20 (S)-PPD phospholipid solid dispersion as well as preparation method and application thereof

Abstract

The invention relates to a composition and a preparation process of a 20 (S)-protopanoxadiol (20 (S)-protopanoxadiol, 20 (S)-PPD) phospholipid / mesoporous silicon solid dispersion, and belongs to the technical field of medicines. The invention provides a 20 (S)-PPD phospholipid / mesoporous silicon solid dispersion preparation. The preparation consists of 20 (S)-PPD, phospholipid, ethanol, a crystal inhibitor and a dispersing agent. The preparation disclosed by the invention is prepared by adopting a solvent evaporation method; the prepared 20 (S)-PPD phospholipid / meso-porous silicon solid dispersion is good in stability; the drug loading capacity is high, and the content of 20 (S)-PPD in the preparation can reach 40% or above; and the absolute bioavailability of oral administration is high and can reach 23.95%. After the preparation is orally taken, the injury of podocyte for maintaining the integrity of a glomerular filtration barrier can be remarkably relieved, and adult chronic nephrotic syndromes such as membranous nephropathy (MN) can be treated.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a 20(S)-PPD phospholipid solid dispersion and a preparation method and application thereof. Background Art

[0002] Chronic kidney disease refers to a progressive disease of kidney structure and function disorders caused by various reasons. It has a high morbidity and mortality rate. It has become an important global public health issue and seriously threatens human health. The global burden of disease (GBD) study shows that the disability and adjusted life years (DALY) caused by chronic kidney disease has risen from 29th in 1990 to 18th in 2019. The burden caused by chronic kidney disease worldwide is increasing rapidly, and it is expected that by 2040, chronic kidney disease will become the fifth most common cause of loss of life in the world. (Collaborators GDaI. Global burden of 369diseases and injuries in 204countries and territories, 1990-2019a systematic analysis for the globalburgen of disease study2019[J].Lancet, 2020, 396(10258): 1204-1222.)

[0003] Membranous nephropathy (MN), also known as membranous glomerulonephritis, is one of the common pathological types of chronic nephrotic syndrome in adults. Those with unknown etiology are called idiopathic membranous nephropathy (IMN). Foreign reports show that IMN accounts for 30% to 40% of primary nephrotic syndrome, while domestic reports show that MN accounts for 24.9% of glomerular diseases nationwide, ranking second and the proportion is increasing year by year, with a proportion as high as 51% in local areas. (Yang Lihong, Su Peiling, Bao Kun. Clinical Practice Guidelines for Idiopathic Membranous Nephropathy in Traditional Chinese Medicine (2021) [J]. Chinese General Practice, 2023, 26(06): 647-659+645.); About 20% of patients have secondary factors such as systemic diseases, infections, drugs or tumors, while the rest have no clear secondary factors, which are called primary membranous nephropathy (pMN).

[0004] It is currently believed that pMN is an autoimmune disease, and the onset is mainly related to the activation of the body's immune system induced by multiple podocyte antigens. (Ronco P, Beck L, Debiec H, et al. Membranous nephropthy [J]. Nat Rev Dis Primers, 2021, 7 (1): 69.)

[0005] Ginseng is one of the traditional Chinese medicines, with pharmacological activities such as anti-inflammatory, anti-tumor, and regulating human immunity. Ginseng extract has unique advantages in the treatment of membranous nephropathy. Ginseng saponins are the main active ingredients extracted from ginseng, which are composed of hydrophilic glycoside chains and hydrophobic aglycones. According to the different skeletons of aglycones, ginsenosides are mainly divided into protopanaxadiol (PPD) type and protopanaxatriol (PPT) type, in which aglycones have a similar steroid structure to cholesterol.

[0006] Phospholipids are amphiphilic molecules composed of a hydrophilic part and a hydrophobic part arranged in a bilayer. They are zwitterionic surfactants and have an emulsifying effect.

[0007] At present, a variety of technical methods have been developed to increase the solubility of 20(S)-PPD to improve its in vivo bioavailability, such as liposomes, solid dispersions, micronization, nano-particles, co-crystals, etc. In oral dosage forms, solid dispersions can form a dispersion system in the form of solid powder by uniformly dispersing the drug in a highly dispersed state in amorphous form in a carrier, forming a supersaturated solution in the gastrointestinal tract to improve drug solubility and bioavailability. It can be obtained by a relatively simple and cost-effective method and is easy to self-administer. However, the relevant solid dispersions studied in the existing research are thermodynamically unstable systems. During the production and storage process, due to the effects of mechanical force, temperature and humidity, the drug may change from an amorphous state to a crystalline state, from a metastable state to a stable state, and phase separation or crystal growth with the carrier may occur, which will lead to a decrease in drug solubility and a slow dissolution rate. (Shen Xiaxia. Preparation and evaluation of nanofibrous solid dispersions based on medicinal acrylic resin [D]. Donghua University, 2015.)

[0008] Therefore, it is of great value to prepare a novel phospholipid solid dispersion preparation to improve the solubility and oral bioavailability of 20(S)-PPD and not to slow down the solubility and bioavailability during production and storage. Summary of the invention

[0009] The present invention discloses a 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation. The two can form self-assembled vesicle-like liposomes. After the obtained preparation is hydrated, a nanosuspension can be spontaneously formed in the gastrointestinal tract, and the solubility and dissolution rate of the drug are significantly improved. It is precisely this superior preparation structure that significantly improves its stability, dissolution rate and oral bioavailability.

[0010] The invention provides a 20(S)-PPD phospholipid / mesoporous silica solid dispersion. The preparation consists of 20(S)-PPD, phospholipid, ethanol, a crystal inhibitor and a dispersant.

[0011] The above-mentioned preparation includes 1 to 6 parts of phospholipids, 1 to 3 parts of PVPK30, and 15 to 20 parts of mesoporous silica; preferably, the content of 20(S)-PPD in the preparation is 10 to 30% (w / w); preferably, the phospholipid is one of soybean phospholipids PC70 or / PC50; preferably, the ratio of the amount of 20(S)-PPD to soybean phospholipid, i.e., the drug-carrying ratio, is 1 to 4:1; preferably, the ethanol concentration is 90 to 100% (w / w).

[0012] In the above-mentioned preparation, the crystal inhibitor is one of polyvinylpyrrolidone K30 (PVPK30) or copolyvidone S630, and the content is 3-10% (w / w).

[0013] In the above-mentioned preparation, the dispersant is one of mesoporous silica or microporous starch, and the content is 50-60% (w / w).

[0014] In the above-mentioned preparation, the function of the dispersant is mesoporous dispersion and makes the 20(S)-PPD phospholipid / mesoporous silica solid dispersion obtain excellent powder properties; the brand and model of the mesoporous silica is preferably XP3050, One of 244FP, with a particle size of 3.5 to 50 μm.

[0015] The preparation method of the preparation comprises the following steps:

[0016] 1) Dissolve 20(S)-PPD, soybean lecithin and PVP K30 in ethanol and stir continuously to fully dissolve.

[0017] 2) Adding an appropriate amount of mesoporous silica as a dispersant to the components in step 1), heating to reflux and curing.

[0018] 3) Ethanol was removed by evaporation under reduced pressure and the solid was collected.

[0019] 4) Place the mixture in a vacuum drying oven and dry it in vacuum for 24 hours to obtain a 20(S)-PPD phospholipid / mesoporous silica solid dispersion.

[0020] In the preparation method, the dissolving method in step 1) is heating and stirring to dissolve, and the heating temperature is preferably 37-60°C.

[0021] In the preparation method, the heating time is 5 to 15 minutes.

[0022] The preparation or the preparation prepared by the preparation method is used in preparing a drug for improving podocyte injury or a drug for treating membranous glomerulonephritis.

[0023] The bioavailability of the preparation or the preparation prepared by the preparation method is significantly improved.

[0024] Schematic diagram of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation is shown in Figure 1 shown.

[0025] In the early stage of the present invention, a large number of experiments were conducted to prepare 20(S)-PPD into various dosage forms, and the pharmacokinetic studies of the various dosage forms were conducted in rats. The absolute bioavailability results are as follows.

[0026] sample Dosage Absolute bioavailability Fat emulsion 1 oral 19.44％ Fat emulsion 2 oral 16.52％ Fat emulsion 3 oral 14.32％ Pellets 1 oral 20.12％ Pellets 2 oral 15.43％ Pellets 3 oral 17.33％ Nanocapsules oral 8.57％ Nanosuspension oral 9.42％

[0027] A large number of experiments were conducted in the early stage of the present invention, and 20(S)-PPD was prepared into various dosage forms. The pharmacokinetic studies of various dosage forms were carried out in rats. The prepared 20(S)-PPD fat emulsion, micropellet, nanosuspension, nanocapsule and other dosage forms still had low absolute oral bioavailability and unstable properties, and had low drug loading, and none of them achieved the expected effect.

[0028] The present invention is based on the above research and further improves to prepare a 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation. The preparation has a high drug loading, high oral absolute bioavailability and significant drug efficacy.

[0029] The 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 was subjected to DSC thermal analysis.

[0030] DSC analysis chart Figure 2 As shown, A: free 20(S)-PPD B: phospholipid C: 20(S)-PPD phospholipid / mesoporous silica solid dispersion D: physical mixture of 20(S)-PPD and phospholipid

[0031] It can be seen from the DSC analysis diagram that free 20(S)-PPD has an endothermic peak at 214.14°C, while the characteristic peak disappears in the DSC curve of 20(S)-PPD phospholipid / mesoporous silica solid dispersion, indicating that there is no crystalline drug component of 20(S)-PPD in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation, and the drug exists in the solid dispersion in an amorphous state.

[0032] Polarization microscopy (PLM) was performed on the 20(S)-PPD-phospholipid-PVPK30 complex prepared in Example 1. The crystallization of the complex was observed using PLM.

[0033] Take appropriate amounts of 20(S)-PPD, phospholipids, and PVPK30 on a glass slide and add silicone oil to disperse them; take 30 μL of the 20(S)-PPD-phospholipid-PVPK30 complex solution prepared in Example 1 on a glass slide, wait for the ethanol to evaporate naturally, observe and take pictures at a magnification of 10×20.

[0034] 20(S)-PPD: has a needle-like crystal structure. For details, see Figure 3 shown.

[0035] Phospholipids: neatly arranged without crystalline structure, see Figure 4 shown.

[0036] 20(S)-PPD-phospholipid-PVPK30 complex: phospholipids are neatly arranged and have no crystalline structure. For the specific structure, see Figure 5 shown.

[0037] Through polarizing microscopy (PLM) observation, it can be determined that 20(S)-PPD is a needle-shaped crystalline structure, and phospholipids are neatly arranged and non-crystalline structures. The absence of crystalline structures in the 20(S)-PPD-phospholipid-PVPK30 complex structure diagram indicates that no 20(S)-PPD drug crystals are precipitated in the complex, and it exists in an amorphous state, which can achieve the maximum drug loading capacity.

[0038] The 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation prepared by the present invention has the appearance of white uniform powder, and can form a nano suspension without obvious insoluble matter after being redissolved in water. Due to the presence of phospholipids, a hydration film can be formed on the surface of the hydrophobic drug 20(S)-PPD particles and charged, thereby reducing the liquid-solid surface tension and increasing the repulsive force between particles, thereby improving the dispersibility of the 20(S)-PPD drug particles and reducing sedimentation.

[0039] The powder properties of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 were measured.

[0040] The solid cone method was used to determine the angle of repose of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion. The preparation prepared in Example 1 was placed in a funnel fixed at the center point of a circular surface dish and allowed to flow out slowly until the 20(S)-PPD phospholipid / mesoporous silica solid dispersion accumulated to the point where it no longer flowed down. The pile height (h) and the surface dish radius (r) were measured three times in parallel, and the angle of repose (θ) was calculated. The results are shown in the following table.

[0041]

[0042] The smaller the repose angle of the powder, the smaller the friction force and the better the fluidity. It is generally believed that when the repose angle is less than 40°, the production fluidity requirements can be met. The results in the above table show that the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation has good fluidity.

[0043] Weigh a certain amount of 20(S)-PPD phospholipid / mesoporous silica solid dispersion and gently put it into a 50ml measuring cylinder to measure the initial loose volume; use the tapping method to make the powder in the tightest state and measure the final volume; calculate the loosest density ρ0 and the tightest density ρ f ; Calculate the compression degree C according to the formula and see the results in the table below.

[0044]

[0045]

[0046] The compressibility of the powder can reflect its fluidity, cohesion and softness. When the compressibility of the powder is below 20%, the fluidity is good, and when the compressibility increases, the fluidity decreases. The results show that the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation has good fluidity.

[0047] The 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation prepared by the present invention utilizes mesoporous silica for dispersion, so that the preparation has good powder properties.

[0048] 40 mg of the sieved powder of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 was weighed into a beaker and added to 20 ml of gastric juice for re-dissolution. A nanosuspension with a Tyndall effect was observed. The particle size of the nanosuspension was about 200 nm after measurement by a Malvern nanoparticle size analyzer. The specific measurement results are shown in Figure 6 shown.

[0049] The pharmacokinetic study of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 and 20(S)-PPD fat emulsion was conducted. The results of the absolute bioavailability comparison between the 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 and the fat emulsion are as follows.

[0050] 1. Pharmacokinetic evaluation in rats

[0051] Male SD rats, 180±20g, were randomly divided into 8 groups, 6 rats in each group. The 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation had a content of 14.42%, a dosage of 35mg / kg, and a volume of 10ml / kg; the 20(S)-PPD fat emulsion preparation had a content of 3.62mg / ml, a dosage of 35mg / kg, and a volume of 10ml / kg were administered by oral gavage.

[0052] Average absolute bioavailability after oral administration F = 23.95%

[0053] Taking oral 20(S)-PPD fat emulsion as reference preparation, the relative bioavailability of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation was 123.22%. Figure 7 (oral medication time curve) and 8 (intravenous injection medication time curve).

[0054] By changing the dosage form of 20(S)-protopanaxadiol and preparing it into a 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation, the absorption capacity of the preparation is promoted and its oral bioavailability is effectively improved.

[0055] The 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 was subjected to a physical stability test.

[0056] An appropriate amount of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation placed under different conditions was dispersed on a glass slide by adding silicone oil, and then placed aside for observation under a polarizing microscope. The results are as follows.

[0057]

[0058] The results of physical stability showed that after being placed at room temperature 25°C and high temperature 40°C, the prepared 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation had no drug precipitation and was in good appearance, indicating that it had good stability and controllable quality; there were small crystals under high humidity conditions, which may be because the 20(S)-PPD phospholipid solid dispersion preparation has a certain hygroscopicity, so care should be taken to keep it dry during storage.

[0059] The purpose of the present invention is to provide a novel solid dispersion preparation and a preparation method thereof which has a simple prescription composition and method, a high drug loading amount, good stability, significant drug efficacy, and can effectively solve the problems of low in vivo oral absorption and utilization of 20(S)-PPD and instability of drug supersaturated solution in the gastrointestinal tract and easy recrystallization.

[0060] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0061] A 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation composition and a preparation method thereof, characterized in that the preparation is composed of 20(S)-PPD, soybean lecithin, ethanol, polyvinylpyrrolidone K30 (PVP K30), and mesoporous silica. The preparation of the present invention is prepared by a solvent volatilization method, specifically comprising the following steps:

[0062] 1) Weigh the prescribed amount of 20(S)-PPD, phospholipids and PVPK30, dissolve them in ethanol and place them in a round-bottom flask. Heat at 40°C and stir continuously to form a clear and transparent solution.

[0063] 2) Weigh a certain amount of mesoporous silica as a dispersant and add it thereto, reflux at 40° C. for 10 minutes and perform dispersion and curing.

[0064] 3) Ethanol was removed by evaporation under reduced pressure and the solid was collected.

[0065] 4) The collected solids are placed in a vacuum drying oven for 24 hours for drying.

[0066] The 20(S)-PPD content is 10-50% (w / w), preferably 20-40% (w / w).

[0067] The soybean lecithin is 1 to 3 parts, preferably 2 parts.

[0068] The content ratio of 20(S)-PPD to soybean lecithin, i.e., the drug-carrying ratio, is 1 to 4:1, preferably 3:1.

[0069] The ethanol concentration is 90-100% (w / w), preferably 95%

[0070] The crystal inhibitor is one of PVPK30 and copolyvidone S630, preferably PVPK30.

[0071] The PVPK30 is used in an amount of 1 to 4 parts, preferably 3 parts; and the content is 3 to 10% (w / w), preferably 8 to 9% (w / w).

[0072] The dispersant is one of mesoporous silica and microporous starch, preferably mesoporous silica.

[0073] The mesoporous silica is XP3050, One of 244FP, with a particle size of 3.5 to 50 μm; preferably XP3050, preferably with a particle size of 50 μm.

[0074] The dispersant is 15 to 20 parts, preferably 16 parts, of one of mesoporous silica and microporous starch; the content is 40 to 60% (w / w), preferably 50 to 55% (w / w).

[0075] The preparation process of the 20(S)-PPD phospholipid solid dispersion preparation is characterized in that the heating temperature of the preparation is 37-60° C., preferably 40° C.; the heating time is 5-15 min, preferably 10 min.

[0076] The advantages of the 20(S)-PPD phospholipid solid dispersion preparation of the present invention are:

[0077] 1) The composition of the invention is simple and easy to obtain, the preparation process is simple, the drug loading capacity is high, and it is suitable for mass production.

[0078] 2) The presence of phospholipids in the present invention can protect the small crystal nuclei of 20(S)-PPD drugs and inhibit their growth under the supersaturated state of the intestine, thereby improving the oral bioavailability of the preparation.

[0079] 3) The polyvinylpyrrolidone K30 (PVP K30) used in the present invention as a crystal inhibitor can not only maintain the amorphous state of the drug in the phospholipid, but also significantly improve the physical stability and oral bioavailability of the preparation.

[0080] 4) The present invention utilizes mesoporous silica as a dispersant, and the complex formed by the drug and the carrier penetrates into the pores of the mesoporous silica by means of capillary action, thereby achieving a uniform and fine solid powder that cannot be achieved by the grinding method. Mesoporous silica has a high affinity with phospholipids, and can be adsorbed on the cell surface and taken up by a variety of cells through endocytosis, thereby improving the solubility and bioavailability of the drug in the small intestine; and the mesoporous silica has a high adsorption amount per unit volume and a large powder density, and has good powder fluidity, and can remain a free-flowing powder after adsorption.

[0081] 5) The present invention has excellent powder properties. Its angle of repose θ is measured to be 39°, and its compression degree is 20%. The powder fluidity is good and meets the production fluidity requirements.

[0082] 6) The nanosuspension can be formed after reconstitution, the drug loading amount is increased and the solubility and dissolution rate of the poorly soluble drugs are significantly improved.

[0083] 7) The present invention has good stability and high drug loading, and the 20(S)-PPD content in the preparation can reach more than 40%; the oral absolute bioavailability is high, which can reach 23.95%. The preparation has significant efficacy, and after oral administration, it can significantly alleviate the damage of podocytes that maintain the integrity of the glomerular filtration barrier, and treat chronic nephrotic syndrome in adults such as membranous nephropathy (MN). BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 Schematic diagram of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation

[0085] Figure 2 DSC thermal analysis diagram of solid dispersion of Example 1

[0086] Figure 3 20(S)-PPD structure diagram

[0087] Figure 4 Phospholipid structure diagram

[0088] Figure 5 Structure of 20(S)-PPD-phospholipid-PVPK30 complex

[0089] Figure 6 Malvern Nano Particle Size Analyzer Determined Particle Size Distribution of Solid Dispersion in Example 1

[0090] Figure 7 Oral medication curve (20(S)-PPD fat emulsion is used as the reference preparation to determine the relative bioavailability of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation)

[0091] Figure 8 Intravenous injection time curve

[0092] Figure 920(S)-PPD phospholipid / mesoporous silica solid dispersion preparation cumulative release curve

[0093] Fig.10 Oral medication time curve

[0094] Fig.11 Intravenous injection time curve

[0095] Fig.12 Body weight changes of rats in each group

[0096] Fig.13 Statistical chart of 24h urine protein excretion in MN model rats

[0097] Fig.14 Statistical chart of BUN and SCr levels in MN model rats (A. BUN level; B. SCr level) Fig.15 Statistical chart of TP and ALB levels in MN model rats (A. TP level; B. ALB level)

[0098] Fig.16 Statistical chart of TG and TC levels in MN model rats (A. TG level; B. TC level)

[0099] Fig.17 Renal tissue of MN model rats observed under light microscope after HE staining (×400)

[0100] Fig.18 Renal tissue of MN model rats observed under light microscope after PAS staining (×400)

[0101] Fig.19 Ultrastructural image of kidney tissue in MN model rats observed under electron microscope

[0102] Fig. 20 Expression of Desmin in glomeruli of MN model rats

[0103] Fig.21 GR expression in glomeruli of MN model rats

[0104] Fig. 22 Expression of GR mRNA in kidney tissue of MN model rats

[0105] Fig.23 Expression of NF-κB mRNA in kidney tissue of MN model rats

[0106] Fig.24 Expression of Desmin in kidney tissue of MN model rats

[0107] Fig.25 Expression of GR in kidney tissue of MN model rats

[0108] Fig.26 Expression of NF-κB P65, p-NF-κB P65 and IL-1β in renal tissue of MN model rats

[0109] Fig. 27 Blood glucose test at rat tail tip

[0110] Fig.28 Glucose detection chart of rat serum

[0111] Fig.29 Expression of TAT and PEPCK mRNA in liver tissues of rats in each group ((A) TAT mRNA expression level; (B) PEPCK mRNA expression level)

[0112] Fig.30 Expression of ANGPTL4 mRNA in renal tissue of rats in each group

[0113] Fig.31 Immune organ index of rats in each group (A. thymus index; B. spleen index; C. adrenal index) DETAILED DESCRIPTION

[0114] The present invention is further described below in conjunction with examples, and embodiments of the present invention include but are not limited to the following examples.

[0115] The preparations in each example were prepared using the following process, and the specific process parameters are shown in each example.

[0116] S1: Weigh the prescribed amount of 20(S)-PPD, phospholipids and PVPK30, dissolve them in ethanol and place them in a round-bottom flask. Heat at 40°C and stir continuously to form a clear and transparent solution.

[0117] S2: Weigh a certain amount of mesoporous silica as a dispersant and add it thereto, reflux at 40° C. for 10 minutes and perform adsorption curing.

[0118] S3: Ethanol was removed by evaporation under reduced pressure and the solid was collected.

[0119] S4: The collected solids are placed in a vacuum drying oven for 24 hours for drying.

[0120] The dissolution of the preparations in each example was determined by the following method, and the specific dissolution results are shown in each example.

[0121] The in vitro dissolution characteristics of the preparation were investigated by the paddle method. 250 mL of 0.5% SDS aqueous solution (dissolution medium) was measured and placed in each dissolution cup. After the temperature of the dissolution medium was constant at 37°C ± 0.5°C, 30 mg of the sample to be tested was placed in each dissolution cup. The instrument was started at a specified speed of 50 r / min and the timing was started. At 10, 15, 20, 30, 45, 60, 75, 90, and 120 min, 5 ml of the dissolution solution was drawn and supplemented with an equal volume (5 ml) of the dissolution medium. 3 ml was discarded and filtered with a 0.45 μm microporous membrane. The filtrate was taken for HPLC analysis to determine the content.

[0122] Embodiment 1:

[0123] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0124] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 92.1% within 120 min.

[0125] Embodiment 2:

[0126] Weigh 600g 20(S)-PPD, 300g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0127] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 89.6% within 120 min.

[0128] Embodiment 3:

[0129] Weigh 600g 20(S)-PPD, 400g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0130] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was non-crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 88.7% within 120 min.

[0131] Embodiment 4:

[0132] Weigh 600g 20(S)-PPD, 500g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0133] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 90.4% within 120 min.

[0134] Embodiment 5:

[0135] Weigh 600g 20(S)-PPD, 600g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0136] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 87.4% within 120 min.

[0137] Embodiment 6:

[0138] Weigh 600g 20(S)-PPD, 600g soybean lecithin (PC70) and 200g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0139] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation was a white uniform powder with a cumulative release of 88.1% within 120 min.

[0140] Embodiment 7:

[0141] Weigh 600g 20(S)-PPD, 500g soybean lecithin (PC70) and 200g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0142] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder with a cumulative release of 85.6% within 120 minutes.

[0143] Embodiment 8:

[0144] Weigh 600g 20(S)-PPD, 600g soybean lecithin (PC70) and 100g PVPK30, dissolve them in 30ml ethanol and place them in a round-bottom flask. Heat at 40℃ and stir continuously to form a clear and transparent solution. Weigh 1600g mesoporous silica. XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0145] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder with a cumulative release of 85.8% within 120 minutes.

[0146] Embodiment 9:

[0147] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC70) and 300g copolyvidone S630, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0148] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder, and the cumulative release within 120 minutes is 84.9%.

[0149] Embodiment 10:

[0150] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 40℃ and stir continuously to form a clear and transparent solution; weigh 1800g mesoporous silica 244FP is added as a dispersant, and the mixture is refluxed at 40°C for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and the solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying to obtain the product.

[0151] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was non-crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation was a white inhomogeneous powder with a cumulative release of 89.6% within 120 min.

[0152] Embodiment 11:

[0153] 600 g 20(S)-PPD, 200 g soybean lecithin (PC70) and 300 g PVPK30 were weighed, dissolved in 30 ml ethanol and placed in a round-bottom flask, and stirred continuously at 40°C to form a clear and transparent solution; 1900 g microporous starch was weighed as a dispersant and added thereto, and refluxed at 40°C for 10 min to solidify; ethanol was evaporated under reduced pressure and the solid matter was collected; the collected solid matter was placed in a vacuum drying oven for 24 hours to obtain the product.

[0154] The prepared 20(S)-PPD-phospholipid-PVPK30 complex was non-crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation was a white inhomogeneous powder with a cumulative release of 85.7% within 120 min.

[0155] Embodiment 12:

[0156] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC50) and 300g PVPK30, dissolve them in 30ml ethanol and place them in a round-bottom flask. Heat at 40℃ and stir continuously to form a clear and transparent solution. Weigh 1600g mesoporous silica. XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0157] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is amorphous, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder with a cumulative release of 90.5% within 120 minutes.

[0158] Embodiment 13:

[0159] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, stir continuously at constant temperature to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, reacted for 10 minutes under constant temperature conditions and solidified; ethanol is evaporated under reduced pressure and solids are collected; the collected solids are placed in a vacuum drying oven for 24 hours for drying to obtain the product.

[0160] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder with a cumulative release of 85.7% within 120 minutes.

[0161] Embodiment 14:

[0162] Weigh 600g 20(S)-PPD, 200g soybean lecithin (PC70) and 300g PVPK30, dissolve in 30ml ethanol and place in a round-bottom flask, heat at 60℃ and stir continuously to form a clear and transparent solution; weigh 1600g mesoporous silica XP3050 is added as a dispersant, and the mixture is refluxed at 40° C. for 10 minutes for solidification; ethanol is evaporated under reduced pressure, and solid matter is collected; the collected solid matter is placed in a vacuum drying oven for 24 hours for drying.

[0163] The prepared 20(S)-PPD-phospholipid-PVPK30 complex is crystalline, and the 20(S)-PPD phospholipid solid dispersion preparation is a white uniform powder, and the cumulative release within 120 minutes is 86.8%.

[0164] The physical parameters of the 20(S)-PPD phospholipid solid dispersion preparations prepared by the respective prescriptions of Examples 1 to 14 are shown in the following table.

[0165]

[0166]

[0167] Embodiment 15

[0168] In vitro dissolution test of the 20(S)-PPD phospholipid solid dispersion preparation of the present invention.

[0169] Test method: This test uses the paddle method to examine the in vitro dissolution characteristics of the preparation. The stirring paddle speed is 50r·min-1, the water bath temperature is (37±0.5)℃, and the dissolution medium is 250ml of 0.5% SDS aqueous solution. The in vitro dissolution results are shown in Fig. 9 shown.

[0170] In the in vitro cumulative release study, the cumulative release of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation was 92.1% within 120 min.

[0171] Example 16

[0172] Pharmacokinetic study of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation of the present invention.

[0173] 1. Pharmacokinetic evaluation of the preparation prepared in Example 1 in rats

[0174] Male SD rats, 180±20g, were randomly divided into 8 groups, 6 rats in each group. The 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation had a content of 14.42%, a dosage of 35mg / kg, and a volume of 10ml / kg; the 20(S)-PPD fat emulsion preparation had a content of 3.62mg / ml, a dosage of 35mg / kg, and a volume of 10ml / kg were administered by oral gavage.

[0175] Average absolute bioavailability after oral administration F = 23.95%

[0176] Taking oral 20(S)-PPD fat emulsion as reference preparation, the relative bioavailability of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation was 123.22%. The specific results are shown in Fig.10 (oral medication curve) and 11 (intravenous injection medication curve).

[0177] Comparative Example 1

[0178] According to the patent specification, the dosage of auxiliary materials of 20(S)-PPD phospholipid / mesoporous silica solid dispersion ranges from 1 to 6 parts of soybean lecithin, 1 to 3 parts of PVPK30, and 15 to 20 parts of mesoporous silica.

[0179] Weigh 600 g of 20(S)-PPD, 7 parts of soybean lecithin (i.e. 700 g), 4 parts of PVPK30 (i.e. 400 g), and 21 parts of mesoporous silica (i.e. 2100 g), and prepare a 20(S)-PPD lecithin / mesoporous silica solid dispersion preparation according to the steps of the above embodiment. The prepared preparation structure has no crystals, but the drug loading is only 18.75%.

[0180] Comparative Example 2

[0181] Weigh 600 g of 20(S)-PPD, 6 parts of soybean lecithin (i.e. 600 g), and 21 parts of mesoporous silica (i.e. 2100 g), and prepare a 20(S)-PPD lecithin / mesoporous silica solid dispersion preparation according to the steps in the above embodiment. The prepared preparation has a large amount of crystalline structure, which does not meet the amorphous requirement, and the actual drug loading is significantly reduced.

[0182] Comparative Example 3

[0183] Weigh 600 g of 20(S)-PPD, 6 parts of soybean lecithin (600 g), 3 parts of PVPK30 (300 g), and 14 parts of mesoporous silica (1400 g), and prepare a 20(S)-PPD lecithin / mesoporous silica solid dispersion preparation according to the steps of the above embodiment. The soybean lecithin cannot be completely solidified, the preparation is viscous, and the fluidity is significantly reduced.

[0184] Experimental Example 1

[0185] Pharmacodynamics study test of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation of the present invention.

[0186] 1. Objective: To investigate the effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation on improving membranous nephritis (MN).

[0187] 2. The pharmacodynamics of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion prepared in Example 1 was studied, and the experimental results are as follows.

[0188] 2.1 General Observations

[0189] During the experiment, the diet, drinking water, defecation, hair color and exercise of each group of rats were observed, and the weight of the rats was weighed. Fig.12 As shown (experimental data The results are shown in Table 1 (n=8). * P<0.05, ** P<0.01vs. blank control group; ## P<0.01vs. model group; & P<0.05, && P<0.01 vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group).

[0190] Summary: Within 1 week after modeling, rats developed different degrees of diarrhea, lack of energy, decreased food intake, etc. After 4 weeks, the rats in the model group showed severe mental depression, dark and messy fur, greatly reduced food intake, and accompanied by edema of the lower limbs and testicles. The mental state of the rats in the normal dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly improved, the food intake was normal, and the edema was also significantly improved; the weight of the rats in the model group decreased significantly after modeling. The experimental results are shown in Figure 3.1. Compared with the blank control group, the body weights of rats in the normal dose Pred group and the 20(S)-PPD phospholipid solid dispersion control group fluctuated slightly, but there was no significant difference. The body weights of rats in the model group decreased significantly. Compared with the blank control group, the body weights of rats in the model group were significantly different from those in the blank control group on the 7th, 14th, 21st and 28th days of the experiment (P<0.01). Compared with the model group, the body weights of rats in each drug-treated group increased to varying degrees. After three weeks of drug intervention, on the 28th day of the experiment, the body weights of rats in each drug-treated group were significantly different from those in the model group (P<0.01).

[0191] 2.2 Effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on 24h urine protein content in MN model rats. The specific excretion amount is shown in Fig.13 As shown (experimental data The results are shown in Table 1 (n=8). * P<0.05, ** P<0.01vs. blank control group; ## P<0.01vs. model group; & P<0.05, && P<0.01 vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group).

[0192] Conclusion: Compared with the blank control group, the 24-hour urinary protein excretion of rats in each group after tail vein injection of ADR modeling continued to increase. On the 7th day of the experiment, the 24-hour urinary protein excretion of the model group was significantly increased (P<0.05), and the urine protein level continued to increase on the 14th, 21st, and 28th days of the experiment (P<0.01). After one week of drug intervention, on the 14th day of the experiment, compared with the model group, the normal dose Pred group, 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred All groups showed significant effects in reducing 24-hour urinary protein excretion in rats (P<0.01). After two and three weeks of drug intervention, on the 21st and 28th days of the experiment, compared with the model group, the 24-hour urinary protein excretion of rats in each drug-treated group was further reduced (P<0.01). The effect of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was better, significantly better than the low-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group (P<0.01), and slightly better than the normal-dose Pred group (P<0.05).

[0193] 2.3 Effects of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on blood biochemical indicators of MN model rats.

[0194] (1) Effects on serum BUN and SCr levels in MN rats. For specific results, see Fig.14 As shown (experimental data The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; ## P<0.01vs. model group; & P<0.05, && P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group. ).

[0195] Summary: Compared with the blank control group, the levels of BUN and SCr in the model group were significantly increased (P<0.01); compared with the model group, the levels of BUN and SCr in the serum of the rats in the normal dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group were significantly decreased (P<0.01), and the levels of BUN and SCr in the serum of the rats in the low dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion group were also decreased to varying degrees (P<0.05, P<0.01); the effect of the PPD+Pred group was significantly better than that of the low dose group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion group (P<0.05, P<0.01), and slightly better than that of the normal dose Pred group (P<0.05).

[0196] (2) Effects on the levels of TP and ALB in the serum of MN rats. For specific results, see Fig.15 As shown (experimental data The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; ## P<0.01vs. model group; & P<0.05, && P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group. ).

[0197] Summary: Compared with the blank control group, the TP and ALB levels of the rats in the model group were significantly decreased, with significant differences (P<0.01, P<0.01), showing hypoalbuminemia; compared with the model group, the TP and ALB levels of the rats in the normal dose Pred group, 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group were significantly increased (P<0.01, P<0.01), and the TP and ALB levels of the rats in the low dose Pred group were also increased to a certain extent (P<0.05); the effect of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of the low dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group, and slightly better than that of the normal dose Pred group, all with significant differences.

[0198] (3) Effects on serum TG and TC levels in MN rats. Fig.16 As shown (experimental data The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; ## P<0.01vs. model group; & P<0.05, && P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group. ).

[0199] Summary: Compared with the blank control group, the TG and TC levels of the rats in the model group were significantly increased (P<0.01, P<0.01), showing hyperlipidemia; compared with the model group, the TG and TC levels of the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group were significantly decreased (P<0.01, P<0.01), and the TG and TC levels of the rats in the Pred group with different doses decreased to varying degrees (P<0.05, P<0.01), but the effect was significantly inferior to that in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group (P<0.01, P<0.05).

[0200] 2.4 Effects of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on renal tissue pathological changes in MN model rats. Specific results are shown in Fig.17 (Image of kidney tissue of MN model rat observed under light microscope after HE staining (×400)), 18 (Image of kidney tissue of MN model rat observed under light microscope after PAS staining (×400)).

[0201] Summary: The renal tissue structure of the rats in the blank control group was normal without any pathological changes. Compared with the blank control group, the rats in the model group showed tubular protein cast lesions (HE staining, indicated by arrows), local thickening of the glomerular basement membrane (PAS staining, indicated by arrows), and widening of the renal capsule cavity, and a small amount of inflammatory cell infiltration in the renal interstitium around some glomeruli. Compared with the model group, the rats in the normal-dose Pred group, 20(S)-PPD phospholipid / mesoporous silica solid dispersion group, and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group had no obvious lesions in the kidneys, the thickness of the glomerular basement membrane was basically uniform, and the kidney damage was significantly improved. The kidney damage of the rats in the low-dose Pred group was improved to a certain extent, but tubular protein casts were still visible.

[0202] 2.5 Effects of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on foot process fusion of glomerular podocytes in MN model rats; see Fig.19 (Ultrastructure of renal tissue of MN model rats observed under electron microscope (scale bar 1 μm)) shown.

[0203] Conclusion: The foot processes of podocytes in the blank control group were intact with obvious slits, while the foot processes of podocytes in the model group were obviously fused and the slits disappeared (marked by black triangles). Compared with the model group, the fusion of foot processes of podocytes in the normal-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly improved, and the fusion of foot processes in the low-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group was also improved to varying degrees. The effect of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of the low-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group.

[0204] 2.6 Effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on the expression of Desmin and GR proteins in the glomeruli of MN model rats.

[0205] (1) Effects on the expression of Desmin protein in the glomeruli of MN rats. For specific results, see Fig. 20 (Expression of Desmin in glomeruli of MN model rats (×400)) It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group. ).

[0206] Summary: Desmin was almost not expressed in the glomeruli of rats in the blank control group; compared with the blank control group, the expression of Desmin in the glomeruli of rats in the model group was significantly increased (P<0.01); compared with the model group, the expression of Desmin in the glomeruli of rats in each drug-treated group was reduced (P<0.05, P<0.01), among which the effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was better than that of low-dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion group (P<0.05).

[0207] (2) Effects on GR protein expression in the glomeruli of MN rats. For specific results, see Fig.21 (GR expression in glomeruli of MN model rats (×400)) (Experimental data are shown in It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group. )

[0208] Summary: Compared with the blank control group, the expression of GR in the glomeruli of the rats in the model group was significantly decreased (P<0.01); compared with the model group, the expression of GR in the glomeruli of the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly increased (P<0.01), and the effect was significantly better than that in the low-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group (P<0.05).

[0209] 2.7 Effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on the expression levels of GR and NF-κB mRNA in the renal tissue of MN model rats.

[0210] (1) Effects on GR mRNA expression levels in renal tissue of MN rats. For specific results, see Fig. 22 (Expression of GR mRNA in renal tissue of MN model rats) (Experimental data are shown in It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group)

[0211] Summary: Compared with the blank control group, the GR mRNA expression level in the renal tissue of the rats in the model group was significantly decreased (P<0.01); compared with the model group, the GR mRNA expression level in the renal tissue of the rats in the normal dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly increased (P<0.01), and the GR mRNA expression level in the renal tissue of the rats in the low dose Pred group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion group also increased to varying degrees (P<0.05); the effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was better than that of 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and low dose Pred group (P<0.05).

[0212] (2) Effects on NF-κB mRNA expression in renal tissue of MN rats. For details, see Fig.23 (Expression of NF-κB mRNA in renal tissue of MN model rats) (Experimental data are shown in It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group).

[0213] Summary: Compared with the blank control group, the expression level of NF-κB mRNA in the renal tissue of the rats in the model group was significantly increased (P<0.01); compared with the model group, the expression level of NF-κB mRNA in the renal tissue of the rats in the normal dose Pred group, 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly decreased (P<0.01); the effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and low dose Pred group (P<0.01), and slightly better than that of the normal dose Pred group (P<0.05).

[0214] 2.8 Effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on the expression levels of Desmin, GR, NF-κB and IL-1β related proteins in MN model rats.

[0215] (1) Effect on the expression level of Desmin protein in the kidney tissue of MN rats. For specific results, see Fig.24 (Expression of Desmin in renal tissue of MN model rats) (Experimental data are shown in **P<0.01 vs. blank control group; #P<0.05, ##P<0.01 vs. model group; &P<0.05 vs. 20(S)-PPD phospholipid solid dispersion + Pred group)

[0216] Summary: Compared with the blank control group, the expression level of Desmin protein in the renal tissue of the rats in the model group was significantly increased (P<0.01); compared with the model group, the expression level of Desmin in the renal tissue of the rats in each drug-treated group decreased to varying degrees, among which the effect of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of the low-dose Pred group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group (P<0.05), and was equivalent to the normal-dose Pred group, but there was no significant difference.

[0217] (2) Effects on GR protein expression in renal tissue of MN rats. For specific results, see Fig.25 (Expression of GR in renal tissue of MN model rats) (Experimental data are shown in **P<0.01 vs. blank control group; #P<0.05, ##P<0.01 vs. model group; &P<0.05, &&P<0.01 vs. 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group)

[0218] Summary: Compared with the blank control group, the protein expression level of GR in the renal tissue of the rats in the model group was significantly decreased (P<0.05); compared with the model group, the expression level of GR in the renal tissue of the rats in each drug-treated group was improved to varying degrees, among which the effect of the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of the low-dose Pred group (P<0.01), and better than that of the single 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the normal dose Pred group (P<0.05).

[0219] (3) Effects on the expression levels of NF-Kb P65, p-NF-κB P65, and IL-1β proteins in the renal tissue of MN rats. The specific results are shown in Fig.26 (Expression of NF-κB P65, p-NF-κB P65 and IL-1β in renal tissue of MN model rats (A) Expression levels of NF-κB P65 and p-NF-κB P65; (B) Expression level of IL-1β) (Experimental data are shown in It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05,&& P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group)

[0220] Summary: Compared with the blank control group, the expression levels of NF-κB P65, p-NF-κB P65 and IL-1β in the renal tissue of rats in the model group were significantly increased (P<0.01, P<0.01 or P<0.01). Compared with the model group, the expression levels of NF-κB P65, p-NF-κB P65 and IL-1β in the renal tissue of rats in each drug-treated group were decreased to varying degrees, among which the effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly better than that of low-dose Pred group and 20(S)-PPD phospholipid solid dispersion group, with significant differences.

[0221] 2.9 Effect of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on the blood glucose levels of rats in each group.

[0222] (1) Effects on blood glucose levels at the tail tip of rats. See Fig. 27 (Rat tail tip blood glucose level) (experimental data The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05, && P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group)

[0223] Summary: Compared with the blank control group, the blood glucose level of the rats in the model group fluctuated slightly at the tip of the tail, and the blood glucose level of the rats on the 7th day of the experiment was significantly increased (P<0.01); after one week of drug intervention, on the 14th day of the experiment, compared with the blank control group, the blood glucose level of the rats in the Pred group was increased (P<0.05), and the blood glucose level of the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation control group was normal. The blood glucose level of the rats in the model group decreased but was still higher than that of the rats in the blank control group, but there was no significant difference; compared with the model group, the blood glucose level of the rats in the normal dose Pred group was slightly increased, but not significantly; after two weeks of drug intervention, on the 21st day of the experiment, compared with the blank control group, the blood glucose level of the rats in the Pred control group was significantly increased (P<0.01), and the blood glucose level of the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation control group was normal. The blood glucose level of the rats in the model group was slightly higher; compared with Compared with the model group, the blood glucose levels of rats in the normal-dose Pred group and the low-dose Pred group increased to varying degrees (P<0.01, P<0.05), while the blood glucose levels of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group did not increase; after three weeks of drug intervention, on the 28th day of the experiment, compared with the blank control group, the blood glucose levels of rats in the Pred control group increased abnormally (P<0.01), while the blood glucose levels of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion control group were still normal, and the blood glucose levels of rats in the model group were slightly higher, but there was no significant difference; compared with the model group, the blood glucose levels of rats in the normal-dose Pred group and the low-dose Pred group increased significantly (P<0.01), while the blood glucose levels of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group were not abnormal.

[0224] (2) Effects on glucose levels in rat serum. For specific results, see Fig.28 (Glucose levels in rat serum) are shown in (experimental data are shown in The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group)

[0225] Summary: Compared with the blank control group, the serum glucose level of rats in the Pred control group was significantly increased (P<0.01), while there was no increase in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion control group. The serum glucose level of rats in the model group was slightly increased, but there was no significant difference. Compared with the model group, the serum glucose levels of rats in the Pred administration groups with different doses increased to varying degrees (P<0.05), while the serum glucose levels of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group decreased, but there was no significant difference. The serum glucose levels of rats in the Pred administration group were significantly different from those in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group (P<0.05).

[0226] 2.10 Effects of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on the expression levels of TAT, PEPCK in liver tissue and ANGPTL4 mRNA in kidney tissue of rats in each group

[0227] (1) Effects on the expression levels of TAT and PEPCK mRNA in rat liver tissue. For specific results, see Fig.29 (Expression levels of TAT and PEPCK mRNA in liver tissues of rats in each group (A) TAT mRNA expression level; (A) PEPCK mRNA expression level) (Experimental data are shown in It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; && P<0.01 vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group).

[0228] Summary: Compared with the blank control group, the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the Pred control group were significantly increased (P<0.01, P<0.01), the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion control group fluctuated slightly, but no abnormality was found, and the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the model group were significantly decreased (P<0.01, P<0.01); compared with the model group, the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the Pred administration groups with different doses were significantly increased (P<0.01, P<0.01), the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group were increased, and the mRNA expression levels of TAT and PEPCK in the liver tissue of rats in the Pred administration groups with different doses were significantly increased. The mRNA expression levels were significantly different compared with the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group (P<0.01, P<0.01).

[0229] (2) Effect on the expression level of ANGPTL4 mRNA in rat kidney tissue. For specific results, see Fig.30 (Expression of ANGPTL4 mRNA in renal tissue of rats in each group) It is represented (n=3). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; && P<0.01vs.20(S)-PPD phospholipid / mesoporous silica solid dispersion+Pred group)

[0230] Summary: Compared with the blank control group, the expression level of ANGPTL4 mRNA in renal tissue of rats in the Pred control group was significantly increased (P<0.01), the expression level of ANGPTL4 mRNA in renal tissue of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion control group was normal, and the expression level of ANGPTL4 mRNA in renal tissue of rats in the model group was increased (P<0.01); compared with the model group, the expression level of ANGPTL4 mRNA in renal tissue of rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion group and the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group was significantly decreased (P<0.01), and the expression levels of ANGPTL4 mRNA in renal tissue of rats in different doses of Pred treatment groups were significantly different from those in the combined treatment group (P<0.01).

[0231] 2.11 Effects of 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred on immune organ indexes of rats in each group. For specific results, see Fig.31 (Immune organ index of each group of rats A. thymus index; B. spleen index; C. adrenal index) (experimental data are shown in The results are shown in Table 1 (n=8). ** P<0.01vs. blank control group; # P<0.05, ## P<0.01vs. model group; & P<0.05, && P<0.01 vs. 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation + Pred group).

[0232] Summary: Compared with the blank control group, the thymus, spleen and adrenal glands of the rats in the Pred control group were atrophied to varying degrees. The thymus, spleen and adrenal gland indexes of the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion control group fluctuated slightly, and no abnormalities were found. The thymus and adrenal glands of the rats in the model group were atrophied, and the spleen was swollen, but there were no significant differences. Compared with the model group, the thymus, spleen and adrenal glands of the rats in the normal dose Pred group were severely atrophied (P<0.01, P<0.01 or P<0.01), the thymus and spleen of the rats in the low dose Pred group were significantly atrophied (P<0.05, P<0.05), and the adrenal glands were also atrophied to a certain extent, but there were no significant differences. No atrophy of immune organs was found in the rats in the 20(S)-PPD phospholipid / mesoporous silica solid dispersion + Pred group.

[0233] Conclusion: Based on the above experimental results, we can draw the following conclusions: 20(S)-PPD phospholipid / mesoporous silica solid dispersion preparation combined with Pred can significantly improve the symptoms of MN model rats. 20(S)-PPD phospholipid / mesoporous silica solid dispersion has a synergistic effect on hormone drugs, reduces the dosage of hormone drugs, and does not produce toxic side effects similar to hormones, such as increased blood sugar levels and atrophy of immune organs. Its mechanism of action may be related to the transcriptional inhibition and transcriptional activation effects on GR.

[0234] The above embodiments are only preferred embodiments of the present invention and are not intended to limit the protection scope of the present invention. Any changes made by adopting the design principles of the present invention and performing non-creative work on this basis should fall within the protection scope of the present invention.

Claims

1. A 20(S)-PPD phospholipid / mesoporous silica solid dispersion, characterized in that: The preparation consists of 20(S)-PPD, phospholipid, ethanol, a crystal inhibitor and a dispersant.

2. The preparation according to claim 1, characterized in that The preparation comprises 1 to 6 parts of phospholipids, 1 to 3 parts of PVPK30, and 15 to 20 parts of mesoporous silica; preferably, the content of 20(S)-PPD in the preparation is 10 to 40% (w / w); preferably, the phospholipid is one of soybean phospholipids PC70 or / PC50; preferably, the ratio of the amount of 20(S)-PPD to soybean phospholipids, i.e., the drug-carrying ratio, is 1 to 4:1; preferably, the concentration of ethanol is 90 to 100% (w / w).

3. The preparation according to claim 1, characterized in that The crystal inhibitor is one of polyvinyl pyrrolidone K30 (PVPK30) and / or copolyvidone S630, and the content is 3-10% (w / w).

4. The preparation according to claim 1, characterized in that The dispersant is one of mesoporous silica and microporous starch, and the content is 50-60% (w / w).

5. The preparation according to claim 4, characterized in that The function of the dispersant is mesoporous dispersion and enables the 20(S)-PPD phospholipid / mesoporous silica solid dispersion to obtain excellent powder properties; the brand and model of the mesoporous silica are preferably XP3050, One of 244FP, with a particle size of 3.5 to 50 μm.

6. A method for preparing the preparation according to any one of claims 1 to 5, comprising the following steps: 1) Dissolve 20(S)-PPD, soybean lecithin and PVP K30 in ethanol and stir continuously to fully dissolve. 2) Adding an appropriate amount of mesoporous silica as a dispersant to the components in step 1), heating to reflux and curing. 3) Ethanol was removed by evaporation under reduced pressure and the solid was collected. 4) Place the mixture in a vacuum drying oven and dry it in vacuum for 24 hours to obtain a 20(S)-PPD phospholipid / mesoporous silica solid dispersion.

7. The preparation method according to claim 6, characterized in that: The dissolving method in step 1) is heating and stirring to dissolve, and the heating temperature is preferably 37-60°C.

8. The preparation method according to claim 7, characterized in that: The heating time is 5 to 15 minutes.

9. Use of the preparation according to any one of claims 1 to 5 or the preparation prepared by the preparation method according to any one of claims 6 to 8 in the preparation of a drug for improving podocyte injury or a drug for treating membranous glomerulonephritis.

10. The bioavailability of the preparation according to any one of claims 1 to 5 or the preparation prepared by the preparation method according to any one of claims 6 to 8 is significantly improved.

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