Caulis kadsurae extract, preparation method thereof, cyclodextrin inclusion compound containing caulis kadsurae extract, pharmaceutical composition containing caulis kadsurae extract and pharmaceutical preparation containing caulis kadsurae extract

Abstract

The invention belongs to the technical field of traditional Chinese medicine extraction and preparations, and relates to a caulis kadsurae extract, a preparation method thereof, a cyclodextrin inclusion compound containing the caulis kadsurae extract, a pharmaceutical composition and a pharmaceutical preparation. The kadsurae caulis extract is prepared by adopting a supercritical carbon dioxide fluid extraction method, and the lignan content of the prepared kadsurae caulis extract is remarkably improved. After the caulis kadsurae extract is prepared into the cyclodextrin inclusion compound, pulverization is realized, the material characteristics are obviously improved, and the cyclodextrin inclusion compound can be conveniently used for preparing granules, tablets, pellets and the like. After the cyclodextrin inclusion compound is further prepared into a pharmaceutical composition and a pharmaceutical preparation (especially pellets), the solubility and the dissolution speed of lignan are obviously improved, and the bioavailability and the drug effect are obviously improved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine extraction and preparation, and relates to a Millettia spatholobi extract and a preparation method thereof, a cyclodextrin inclusion compound, a pharmaceutical composition and a pharmaceutical preparation thereof. Background Art

[0002] Kadsurae Caulis is the dried vine stem of Kadsura interior A.C. Smith, a plant of the Magnoliaceae family. It can invigorate blood circulation, relax tendons and dredge collaterals, regulate menstruation and relieve pain, and is used to treat irregular menstruation, dysmenorrhea, and deficiency of qi and blood. Kadsurae Caulis contains chemical components such as lignans, triterpenes, and volatile oils. Lignans are its main biologically active components, showing a variety of good biological activities, including inhibition of platelet aggregation, anti-oxidation, anti-tumor, and anti-human immunodeficiency virus (HIV).

[0003] Lignans are a class of polyphenolic compounds. The lignans isolated and identified from Millettia yunnanensis include: biphenylcyclooctenes lignans (such as schisandrin ester A, schisandrin ester B, schisandrin ketone, schisandrin, iso-schisandrin C, etc.), spirobenzofuran-type biphenylcyclooctenes lignans (such as schisandrin ester C, schisandrin ester, iso-schisandrin D, endo-schisandrin, etc.), and other lignans (such as gomisin D, etc.). Among them, iso-schisandrin D is considered to be one of the important basic substances for Millettia yunnanensis's blood circulation and blood stasis function.

[0004] At present, the main method of extracting Millettia yunnanensis from Yunnan is to make Millettia yunnanensis paste powder, which is used to treat blood diseases and is also the main ingredient of the traditional gynecological compound Millettia yunnanensis paste. However, the extraction rate of lignan components in the water extraction process is low, and the loss of effective components is large. Huang Furong et al. (Huang Furong, Chen Zhen, Ma Chunni. Comparison of total lignan content in water and alcohol extracts of Millettia yunnanensis [J]. Science and Technology Wind, 2015 (16): 1) studied the water and alcohol extraction process of Millettia yunnanensis. The results showed that the extraction rate of lignans by percolation with ethanol as the solvent was significantly higher than that of water extraction. Lu Yan (Lu Yan. Study on the chemical composition of Millettia yunnanensis and the preparation process and quality standard of compound Millettia yunnanensis preparation [D]. Fudan University, 2006.) compared the content of total lignans in Millettia yunnanensis extracted by percolation and reflux methods. The result was that the content of total lignans in the percolation method was also higher. The content of Millettia yunnanensis in the Chinese Pharmacopoeia (2020 edition) was determined using ultrasonic extraction method, and the extraction solvent was cyclohexane.

[0005] At present, there is no report on the extraction of lignans from Millettia yunnanensis by supercritical CO2 fluid extraction. However, the product extracted by supercritical CO2 fluid extraction is often a viscous liquid, which causes many difficulties in the preparation of drugs.

[0006] In addition, the utilization of lignans in the human body is low, and there are great difficulties in improving their bioavailability and efficacy.

[0007] Pellets are spherical or quasi-spherical solid preparations with a diameter of less than 2.5 mm, generally between 0.5 and 1.5 mm, and belong to a multi-particle drug delivery system. The pellets have uniform particle size and good fluidity, which can reduce the irritation to the gastric mucosa caused by excessive local drug concentration. In addition, the pellets are less affected by the gastric emptying rate, which can avoid being affected by the patient's dietary conditions; the effect of the gastric emptying rate on the retention time of the pellets in the upper gastrointestinal tract can be reduced, thereby reducing the intra-individual and inter-individual differences in blood drug concentration, the reproducibility of blood drug concentration between doses is good, and the drug absorption rate is relatively uniform. The active ingredients concentrated in the pellets can also make the blood drug concentration quickly reach the therapeutic concentration, so as to improve the absorption rate of the drug and maintain the drug release rate, thereby improving the bioavailability. The pellets have a large surface area, a wide contact surface with the gastrointestinal mucosa, a good absorption percentage, high bioavailability, and good safety. Therefore, if the extract of Millettia yunnanensis is prepared into pellets, it will be more conducive to the full utilization of the medicinal materials and the exertion of the efficacy. Summary of the invention

[0008] Problem that the invention aims to solve

[0009] The present invention aims to provide a Millettia spatholobi extract and a preparation method thereof, including a cyclodextrin inclusion compound, a pharmaceutical composition and a pharmaceutical preparation thereof. The Millettia spatholobi extract has a high lignan content, and the preparation method is simple, time-saving and does not use organic solvents. After the Millettia spatholobi extract is prepared into a cyclodextrin inclusion compound and further prepared into a pharmaceutical composition and a pharmaceutical preparation (especially micropills), the material properties and the solubility and dissolution rate of the lignan components can be significantly improved, thereby improving the bioavailability.

[0010] Solutions for solving problems

[0011] In the first aspect, the present invention provides a method for preparing a Millettia spatholobi extract, which comprises the following steps: subjecting Millettia spatholobi to supercritical carbon dioxide fluid extraction to obtain the Millettia spatholobi extract; wherein the pressure of the extraction kettle is 25-35MPa, the temperature of the extraction kettle is 40-50°C, the pressure of the separation kettle is 5-6.5MPa, and the temperature of the separation kettle is 30-40°C.

[0012] Preferably, the flow rate of the carbon dioxide is 25-35 L / h, and the extraction time is 1.5-3 h.

[0013] Preferably, after subjecting Millettia spatholobi to supercritical carbon dioxide fluid extraction to obtain an extract, 45%-98% ethanol aqueous solution is added to the extract, and the extract is extracted and purified to obtain the Millettia spatholobi extract; more preferably, the extraction and purification method is ultrasonic extraction, and after ultrasonic extraction, the supernatant is taken, and ethanol and water are removed to obtain the Millettia spatholobi extract.

[0014] In a second aspect, the present invention provides a Millettia spatholobi extract prepared by the preparation method of the Millettia spatholobi extract described in the first aspect.

[0015] Preferably, the total content of lignans in the Millettia spatholobi extract is above 10 wt%.

[0016] More preferably, the lignans comprise gomisin D, endoschizandrin, isoschizandrin and schizandrin.

[0017] In the third aspect, the present invention provides a method for preparing a cyclodextrin inclusion complex of Millettia spatholobi, which comprises the following steps: dissolving the Millettia spatholobi extract described in the second aspect in anhydrous ethanol, adding dropwise to a saturated aqueous solution of cyclodextrin, stirring while keeping warm, and cooling to obtain the cyclodextrin inclusion complex of Millettia spatholobi.

[0018] Preferably, the mass ratio of the Millettia spatholobi extract to cyclodextrin is 1:6-1:3.

[0019] Preferably, the temperature of the cyclodextrin saturated aqueous solution is 50-90°C.

[0020] Preferably, the stirring time is 1-4h.

[0021] Preferably, the cooling temperature is 0-15°C.

[0022] Preferably, the cooling time is 10-24 hours.

[0023] Preferably, the cyclodextrin is β-cyclodextrin.

[0024] In a fourth aspect, the present invention provides a Millettia spatholobi cyclodextrin inclusion complex prepared by the preparation method of Millettia spatholobi cyclodextrin inclusion complex described in the third aspect.

[0025] In a fifth aspect, the present invention provides a pharmaceutical composition comprising the Millettia spatholobi cyclodextrin inclusion complex described in the fourth aspect.

[0026] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0027] More preferably, the pharmaceutically acceptable carrier comprises one or more of a filler, a lubricant, a disintegrant and a binder.

[0028] Preferably, the filler is a combination of microcrystalline cellulose and lactose, and the mass ratio is preferably 1:1.2-1:2.5; the lubricant is micropowder silica gel; the disintegrant is one or more of cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and low-substituted hydroxypropyl cellulose; the binder is one or a combination of povidone K30 and hydroxypropyl methylcellulose.

[0029] Preferably, based on the total mass of the pharmaceutical composition, the content of the Millettia spatholobi cyclodextrin inclusion complex is 45%-50%, and the content of the lubricant is 1%-3%.

[0030] In a sixth aspect, the present invention provides a pharmaceutical preparation comprising the pharmaceutical composition described in the fifth aspect.

[0031] Preferably, the pharmaceutical preparation is a pellet, a drop pill, a granule, a suppository, a tablet or a capsule.

[0032] Preferably, the pharmaceutical preparation is pellets.

[0033] More preferably, the micropellets can pass through a 16-mesh sieve but cannot pass through a 24-mesh sieve.

[0034] Effects of the Invention

[0035] The present invention has the following excellent technical effects:

[0036] The preparation method of the Millettia yunnanensis extract provided by the present invention significantly improves the lignan content in the extract, and the lignan components are highly enriched. The supercritical CO2 fluid extraction process is simple, takes a short time, and does not use organic solvents.

[0037] After the Millettia yunnanensis extract of the present invention is prepared into a cyclodextrin inclusion compound, powderization is achieved, the material properties are significantly improved, and it can be conveniently used to prepare granules, tablets, pellets, etc. After the cyclodextrin inclusion compound is further prepared into a pharmaceutical composition and a pharmaceutical preparation (especially pellets), the solubility and dissolution rate of the lignan components are significantly improved, the bioavailability of the lignan components is significantly increased, and the drug efficacy is improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is a high performance liquid chromatogram of the supercritical CO2 fluid extract in Example 1; wherein, peak 1 is gomisin D, peak 2 is endoschizandrin, peak 3 is isoschisandrin, and peak 4 is schisandrin.

[0039] Figure 2The differential scanning calorimetry analysis diagram of β-cyclodextrin, total lignans thick extract, a mixture of β-cyclodextrin and total lignans thick extract, and the cyclodextrin inclusion complex obtained in Experimental Group 10 in Example 3; wherein: A is β-cyclodextrin, B is total lignans thick extract, C is a mixture of β-cyclodextrin and total lignans thick extract, and D is the cyclodextrin inclusion complex obtained in Experimental Group 10 in Example 3.

[0040] Figure 3 The infrared spectra are of β-cyclodextrin, total lignans thick extract, a mixture of β-cyclodextrin and total lignans thick extract, and the cyclodextrin inclusion complex obtained in Experimental Group 10 in Example 3; wherein: A is β-cyclodextrin, B is total lignans thick extract, C is a mixture of β-cyclodextrin and total lignans thick extract, and D is the cyclodextrin inclusion complex obtained in Experimental Group 10 in Example 3.

[0041] Figure 4 This is a scanning electron micrograph of β-cyclodextrin (magnified 1000 times).

[0042] Figure 5 This is a scanning electron micrograph of a mixture of β-cyclodextrin and total lignan thick extract (magnified 1000 times).

[0043] Figure 6 This is a scanning electron micrograph of the cyclodextrin inclusion complex obtained from Experimental Group 10 in Example 3 (magnified 10,000 times).

[0044] Figure 7 The dissolution curves of gomisin D (GD), inner schisandrin (IN), heteromorphic schisandrin D (HD), and schisandrin (KA) in the micropellets prepared in Example 9 are shown.

[0045] Figure 8 This is the dissolution curve of gomisin D (GD), endonic schisandrin (IN), heterotype schisandrin D (HD), and southern schisandrin (KA) in the thick extract of total lignans from Millettia spatholobi in Comparative Example 2. DETAILED DESCRIPTION

[0046] Various exemplary embodiments, features and aspects of the present invention will be described in detail below. The word "exemplary" used here means "used as an example, embodiment or illustrative". Any embodiment described here as "exemplary" is not necessarily interpreted as being superior or better than other embodiments.

[0047] In addition, in order to better illustrate the present invention, numerous specific details are provided in the following specific embodiments. It should be understood by those skilled in the art that the present invention can be implemented without certain specific details. In other examples, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the subject matter of the present invention.

[0048] Unless otherwise stated, the units used in this specification are all international standard units, and the numerical values ​​and numerical ranges appearing in the present invention should be understood to include the inevitable systematic errors in industrial production.

[0049] In this specification, the word "may" means both performing a certain process and not performing a certain process.

[0050] In this specification, the references to "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc., mean that the specific elements (e.g., features, structures, properties and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not exist in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0051] In this specification, the numerical range expressed using "a numerical value A to a numerical value B" means a range including the endpoints numerical values ​​A and B.

[0052] In this specification, "X times the amount of solvent" means that the mass of the added solvent is X times the mass of the solute. For example, "add 10 times the amount of 80% ethanol to the Millettia spatholobi crude powder", if the mass of the Millettia spatholobi crude powder is 1g, the mass of the added 80% ethanol is 10g.

[0053] In this specification, "ethanol concentration" refers to the volume concentration of ethanol in an ethanol aqueous solution. For example, "80% ethanol" is prepared by mixing ethanol and water in a volume ratio of 80:20.

[0054] Preparation method of Millettia yunnanensis extract

[0055] The present invention proposes for the first time a method for extracting lignan components in Millettia spatholobi using supercritical carbon dioxide fluid. Specifically, the preparation method of Millettia spatholobi extract provided by the present invention comprises the following steps: extracting Millettia spatholobi with supercritical carbon dioxide fluid to obtain the Millettia spatholobi extract; wherein the pressure of the extraction kettle is 25-35MPa, the temperature of the extraction kettle is 40-50°C, the pressure of the separation kettle is 5-6.5MPa, and the temperature of the separation kettle is 30-40°C.

[0056] In some embodiments, the extraction kettle pressure is 28-32 MPa, such as 30 MPa.

[0057] In some embodiments, the extraction kettle temperature is 42-55°C, such as 45°C.

[0058] In some embodiments, the separation tank pressure is 5.5-6 MPa, such as 5.8 MPa.

[0059] In some embodiments, the separation tank temperature is 32-38°C, such as 35°C.

[0060] In some embodiments, the flow rate of carbon dioxide is 25-35 L / h, such as 28 L / h, 30 L / h, 32 L / h, etc.

[0061] In some embodiments, the frequency of the carbon dioxide is 10-20 Hz, such as 12 Hz, 15 Hz, 18 Hz, etc.

[0062] In some embodiments, the extraction time is 1.5-3 h, preferably 1.5-2.5 h, such as 2 h, 2.5 h.

[0063] In some embodiments, after subjecting Millettia yunnanensis to supercritical carbon dioxide fluid extraction to obtain an extract, 45%-98% ethanol aqueous solution is added to the extract, and the extract is extracted and purified to obtain Millettia yunnanensis extract.

[0064] In some specific embodiments, the extraction and purification method is ultrasonic extraction. After ultrasonic extraction, the supernatant is taken, and ethanol and water are removed (for example, ethanol and water are removed by rotary evaporation) to obtain the Millettia spatholobi extract.

[0065] In some specific embodiments, after removing ethanol and water, drying is performed (for example, drying under reduced pressure at 50-80° C.) to constant weight to obtain the Millettia spatholobi extract.

[0066] In some specific embodiments, the concentration of the added ethanol aqueous solution is 50%-95%, preferably 80%-95%.

[0067] In some specific embodiments, the mass of the added ethanol aqueous solution is 80-200 times, preferably 120-200 times, such as 150 times, 180 times, etc., the mass of the extract.

[0068] In some specific embodiments, the extraction and purification method is ultrasonic extraction, and a supernatant and a precipitate are obtained after ultrasonic extraction; the precipitate is rinsed with a 90%-98% ethanol aqueous solution, and the rinsing liquid is collected; the supernatant and the rinsing liquid are combined, and ethanol and water are removed to obtain the Millettia spatholobi extract.

[0069] In some embodiments, the method for preparing the Millettia spatholobi extract further comprises the step of pretreating the Millettia spatholobi.

[0070] In some embodiments, the pretreatment step includes drying and crushing the Millettia spatholobi.

[0071] In some specific embodiments, the pretreatment step is: drying the Millettia spatholobi, crushing it, passing it through a No. 2 sieve, and retaining the sieve residue.

[0072] In some specific embodiments, the drying temperature is below 60°C, such as 40-60°C.

[0073] In some specific embodiments, the drying is performed until the moisture content of Millettia yunnanensis is less than 10%, such as 1%-10% or 1%-5%.

[0074] Millettia yunnanensis extract

[0075] The present invention also provides a Millettia spatholobi extract prepared by the Millettia spatholobi extract preparation method.

[0076] In some embodiments, the total content of lignans in the Millettia spatholobi extract is above 10 wt %.

[0077] In some embodiments, the lignans comprise gomisin D, endoschizandrin, isoschizandrin, and schizandrin.

[0078] In some embodiments, the content of gomisin D in the Millettia yunnanensis extract is above 0.5wt%, the content of endoschizandrin is above 1.5wt%, the content of isoschizandrin is above 8wt%, and the content of schisandrin is above 1wt%.

[0079] In some embodiments, the Millettia spatholobi extract provided by the present invention is an extract.

[0080] Preparation method of Millettia yunnanensis cyclodextrin inclusion compound

[0081] The extract of Millettia spatholobi has high viscosity, poor solubility, and is in a semi-solid state, which is not conducive to further preparation into solid or liquid preparations. The present invention adopts a cyclodextrin inclusion process to process the obtained extract to obtain a Millettia spatholobi cyclodextrin inclusion compound, which realizes powderization on the one hand, significantly improves the material properties, and can be conveniently used to prepare granules, tablets, pellets, etc. On the other hand, it significantly improves the solubility and dissolution rate of lignan components, enhances its solubility and stability in water, reduces gastrointestinal irritation, and improves the bioavailability of lignan compounds.

[0082] Specifically, the preparation method of the Millettia spatholobi cyclodextrin inclusion complex provided by the present invention comprises the following steps: dissolving the Millettia spatholobi extract in anhydrous ethanol, adding dropwise to a saturated aqueous solution of cyclodextrin, stirring at a constant temperature, and cooling to obtain the Millettia spatholobi cyclodextrin inclusion complex.

[0083] In some embodiments, the mass ratio of the Millettia spatholobi extract to cyclodextrin is 1:6-1:3, preferably 1:5-1:3.75, for example 1:4, 1:3.5, etc.

[0084] In some embodiments, the temperature of the cyclodextrin saturated aqueous solution is 50-90°C, preferably 60-80°C.

[0085] In some embodiments, the stirring time is 1-4 h, preferably 1-3 h.

[0086] In some embodiments, the cooling temperature is 0-15°C, preferably 0-10°C.

[0087] In some embodiments, the cooling time is 10-24 h, preferably 12-24 h, such as 15 h, 20 h, etc.

[0088] In some embodiments, the cyclodextrin is β-cyclodextrin.

[0089] In some specific embodiments, the mass ratio of the anhydrous ethanol to the Millettia spatholobi extract is 5-10, preferably 7-8.

[0090] In some specific embodiments, during the dropwise addition, the cyclodextrin saturated aqueous solution is in a stirring state.

[0091] In some specific embodiments, before the dropwise addition, the temperature of the cyclodextrin saturated aqueous solution is 50-90°C, preferably 60-80°C.

[0092] In some specific embodiments, after cooling, the solid and liquid are separated, and the obtained solid is washed alternately with anhydrous ethanol and water, and then dried under reduced pressure to obtain the Millettia spatholobi cyclodextrin inclusion complex.

[0093] Millettia yunnanensis cyclodextrin inclusion complex

[0094] The present invention also provides a Millettia spatholobi cyclodextrin inclusion compound prepared by using the Millettia spatholobi cyclodextrin inclusion compound preparation method.

[0095] Pharmaceutical composition

[0096] The present invention also provides a pharmaceutical composition, which comprises the above-mentioned Millettia yunnanensis cyclodextrin inclusion compound.

[0097] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0098] In some specific embodiments, the pharmaceutically acceptable carrier comprises one or more of a filler, a lubricant, a disintegrant, and a binder.

[0099] In some specific embodiments, the filler is a combination of microcrystalline cellulose (e.g., MCC 102) and lactose; the lubricant is micropowdered silica gel; the disintegrant is one or more of cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose; and the binder is one or a combination of povidone K30 and hydroxypropyl methylcellulose.

[0100] In some specific embodiments, the filler is a combination of microcrystalline cellulose (eg, MCC 102) and lactose; the lubricant is micronized silica gel; the disintegrant is cross-linked polyvinylpyrrolidone; and the binder is polyvinylpyrrolidone K30.

[0101] In some specific embodiments, the filler is a combination of microcrystalline cellulose and lactose, and the mass ratio of microcrystalline cellulose to lactose can be 1:1.2-1:2.5, preferably 1:1.5-1:2, such as 1:1.3, 1:1.4, 1:1.8, 1:2.1, 1:2.2, etc.

[0102] In some specific embodiments, based on the total mass of the pharmaceutical composition, the content of the Millettia spatholobi cyclodextrin inclusion complex is 45%-50%, and the content of the lubricant is 1%-3%.

[0103] In some specific embodiments, based on the total mass of the pharmaceutical composition, the content of the filler is 40%-55% (preferably 40%-50%), the content of the disintegrant is 3%-5%, and the content of the binder is 0%-5% (preferably 0.5%-3%).

[0104] Pharmaceutical preparations

[0105] The present invention also provides a pharmaceutical preparation, which comprises the above pharmaceutical composition.

[0106] In some embodiments, the pharmaceutical preparation is micropellets, pellets, granules, suppositories, tablets or capsules.

[0107] In some specific embodiments, the pharmaceutical preparation is a pellet.

[0108] In some preferred embodiments, the pellets can pass through a 16-mesh sieve but cannot pass through a 24-mesh sieve. The aperture of the "16-mesh sieve" is 1 mm, and the aperture of the "24-mesh sieve" is 0.85 mm. Therefore, the diameter of the pellets of the present invention is 0.85-1 mm, or the diameter of the pellets of the present invention is: 0.85 mm < diameter ≤ 1 mm.

[0109] abbreviation

[0110] The English abbreviations and Chinese full names involved in the present invention are shown in the following table:

[0111] Abbreviations Chinese full name GD Gomisin D IN Schisandrin HD Schisandrin KA Schisandrin PVP K30 Povidone K30 HPMC Hydroxypropyl methylcellulose MCC 102 Microcrystalline cellulose 102 CMS-Na Sodium carboxymethyl starch PVPP Crospovidone L-HPC Low substituted hydroxypropyl cellulose

[0112] High performance liquid chromatography conditions used in the present invention

[0113] Chromatographic column: CAPCELL PAK MG C18 chromatographic column (4.6 mm × 250 mm, 5 μm), column temperature: 35 ° C; mobile phase: methanol-acetonitrile-water continuous gradient elution, 10:40:50 (0 min) → 10:47:43 (23 min) → 10:50:40 (35 min) → 10:65:25 (50 min), a total of 55 min; flow rate 1 mL / min; detection wavelength 230 nm.

[0114] Drug loading rate

[0115] In the present invention, the calculation formula of the "drug loading rate" of the inclusion complex is: the amount of the included Millettia spatholobi extract / (the amount of the included Millettia spatholobi extract+the amount of β-cyclodextrin)*100%.

[0116] Example

[0117] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0118] The Millettia spatholobi medicinal material used in the following examples and comparative examples is Millettia spatholobi decoction pieces, provided by Yunnan Guangfu Pharmaceutical Co., Ltd., with batch number 20210325.

[0119] The schizandrin standard (purity: 98.11%, batch number: 23030106) and endospermum schisandrin standard (purity: 99.69%, batch number: 23030105) used in the determination of lignan content in the following examples and comparative examples were provided by Chengdu Gelip Biotechnology Co., Ltd.; the gomisin D standard (purity: 98%, batch number: J26HB1860531) and isospermum schisandrin standard (purity: 98%, batch number: M06GB140638) were provided by Shanghai Yuanye Biotechnology Co., Ltd.

[0120] Example 1: Extraction of Millettia repens lignans by supercritical CO2 fluid extraction

[0121] The Yunnan Millettia spatholobi medicinal material was crushed to obtain Yunnan Millettia spatholobi coarse powder. After passing the Yunnan Millettia spatholobi coarse powder through a No. 2 sieve, 1050g was taken and placed in a 5L extraction kettle, with an extraction pressure of 30MPa, an extraction temperature of 45°C, a pressure of 5.8MPa in separation kettles I and II, a temperature of 35°C, a CO2 flow rate of 29L / h, a CO2 frequency of 15Hz, and an extraction time of 2h to obtain an extract, namely, Yunnan Millettia spatholobi extract. The above extract was subjected to liquid chromatography analysis, and its liquid chromatogram was as follows Figure 1 As shown ( Figure 1 The liquid chromatogram in the figure is the sum of the elution time of 55 min and the equilibrium needle time in the chromatographic conditions of the present invention, which contains the characteristic peaks of four lignan components, namely, gomisin D (GD), endonic schisandrin (IN), heterotype schisandrin (HD), and ka, and the content of each component is calculated.

[0122] Comparative Example 1: Extraction of Millettia spatholobi lignans by ethanol percolation extraction, ethanol reflux extraction, and traditional water extraction

[0123] Percolation group: Crush the medicinal materials of Millettia spatholobi to obtain coarse Millettia spatholobi powder. Take the coarse Millettia spatholobi powder, add 10 times the amount of 80% ethanol and cold soak for 48 hours, and collect 30 times the volume of percolation liquid by percolation at a rate of 1 to 3 times volume / h. The percolation liquid is concentrated under reduced pressure or by rotary evaporation until there is no alcohol taste to recover ethanol and obtain the ethanol extract of Millettia spatholobi.

[0124] Reflux group: Crush the medicinal materials of Millettia spatholobi to obtain coarse Millettia spatholobi powder. Take the coarse Millettia spatholobi powder, weigh it accurately, add 5 times the amount of 80% ethanol, reflux and extract it in a 90℃ water bath for 3 times, add 5 times the amount of 80% ethanol each time, reflux, and filter. Combine the three filtrates and evaporate to dryness to obtain the Millettia spatholobi extract.

[0125] Water extraction group: Take the cleaned Millettia yunnanensis, put it in a round-bottom flask, add water and boil it three times: add 6 times the amount of water for the first time, boil it and cook it for 4 hours, filter it; add 4 times the amount of water for the second time, boil it slightly and cook it for 3 hours, filter it; add 4 times the amount of water for the third time, boil it and cook it for 2 hours, filter it. Combine the three filtrates and concentrate them into a thick extract with a specific gravity of 1.21-1.40 (80℃).

[0126] The total lignans content of Millettia spatholobi was represented by gomisin D (GD), inner schisandrin (IN), heteromorphic schisandrin (HD), and southern schisandrin (KA). The total lignans content in the Millettia spatholobi extract obtained in Example 1 and Comparative Example 1 was determined by high performance liquid chromatography, and the results are shown in Table 1.

[0127] Table 1 Results of lignan content under various extraction methods

[0128]

[0129] As shown in Table 1, the total lignan extraction rate of the supercritical group was the highest, which was 517 times that of the traditional water extraction method, 17 times that of the ethanol percolation extraction method, and 24 times that of the ethanol reflux extraction method. In addition, the extraction rate of each lignan component in the supercritical group also far exceeded that of other experimental groups.

[0130] Example 2: Purification of supercritical extract

[0131] The ethanol ultrasonic extraction method was used to remove some resin impurities in the supercritical extract.

[0132] Take the extract obtained in Example 1, add 150 times of ethanol aqueous solution of different concentrations: 30%, 50%, 70%, 80%, 95%. Ultrasonic treatment (power 360W, frequency 40kHz) for 50min, cool, shake well, centrifuge at 3000r / min, 5min, and separate the supernatant and precipitate. Add distilled water to the precipitate, centrifuge and wash 3 times, evaporate to dryness, weigh, and measure the content of total lignans in the precipitate and supernatant. The results are shown in Table 2. Collect the supernatant, evaporate at 60℃ until there is no alcohol taste, and dry the thick paste at 60℃ under reduced pressure to constant weight to obtain the thick extract of total lignans of Millettia yunnanensis.

[0133] Table 2 Content of total lignans in precipitate and supernatant

[0134]

[0135] The results showed that except for 30% ethanol, the total lignan content in the precipitate at other concentrations was low and could be removed. The total lignan content in the supernatant of 95% ethanol was high, the total lignan content in the precipitate was low, and the loss of total lignans was low after removing the precipitate.

[0136] Example 3: Preparation of cyclodextrin inclusion complex

[0137] β-cyclodextrin was added to deionized water and heated to different temperatures until it was completely dissolved and reached a saturated state. The solution was kept warm. The total lignan thick extract (purified from 95% ethanol) prepared in Example 2 was dissolved in 7.5 times of anhydrous ethanol and slowly added dropwise to the saturated aqueous solution of cyclodextrin. After stirring for different periods of time, the resulting material suspension was placed in a refrigerator at 4°C for 12 hours, filtered, and rinsed alternately with anhydrous ethanol and deionized water for 3 times. The wet material was dried under reduced pressure at 60°C to obtain a total lignan β-cyclodextrin inclusion complex.

[0138] Different stirring times, heating temperatures, and ratios (mass ratios) of extract to β-cyclodextrin were selected for the experiments, and the inclusion rate, yield, and comprehensive score obtained in each group of experiments were calculated. The results are shown in Table 3.

[0139]

[0140] E——inclusion rate, %; F——yield, %; G——total lignan content, %; H——comprehensive score;

[0141] m1——the actual drug loading mass of the inclusion compound, g; m2——the mass of the drug added when preparing the inclusion compound, g;

[0142] c——blank recovery rate%;

[0143] m3——mass of inclusion compound, g; m4——mass of β-cyclodextrin added when preparing inclusion compound, g;

[0144] G——total lignan content, %, E max ——The maximum value of inclusion rate, %;

[0145] F max ——The maximum value of yield, %;

[0146] G max ——The maximum value of total lignan content, %.

[0147] Table 3 Inclusion rate, yield and comprehensive score of cyclodextrin inclusion complexes obtained under different parameter conditions

[0148] Experimental groups Time / h Temperature / ℃ Drug-excipient ratio Inclusion rate / % Yield / % Overall Rating 1 1 60 1:7.5 56.44 43.28 40.51 2 1 70 1:5 89.88 63.33 61.81 3 1 80 1:3.75 81.29 54.35 56.96 4 2 60 1:5 81.38 54.56 54.08 5 2 70 1:3.75 82.63 58.03 60.16 6 2 80 1:7.5 74.33 63.95 58.16 7 3 60 1:3.75 83.57 63.59 64.88 8 3 70 1:7.5 65.75 43.34 41.13 9 3 80 1:5 82.01 58.79 57.56 10 2 80 1:3.75 86.86 65.43 66.69

[0149] β-cyclodextrin, total lignans thick extract, a mixture of β-cyclodextrin and total lignans thick extract (mass ratio of 3.75:1, physical mixing), and the cyclodextrin inclusion complex obtained from experimental group 10 were subjected to differential scanning calorimetry (DSC) and infrared spectroscopy, respectively. The results are shown in Figure 2 , Figure 3 β-cyclodextrin, a mixture of β-cyclodextrin and total lignan thick extract (mass ratio of 3.75:1, physical mixing), and the cyclodextrin inclusion complex obtained from experimental group 10 were observed by scanning electron microscopy. The results are shown in Figure 4 , Figure 5 , Figure 6 .

[0150] Depend on Figures 4 to 6 It can be seen that under a magnification of 10,000 times, the surface of the Millettia spatholobi cyclodextrin inclusion complex is smooth and diamond-shaped particles, and the apparent morphology has changed significantly, indicating that the Millettia spatholobi extract has been successfully encapsulated.

[0151] Depend on Figure 3 It can be seen that the characteristic peak of Millettia spatholobi of the inclusion complex is weakened, indicating that Millettia spatholobi lignans have entered the cavity of β-cyclodextrin and the inclusion complex is formed, which is consistent with the results of scanning electron microscopy observation.

[0152] Depend on Figure 2It can be seen that the endothermic peak of the inclusion complex at 110°C is not obvious, and the endothermic peak at 330°C disappears, indicating that the Millettia reticulata lignans have replaced the crystalline water in the cavity of β-cyclodextrin, indicating that the inclusion complex has been formed.

[0153] As shown in Table 3, the inclusion rate of each experimental group was above 55%, the comprehensive score was above 40, and the inclusion effect was good. Among them, when the drug-excipient ratio was 1:5 or 1:3.75, the inclusion rate could even reach above 80%, and the inclusion effect was better.

[0154] Example 4: Screening of micropellet formulation: Investigating the effect of drug loading on micropellets

[0155] MCC 102 and lactose were used as fillers, the ratio of MCC 102 to lactose was 4:1 (mass ratio), PVP K30 was used as a binder, the cyclodextrin inclusion complex obtained in Experimental Group 10 of Example 3, the filler, and the 0.5% PVP K30 aqueous solution were mixed, and a 1.0×1.0mm aperture sieve plate was used, the extrusion speed was 30r / min, the spheronization speed was 3000r / min, and the spheronization time was 5min to prepare wet micropills. The wet micropills were dried at 60°C for 1h, sieved, and the micropills that passed through the 16-mesh sieve but not through the 24-mesh sieve were used as the finished micropills. The micropill yield was calculated according to the formula. The micropill yield and appearance were used as evaluation indicators to investigate the effects of 35%, 40%, 45%, 50% and 55% of the drug loading (based on the cyclodextrin inclusion complex of Millettia yunnanensis) on the micropills.

[0156] Pellets yield = (16-24 mesh pellets weight / total pellets weight) × 100%

[0157] Table 4 Effect of drug loading on pellets

[0158]

[0159] Note: The proportion of each component in the formula is: the mass proportion of each component based on the total mass of the composition.

[0160] It can be seen from Table 4 that when the drug loading is 45%-50%, the pellet yield is high, the properties are good, the particle size is relatively uniform, and there is no fine powder or a small amount of fine powder.

[0161] Example 5: Screening of micropellet formulation: Investigating the effect of binders on micropellets

[0162] The type or concentration of the binder in Formulation 4 of Example 4 was replaced, and other parameters remained unchanged. The effect of the binder on the yield and appearance of the micropellets was investigated. The results are shown in Table 5.

[0163] Table 5 Effect of adhesive on micropellets

[0164] Adhesive Yield % Microparticle properties 0.5% PVP K30 93.58 The pellets are short cylindrical, with uniform particle size and no fine powder. 2.5% PVP K30 96.98 The pellets are short cylindrical, with uniform particle size and no fine powder. 2% HPMC 92.10 The pellets are short cylindrical, with uniform particle size and no fine powder. 50% ethanol / The extrudate is not sticky and cannot be extruded 70% ethanol / The extrudate is not sticky and cannot be extruded

[0165] It can be seen from Table 5 that when the binder is 0.5% PVP K30, 2.5% PVP K30, and 2% HPMC, the yield of the micropellets is high, and the properties of the micropellets are good, the particle size is more uniform, and there is no fine powder.

[0166] Example 6: Screening of micropellet formulation: Investigating the effect of fillers on micropellets

[0167] The type or proportion of the filler in Formulation 4 of Example 4 was replaced, and other parameters remained unchanged. The effect of the filler on the yield and appearance of the pellets was investigated. The results are shown in Table 6.

[0168] Table 6 Effect of fillers on pellets

[0169] Filler type and proportion Yield % Microparticle properties MCC 102 93.05 The pellets are short cylindrical, with uniform particle size and a small amount of fine powder. lactose / Too sticky, can't be squeezed out MCC 102: lactose = 4:1 96.98 The pellets are short cylindrical, with uniform particle size and no fine powder. MCC 102: lactose = 2:1 95.64 The pellets are long dumbbell-shaped, with uniform particle size and no fine powder. MCC 102: lactose = 1:1 90.19 The pellets are short dumbbell-shaped, with uniform particle size and no fine powder. MCC 102: lactose = 1:1.5 95.08 The pellets are spherical, uniform in size, and free of fine powder. MCC 102: lactose = 1:2 94.27 The pellets are spherical, with different particle sizes and no fine powder. MCC 102: lactose = 1:4 / Too sticky, the balls merge to form a big ball

[0170] As shown in Table 6, when the filler is MCC 102 and MCC 102: lactose (4:1-1:2), the yield of the micropellets is high, and the properties of the micropellets are good, the particle size is relatively uniform or slightly different, and there is no fine powder or a small amount of fine powder. Among them, when the ratio of MCC 102: lactose is 1:1.5-1:2, the micropellets are spherical and have a better appearance.

[0171] Example 7: Screening of micro-pellet formulation: Investigating the effect of micro-powder silica gel dosage on micro-pellets

[0172] Micro-powder silica gel was used to replace part of the filler in Formulation 4 of Example 4. For example, if the filler in the original formulation was 50%, the filler after replacement was 2% micro-powder silica gel and 48% filler. Other parameters remained unchanged. The effect of the amount of micro-powder silica gel on the yield and appearance of the micropills was investigated. The results are shown in Table 7.

[0173] Table 7 Effect of the amount of micropowder silica gel on the micropellets

[0174] Micro powder silica gel dosage Yield % Microparticle properties 0 94.53 The pellets are short cylindrical, with uniform particle size, no fine powder, and high viscosity and hardness. 1％ 97.02 The pellets are spherical, uniform in size, free of fine powder, and moderate in viscosity and hardness. 2％ 98.62 The pellets are spherical, uniform in size, free of fine powder, and moderate in viscosity and hardness. 3％ 98.33 The pellets are spherical, uniform in size, free of fine powder, and moderate in viscosity and hardness. 4％ 98.29 The pellets are gourd-shaped, uniform in size, free of fine powder, and moderate in viscosity and hardness.

[0175] It can be seen from Table 7 that the micro-pellets have greater viscosity and hardness without using micro-powder silica gel; when the amount of micro-powder silica gel is 4%, the micro-pellets have a gourd-shaped appearance, poor plasticity, and a sandy feel; when the amount of micro-powder silica gel is 1%-3%, the micro-pellets have a spherical appearance, uniform particle size, no fine powder, moderate viscosity and hardness, and a better effect.

[0176] Example 8: Screening of micropellet formulation: Investigating the effect of disintegrant on micropellets

[0177] Part of the filler in Formulation 4 of Example 4 was replaced with 2% micro-powder silica gel and 4% disintegrant. For example, if the filler in the original formulation was 50%, it would be 2% micro-powder silica gel, 4% disintegrant and 44% filler after replacement. The type of disintegrant is shown in Table 8. Other parameters remain unchanged. The effect of disintegrant on the yield and appearance of pellets was investigated. The results are shown in Table 8.

[0178] Table 8 Effect of disintegrant on pellets

[0179] 4% disintegrant Yield % Microparticle properties CMS-Na 95.31 The pellets are spherical, with large and uniform particle size and no fine powder. PVPP 97.03 The pellets are spherical, uniform in size, and free of fine powder. L-HPC 93.22 The pellets are spherical, uniform in size, and contain a small amount of fine powder.

[0180] It can be seen from Table 8 that when the disintegrants are CMS-Na, PVPP, and L-HPC, the yields of the micropellets are all high, and the properties of the micropellets are all good, the particle size is relatively uniform, and there is no fine powder or a small amount of fine powder.

[0181] Example 9: Preparation of micropellets

[0182] Take 1050g of Millettia yunnanensis medicinal material, dry it with hot air below 60°C until the moisture content is less than 10%, then crush it and pass it through a No. 2 sieve, retain the sieved material, and obtain Millettia yunnanensis powder.

[0183] 1050 g of Millettia spatholobi powder was added into a 5 L supercritical CO2 extraction kettle. The extraction experimental conditions were: extraction pressure 30 MPa, extraction temperature 45 °C, separation kettle I and II pressure 5.8 MPa, separation kettle temperature: 35 °C, CO2 flow rate 29 L / h, CO2 frequency 15 Hz, and extraction time 2.5 h.

[0184] The extract was collected, 3L of 95% ethanol was added, and ultrasonic treatment (power 360W, frequency 40kHz) was performed for 30min, cooled, shaken, and centrifuged at 3000r / min for 5min to obtain the supernatant and precipitate. The precipitate was repeatedly washed with 95% ethanol until it turned white, and the ethanol solution was collected.

[0185] The supernatant and ethanol solution were combined, and rotary evaporated at 60°C until there was no alcohol smell to obtain a thick paste. The thick paste was dried at 60°C under reduced pressure to constant weight to obtain 21.00 g of total lignans thick extract of Millettia yunnanensis.

[0186] 15g of β-cyclodextrin was added to 75mL of deionized water and heated to 80°C. It was heated for 10min until it was completely dissolved and reached saturation. The temperature was kept at 80°C and the speed was 850r / min with magnetic stirring. 4g of the above-mentioned total lignan thick extract of Millettia yunnanensis was dissolved in 30mL of anhydrous ethanol and slowly added dropwise to the saturated aqueous solution of cyclodextrin and stirred for 2h. The resulting material suspension was placed in a 4°C refrigerator for 12h, filtered, and rinsed alternately with anhydrous ethanol and deionized water 3 times, 5mL each time. The wet material was dried under reduced pressure at 60°C to obtain a total lignan β-cyclodextrin inclusion complex, a total of 14g. The inclusion complex was a light yellow powder with no viscosity, a drug loading rate of 22.03%, and an inclusion rate of 87.71%.

[0187] Take 4.7g of total lignan β-cyclodextrin inclusion complex and 0.2g of micropowder silica gel, grind the micropowder silica gel and cyclodextrin inclusion complex in a mortar to make a soft material. Then add 1.88g of microcrystalline cellulose, 2.82g of lactose, 0.4g of cross-linked polyvinylpyrrolidone, and 3.6mL of 2.5% polyvinylpyrrolidone K30 solution, use a 1.0×1.0mm aperture sieve plate, extrusion speed of 30r / min, spheronization speed of 3000r / min, spheronization time of 5min, and make wet micropills. The wet micropills are hot-air dried below 60°C and sieved through 16-24 mesh to obtain finished micropills. The micropills are spherical, relatively uniform in size, light yellow in overall color, and relatively smooth on the surface.

[0188] Take 4g of micro-pellets and add 900mL of 0.5wt% sodium dodecyl sulfate as a surfactant for dissolution test. The dissolution was carried out by paddle method, the dissolution temperature was 37.0℃, and the rotation speed was 50r / min. The dissolution curves of gomisin D (GD), inner schisandrin (IN), iso-schisandrin (HD), and schisandrin (KA) are shown in Figure 2. Figure 7 The cumulative dissolution results of each lignan component are shown in Table 9 below.

[0189] Table 9 Cumulative dissolution of each lignan component of Example 9

[0190]

[0191] The experimental results show that the Millettia spatholobi thick extract of the present invention is prepared into an inclusion compound and further prepared into micropills, and the dissolution effect is excellent. At 45 minutes, the cumulative solubility of each lignan component reaches more than 80%, at 90 minutes, it reaches more than 90%, and at 240 minutes, it reaches or approaches 100%.

[0192] Example 10: Preparation of micropellets

[0193] 15gβ-cyclodextrin was added to 120mL deionized water, heated to 70°C, heated for 10min until completely dissolved and saturated, kept at 70°C, and stirred at a speed of 1100r / min with magnetic stirring. 4g total lignan thick extract (same as the total lignan thick extract of Millettia spatholobi in Example 9) was dissolved in 50mL anhydrous ethanol, slowly added dropwise to the saturated aqueous solution of cyclodextrin, and stirred for 3h. The obtained material suspension was placed in a 4°C refrigerator for 12h, filtered, and rinsed alternately with anhydrous ethanol and deionized water 3 times, 5mL each time, and the wet material was dried under reduced pressure at 60°C to obtain a total lignan β-cyclodextrin inclusion complex, a total of 11g. The inclusion complex was a light yellow powder, non-sticky, with a drug loading rate of 23.07% and an inclusion rate of 82.63%.

[0194] Take 4.65g of total lignan β-cyclodextrin inclusion complex and 0.2g of micropowder silica gel, grind the micropowder silica gel and cyclodextrin inclusion complex in a mortar to make a soft material. Then add 1.55g of microcrystalline cellulose, 3.1g of lactose, 0.5g of cross-linked polyvinylpyrrolidone, and 3.5mL of 2.5% polyvinylpyrrolidone K30 solution, use a 1.0×1.0mm aperture sieve plate, extrusion speed of 30r / min, spheronization speed of 2000r / min, spheronization time of 10min, and make wet micropills. The wet micropills are hot-air dried at below 60°C and sieved through 16-24 mesh to obtain finished micropills. The micropills are spherical, relatively uniform in size, light yellow in overall color, and relatively smooth on the surface.

[0195] Comparative Example 2: Direct determination of dissolution effect of total lignans from Millettia spatholobi thick extract

[0196] No inclusion compound was prepared. The total lignans thick extract of Millettia spatholobi in Example 9 was used as the main drug. The auxiliary materials and the main drug could not be mixed evenly, and the micropills could not be prepared. The above-mentioned total lignans thick extract of Millettia spatholobi (equivalent dose to the extract in the micropills of Example 9) was directly taken to measure the dissolution effect. The dissolution conditions were the same as those in Example 9. The dissolution curves of gomisin D (GD), inner schisandrin (IN), heteromorphic schisandrin (HD), and schisandrin (KA) are shown in Figure 2. Figure 8 The cumulative dissolution results of each lignan component are shown in Table 10 below.

[0197] Table 10 Cumulative dissolution of each lignan component of Comparative Example 2

[0198]

[0199] The results show that the Millettia spatholobi thick extract of the present invention is not prepared into an inclusion compound but is directly used as the main drug, and its dissolution effect is poor. At 360 minutes, the cumulative dissolution rate of each lignan component is only about 30%, and even does not reach 30%.

[0200] Effect example: Antithrombotic effect of Millettia yunnanensis inclusion compound

[0201] (1) Establishment of mouse tail thrombosis model

[0202] ICR male mice were taken and then divided into 3 groups, namely, normal group, model group, and Millettia spatholobi inclusion compound group (the inclusion compound prepared from experimental group 10 in Example 3), with 8 mice in each group. The Millettia spatholobi group was intragastrically administered with 255.28 mg·kg daily. -1 (equivalent to 1 times the human equivalent dose), the normal group and the model group were gavaged with the same amount of 0.5% sodium carboxymethyl cellulose solution. Each mouse was gavaged once a day for 7 consecutive days.

[0203] After 7 days of administration, 1% carrageenan was injected intraperitoneally 1 hour after the last administration to establish the model, 50 mg / mL, and placed at room temperature for 24 hours. Method for determining the success of the tail thrombosis model: dark red thrombus is formed in the tail.

[0204] (2) Detection of mouse black tail length

[0205] The survival status of the mice was observed 24 hours after the model was established. The mice were anesthetized intraperitoneally with sodium pentobarbital, and the total tail length and thrombus length of each group of mice were measured with a vernier caliper. The relative black tail length of each group of mice was calculated. The results are shown in Table 11. Relative black tail length = tail thrombus length / total tail length × 100%.

[0206] Table 11 Comparison of tail thrombus length and relative tail thrombus length in mice (n=8)

[0207]

[0208] Except for the normal group, the tails of mice in other groups showed dark red thrombi to varying degrees. Compared with the modeling group, the tail thrombus length and relative length of tail thrombus in the Millettia spatholobi inclusion compound group were reduced, indicating that the Millettia spatholobi inclusion compound of the present invention has an inhibitory effect on the formation of thrombi.

[0209] It should be noted that, although the technical solution of the present invention is introduced with specific examples, those skilled in the art will appreciate that the present invention should not be limited thereto.

[0210] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or technical improvements in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing a Millettia spatholobi extract, comprising the following steps: The Millettia spatholobi extract is obtained by subjecting Millettia spatholobi to supercritical carbon dioxide fluid extraction; wherein the pressure of the extraction kettle is 25-35MPa, the temperature of the extraction kettle is 40-50°C, the pressure of the separation kettle is 5-6.5MPa, and the temperature of the separation kettle is 30-40°C.

2. The preparation method according to claim 1, characterized in that: The flow rate of the carbon dioxide is 25-35 L / h, and the extraction time is 1.5-3 h.

3. The preparation method according to claim 1 or 2, characterized in that: After subjecting Millettia spatholobi to supercritical carbon dioxide fluid extraction, an extract is obtained, and a 45%-98% ethanol aqueous solution is added to the extract to extract and purify the extract to obtain Millettia spatholobi extract; preferably, the extraction and purification method is ultrasonic extraction, and after ultrasonic extraction, the supernatant is taken, and ethanol and water are removed to obtain the Millettia spatholobi extract.

4. A Millettia spatholobi extract prepared by the preparation method according to any one of claims 1 to 3; Preferably, the total content of lignans in the Millettia spatholobi extract is above 10wt%; More preferably, the lignans comprise gomisin D, endoschizandrin, isoschizandrin and schizandrin.

5. A method for preparing a Millettia yunnanensis cyclodextrin inclusion compound, comprising the following steps: Dissolve the Millettia spatholobi extract according to claim 4 in anhydrous ethanol, add dropwise to a saturated aqueous solution of cyclodextrin, stir at room temperature, and cool to obtain the Millettia spatholobi cyclodextrin inclusion compound; Preferably, the mass ratio of the Millettia spatholobi extract to cyclodextrin is 1:6-1:3; Preferably, the temperature of the cyclodextrin saturated aqueous solution is 50-90°C; Preferably, the stirring time is 1-4h; Preferably, the cooling temperature is 0-15°C; Preferably, the cooling time is 10-24h; Preferably, the cyclodextrin is β-cyclodextrin.

6. A Millettia spatholobi cyclodextrin inclusion complex prepared by the preparation method according to claim 5.

7. A pharmaceutical composition comprising the Millettia spatholobi cyclodextrin inclusion compound according to claim 6; Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier; More preferably, the pharmaceutically acceptable carrier comprises one or more of a filler, a lubricant, a disintegrant and a binder.

8. The pharmaceutical composition according to claim 7, characterized in that The filler is a combination of microcrystalline cellulose and lactose, and the mass ratio is preferably 1:1.2-1:2.5; the lubricant is micropowder silica gel; the disintegrant is one or more of cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch and low-substituted hydroxypropyl cellulose; the binder is one or a combination of povidone K30 and hypromellose; Preferably, based on the total mass of the pharmaceutical composition, the content of the Millettia spatholobi cyclodextrin inclusion complex is 45%-50%, and the content of the lubricant is 1%-3%.

9. A pharmaceutical preparation comprising the pharmaceutical composition according to claim 8; preferably, the pharmaceutical preparation is a micropill, a dripping pill, a granule, a suppository, a tablet or a capsule.

10. The pharmaceutical preparation according to claim 9, which is micropellets; preferably, the micropellets can pass through a 16-mesh sieve but cannot pass through a 24-mesh sieve.

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