A solid dispersion of nobiletin and its application in preparing medicine for preventing and treating liver damage

Abstract

The present invention belongs to the technical field of natural products, and particularly relates to a solid dispersion of Nobiletin and a preparation method thereof and an application thereof in the preparation of a drug for preventing and treating liver damage. The solid dispersion of Nobiletin of the present invention comprises the following components in mass percentage: Soluplus 70-80%, PVP / VA64 10-15%, and the content of Nobiletin is 10-15%. The present invention also provides a method for preparing the above dispersion. The solid dispersion of Nobiletin of the present invention is compounded by Nobiletin and carrier materials Soluplus and PVP / VA64, and has the advantages of accurate content, small particle size, stable functional substance properties, not easy to oxidize, and high bioavailability. The solid dispersion of Nobiletin of the present invention exhibits obvious liver protection, can prevent and / or reduce and / or reverse the liver damage, and can be used in the preparation of drugs / health products for treating or preventing or reversing or reducing liver damage.

Description

Technical Field

[0001] The invention belongs to the technical field of natural products, and particularly relates to a nobiletin solid dispersion and a preparation method thereof and application thereof in preparing a drug for preventing and treating liver damage. Background Art

[0002] Nobiletin (NOB) is a low-polar polymethoxyflavonoid natural product with a planar structure. It is found in high concentrations in traditional Chinese medicines such as dried orange peel and dried orange peel of the Rutaceae family, and has good antioxidant properties. Therefore, it is speculated that developing Nobiletin as a liver-protecting drug for preventing and treating liver damage has strong feasibility. However, since NOB is a non-polar molecule, it has problems such as low water solubility, poor oral absorption, and low bioavailability (only about 4%), so the clinical application of NOB is greatly limited. In order to improve the antioxidant properties of Nobiletin, it is necessary to adopt appropriate formulation technology to improve its solubility so as to better exert the effect of NOB in preventing and treating liver damage.

[0003] Amorphous solid dispersion (ASD) technology is a new technology for highly dispersing drugs in solid carriers, and is an effective method to improve the solubility of poorly soluble drugs. Poorly soluble drugs are prepared into solid dispersions, and the drugs can be highly dispersed in the carrier in molecular, colloidal, microcrystalline or amorphous forms. The high dispersibility of the drug and the high solubility of the carrier can effectively improve the solubility and dissolution of the poorly soluble drug, thereby improving the bioavailability of the poorly soluble drug and enhancing its pharmacological activity.

[0004] Hot melt extrusion (HME) is a sustainable formulation technology for preparing solid dispersions. When using HME technology to prepare solid dispersions, the drug and carrier are evenly mixed in a molten state, and the mixture is extruded into shape using a rotating screw, cooled and pelletized. Compared with other solid dispersion preparation technologies, HME technology integrates multiple unit operations such as solid conveying, material melting, shear mixing, melt conveying, and extrusion molding, and has significant advantages such as no organic solvent residue and sustainable industrial production.

[0005] The liver is the main organ of human metabolism. Many factors such as drugs, poisons, and ethanol can induce liver damage. Severe or persistent liver damage will lead to various chronic liver diseases and even liver failure. Liver damage can be divided into two categories: chemical liver damage and immune liver damage. Chemical liver damage can be caused by a variety of chemical hepatotoxic substances, including carbon tetrachloride, acetaminophen, acetaldehyde, etc. Among them, acetaminophen is a classic hepatotoxic poison that can cause acute liver damage. In recent years, the incidence of acute liver damage has increased year by year, and various liver diseases have seriously threatened human health. Therefore, it is of great significance to study natural products or drugs for the prevention and treatment of liver damage. Summary of the invention

[0006] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide a solid dispersion of nobiletin.

[0007] Another object of the present invention is to provide a method for preparing the above-mentioned Nobiletin solid dispersion.

[0008] Another object of the present invention is to provide the use of the nobiletin solid dispersion in the preparation of drugs for preventing and treating liver damage.

[0009] The purpose of the present invention is achieved through the following solutions:

[0010] A nobiletin solid dispersion (NOB / SD) comprises the following components in mass percentage: Soluplus 70-80%, PVP / VA64 10-15%, and the content of nobiletin is 10-15%.

[0011] As one embodiment of the present invention, the nobiletin is nobiletin from citrus sources such as tangerine peel.

[0012] As a scheme of the present invention, the mass ratio of PVP / VA64 to Soluplus is 1:8-3:14. The present invention adopts Soluplus, PVP / VA64 and Nobiletin to compound to obtain Nobiletin solid dispersion, which has high solubility, large drug loading and good stability; the Nobiletin solid dispersion of the present invention is added into water, and its particle size is less than or equal to 200nm; its equilibrium solubility is 5.8 times that of free NOB (Crystalline NOB), and the cumulative dissolution is more than 70%, which effectively improves the solubility, bioavailability and biological activity of Nobiletin, and effectively overcomes the technical problems of poor water solubility and low bioavailability of Nobiletin in the prior art.

[0013] The present invention also provides a method for preparing the above-mentioned Nobiletin solid dispersion, which is a hot melt extrusion method, specifically comprising uniformly mixing the components in proportion and extruding the mixture through a hot melt extruder.

[0014] As a solution of the present invention, the extrusion temperature is less than or equal to 170°C.

[0015] As a solution of the present invention, the feeding speed of the hot melt extruder is less than or equal to 50 mg / min.

[0016] As one solution of the present invention, the screw speed of the hot melt extruder may be 40-80 rpm.

[0017] As one solution of the present invention, the indoor humidity during the preparation process is preferably 40-55%.

[0018] As one solution of the present invention, the solid dispersion of Nobiletin obtained by extrusion is in the form of a wire, and a powdered product can be obtained by pulverization or the like.

[0019] As a solution of the present invention, the crushed material may be processed by screening.

[0020] The nobiletin solid dispersion of the present invention is prepared by compounding nobiletin and carrier materials Soluplus and PVP / VA64 through hot melt extrusion, and has the advantages of accurate content, small particle size, stable functional substance properties, low oxidation resistance, high bioavailability and the like.

[0021] The present invention also provides a method for preparing the above-mentioned Nobiletin solid dispersion, which is a solvent volatilization method, specifically comprising dissolving each component in a solvent in proportion, mixing them evenly, and removing the solvent to obtain a product.

[0022] As one embodiment of the present invention, the solvent may be anhydrous ethanol, water, methanol or the like.

[0023] As one solution of the present invention, ultrasound or stirring can be used to accelerate the dissolution rate during dissolution.

[0024] As one solution of the present invention, the uniform mixing can be achieved by stirring or ultrasonic methods.

[0025] As one embodiment of the present invention, the solvent can be removed by heating, rotary evaporation, vacuum drying, etc.

[0026] As one solution of the present invention, after the solvent is removed, the product can also be placed in a drying oven to dry to remove the influence of moisture and the like.

[0027] The invention also provides application of the nobiletin solid dispersion in preparing a medicine for preventing and treating liver damage.

[0028] The present invention also provides the use of the nobiletin solid dispersion in the preparation of a drug for treating, preventing, reversing or reducing liver damage.

[0029] As one embodiment of the present invention, the liver damage includes liver tissue and cell damage.

[0030] As one embodiment of the present invention, the liver damage is caused by chemical hepatotoxic substances.

[0031] As one embodiment of the present invention, the chemical hepatotoxic substances include alcohol, carbon tetrachloride, acetaminophen, acetaldehyde and the like.

[0032] As one embodiment of the present invention, the liver injury includes toxic hepatitis, cirrhosis and liver fibrosis.

[0033] The drugs are the same or different and include a therapeutically effective amount of nobiletin solid dispersion.

[0034] The drugs described above may be the same or different and may be made into various pharmaceutical dosage forms by conventional methods, including tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, lozenges, granules, granules, pills, elixirs, suspensions, wine preparations, tinctures, drops and other oral dosage forms, as well as injections and other dosage forms other than oral administration, such as injections.

[0035] The drugs, which are the same or different, may also contain one or more pharmaceutically acceptable carriers or excipients.

[0036] The excipients may include diluents, wetting agents, lubricants, fillers, preservatives and the like.

[0037] As a solution of the present invention, the Nobiletin solid dispersion comprises the following components in mass percentage: Soluplus 70-80%, PVP / VA64 10-15%, and Nobiletin 10-15%.

[0038] As one scheme of the present invention, the solid dispersion of Nobiletin of the present invention is applied to damaged liver tissue and cells, and tissue pathological sections, ALT and AST are used as detection indicators to verify the protective effect of the solid dispersion of Nobiletin on liver damage, showing obvious liver protective effect, which can prevent and / or reduce and / or reverse the liver damage.

[0039] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0040] (1) The solid dispersion of Nobiletin of the present invention has high solubility, large drug loading, good stability, equilibrium solubility of 5.8 times that of free NOB, and cumulative dissolution of more than 70%, which effectively improves the solubility, bioavailability and biological activity of Nobiletin, and effectively overcomes the technical problems of poor water solubility and low bioavailability of Nobiletin in the prior art;

[0041] (2) The solid dispersion of Nobiletin of the present invention has a simple formula and a convenient preparation process, and has the advantages of accurate content, small particle size, stable functional substance properties, non-oxidation, and high bioavailability.

[0042] (3) The solid dispersion of nobiletin of the present invention exhibits significant liver protection effects, can prevent and / or reduce and / or reverse liver damage, and can be used in the preparation of drugs for treating, preventing, reversing or reducing liver damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0044] Figure 1 This is the particle size distribution diagram of the solid dispersion of Nobiletin and free Nobiletin.

[0045] Figure 2 The scanning electron micrographs of the solid dispersion of nobiletin and free nobiletin.

[0046] Figure 3 These are the DSC curve, FTIR curve and XRD curve of the solid dispersion of Nobiletin.

[0047] Figure 4-Figure 6 This is the dissolution diagram of Nobiletin solid dispersion.

[0048] Figure 7 This is the DSC curve of the Nobiletin solid dispersion after storage for 3 months.

[0049] Figure 8 The activity statistics of ALT and AST in mouse serum.

[0050] Fig. 9 This is a HE-stained section of mouse liver.

[0051] Fig.10 This is a graph showing the liver damage area in mice.

[0052] Fig.11 The blood concentration-time curves of nobiletin solid dispersion and free nobiletin. DETAILED DESCRIPTION

[0053] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. The materials involved in the following examples can be obtained from commercial channels unless otherwise specified. The methods described are conventional methods unless otherwise specified.

[0054] In one embodiment, a solid dispersion of nobiletin comprises the following components in mass percentages: Soluplus 70-80%, PVP / VA64 10-15%, and nobiletin 10-15%.

[0055] In one embodiment, the nobiletin is derived from dried orange peel.

[0056] In one embodiment, the mass ratio of nobiletin:Soluplus:PVP / VA64 is 1:5:1.

[0057] In another embodiment, the mass ratio of Nobiletin:Soluplus:PVP / VA64 is 3:15:2.

[0058] In another embodiment, the mass ratio of Nobiletin:Soluplus:PVP / VA64 is 3:14:3.

[0059] In yet another embodiment, the mass ratio of Nobiletin:Soluplus:PVP / VA64 is 2:15:3.

[0060] In one embodiment, the mass ratio of PVP / VA64 to Soluplus is 1:8-3:14.

[0061] The invention adopts the above-mentioned specific ratio of Soluplus, PVP / VA64 and Nobiletin to compound to obtain Nobiletin solid dispersion, which has high solubility, large drug loading and good stability; when the Nobiletin solid dispersion of the invention is added into water, its particle size is less than or equal to 200nm; its equilibrium solubility is 5.8 times of free NOB (Free NOB), and the cumulative solubility is more than 70%, which effectively improves the solubility, bioavailability and biological activity of Nobiletin, and effectively overcomes the technical problems of poor water solubility and low bioavailability of Nobiletin in the prior art.

[0062] In one embodiment, the solid dispersion of nobiletin of the present invention is added into water, and the particle size thereof is less than or equal to 200 nm.

[0063] In another embodiment, the D50 of the nobiletin solid dispersion of the present invention is 60-98 nm.

[0064] In another embodiment, the D90 of the nobiletin solid dispersion of the present invention is 32-156 nm.

[0065] In another embodiment, a method for preparing a solid dispersion of Nobiletin is a hot melt extrusion method, which specifically includes uniformly mixing the components in proportion and extruding the mixture through a hot melt extruder.

[0066] In one embodiment, the extrusion temperature is less than or equal to 170°C.

[0067] In one embodiment, the feeding speed of the hot melt extruder is less than or equal to 50 mg / min.

[0068] In one embodiment, the screw speed of the hot melt extruder may be 40-80 rpm.

[0069] In one embodiment, the indoor humidity during the preparation process is preferably less than or equal to 40-55%.

[0070] In one embodiment, the solid dispersion of nobiletin obtained by extrusion is in the form of a wire, and a powder product can be obtained by pulverization or the like.

[0071] In one embodiment, the crushed material may be processed by screening.

[0072] The nobiletin solid dispersion of the present invention is prepared by compounding nobiletin and carrier materials Soluplus and PVP / VA64 through hot melt extrusion, and has the advantages of accurate content, small particle size, stable functional substance properties, low oxidation resistance, high bioavailability and the like.

[0073] In another embodiment, a method for preparing a solid dispersion of Nobiletin is a solvent volatilization method, which specifically includes dissolving each component in a solvent in proportion, mixing them evenly, and removing the solvent to obtain a product.

[0074] In one embodiment, the solvent is anhydrous ethanol, water, methanol or the like.

[0075] In one embodiment, ultrasound or stirring is used to accelerate the dissolution rate during dissolution.

[0076] In one embodiment, the mixing is carried out uniformly by stirring or ultrasonic method.

[0077] In one embodiment, the solvent is removed by heating, rotary evaporation, vacuum drying, etc.

[0078] In one embodiment, after the solvent is removed, the product can be placed in a drying oven to remove moisture and other effects.

[0079] One embodiment is the use of a solid dispersion of nobiletin in the preparation of a drug for preventing and treating liver damage.

[0080] Another embodiment is the use of the nobiletin solid dispersion in the preparation of a drug for treating, preventing, reversing or reducing liver damage.

[0081] In one embodiment, the liver damage includes liver tissue and cell damage.

[0082] In one embodiment, the liver damage is caused by chemical hepatotoxic substances.

[0083] In one embodiment, the chemical hepatotoxic substances include alcohol, carbon tetrachloride, acetaminophen, acetaldehyde, etc.

[0084] In one embodiment, the liver injury includes toxic hepatitis, cirrhosis and liver fibrosis.

[0085] In one embodiment, the drugs are the same or different and each comprises a therapeutically effective amount of a solid dispersion of nobiletin.

[0086] In one embodiment, the drugs that are the same or different can be made into various pharmaceutical dosage forms by conventional methods, including: tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, drops and other oral dosage forms and injections and other dosage forms other than oral administration, such as injections.

[0087] In one embodiment, the drugs, whether the same or different, may further contain one or more pharmaceutically acceptable carriers or excipients.

[0088] In one embodiment, the excipient may include a diluent, a wetting agent, a lubricant, a filler, a preservative, etc.

[0089] In one embodiment, the Nobiletin solid dispersion comprises the following components in percentage by mass: Soluplus 70-80%, PVP / VA64 10-15%, and Nobiletin 10-15%.

[0090] In one embodiment, the solid dispersion of Nobiletin of the present invention is applied to damaged liver tissue and cells, and histopathological sections, ALT and AST are used as detection indicators to verify the protective effect of the solid dispersion of Nobiletin on liver damage, showing obvious liver protective effect, which can prevent and / or reduce and / or reverse the liver damage. Specific embodiments

[0092] Example 1

[0093] A nobiletin solid dispersion comprises the following components in percentage by mass: 70-80% Soluplus, 10-15% PVP / VA64 and 10-15% nobiletin.

[0094] Specifically, the mass ratio of nobiletin:Soluplus:PVP / VA64 can be 1:5:1;

[0095] Or, the mass ratio of nobiletin:Soluplus:PVP / VA64 may be 3:15:2;

[0096] Or, the mass ratio of nobiletin:Soluplus:PVP / VA64 may be 3:14:3;

[0097] Or, the mass ratio of nobiletin:Soluplus:PVP / VA64 may be 2:16:2;

[0098] Alternatively, the mass ratio of nobiletin:Soluplus:PVP / VA64 may be 2:15:3.

[0099] Example 2: Preparation of Nobiletin Solid Dispersion

[0100] Hot melt extrusion method: After mixing the components in proportion, add them to the hot melt extruder, feed rate is less than or equal to 50 mg / min, screw speed is 40-80 rpm, extrusion temperature is less than or equal to 170 ° C, and linear Nobiletin solid dispersion is obtained, crushed and passed through a 40 mesh sieve to obtain a powdered Nobiletin solid dispersion (NOB / SD) for standby use. Among them, the mass ratio of Nobiletin: Soluplus: PVP / VA64 is 1:5:1 and the product is prepared by hot melt extrusion method, marked as NOB / SDⅠ; the mass ratio of Nobiletin: Soluplus: PVP / VA64 is 3:15:2 and the product is prepared by hot melt extrusion method, marked as NOB / SDⅡ;

[0101] Solvent evaporation method: weigh the Nobiletin API and carrier in proportion, dissolve NOB and carrier in appropriate amount of anhydrous ethanol respectively, stir and ultrasonicate for 5 minutes to fully dissolve, then mix them evenly and ultrasonicate for 10 minutes, continue stirring and heating in a constant temperature water bath at 80℃ until the anhydrous ethanol is completely evaporated, quickly transfer to a -20℃ refrigerator to cool and solidify for 12 hours, then transfer to a drying oven to dry for 48 hours to obtain Nobiletin solid dispersion. Among them, the product prepared by solvent evaporation method with a mass ratio of Nobiletin: Soluplus: PVP / VA64 of 1:5:1 was marked as NOB / SDⅢ.

[0102] The solid dispersion prepared by grinding method was used as the control group, which can be prepared by the following method: After mixing the components in proportion, adding them into a ball mill, grinding (revolution: 30-200rpm, rotation: 60-200rpm) for 20min, a mixture (NOB+Soluplus+PVP / VA64, or noted as Mixture NOB) was obtained. Among them, the product was prepared with a mass ratio of nobiletin: Soluplus: PVP / VA64 of 1:5:1, and was marked as Mixture NOBⅠ.

[0103] Example 3

[0104] (1) Characterization of physical and chemical properties

[0105] The NOB / SD was analyzed by particle size analyzer, scanning electron microscope (SEM), differential scanning calorimeter (DSC), Fourier transform infrared spectrometer (FTIR), and multi-crystal diffractometer (XRD) to evaluate whether NOB / SD was formed and to determine whether the dispersion state of NOB / SD promoted the presence of nobiletin in the matrix in an amorphous state. Figure 1-Figure 3 .

[0106] Figure 1 The figure is a particle size distribution diagram of the solid dispersion NOB / SDⅠ of Nobiletin and free Nobiletin. As can be seen from the figure, when the solid dispersion of Nobiletin of the present invention is added into water, its particle size is less than or equal to 200nm, D50 is 60-98nm, and D90 is 32-156nm. However, due to its extremely poor water solubility, the particle size distribution of free Nobiletin is 1000-10000nm.

[0107] Figure 2 The scanning electron microscope images of the solid dispersion of Nobiletin and free Nobiletin are shown in Figure 1. As can be seen from the figure, free Nobiletin exists in a crystalline state (Crystalline NOB); in the mixture of NOB+Soluplus+PVP / VA64 Mixture NOBⅠ, Nobiletin is attached to the carrier surface in the form of crystals; and in the solid dispersions of Nobiletin NOB / SDⅠ and NOB / SDⅢ of the present invention, Nobiletin is uniformly dispersed in the matrix in an amorphous state.

[0108] Figure 3 The DSC curve, FTIR curve and XRD curve of the solid dispersion of Nobiletin NOB / SDⅠ are shown in the DSC curve (see Figure 3 A), free NOB and simply mixed NOB matrix components have sharp melting peaks at 140°C; while the melting peak of NOB / SD at 140°C completely disappears, indicating that preparing NOB into a solid dispersion form changes its crystal shape in the matrix. FTIR curve (see Figure 3 B) shows the free NOB and the NOB matrix components in simple mixtures at 1646 cm -1 There are C=O stretching vibration absorption peaks in all places, but no obvious peaks in NOB / SD, indicating that NOB and the matrix have formed an amorphous state. XRD curve (see Figure 3 C) shows that the sharp peak of NOB / SD at 5-30° completely disappears, indicating that the compound is converted into an amorphous complex.

[0109] The test results of other ratios of Nobiletin solid dispersions are similar or consistent with the above, and are not described here one by one.

[0110] (2) In vitro release

[0111] PBS was selected as the release medium for the dissolution experiment. NOB / SD with a drug content of 54 mg and free NOB with a drug content of 54 mg were respectively placed in 900 mL of PBS release medium with a pH of 6.5 and placed in a 37°C constant temperature shaker. At different time points, 4 mL was taken from the release medium for testing and supplemented with an equal volume of PBS (n=3). After the sample was filtered through a 0.45 μm microporous membrane, the NOB concentration was measured at 330 nm by a UV-visible spectrophotometer, and the dissolution was calculated. The results are shown in Figure 4 and Figure 5 .in, Figure 4 This is the dissolution diagram of NOB / SDⅠ. Figure 5 These are the dissolution graphs of NOB / SDⅠ and NOB / SDⅡ.

[0112] From the dissolution curve ( Figure 4 ) It can be seen that the maximum cumulative dissolution of NOB / SDⅠ is 94.35%, and the equilibrium solubility is 71.65±6.79%, which is 5.8 times higher than that of free NOB. Figure 4 It can be seen that within 5 minutes, NOB / SDⅠ rapidly diffused in the PBS release medium, with a release rate of 94.35±1.39%, while the release rate of free NOB was only 1.21±0.10%, and the release rate of NOB / SD was 78 times that of free NOB; within 2 hours, the release behavior of NOB / SDⅠ decreased, with a release rate of 71.65±6.79%, while the release rate of free NOB gradually increased, and finally reached 12.28±2.5%. In general, the dissolution rate of NOB / SDⅠ was 5.8 times higher than that of free NOB at 2 hours. These phenomena indicate that compared with free NOB, the NOB / SD release of the present invention has an obvious rapid release effect, a large release amount, and a high total amount. Evidence from the characterization of physicochemical properties and in vitro release studies shows that the NOB / SD of the present invention has significant advantages in oral absorption.

[0113] Depend on Figure 5It can be seen that the maximum dissolution rate of NOB / SDⅡ is 87.20±10.92%, and the dissolution rate at 2h is 72.34±3.91%, which is 5.9 times the maximum release rate of free NOB, similar to the dissolution behavior of NOB / SDⅠ, indicating that NOB / SD in different ratios of the present invention has a higher release rate than free drugs.

[0114] The test results of other ratios of Nobiletin solid dispersions are similar or consistent with the above, and are not described here one by one.

[0115] The release test results of NOB / SDⅠ and NOB / SDⅢ are shown in Figure 6 As can be seen from the figure, the maximum release of the solid dispersion of Nobiletin prepared by hot melt extrusion is 94.35±1.39%, while the maximum release of the solid dispersion prepared by solvent evaporation is 66.24±2.20%, indicating that the solid dispersion prepared by hot melt extrusion has a higher degree of dispersion and better release effect than that prepared by solvent evaporation. The product was sealed and stored at room temperature for 3 months before the release test was carried out. The results are as follows Figure 6 As shown in B, the release rate of NOB / SDⅠ was still 69.05±5.71% at 2h release, while the release rate was only 41.61%±2.03% at the ratio of NOB:Soluplus=1:6. Figure 7 The DSC curve shows that after being sealed and stored at room temperature for 3 months, the NOB / SDⅠ( Figure 7 A) There is no obvious endothermic peak of nobiletin at 140℃; while NOB / SD (NOB:Soluplus=1:6) ( Figure 7 B) The endothermic peak of Nobiletin appeared, indicating the presence of Nobiletin crystals, and the aging of the solid dispersion caused some Nobiletin crystals to precipitate. This result shows that the use of soluplus+PVP / VA64 composite dispersion can effectively slow down the aging of the solid dispersion.

[0116] Example 4: Protective effect of nobiletin solid dispersion on acute liver injury in vivo

[0117] (1) Experimental subjects: C57 mice

[0118] (2) Experimental groups: ① blank control group (oral administration of saline); ② model group (oral administration of saline); ③ free NOB group (oral administration dose: 50 mg / kg); ④ NOB / SD group (oral administration dose: NOB 50 mg / kg); 3 mice in each group. NOB / SD group was treated with NOB / SDⅠ.

[0119] (3) Experimental procedures: The rats were given drugs for one week in each group, and the acute liver injury animal model was induced by intraperitoneal injection of acetaminophen (350 mg / kg). The blank control group was not injected. 48 hours after the injection, 4% tribromoethanol was injected intraperitoneally for anesthesia. The eyeballs were removed to obtain whole blood. After being placed at room temperature for 2 hours, the blood was centrifuged at 3000 rpm for 10 minutes to obtain serum. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the rat serum were detected by biochemical analyzer. Results are shown in Figure 8 .

[0120] The mice were killed by cervical dislocation and the livers were removed and fixed in 4% paraformaldehyde solution for more than 48 hours for HE staining. Figure 9-10 .

[0121] Depend on Figure 8 It can be seen that the ALT and AST activities of mice in the free NOB group (Free NOB) were 819.22±39.2 and 535.5±60.42U / L, respectively; the ALT and AST activities of mice in the NOB / SD group were 253.88±135.98 and 131.36±26.08U / L, respectively. The liver function enzyme activities of mice in the NOB / SD group were significantly reduced (p<0.05), indicating that the bioavailability of NOB / SD is higher than that of free NOB.

[0122] Depend on Figure 9-10 It can be seen that the surface of the liver of the blank control group (CTL) mice was smooth and delicate, while the surface of the liver of the model group (normal saline) was rough, the overall color was whitish, and a large number of granular white fat droplets and dot-like red inflammatory lesions appeared. Compared with the model group, the roughness of the liver surface in the free NOB group was improved, but there were still granular white fat droplets and dot-like red inflammatory lesions. The liver surface of the NOB / SD group tended to be smooth, and the area of ​​granular white fat droplets and dot-like red inflammatory lesions was significantly reduced. The results of HE staining sections of mouse livers showed that the liver cell structure of the model group mice was loose, the liver cells were obviously necrotic, and more fibrous connective tissues were abnormally proliferated, and obvious inflammatory cell infiltration was visible in the tissue (pathological area 41.82% ± 4.86). The area of ​​liver cell necrosis and inflammatory cell infiltration in the free NOB group was reduced, which had a certain improvement effect on liver damage (pathological area 26.03% ± 8.02). In the NOB / SD group, the area of ​​liver cell necrosis and inflammatory cell infiltration was greatly reduced (pathological area 6.91% ± 1.33), showing a significant protective effect on liver cells. The above results indicate that NOB / SD of the present invention exhibits excellent liver protection effect in treating acute liver injury caused by acetaminophen.

[0123] Example 5: In vivo pharmacokinetic study of nobiletin solid dispersion

[0124] (1) Experimental subjects: Male Sprague-Dawley (SD) rats (SPF, 200-220 g) were provided by Beijing Huafukang Biotechnology Co., Ltd. (Beijing, China).

[0125] (2) Experimental groups: ① NOB / SD group (aqueous solution, oral dose: 50 mg / kg); ② free NOB group (aqueous solution, oral dose: 50 mg / kg); 3 mice in each group; the concentration of NOB in the two groups was 2.5 μg / mL. In the NOB / SD group, the mass ratio of nobiletin: Soluplus: PVP / VA64 was 1:5:1, and the NOB / SD group was prepared by hot melt extrusion.

[0126] (3) Experimental procedures: Rats were fed and maintained under constant conditions of room temperature, 50±5% humidity and 12h light / dark cycle. All rats were fasted for 12h during the experiment and had free access to water. After administration, 200μL of blood samples were collected from the tail vein of each rat at 0, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 8 and 12h, respectively, and placed in heparinized tubes. The samples were then immediately centrifuged at 6000rpm for 10min, 100μL of plasma was placed in a 1.5mL sterile centrifuge tube, 10μL of tangeretin internal standard working solution (20μg / mL) was added, 250μL of acetonitrile was added to precipitate protein, and then vortexed for 2min, centrifuged at 13000rpm for 10min at 4°C, and the supernatant was sampled for detection and analysis. Results are shown in Fig.11 .

[0127] Depend on Fig.11 It can be seen that the peak time of NOB in the NOB / SD group was 0.67±0.14h, and the peak blood concentration was 7.23±1.62μg / mL, while the free NOB reached the highest blood concentration at 0.83±0.14h, which was 1.26±0.51μg / mL. The highest blood concentration of NOB / SD was 5.74 times that of free NOB. After one oral administration, the AUC0–12 of NOB / SD was 9.68±1.73μg·h / mL, while that of free NOB was only 2.89±1.28μg·h / mL. The relative bioavailability of oral NOB / SD was 3.25 times that of free NOB. After administration, the NOB blood concentration in the NOB / SD group increased rapidly, achieving a higher effective blood concentration level, thereby achieving a rapid and effective therapeutic effect. In addition, within the first 3 hours, the NOB blood concentration in the NOB / SD group was significantly higher than that in the free NOB group, indicating that the NOB / SD group had higher bioavailability.

[0128] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A solid dispersion of nobiletin, characterized in that The composition comprises the following components in mass percentage: Soluplus 70-80%, PVP / VA64 10-15%, and nobiletin 10-15%; The mass ratio of PVP / VA64 to Soluplus is 1:8-3:14; The nobiletin solid dispersion is prepared by a hot melt extrusion method, which specifically includes uniformly mixing the components in proportion and extruding the mixture through a hot melt extruder.

2. A method for preparing the solid dispersion of Nobiletin according to claim 1, characterized in that The method is a hot melt extrusion method, which specifically includes uniformly mixing the components in proportion and extruding the mixture through a hot melt extruder.

3. The preparation method according to claim 2, characterized in that: The extrusion temperature is less than or equal to 170°C.

4. Use of the nobiletin solid dispersion according to claim 1 in the preparation of drugs for preventing and treating liver damage.

5. The use according to claim 4, characterized in that The medicine also contains one or more pharmaceutically acceptable carriers or excipients.

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