigh 10 mg of Rh2 and 10 mg of TPGS, dissolve them in 0.5 mL of ethanol, and dropwise add the solution to 4 ml of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0211] Result: Turbidity appears immediately after injection, and aggregated precipitation appears after removing ethanol.

[0212] Comparative Example 3: Rh2:TPGS = 1:5, Rh2 = 2.5 mg / mL

[0213] Method: Weigh 10 mg of Rh2 and 50 mg of TPGS, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate the ethanol under reduced pressure at 45°C.

[0214] Result: The average particle size is (353.7 ± 12.6) nm, and the PDI value is 0.379 ± 0.086. However, irreversible precipitation occurs after standing overnight.

[0215] Comparative Example 4: Rh2:PLGA2000 - PEG2000 = 1:3, Rh2 = 5 mg / mL

[0216] Method: Weigh 10 mg of Rh2 and 30 mg of PLGA2000 - PEG2000, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate the ethanol under reduced pressure at 45°C.

[0217] Result: Turbidity appears immediately after injection, and aggregation precipitation occurs after removing ethanol.

[0218] Comparative Example 5: Rh2:PCL2000 - PEG2000 = 1:3, Rh2 = 5 mg / mL

[0219] Method: Weigh 10 mg of Rh2 and 30 mg of PCL2000 - PEG2000, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate the ethanol under reduced pressure at 45°C.

[0220] Result: Turbidity appears immediately after injection, and aggregation precipitation occurs after removing ethanol.

[0221] Comparative Example 6: Rh2:DSPE - mPEG2000 = 1:3, Rh2 = 5 mg / mL

[0222] Method: Weigh 10 mg of Rh2 and 30 mg of DSPE - PEG2000, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate the ethanol under reduced pressure at 45°C.

[0223] Result: Turbidity appears immediately after injection, and aggregation precipitation occurs after removing ethanol.

[0224] Comparative Example 7: Rh2:P188 = 1:3, Rh2 = 5 mg / mL

[0225] Method: Weigh 10 mg of Rh2 and 30 mg of P188, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate the ethanol under reduced pressure at 45°C.

[0226] Result: A large amount of turbidity appeared immediately after injection, and a large amount of precipitation appeared after removing ethanol.

[0227] Comparative Example 8 Rh2:DSPE-mPEG2000 = 1:3, Rh2 = 5 mg / mL

[0228] Method: Weigh 10 mg of Rh2 and 30 mg of DSPE-PEG2000, dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0229] Result: Turbidity appeared immediately after injection, and aggregated precipitation appeared after removing ethanol.

[0230] Comparative Example 9 Rh2:SDS = 1:3, Rh2 = 5 mg / mL

[0231] Method: Weigh 10 mg of Rh2 and 30 mg of sodium dodecyl sulfate (SDS), dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0232] Result: A large amount of turbidity appeared immediately after injection, and a large amount of precipitation appeared after removing ethanol.

[0233] Comparative Example 10 Rh2:SPC = 1:3, Rh2 = 5 mg / mL

[0234] Method: Weigh 10 mg of Rh2 and 30 mg of soy lecithin (SPC), dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0235] Result: A large amount of turbidity appeared immediately after injection, and a large amount of precipitation appeared after removing ethanol.

[0236] Comparative Example 11 Rh2:EPC = 1:3, Rh2 = 5 mg / mL

[0237] Method: Weigh 10 mg of Rh2 and 30 mg of egg yolk lecithin (EPC), dissolve them in 0.5 mL of ethanol, and drip the solution into 4 mL of water under stirring at 1000 rpm. Evaporate ethanol under reduced pressure at 45 °C.

[0238] Result: A large amount of turbidity appeared immediately after injection, and a large amount of precipitation appeared after removing ethanol.

[0239] Comparative Example 12 Rh2:refined soybean oil:TPGS = 2:2:1, Rh2 = 2.5 mg / mL, soybean oil

[0240] Method: Weigh 10 mg of Rh2, 10 mg of refined soybean oil, and 5 mg of TPGS and dissolve them in 0.5 mL of ethanol. While stirring at 500 rpm, slowly drip the solution into 4 mL of water, and then remove the ethanol by evaporation under reduced pressure at 45 °C.

[0241] Result: Turbidity occurred immediately after injection, and aggregated precipitation appeared after removing the ethanol.

[0242] Comparative Example 13 Rh2:refined soybean oil:TPGS = 2:2:1, Rh2 = 5 mg / mL, soybean oil

[0243] Method: Weigh 15 mg of Rh2, 15 mg of refined soybean oil, and 7.5 mg of TPGS and dissolve them in 0.6 mL of ethanol. While stirring at 1000 rpm, slowly drip the solution into 3 mL of water, and then remove the ethanol by evaporation under reduced pressure at 45 °C. Measure the particle size.

[0244] Result: Turbidity occurred immediately during injection, and aggregated precipitation appeared after removing the ethanol.

[0245] Comparative Example 14 Rh2:refined soybean oil:TPGS:sodium oleate = 4:4:2:1, soybean oil

[0246] Method: Weigh 20 mg of ginsenoside Rh2 and 10 mg of refined soybean oil and dissolve them in 0.6 mL of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate and dissolve them in 4 mL of water. While under 250 W ultrasonic treatment, slowly drip the ethanol solution into the aqueous phase, and then remove the ethanol by rotary evaporation at 45 °C.

[0247] Result: Turbidity occurred immediately after injection, and aggregated precipitation appeared after removing the ethanol.

[0248] Comparative Example 15 Rh2:corn oil:TPGS = 2:2:1, Rh2 = 2.5 mg / mL, corn oil

[0249] Method: Weigh 10 mg of Rh2, 10 mg of corn oil, and 5 mg of TPGS and dissolve them in 0.5 mL of ethanol. While stirring at 500 rpm, slowly drip the solution into 4 mL of water, and then remove the ethanol by evaporation under reduced pressure at 45 °C.

[0250] Result: Turbidity occurred immediately after injection, and aggregated precipitation appeared after removing the ethanol.

[0251] Comparative Example 16 Rh2:corn oil:TPGS = 2:2:1, Rh2 = 5 mg / mL, corn oil

[0252] Method: Weigh 15 mg of Rh2, 15 mg of corn oil, and 7.5 mg of TPGS and dissolve them in 0.6 mL of ethanol. While stirring at 1000 rpm, slowly drip the solution into 3 mL of water, and then remove the ethanol by evaporation under reduced pressure at 45 °C.

[0253] Results: Turbidity occurred immediately upon injection, and aggregation precipitation occurred after removing ethanol.

[0254] Comparative Example 17 Rh2: Corn oil: TPGS: Sodium oleate = 4:4:2:1, corn oil

[0255] Method: Weigh 20 mg of ginsenoside Rh2 and 10 mg of corn oil and dissolve them in 0.6 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate and dissolve them in 4 mL of water as the aqueous phase. Under stirring at 1500 rpm, drop the organic phase into the 4 ml of aqueous phase, and remove ethanol by rotary evaporation at 45°C.

[0256] Results: Turbidity occurred immediately after injection, and aggregation precipitation occurred after removing ethanol.

[0257] Comparative Example 18 Rh2: Olive oil: TPGS = 2:2:1, Rh2 = 2.5 mg / mL, olive oil

[0258] Method: Weigh 10 mg of Rh2, 10 mg of olive oil, and 5 mg of TPGS and dissolve them in 0.5 mL of ethanol. Under stirring at 500 rpm, drop it into 4 ml of water, and remove ethanol by reduced pressure evaporation at 45°C.

[0259] Results: Turbidity occurred immediately after injection, and aggregation precipitation occurred after removing ethanol.

[0260] Comparative Example 19 Rh2: Olive oil: TPGS = 2:2:1, Rh2 = 5 mg / mL, olive oil

[0261] Method: Weigh 15 mg of Rh2, 15 mg of corn oil, and 7.5 mg of TPGS and dissolve them in 0.6 ml of ethanol. Under stirring at 1000 rpm, drop it into 3 ml of water, and remove ethanol by reduced pressure evaporation at 45°C.

[0262] Results: Turbidity occurred immediately upon injection, and aggregation precipitation occurred after removing ethanol.

[0263] Comparative Example 20 Rh2: Olive oil: TPGS: Sodium oleate = 4:4:2:1, olive oil

[0264] Method: Weigh 20 mg of ginsenoside Rh2 and 10 mg of olive oil and dissolve them in 0.6 ml of ethanol. Weigh 10 mg of TPGS and 5 mg of sodium oleate and dissolve them in 4 mL of water as the aqueous phase. Under stirring at 1500 rpm, drop the organic phase into the 4 ml of aqueous phase, and remove ethanol by rotary evaporation at 45°C.

[0265] Results: Turbidity occurred immediately after injection, and aggregation precipitation occurred after removing ethanol.

[0266] Experimental Example 1 Investigation of particle size stability in physiological medium:

[0267] The prepared ginsenoside Rh2 nanoparticles were mixed with 2×PBS (PBS with twice the concentration), 1.8% NaCl solution, 10% glucose solution, etc. in equal volumes, or mixed with 4 times the volume of artificial gastric juice and artificial intestinal juice, incubated at 37°C, and the particle size and particle size distribution were measured at specific time points to observe whether there was turbidity, aggregation or precipitation. The specific data and results are shown in the corresponding examples.

[0268] Experimental Example 2 Investigation of stability at room temperature:

[0269] The newly prepared ginsenoside Rh2 nanoparticles were placed at room temperature for 30 days, and samples were taken at different time points (such as 0, 7, 14, 30 days, or within one week) to measure the particle size and particle size distribution, and at the same time, carefully observe whether there was turbidity, aggregation or precipitation. The specific data and results are shown in the corresponding examples.

[0270] Experimental Example 3 HPLC content determination method of ginsenoside Rh2:

[0271] Chromatographic conditions: The chromatographic column was a C18 column (4.6 mm×250 mm, 5 μm), the mobile phase was acetonitrile - water (63:37), the flow rate was 1.0 mL / min, the injection volume was 20 μL, the detection wavelength was 203 nm, and the column temperature was 25°C.

[0272] Establishment of the Rh2 standard curve: Weigh a certain amount of Rh2 precisely in a volumetric flask, add an appropriate amount of chromatographic methanol, sonicate to fully dissolve the drug, and make up the volume with chromatographic methanol and shake well to obtain a stock solution with a concentration of 1 mg / mL. Take the above stock solution and dilute it with chromatographic methanol to a series of standard solutions with concentrations of 0.5, 1, 2, 5, 10, 25, 50, 100, 200, 500 μg / mL. Inject the samples for HPLC detection, perform linear regression with the peak area (Y) against the drug concentration (X), and obtain the standard curve equation y = 0.1299x - 0.0992, R 2 = 0.9992, and the linear range was 5 - 500 μg / mL.

[0273] Experimental Example 4 Determination of the drug loading of ginsenoside Rh2 nano - composition (nanoparticles similar to nano - suspension):

[0274] Take the freshly prepared Rh2 nano - composition, directly freeze - dry it without adding any protective agent. Weigh 10 mg of the freeze - dried powder precisely (denoted as W) into a 10 mL volumetric flask, add chromatographic methanol to dissolve and make up the volume to the scale and shake well. Precisely pipette 1 mL from it into another 10 mL volumetric flask, add chromatographic methanol to dissolve and make up the volume to the scale and shake well.

[0275] Take 1 mL of the diluted solution (dilute with chromatographic methanol if necessary) and place it in an HPLC sample vial. Perform HPLC analysis according to the chromatographic conditions in Experimental Example 3, record the peak area, substitute the obtained peak area into the regression equation y = 0.1299x - 0.0992 to calculate the drug concentration, and calculate the concentration C (mg / mL) of Rh2 in the first volumetric flask according to the dilution factor. Then, the drug loading content of ginsenoside Rh2 in the nano - composition = 10C / W × 100%.

[0276] Experimental Example 5 Freeze - drying, reconstitution and screening of lyoprotectants:

[0277] The newly prepared ginsenoside Rh2 nanoparticles (Example 3) were freeze - dried without adding anything, adding 1% trehalose, adding 1% glucose, and adding 1% mannitol. The obtained freeze - dried powder was added with an equal volume of pure water and shaken. The particle size and particle size distribution after reconstitution were measured. The results are shown in the following table, indicating that mannitol is an excellent lyoprotectant for the ginsenoside Rh2 nano - composition. For the freeze - dried powder of Example 3 after adding 1% mannitol, the particle size after reconstitution in PBS was (140.4 ± 2.67) nm, and the PDI was 0.251 ± 0.108.

[0278] Table 1 Screening of lyoprotectants in Example 3 (particle size and PDI after adding different lyoprotectants)

[0279]

[0280] The Rh2 nano - compositions obtained from other examples were freeze - dried after adding 1% mannitol. The freeze - dried powder was shaken with water for reconstitution. The results are shown in Table 2, indicating that mannitol has good lyoprotective effect.

[0281] Table 2 Particle size and PDI after reconstitution of samples from different Example 3 after adding 1% mannitol

[0282]

[0283] Experimental Example 6 Establishment of an LC - MS / MS analysis method for Rh2 in rat plasma samples

[0284] Preparation of reference solution: Weigh an appropriate amount of the reference standard of ginsenoside Rh2 precisely, and prepare stock solutions of 5 mg / mL with chromatographic methanol and DMSO respectively. Then dilute them with methanol to a reserve solution of 50 μg / mL, and further dilute step by step with 50% methanol solution to prepare a series of standard solutions with concentrations of 10 μg / mL, 5 μg / mL, 2 μg / mL, 1 μg / mL, 500 ng / mL, 200 ng / mL, 100 ng / mL, 50 ng / mL, 20 ng / mL, 10 ng / mL, 5 ng / mL, 2 ng / mL, 1 ng / mL. Store them at -20 °C for future use. Weigh an appropriate amount of nimodipine (IS) precisely, prepare a reserve solution of 50 μg / mL with methanol, and prepare a single standard solution of 100 ng / mL with 50% methanol. Store it at -20 °C for future use.

[0285] Preparation and treatment of standard samples: Take three SD rats' blank plasma after fasting for 18 h. Precisely pipette 45 μL of blank plasma, add 5 μL of Rh2 standard solution with concentrations from 1 ng / mL to 10 μg / mL respectively, then add 450 μL of methanol and acetonitrile (1:1) precipitation solution, vortex at 2500 speed for 5 min, centrifuge at 13000 rpm for 10 min, pipette 450 μL of the supernatant, and concentrate it to dryness under vacuum. Add 50 μL of 50% methanol containing 5 ng of nimodipine to dissolve the residue, vortex at 2500 speed for 5 min, sonicate at 100 w for 10 min, vortex again for 30 s, centrifuge at 13000 rpm for 10 min, pipette 40 μL of the supernatant, centrifuge the supernatant again at 13000 rpm for 10 min, and transfer 35 μL of the supernatant to a sample bottle for determination. The concentrations of the standard samples for the final standard curve are 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000 ng / mL respectively.

[0286] Preparation and treatment of biological samples: Precisely pipette 50 μL of the plasma to be determined, add 450 μL of methanol and acetonitrile (1:1) precipitation solution, vortex at 2500 speed for 5 min, centrifuge at 13000 rpm for 10 min, pipette 450 μL of the supernatant, and concentrate it to dryness under vacuum. Add 50 μL of 50% methanol solution containing 5 ng of nimodipine to dissolve the residue, vortex at 2500 speed for 5 min, sonicate at 100 w for 10 min, vortex again for 30 s, centrifuge at 13000 rpm for 10 min, pipette 40 μL of the supernatant, centrifuge the supernatant again at 13000 rpm for 10 min, and transfer 35 μL of the supernatant to a sample bottle for determination.

[0287] Preparation and treatment of quality control samples: Prepare standard solutions with the lowest quantitation limit, low, medium and high concentrations containing ginsenoside Rh2 respectively. Precisely pipette blank plasma and add the above standard solutions with different concentrations respectively, and the treatment method is the same as above.

[0288] Chromatographic and mass spectrometric conditions: Chromatographic column: XSelect HSS T3 C18 2.5 μm, 2.1×100 mm (Waters Co.). Mobile phase: A is acetonitrile / methanol (1:1 v / v) + 2 mM ammonium acetate solution, B is water + 0.05% formic acid solution. Flow rate is 0.2 mL / min, sample cell temperature is 10 °C, injection volume is 5 μL. Negative ion scanning mode of electrospray ionization source (ESI).

[0289] Standard curve and linear range: Prepare a mixed reference solution with a concentration of 1000 ng / mL, and then perform gradient dilution to obtain a series of mixed reference solutions with different concentrations. Take 50 mL of blank rat plasma and place it in a centrifuge tube. Add the mixed reference solutions and internal standard solution at each concentration, and perform LC-MS / MS analysis after processing according to the above biological sample treatment method. Plot the standard curve with the concentration as the abscissa (X) and the ratio of the peak area of each compound to the internal standard as the ordinate (Y). The results show that all compounds have a good linear relationship within the set concentration range. The regression equation of Rh2 is y = 3.68377e-5x + 2.95485e-4, the R2 value is 0.99906, and the linear range is 1 - 1000 ng / mL.

[0290] Precision: Take the quality control plasma samples at different concentrations above, 5 samples for each concentration, and analyze them by HPLC-MS method. Measure the concentration of the saponin components to be measured in each sample according to the standard curve on the same day, and measure continuously for three days. Calculate the intra-day precision and accuracy according to the measurement results within one day. Calculate the inter-day precision and accuracy according to the measurement results for three days. Precision is the degree of closeness of multiple measurement results, expressed as RSD (%). It is required that the RSD of LQC is less than or equal to 20%, and the RSD of the three quality control samples at low, medium, and high concentrations is less than or equal to 15%. Accuracy is the degree of closeness between the measured value and the true value (%), and generally requires the accuracy of biological sample determination to be between 85% and 115%.

[0291] Take the quality control plasma samples at different concentrations in the merchant, 4 samples for each concentration. Additionally, prepare a mixed standard solution with the same concentration, 5 samples for each concentration, and analyze them by LC-MS / MS method. Take the ratio of the peak area of Rh2, Rb1, Rg1, R1, Re, and Rd in the quality control samples to the corresponding peak area in the standard solution as the recovery rate, and require the recovery rate of biological samples to be between 80% and 120%.

[0292] Table 3 Intra- (inter-) day precision and accuracy of detecting Rh2 in rat plasma by LC-MS / MS method

[0293]

[0294]

[0295] Stability: Take 50 μL of blank rat plasma, add reference substance solutions at low, medium, and high three mass concentrations to obtain quality control samples (QC) at low, medium, and high three mass concentrations containing ginsenoside Rh2. Place them in a -40 °C refrigerator for freezing, then take them out and thaw at room temperature, and repeat the freeze-thaw cycle 3 times. After the last thawing at room temperature, process the plasma samples according to the method in item 1.3 and perform LC-MS / MS determination.

[0296] Take 50 μL of blank rat plasma, add reference substance solutions at low, medium, and high three mass concentrations to obtain quality control samples (QC) at low, medium, and high three mass concentrations containing ginsenoside Rh2. Place them in a -40 °C refrigerator for freezing for 15 days, thaw at room temperature, then process the plasma samples according to the method in item 1.3 and perform LC-MS / MS determination.

[0297] Take 50 μL of blank rat plasma, add reference substance solutions at low, medium, and high three mass concentrations to obtain quality control samples (QC) at low, medium, and high three mass concentrations containing ginsenoside Rh2. Process the plasma samples according to the method in item 1.3, place them at 4 °C for 24 h, and then perform LC-MS / MS determination. The results are shown in Table 3.

[0298] Table 4 Stability of Rh2 in rat plasma detected by LC-MS / MS under 4 °C conditions, long-term storage, and repeated freeze-thaw

[0299]

[0300] Recovery rate and matrix effect: Take the above QC samples at low, medium, and high three mass concentrations, prepare 6 replicates for each mass concentration, process the plasma samples according to the method in item 1.3 and perform LC-MS / MS determination, and record the ratio (A) of the component peak area to the internal standard peak area. Take another blank rat plasma, process it according to the method under item 4.2.1.3.1.2 and then add reference substance solutions at low, medium, and high three mass concentrations, perform LC-MS / MS determination, and record the ratio (B) of the component peak area to the internal standard peak area. Calculate the extraction recovery rate according to the formula extraction recovery rate = A / B × 100%. Take an aqueous solution instead of blank plasma, process the samples according to the method in item 1.3 and perform LC-MS / MS determination to obtain the ratio (C) of the component peak area to the internal standard peak area, and calculate the matrix effect according to the formula matrix effect = A / C × 100%.

[0301] Experimental example 7 Experimental method for the oral bioavailability of rats

[0302] Preparation of blank SD rat plasma: Fast the rats for 24 h (only normal drinking water, all bedding and feed removed), anesthetize the rats, collect blood by jugular vein intubation, collect it into an EDTA anticoagulant tube, centrifuge at 3000 rpm for 15 min at 10 °C. Transfer the upper-layer plasma after mixing to a 2.0 mL or 5.0 mL EP tube and store at -80 °C.

[0303] Surgery for jugular vein intubation in rats: After fasting, anesthetize the SD rats by intraperitoneal injection of 7% chloral hydrate, perform jugular vein intubation surgery, and suture the wound after the surgery.

[0304] Dosing regimen and grouping: The grouping for oral bioavailability study is as follows, with 8 rats in each group (n = 8). After intubation, fast for 18 h and then administer the drug by gavage. The following nanoparticle formulations are all administered by gavage (the dosing dose is 50 mg / kg):

[0305] ① Rh2 nanoparticles (Rh2-oleic acid-TPGS NPs, Example 3)

[0306] ② Rh2 suspension (suspended in 0.5% CMC-Na, i.g)

[0307] ③ Rh2 nanoparticles containing borneol (Rh2-linoleic acid-borneol-TPGS NPs, Example 47)

[0308] Drug administration and sample collection: First, aspirate 0.3 mL of waste blood before sampling. At 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h after drug administration, use a 1 mL flat-tip sampler to aspirate 0.4 mL of blood samples from the silicone tube and place them in heparinized centrifuge tubes. Centrifuge the blood samples at 5000 rpm for 10 min to separate the plasma, and store at -80 °C for further measurement.

[0309] Treatment of plasma samples: Precisely aspirate 50 μL of the plasma to be measured, add 450 μL of a precipitation solution of methanol and acetonitrile (1:1), vortex at 2500 speed for 5 min, centrifuge at 13000 rpm for 10 min, aspirate 450 μL of the supernatant, and concentrate it to dryness under vacuum. Dissolve the residue in 50 μL of a 50% methanol solution containing 5 ng of nimodipine, vortex at 2500 speed for 5 min, sonicate at 100 w for 10 min, vortex again for 30 s, centrifuge at 13000 rpm for 10 min, aspirate 40 μL of the supernatant, centrifuge the supernatant again at 13000 rpm for 10 min, and transfer 35 μL of the supernatant to a sample vial for measuring the sample concentration by UPLC-MS.

[0310] Results: To study the effect of Rh2 nano-composite on improving the bioavailability of ginsenoside Rh2, Rh2 aqueous suspension was selected as the control group by intragastric administration, and the results of pharmacokinetic parameters in rats are shown in Table 5. The Cmax of the Rh2 nano-composite group (Rh2 NPs) and the Rh2-borneol nano-composite group (Rh2-borneol NPs) was significantly higher than that of the Rh2 oral suspension control group. Compared with the Rh2 aqueous suspension, the Tmax of the two experimental groups was delayed and the mean residence time MRT was prolonged. The areas under the drug-time curves AUC of the Rh2 nano-composite group (Rh2 NPs) and the Rh2-borneol nano-composite group (Rh2-borneol NPs) were 2.67 and 5.10 times that of the Rh2 physical suspension, respectively, indicating that the nano-composite significantly improved the oral bioavailability of ginsenoside Rh2, and the incorporation of borneol into the nanoparticles could further improve the oral bioavailability of ginsenoside Rh2.

[0311] Table 5 Pharmacokinetic parameters of Rh2 in Rh2 nanoparticles (n = 8)

[0312]

[0313] Note: Maximum blood drug concentration (peak concentration): Cmax; Time to peak concentration: Tmax; Area under the curve from time zero to infinity: AUC0→∞; Mean residence time: MRTlast.

[0314] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A ginsenoside Rh2 composition, characterized in that, The composition comprises ginsenoside Rh2, oleic acid / linoleic acid, and an amphiphilic excipient; wherein, The weight ratio of ginsenoside Rh2, oleic acid / linoleic acid to the amphiphilic excipient is 4:2 - 8:1 - 8; Moreover, the preparation method of the ginsenoside Rh2 composition comprises the following steps: Dissolve ginsenoside Rh2, the amphiphilic excipient, and oleic acid / linoleic acid in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution to water, and remove the organic solvent by reduced pressure evaporation to finally obtain the ginsenoside Rh2 composition; Or, the preparation method of the ginsenoside Rh2 composition comprises the following steps: Dissolve ginsenoside Rh2 and oleic acid / linoleic acid in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution to an aqueous solution containing the amphiphilic excipient and sodium oleate, and remove the organic solvent by reduced pressure evaporation to finally obtain the ginsenoside Rh2 composition; The amphiphilic excipient is one or a combination of more than one of TPGS, PLA-PEG, PLGA-PEG, PCL-PEG, DSPE-PEG, and phospholipid; The organic solvent is one or a combination of more than one of ethanol, methanol, and acetone, and the antisolvent is water; the oleic acid / linoleic acid is a mixed solution of oleic acid and linoleic acid in any ratio.

2. The ginsenoside Rh2 composition according to claim 1, characterized in that, The composition comprises pharmaceutically acceptable excipients, and the excipients include excipients for adjusting osmotic pressure and pH, specifically including glucose, sodium chloride, phosphate buffer solution, glycerol, and mannitol.

3. A preparation method of the ginsenoside Rh2 composition as described in claim 1, characterized in that, The method comprises the following steps: Dissolve ginsenoside Rh2, the amphiphilic excipient, and oleic acid / linoleic acid in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution to water, and remove the organic solvent by reduced pressure evaporation to finally obtain the ginsenoside Rh2 composition; The amphiphilic excipient is one or a combination of more than one of TPGS, PLA-PEG, PLGA-PEG, PCL-PEG, DSPE-PEG, and phospholipid; The organic solvent is one or a combination of more than one of ethanol, methanol, and acetone.

4. A method for preparing the ginsenoside Rh2 composition according to claim 1, characterized in that, The method comprises the following steps: Dissolve ginsenoside Rh2 and oleic acid / linoleic acid in an organic solvent to obtain a mixed solution; under stirring or ultrasonic treatment, add the mixed solution to an aqueous solution containing the amphiphilic excipient and sodium oleate, and remove the organic solvent by reduced pressure evaporation to finally obtain the ginsenoside Rh2 composition; The amphiphilic excipient is one or a combination of more than one of TPGS, PLA-PEG, PLGA-PEG, PCL-PEG, DSPE-PEG, and phospholipid; The organic solvent is one or a combination of more than one of ethanol, methanol, and acetone.

5. The preparation method of the ginsenoside Rh2 composition according to claim 3 or 4, characterized in that, The volume of water is 3 - 50 times the volume of the organic solvent, the stirring speed is 300 - 2000 rpm, and the water bath ultrasonic power is 100 - 500 W.

6. The preparation method of the ginsenoside Rh2 composition according to claim 3 or 4, characterized in that, For the aqueous dispersion system prepared by the method, after adding 1% mannitol and mixing evenly, it is freeze-dried to obtain the solid powder of the ginsenoside Rh2 composition.

7. Use of the ginsenoside Rh2 composition as claimed in claim 1 or the ginsenoside Rh2 composition prepared by the method as claimed in claim 3 or 4 in pharmaceutical preparations.

Images