A glycyrrhetinic acid and cyclodextrin-metal organic framework complex

Abstract

The application provides a complex of glycyrrhetinic acid and a cyclodextrin-metal organic framework, specifically, the complex takes the cyclodextrin-metal organic framework as a carrier, and the glycyrrhetinic acid enters the cavity of the cyclodextrin-metal organic framework to form the complex. The complex can greatly improve the solubility of the glycyrrhetinic acid, has a biphasic release characteristic, can improve the biological activity of the glycyrrhetinic acid, reduces the administration frequency, and increases the medication compliance. In addition, the complex can effectively deliver the glycyrrhetinic acid to the lung for treating lung diseases, provides a good intermediate for inhalation preparation and oral administration of the glycyrrhetinic acid, and has a simple and safe preparation process.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical preparations, and particularly relates to a glycyrrhetinic acid complex with a cyclodextrin-metal organic framework as a carrier. BACKGROUND

[0002] 18-β-glycyrrhetinic acid (GA) is the most important active ingredient in licorice and also the metabolite and active substance of glycyrrhizic acid in vivo, which has the effects of anti-inflammatory, antiviral, liver protection and detoxification, and immune function enhancement. The structure of glycyrrhetinic acid belongs to pentacyclic triterpenes, which has a steroid-like anti-inflammatory effect and an adrenal cortex hormone-like pharmacological effect, and is widely used in the treatment of various chronic hepatitis, urticaria and the like in clinical practice.

[0003] In recent years, the effect of glycyrrhetinic acid in preventing and treating lung diseases has been frequently reported, including pulmonary fibrosis, lung injury, lung tumors, and new coronavirus. Glycyrrhetinic acid can effectively reduce lung injury caused by X-ray irradiation, and the inflammatory factor level of the treatment group is consistent with that of the normal group, and can also treat acute lung injury caused by lipopolysaccharide. Compared with the lipopolysaccharide modeling group, glycyrrhetinic acid inhibits the formation and activation of Nlrp3, down-regulates the production of reactive oxygen species (ROS) in macrophages and the phosphorylation level of PI3K and AKT, thereby inhibiting the occurrence of inflammation. Pulmonary fibrosis is caused by inflammation damage to the normal alveolar structure due to various reasons, and collagen scar tissue accumulates to repair the damage, i.e., fibrosis occurs, so that the lung tissue gradually loses normal respiratory function, resulting in symptoms such as dyspnea and hypoxia, and eventually leading to respiratory failure. Therefore, glycyrrhetinic acid can effectively prevent the formation of pulmonary fibrosis on the basis of treating inflammation. In addition, glycyrrhetinic acid can also reduce protein synthesis in cells and block the cycle of cells in the G1 phase, thereby treating lung non-small cell carcinoma. In the past two years, the new coronavirus has spread globally, causing great losses to people's safety and economic development. Glycyrrhetinic acid has the ability to kill a variety of viruses, including the new coronavirus. The main mechanism is to induce the formation of cholesterol-dependent lipid rafts on the cell membrane, thereby preventing the virus from entering the cell, and capturing high mobility group box protein B1 in the cell interior and proliferation process, blocking the alarm function of HMGB1, and further blocking the proliferation of the virus.

[0004] However, glycyrrhizinic acid has poor water solubility and low bioavailability, and the amount reaching the lungs is low after systemic administration, which brings challenges to its treatment of lung diseases, and how to develop a preparation method capable of improving the water solubility of glycyrrhizinic acid, increasing the bioavailability, and targeting the lungs has become a research hotspot. For example, CN110787130A discloses "18β-glycyrrhizinic acid solid dispersion and a preparation method thereof", which is obtained by mixing 18β-glycyrrhizinic acid, a hydrophilic carrier and a bonding agent, and then processing by a solid dispersion technology, so as to improve the in-vitro dissolution rate of the drug and improve the pH-dependent solubility of glycyrrhizinic acid. However, this technology needs a hot melt extruder, and is extruded at a temperature above 150°C, and the amorphous powder obtained may have a crystal transformation phenomenon, affecting the stability of the particles.

[0005] CN1022552982 discloses "a glycyrrhizinic acid temperature-sensitive gel and a preparation method thereof", which can overcome the problems of low absorption and liver first-pass effect caused by oral administration, and gel preparations can only be used for skin and body cavity, and are not suitable for lung drug delivery. It has been reported in the literature that the solubility of GA can be increased by material modification, and the solubility of GA in water and phosphate buffer is greatly improved after synthesis of chitosan derivatives; in addition, the solubility of GA can be increased by nanocrystallization and preparation of liposomes by freeze-drying single-phase solution method, but the above-mentioned preparation process is relatively complex, which brings challenges to commercial development. At present, it is reported that glycyrrhizinic acid is used for treating pulmonary fibrosis, lung injury, pulmonary hypertension and lung cancer by directly administering raw materials or extracts of licorice, and therefore, there is a need to develop a preparation process for improving the bioavailability and lung targeting of glycyrrhizinic acid.

[0006] Therefore, there is an urgent need in the art to develop a glycyrrhizinic acid complex with high solubility, high bioavailability and excellent in-vivo absorption effect. SUMMARY

[0007] The purpose of the present application is to provide a glycyrrhizinic acid complex, which has significantly improved water solubility, has a biphasic release characteristic, can significantly improve the bioavailability of GA after oral administration, and can be used as a dry powder inhalation powder to realize lung local administration, improve the effective absorption amount of the drug in the lungs, and thus increase the treatment effect of GA on lung diseases.

[0008] In a first aspect of the present application, a complex is provided, which comprises components (a) glycyrrhizinic acid; (b) cyclodextrin-metal organic framework.

[0009] In another preferred embodiment, the complex is a glycyrrhizinic acid biphasic release complex.

[0010] In another preferred embodiment, the biphasic release means that the complex releases more than 40% (preferably more than 50%) of the glycyrrhetinic acid rapidly in water within 0.5 h, and the release is stable for 10-16 h; then the glycyrrhetinic acid is released slowly after 10-16 h, and reaches about 100% after about 96 h.

[0011] In another preferred embodiment, the complex has one or more features selected from the group consisting of:

[0012] (1) the complex is a crystal, preferably a square crystal, more preferably a cubic crystal;

[0013] (2) the particle size of the complex is 0.3-10 μm, for example 0.3, 0.5, 0.7, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9 μm;

[0014] (3) the complexation mode of (a) glycyrrhetinic acid; (b) cyclodextrin-metal organic framework in the complex is that the glycyrrhetinic acid enters the cavity of the cyclodextrin-metal organic framework, and / or the glycyrrhetinic acid is semi-included by the cyclodextrin;

[0015] (4) the drug loading (wt%) of the glycyrrhetinic acid in the complex is 5%-30%, preferably 8%-20%, for example 8%, 10%, 12%, 15%, 18%;

[0016] (5) the solubility of the complex in water (25°C) is 0.5-5 mg / mL, preferably 1-4 mg / mL, more preferably 2-4 mg / mL, more preferably 2-3 mg / mL;

[0017] (6) the solubility of the complex in water (25°C) is increased by 3000-10000 times, preferably 5000-10000 times, more preferably 5500-8000 times, compared with the raw drug;

[0018] (7) the drug loading molar ratio of the glycyrrhetinic acid to the cyclodextrin-metal organic framework in the complex is 0.02-0.8:1, preferably 0.3-0.8:1, more preferably 0.5-0.8:1;

[0019] (8) the particle size of the cyclodextrin-metal organic framework in the complex is 0.3-10 μm, for example 0.3, 0.5, 0.7, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9 μm.

[0020] In another preferred embodiment, the cyclodextrin is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or a combination thereof, preferably γ-cyclodextrin.

[0021] In another preferred embodiment, the complex is obtained by incubating glycyrrhetinic acid in a solution of cyclodextrin-metal organic framework using an incubation method.

[0022] In another preferred embodiment, the complex is obtained by the method of the second aspect of the present application.

[0023] In a second aspect of the present application, there is provided a method for preparing a complex as described in the first aspect of the present application, wherein the complex is prepared by the following method:

[0024] (s1) providing a cyclodextrin-metal organic framework;

[0025] (s2) mixing glycyrrhetinic acid with the cyclodextrin-metal organic framework and incubating for a time t1 at a temperature T1 to obtain the complex.

[0026] In another preferred embodiment, the cyclodextrin-metal organic framework is a micron-sized cyclodextrin-metal organic framework or a nanometer-sized cyclodextrin-metal organic framework, preferably a nanometer-sized cyclodextrin-metal organic framework.

[0027] In another preferred embodiment, the micron-sized cyclodextrin-metal organic framework has a size of 1-10 μm, for example 1-5 μm or 2-7 μm.

[0028] In another preferred embodiment, the nanometer-sized cyclodextrin-metal organic framework has a size of 0.1-1 μm, preferably 0.3-1 μm.

[0029] In another preferred embodiment, the reaction conditions of step (s2) have one or more features selected from the group consisting of:

[0030] (1) the molar ratio of glycyrrhetinic acid to cyclodextrin-metal organic framework is 1-15:1, preferably 5-15:1, preferably 5-10:1;

[0031] (2) the incubation temperature T1 is 40-70 °C, preferably 50-60 °C;

[0032] (3) the incubation is carried out under water bath conditions;

[0033] (4) the incubation time t1 is 1-12 h, preferably 1-6 h;

[0034] (5) the incubation is carried out in the presence of a solvent selected from the group consisting of: methanol, ethanol, or a combination thereof;

[0035] (6) the mass volume ratio of glycyrrhetinic acid to solvent is 0.2-0.5 mg / mL, preferably 0.25 mg / mL.

[0036] In another preferred embodiment, the reaction is carried out under stirring, and the stirring speed is 200-500 rpm, preferably 400 rpm.

[0037] In another preferred embodiment, the cyclodextrin-metal organic framework is a neutral cyclodextrin-metal organic framework.

[0038] In another preferred embodiment, the cyclodextrin-metal organic framework is prepared by the following method:

[0039] (a) mixing cyclodextrin with a base in a first solvent to obtain a first mixed solution;

[0040] (b) adding a second solvent to the first mixed solution and mixing to obtain a second mixture;

[0041] Optionally, (c) adding a size adjusting agent to the second mixture to crystallize and form a third mixture containing the cyclodextrin-metal organic framework material; and

[0042] (d) separating the third mixture to obtain the cyclodextrin-metal organic framework material.

[0043] In another preferred embodiment, in step (a), the cyclodextrin and the base are heated or ultrasonically dissolved in the first solvent to obtain the first mixed solution.

[0044] In another preferred embodiment, step (a) has one or more of the following characteristics:

[0045] (1) the first solvent is water, preferably pure water;

[0046] (2) the molar ratio of the cyclodextrin to the base is 1:1-10, preferably 1:2-8, more preferably 1:5-8;

[0047] (3) the concentration of the cyclodextrin in the first solvent is 20-1000 mM, preferably 20-100 mM, more preferably 20-50 mM, more preferably 25 mM;

[0048] (4) the base contains (a) metal ions and (b) acid radicals, wherein the metal ions are selected from K + , Li + , Na + , Mg 2+ , Ca 2+ , or a combination thereof; and the acid radicals are selected from OH - , CH3COO - , halide ions, or a combination thereof, preferably the base is KOH or CH3COOK.

[0049] In another preferred embodiment, the step (a) optionally further comprises a post-treatment step of filtering the product of mixing the cyclodextrin with the base in the first solvent, thereby obtaining a first mixed solution.

[0050] In another preferred embodiment, the step (b) has one or more of the following features:

[0051] (1) the second solvent is selected from ethanol, methanol, or a combination thereof;

[0052] (2) the volume ratio of the first mixed solution to the second solvent is 1:2-0.5, preferably 1:1-0.6;

[0053] (3) the reaction is carried out at 50-70°C;

[0054] (4) the reaction time is 0.25-1h, preferably 0.3h.

[0055] In another preferred embodiment, the step (b) optionally further comprises mixing the first mixed solution and the second solvent and heating to clarify, and the reaction obtains a second mixture.

[0056] In another preferred embodiment, the step (c) has one or more of the following features:

[0057] (1) the size adjusting agent is polyethylene glycol;

[0058] (2) the molecular weight of the polyethylene glycol is 15000-25000, preferably 20000;

[0059] (3) the mass ratio of the size adjusting agent to the cyclodextrin is 0.1-0.5, preferably 0.1-0.3;

[0060] (4) the size adjusting agent is added in the form of a solution dissolved in a third solvent; wherein the third solvent is selected from methanol, ethanol.

[0061] In another preferred embodiment, the step (c) comprises mixing the second mixture with the size adjusting agent, and standing in a cold water bath to crystallize, and the crystallization obtains a third mixture.

[0062] In another preferred embodiment, the crystallization is carried out at 2-10°C.

[0063] In another preferred embodiment, the crystallization time is 10-18h.

[0064] In another preferred embodiment, the step (c) comprises mixing the second mixture with the size adjusting agent, heating to react, and then standing to crystallize, thereby obtaining a third mixture.

[0065] In another preferred embodiment, the heating is carried out at 50-70°C.

[0066] In another preferred embodiment, the heating is for a time period of 0.2-0.5 h.

[0067] In another preferred embodiment, the crystallization is performed at 10-40 °C.

[0068] In another preferred embodiment, the crystallization is for a time period of 1-3 h.

[0069] In another preferred embodiment, the step (d) comprises a post-treatment step of centrifugation, washing and / or drying.

[0070] In another preferred embodiment, the washing comprises washing with methanol to a pH of 6.5-7.5.

[0071] In another preferred embodiment, the washing comprises washing once with absolute ethanol (containing 4% acetic acid), washing twice with absolute ethanol, and finally washing with methanol to a pH of 6.5-7.5.

[0072] In another preferred embodiment, the step (b) optionally further comprises centrifuging the product of mixing the first mixed solution and the second solvent, discarding the supernatant to obtain a second mixture.

[0073] In another preferred embodiment, the step (b) is repeated 0-2 (e.g. 1, 2) times.

[0074] In a third aspect of the present application, there is provided a formulation comprising the complex of the first aspect of the present application.

[0075] In another preferred embodiment, the formulation is for administration by inhalation or oral administration, preferably by inhalation.

[0076] In another preferred embodiment, the formulation is an inhalation formulation or an oral formulation.

[0077] In another preferred embodiment, the inhalation formulation comprises a dry powder inhaler, a spray, a powder mist or an aerosol.

[0078] In another preferred embodiment, the formulation further comprises a pharmaceutically acceptable carrier.

[0079] In another preferred embodiment, the pharmaceutically acceptable carrier is selected from the group consisting of lactose, cholesterol, magnesium stearate, or a combination thereof.

[0080] In another preferred embodiment, the lactose comprises coarse lactose and fine lactose.

[0081] In another preferred embodiment, the fine lactose is selected from the group consisting of lactose having a D50 particle size of 0-5 μm, lactose having a D50 particle size of 4-11 μm, or a combination thereof.

[0082] In another preferred embodiment, the coarse lactose is selected from the group consisting of lactose with a D50 particle size of 40-70 μm, lactose with a D50 particle size of 70-130 μm, lactose with a D50 particle size of 53-66 μm, lactose with a D50 particle size of 95-125 μm, or a combination thereof.

[0083] In another preferred embodiment, the pharmaceutically acceptable carrier is a mixture of coarse lactose and fine lactose, wherein the mass ratio of the fine lactose to the coarse lactose is 0.1-1:10, preferably 0.2, 0.5, 0.8:10.

[0084] In another preferred embodiment, the mass ratio of the complex to the pharmaceutically acceptable carrier is 1:0.01-2, preferably 1:1.

[0085] In another preferred embodiment, the preparation is prepared by premixing the complex and the pharmaceutically acceptable carrier in equal amounts.

[0086] In another preferred embodiment, the preparation is prepared by premixing the complex and the pharmaceutically acceptable carrier in equal amounts, and then adding the premixed powder into a V-type mixer, mixing at 40 r / min for 15 min, and continuing to mix at the same speed for 15 min to obtain the preparation.

[0087] In another preferred embodiment, the preparation (preferably a dry powder inhalation preparation) has one or more characteristics selected from the group consisting of:

[0088] (1) the fine particle fraction of the preparation is 10%-50%, preferably 20%-40%;

[0089] (2) the emptying rate of the preparation is more than 98%, preferably more than 99%, more preferably more than 99.5%;

[0090] (3) the in vivo absorption of glycyrrhetic acid in the preparation in rats is biphasic, i.e., more than 25% is rapidly absorbed within 5 min, and then slowly absorbed, with more than 40% absorbed after 8 h;

[0091] (4) the in vivo absorption of glycyrrhetic acid in the preparation in rats is increased by more than 3 times compared to the absorption of the raw material drug;

[0092] (5) the tissue distribution of glycyrrhetic acid in the preparation in rats is more than 50 times that in other tissues, and the concentration of glycyrrhetic acid in the lung is more than 70 ng / mg after 10 min;

[0093] (6) the lung coefficient of the preparation is reduced by more than 30% compared to directly administering the raw material drug, and the Smad3 concentration is reduced by 50%.

[0094] In a fourth aspect of the present application, there is provided an oral inhalation device, said device comprising an inhalation device and a formulation according to the third aspect of the present application.

[0095] In another preferred embodiment, the inhalation device comprises a dry powder inhalation device, a spray device, a powder mist device or an aerosol device.

[0096] In another preferred embodiment, the inhalation device is a single dose capsule inhalation device.

[0097] In a fifth aspect of the present application, there is provided the use of a complex according to the first aspect of the present application and / or a formulation according to the third aspect of the present application for biphasic release of glycyrrhetinic acid in solution.

[0098] In another preferred embodiment, the complex according to the first aspect of the present application and / or the formulation according to the third aspect of the present application is used for the treatment and / or prevention of a lung disease.

[0099] In another preferred embodiment, the lung disease is selected from the group consisting of pulmonary fibrosis, lung injury, lung tumor and COVID-19 virus.

[0100] In a sixth aspect of the present application, there is provided a method for biphasic release of glycyrrhetinic acid in solution, comprising administering a complex according to the first aspect of the present application and / or a formulation according to the third aspect of the present application to a patient in need thereof.

[0101] In another preferred embodiment, the method is performed in vitro.

[0102] In a seventh aspect of the present application, there is provided a method for the treatment and / or prevention of a lung disease, comprising administering a complex according to the first aspect of the present application and / or a formulation according to the third aspect of the present application to a patient in need thereof.

[0103] In another preferred embodiment, the lung disease is selected from the group consisting of pulmonary fibrosis, lung injury, lung tumor and COVID-19 virus.

[0104] In another preferred embodiment, the method is performed in vitro.

[0105] It should be understood that, within the scope of the present application, all combinations of the above-described technical features of the present application and the technical features specifically described hereinafter (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they are not listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0106] Figure 1 SEM images of nano-CD-MOF and GA@nano-CD-MOF in Example 1.

[0107] Figure 2PXRD pattern of nano-CD-MOF and GA@nano-CD-MOF in Example 1.

[0108] Figure 3 DSC pattern of nano-CD-MOF and GA@nano-CD-MOF in Example 1.

[0109] Figure 4 TGA pattern of nano-CD-MOF and GA@nano-CD-MOF in Example 1.

[0110] Figure 5 NGI result of GA@nano-CD-MOF in Example 1.

[0111] Figure 6 In vitro release of GA and γ-CD in GA@nano-CD-MOF in Example 1.

[0112] Figure 7 In vivo absorption curve of GA@nano-CD-MOF after inhalation and oral administration of GA in Example 1.

[0113] Figure 8 Molecular simulation diagram of GA@nano-CD-MOF in Example 1.

[0114] Figure 9 Mean blood concentration-time curve of different administration groups in Example 1.

[0115] Figure 10 Drug distribution concentration in different tissues of rats after oral and pulmonary inhalation of GA@nano-CD-MOF in Example 1.

[0116] Figure 11 HE and Masson staining diagram, HYP, TGF-β1 and Smad3 content of rat lung in Example 1. DETAILED DESCRIPTION

[0117] The present inventors have made extensive and in-depth studies and unexpectedly found a complex obtained by compounding glycyrrhetinic acid and cyclodextrin-metal organic framework, wherein the glycyrrhetinic acid enters the cavity of the cyclodextrin metal framework; the complex can significantly improve the water solubility of the glycyrrhetinic acid raw material drug, and the complex has a sustained-release feature and a biphasic release feature, which can make the drug amount in the body immediately reach a therapeutic concentration, and then slowly release to maintain the drug concentration in the body, reduce the administration frequency, and improve the medication compliance.

[0118] The present application also provides a preparation containing the complex, especially a dry powder inhaler, which can be directly delivered to the lung, increase the deposition amount of the drug in the lung, reduce the dosage, and reduce side effects. On this basis, the inventors have completed the present application.

[0119] Definitions

[0120] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0121] As used herein, the terms "comprising", "including", "containing", are interchangeable and include both open- and semi- closed definitions. In other words, the terms include "consisting of" and "consisting essentially of".

[0122] As used herein, the terms "cyclodextrin" and "CD" are interchangeable.

[0123] As used herein, the terms "cyclodextrin-metal organic framework" and "CD-MOF" are interchangeable.

[0124] Glycyrrhetinic acid

[0125] Glycyrrhetinic acid is the most important active ingredient in licorice and is also the metabolite and active substance of glycyrrhizic acid in the body. It has the effects of anti-inflammatory, antiviral, liver protection and detoxification, and immune function enhancement. The structure of glycyrrhetinic acid belongs to pentacyclic triterpenes, which has a steroid-like anti-inflammatory effect and an adrenal cortex hormone-like pharmacological effect, and is widely used in the treatment of various chronic hepatitis, urticaria, etc.

[0126] In recent years, the role of glycyrrhetinic acid in preventing and treating lung diseases has been frequently reported, including pulmonary fibrosis, lung injury, lung tumors, and new coronavirus pneumonia virus, etc. However, glycyrrhetinic acid has poor water solubility, and is basically not absorbed by the body through conventional administration methods such as oral administration and subcutaneous injection, resulting in low bioavailability, which greatly reduces the clinical application value of glycyrrhetinic acid. Moreover, long-term use of the drug can cause adverse reactions such as hyperaldosteronism, which manifests as edema, eczema, hypokalemia, hypertension related to electrolyte disturbance, and sodium retention in patients after long-term medication. The existence of these side effects may be due to the enterohepatic circulation of glycyrrhetinic acid, which leads to long-term accumulation in the body and cannot be excluded from the outside. Therefore, exploring new dosage forms of glycyrrhetinic acid, improving bioavailability, and reducing the dosage to eliminate adverse reactions have clinical practical application value.

[0127] Cyclodextrin-metal organic framework

[0128] The present application provides a cyclodextrin-metal organic framework having a suitable pore size inside for glycyrrhetinic acid to enter the internal pore or to be included.

[0129] The cyclodextrin-metal organic framework of the present application includes a cyclodextrin-metal organic framework (CD-MOF) with a pore size in the range of 2-7 μm, a micro-sized cyclodextrin-metal organic framework (micro-CD-MOF) with a pore size in the range of 1-5 μm, and a nano-sized cyclodextrin-metal organic framework (nano-CD-MOF) with a pore size in the range of 0.3-1 μm.

[0130] Preferably, the cyclodextrin-metal organic framework of the present application is a micro-sized cyclodextrin-metal organic framework (micro-CD-MOF) with a pore size in the range of 1-5 μm and a nano-sized cyclodextrin-metal organic framework (nano-CD-MOF) with a pore size in the range of 0.3-1 μm, both of which can be used for pulmonary administration.

[0131] The complex of the present application

[0132] The present application provides a complex comprising glycyrrhetinic acid and a cyclodextrin-metal organic framework.

[0133] As used herein, the term "the complex of the present application", "GA@CD-MOF" refers to the glycyrrhetinic acid-cyclodextrin-metal organic framework complex obtained by the present application, in which glycyrrhetinic acid enters the cavity of the cyclodextrin-metal framework to form a glycyrrhetinic acid-cyclodextrin-metal organic framework complex. The complex has a sustained-release feature and a biphasic release feature.

[0134] Preferably, the biphasic release refers to the rapid release of more than 40% (preferably more than 50%) of glycyrrhetinic acid in the complex within 0.5 h, which is stable and lasts for 10-16 hours; then the slow release of glycyrrhetinic acid after 10-16 hours, which reaches about 100% around 120 hours.

[0135] The complex of the present application can significantly improve the water solubility of glycyrrhetinic acid, and the solubility of the complex is increased by 3000-10000 times compared with the raw drug.

[0136] The present application uses a cyclodextrin-metal organic framework as a carrier to prepare a glycyrrhetinic acid complex, improves the bioavailability and achieves the purpose of pulmonary delivery, especially referring to the loading of glycyrrhetinic acid into a cyclodextrin organic framework, which increases the water solubility by thousands of times, and produces a biphasic release mechanism, so as to achieve the purpose of treating pulmonary diseases after oral and inhalation administration.

[0137] The cyclodextrin metal organic framework particle size is controllable, and after loading glycyrrhetic acid, a dry powder inhalation powder can be prepared, which is effectively delivered to the alveolar part, greatly increasing the deposition amount of glycyrrhetic acid in the lung. In addition, the feature of biphasic release can make glycyrrhetic acid be released rapidly in the body to reach the therapeutic concentration, and the remaining amount is slowly released to maintain the drug concentration in the body, thereby reducing the amount and frequency of administration, improving the medication compliance of patients, and reducing side effects.

[0138] Biphasic release

[0139] Biphasic release is a unique sustained-release technology, which refers to that the drug amount in the body reaches the therapeutic concentration (preferably within 30 min) immediately after administration, and then slowly releases to maintain the drug concentration in the body.

[0140] The complex of the present application has the feature of biphasic release, and the complex of the present application releases more than 40% (preferably more than 50%) of glycyrrhetic acid in water within 0.5 h, and is stable and lasts for 10-16 hours; then glycyrrhetic acid is slowly released after 10-16 hours, and about 100% is reached after about 96 hours.

[0141] Biphasic release can significantly reduce the frequency of administration and improve medication compliance.

[0142] Preparation

[0143] The present application provides a preparation, which comprises the complex of the present application.

[0144] In another preferred embodiment, the preparation of the present application is an oral preparation or an inhalation preparation, preferably an inhalation preparation.

[0145] In another preferred embodiment, the preparation of the present application further comprises a pharmaceutically acceptable carrier.

[0146] In the present application, the components of the term "pharmaceutically acceptable carrier" refer to substances suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e. with a reasonable benefit / risk ratio.

[0147] It should be understood that in the present application, the pharmaceutically acceptable carrier is not particularly limited, and is a material commonly used in the art, the types, methods of use, and sources of which are well known to those skilled in the art.

[0148] Examples of the pharmaceutically acceptable carrier include (but are not limited to) sugars, amino acids, propellants, polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium dodecyl sulfate), buffers, chelating agents, thickening agents, pH adjusting agents, transdermal enhancers, stabilizers, antioxidants, preservatives, bacteriostatic agents, pure water, pyrogen-free water, etc.

[0149] The inhalation preparation of the present application can be a dry powder inhalant, a spray, a powder mist or an aerosol.

[0150] A typical inhalation preparation is a dry powder inhalant.

[0151] In another preferred embodiment, the weight ratio of the complex to the pharmaceutically acceptable carrier is 0.2-2:0.2-2, preferably 0.5-1.5:0.5-1.5, more preferably 0.8-1.2:0.8-1.2.

[0152] In another preferred embodiment, the pharmaceutically acceptable carrier is selected from the group consisting of a sugar, an amino acid, lecithin and phosphatidylcholine, or a combination thereof.

[0153] In another preferred embodiment, the pharmaceutically acceptable carrier is lactose, preferably a combination of coarse lactose and fine lactose.

[0154] In another preferred embodiment, the pharmaceutically acceptable carrier is selected from the group consisting of lactose with a D50 particle size of 0-5 μm, lactose with a D50 particle size of 4-11 μm, lactose with a D50 particle size of 40-70 μm, lactose with a D50 particle size of 70-130 μm, lactose with a D50 particle size of 53-66 μm, lactose with a D50 particle size of 95-125 μm, or a combination thereof.

[0155] The main advantages of the present application include:

[0156] (1) The cyclodextrin-metal organic framework of the present application can significantly improve the solubility of GA;

[0157] (2) The complex of the present application has a biphasic release characteristic, the first phase releases more than 40% (preferably more than 50%) rapidly within 0.5 h, and is stable for 10-16 hours; the second phase is a slow release of glycyrrhetinic acid after 10-16 hours, reaching about 100% after about 96 hours, and having a sustained release effect after inhalation;

[0158] (3) Based on the particle size and morphology advantages of the CD-MOF particles, the GA is prepared into an inhalable dry powder, which is directly delivered to the lungs, increasing the drug deposition in the lungs, reducing the drug dosage, and reducing side effects;

[0159] (4) After mixing GA@CD-MOF with lactose, cholesterol or magnesium stearate, the flowability of the powder and the aerodynamic effect can be improved.

[0160] The application will be further described in conjunction with specific examples. It should be understood that these examples are only used to illustrate but not to limit the scope of the application. The experimental methods in the following examples, if not otherwise specified, are generally carried out according to the conventional conditions or the conditions recommended by the manufacturers. Unless otherwise specified, percentages and parts are by weight.

[0161] Example 1

[0162] (1) Preparation of GA@nano-CD-MOF

[0163] Preparation of nano-CD-MOF: γ-CD and KOH were prepared according to the molar ratio of n γ-CD : n KOH = 1:8, 12.96 g of γ-CD and 4.48 g of potassium hydroxide were weighed into 400 mL of pure water, ultrasonically dissolved, and filtered through a 0.8 μm filter membrane to obtain a reaction mother liquor. 100 mL of the mother liquor was measured into a 500 mL reagent bottle (4 bottles in total), 60 mL of methanol was added to the mother liquor, the solution system became turbid, and was fully shaken and mixed. The reaction liquid was placed in a water bath (60°C) for heating. After the precipitate was completely redissolved and the system was clear, it was placed in a 60°C water bath for 20 min, and then 80 mL of 8 mg / mL PEG 20000 methanol solution was added. The solution system became turbid again. It was fully shaken and mixed, and the reaction liquid was placed in a 60°C water bath for heating for 20 min. After being taken out, it was allowed to stand at room temperature for 2 h to allow the crystals to precipitate. After centrifugation, the supernatant was removed, and the precipitate was washed with anhydrous ethanol (containing 4% acetic acid), ultrasonically dispersed, centrifuged to remove the supernatant, and the lower precipitate was washed once with anhydrous ethanol. Finally, it was washed with methanol until the pH was 6.5-7.5, and then dried in a 60°C oven for 5 h to obtain neutralized nano-KOH-CD-MOF.

[0164] Preparation of GA@nano-CD-MOF: Drug glycyrrhetic acid (GA) was loaded by incubation method. The steps include: 1.18 g of GA was added to a 10 mL test tube, 10 mL of ethanol was added, and ultrasonication was performed until the drug was completely dissolved. GA and CD-MOF were added to 0.36 g of nano-CD-MOF prepared in Example 1 at a molar ratio of 10:1, and were added to a test tube. A magnetic stirrer was used to stir at 60°C in a water bath at 400 rpm for 6 h. The suspension was filtered with a Buchner funnel, and the filter cake was dried in a 60°C vacuum drying oven overnight to obtain GA@nano-CD-MOF.

[0165] Determination of solubility: 50 mg of 18β-glycyrrhetic acid raw material and GA@nano-CD-MOF were precisely weighed into centrifuge tubes, 1 mL of pure water was added for dissolution, and the tubes were placed in a constant temperature incubation shaker at 200 rpm and room temperature for 1 h. The supernatant was filtered through a 0.22 μm microporous filter and then subjected to high performance liquid chromatography determination.

[0166] High performance liquid chromatography (HPLC) conditions: The chromatographic column used was Spursil C10. 18 The chromatographic column (150×4.6mm, 5μm) was used, the mobile phase was 0.1% phosphoric acid solution-acetonitrile (15:85, v / v), the flow rate was 1mL / min, the UV detector was used, the detection wavelength was 254nm, the column temperature was 30℃, and the injection volume was 10μL.

[0167] Drug loading is calculated using the following formula: W% = M GA / M GA@nano-CD-MOF *100, where M GA To measure the GA quality, M GA@nano-CD-MOF The mass of the GA@nano-CD-MOF sample is measured.

[0168] The molar ratio of glycyrrhetinic acid to CD-MOF in GA@nano-CD-MOF was determined to be 0.64:1, and the drug loading was 17.2% (w% = M). GA / M GA@nano-CD-MOF The water solubility of the active pharmaceutical ingredient is 0.5 μg / mL, while the water solubility of GA@nano-CD-MOF is 3.92 mg / mL (both calculated based on the GA content), which is 7840 times higher.

[0169] (2) Characterization

[0170] The morphology of CD-MOF and GA@CD-MOF was evaluated using scanning electron microscopy (SEM). Figure 1 As can be seen, GA@CD-MOF has uniform particle size and regular cubic shape, with a geometric particle size of 0.3-1μm.

[0171] The crystallinity of the samples was characterized using powder X-ray diffraction (PXRD). Figure 2 GA exhibits characteristic peaks at 6°, 7°, 10°, 14°-16°, and 20°; nano-CD-MOF exhibits characteristic peaks at 4°, 6°, 7°, 14°, and 16°. The characteristic peaks of GA@nano-CD-MOF after drug loading are consistent with those of nano-CD-MOF, and the characteristic peaks of GA largely disappear after drug loading, indicating to some extent that GA was loaded into the cavity of nano-CD-MOF.

[0172] Figure 3GA exhibits a significant exothermic peak at 307℃, while nano-CD-MOF shows exothermic peaks at 251℃ and 368℃. In the DSC curve of GA@nano-CD-MOF, the characteristic peaks of both nano-CD-MOF and GA disappear, indicating to some extent that GA loading into the MOF cavity alters the properties of the MOF. GA initially shows a significant exothermic peak at 307℃, but when GA is loaded into MOF and γ-CD, the exothermic peak only appears after 307℃, indicating that the thermal stability of GA is improved after drug loading.

[0173] TGA results of nano-CD-MOF and GA@nano-CD-MOF ( Figure 4 The results showed that GA rapidly lost about 60% of its weight at 300-400℃, and showed a significant weight loss trend at 368℃. nano-CD-MOF lost about 33% of its weight at 236-300℃, and showed a significant weight loss trend starting at 236℃. The TGA curve of GA@nano-CD-MOF was similar to that of nano-CD-MOF, indicating that GA was loaded into the cavity of nano-CD-MOF.

[0174] (3) Aerodynamic evaluation

[0175] GA@nano-CD-MOF was passed through a 200-mesh sieve and filled into HPMC second-generation capsules, forming the final dry powder inhaler with a single-dose capsule-type inhalation device. The device was used to determine NGI at a flow rate of 65 L / min. Figure 5 Samples were collected from the device, adapter, artificial larynx, pre-separator, impactor body (stages 1-7), and microporous collector (MOC) respectively. The content of each stage was determined, and the fine particle fraction (FPF%) was calculated using CITDAS software (Copley, UK). Particles smaller than 5 μm were defined as fine particles. The FPF% of GA@nano-CD-MOF dry powder inhaler was 36.44%, indicating that more than 35% of the drug powder could be deposited deep into the lungs to exert its effect. The emptying rate of GA@nano-CD-MOF was greater than 98%, indicating that the prepared inhaled powder capsule had a good emptying rate.

[0176] (4) In vitro release assay

[0177] Approximately 650 mg of GA@nano-CD-MOF prepared in Example 1 was accurately weighed and dissolved in 500 mL of ultrasonically degassed pure water. Dissolution was determined using a paddle method at 37°C and 70 r / min. 5 mL of the suspension was collected at 0.08, 0.5, 1, 1.5, 2, 4, 6, 8, 10, 24, 30, 48, 72, 97, and 120 h, filtered through a 0.22 μm filter membrane, and the release of GA and γ-CD was measured. In the in vitro GA release assay, as...Figure 6 As shown, GA@nano-CD-MOF presents biphasic release of the drug over time. The first phase is a rapid release of more than 40% (preferably more than 50%) within 0.5 h, which is stable for 10-16 hours. The second phase is a slow release of glycyrrhetinic acid after 10-16 hours, which reaches 100% after about 96 hours. The first phase of the release of GA@CD-MOF calculated in terms of GA is the disintegration of CD-MOF, and the content of free γ-CD reaches 82% and is stable for 12 hours. The second phase is the slow disintegration of CD-MOF after 12 hours, which reaches about 100% after about 120 hours.

[0178] (5) Pharmacokinetics

[0179] Twenty-four healthy male rats weighing 250±50 g were divided into four groups, A, B, C and D, with six rats in each group. Group A was intravenously injected with GA@nano-CD-MOF (i.v.GA@nano-CD-MOF); group B was orally administered with a suspension of 18β-glycyrrhetinic acid raw material (i.g.GA); group C was orally administered with a suspension of GA@nano-CD-MOF (i.g.GA@nano-CD-MOF); and group D was administered with a dry powder inhalant of GA@nano-CD-MOF prepared in Example 1 (inhal.GA@nano-CD-MOF) by pulmonary administration. The administration dose was 10 mg / kg (calculated in terms of glycyrrhetinic acid content), and the blood samples were taken from the canthus at 5, 10, 15, 30 min and 1, 2, 4, 6, 8, 24 h after administration to determine the blood drug concentration of glycyrrhetinic acid. The pharmacokinetic parameters of the experimental data of different administration groups were analyzed by DAS 2.0 software, and the results showed that the in vivo bioavailability of GA could be significantly improved after CD-MOF was loaded with GA, and the absorption effect of GA in vivo was significantly improved. As shown in the blood drug concentration-time curves of different administration groups, Figure 7 ), the oral and inhalation administration of GA@CD-MOF can improve the bioavailability of GA in vivo, and the pulmonary administration has a more significant effect on improving the bioavailability of GA. The AUC of intravenous injection of GA@CD-MOF is 15.81±4.13 μg / mL*h, and the inhalation administration group improves the GA concentration from 2.81±1.32 μg / mL*h to 10.47±2.06 μg / mL*h. At the same time, the MRT (0-t) of oral administration of GA@CD-MOF and pulmonary administration of GA@CD-MOF is greater than that of intravenous administration, indicating that the absorption of GA in vivo is prolonged and the metabolism of GA in vivo is slowed down after CD-MOF is loaded with GA, which has a significant sustained-release effect. (The GA concentration data in the plasma of the four groups are shown in Table 1).

[0180] Table 1 Pharmacokinetic results of each group (n=6)

[0181]

[0182] (6) GA@nano-CD-MOF in vivo absorption curve

[0183] To illustrate the absorption process of GA@nano-CD-MOF in vivo, the absorption curves of inhaled administration and oral administration of raw materials were calculated according to the residual method (Figure 8). Figure 8 Both inhaled administration and oral administration of raw materials showed significant absorption process in 0-8 hours, but the absorption amount of inhaled administration was significantly improved. Compared with intravenous injection, GA@nano-CD-MOF inhalation had obvious sustained release process, similar to in vitro release, there was a two-phase absorption process, first 27.7% was rapidly absorbed within 5 min, then slowly absorbed, and 42.0% was absorbed at 8 hours. After oral administration of raw materials, it was slowly absorbed at a low level all the time, and finally only reached 13%.

[0184] (7) Molecular simulation to explain the two-phase release behavior of GA@nano-CD-MOF

[0185] Molecular simulation of the position of GA in nano-CD-MOF was performed using AutoDock Vina 1.1.2. According to the drug loading molar ratio of GA to nano-CD-MOF (GA:CD=0.7:1), 20 combination models were generated, and the most likely structure-activity relationship was obtained after evaluation with energy score graph (Figure 9). Figure 9 The blue GA represents in the double-cyclodextrin pair of CD-MOF, with a binding free energy of -9.3 kcal / mol; while the green GA represents in the large cavity of CD-MOF, with a binding free energy of -7.8 kcal / mol, the low binding free energy indicates that GA tends to be in the external double-cyclodextrin pair. The binding free energy of surface adsorbed free is -5.6 kcal / mol, which is much higher than -9.3 kcal / mol, so this hypothesis can be ignored, that is, the possibility of surface adsorbed GA is extremely small.

[0186] This also explains the biphasic release of GA. The process of GA being loaded into CD-MOF involves micronizing GA to the nanoscale, or even smaller. GA molecules distributed within the bicyclodextrin pairs are already in a semi-encapsulated state by the cyclodextrins, while the molecules entering the CD-MOF cavity remain in a free state. Therefore, in water, CD-MOF disintegrates into cyclodextrins, and the GA in the bicyclodextrin pairs forms cyclodextrin inclusion complexes during the dynamic equilibrium reaction, dissolving in the water. Meanwhile, the free GA molecules in the large cavity, if not rapidly encapsulated, aggregate due to similar compatibility, increasing their surface area. Furthermore, since CD is already dispersed in the aqueous solution, the inclusion conditions are not met, slowing down the inclusion rate. Encapsulation and dissolution in water can only occur slowly through accidental encounters and collisions between GA molecules and CD. However, even if GA molecules aggregate, their particle size is still smaller than the unencapsulated drug substance, and with prolonged exposure time, they will still be encapsulated and dissolved by CD.

[0187] (8) Tissue distribution experiment

[0188] Twenty-four male SD rats, weighing (250±50) g, were randomly divided into two groups, A and B, with 12 rats in each group. Group A received GA raw material suspension via gavage (igGA); Group B received GA@nano-CD-MOF via pulmonary delivery (inhal.GA@nano-CD-MOF). The dosage for the rats was 10 mg / kg (calculated as GA). Rats were fasted for 12 hours before and 4 hours after administration, but had free access to water. Blood was excised from the abdominal aorta at 10, 30 min, 1, and 4 h after administration, and the heart, liver, spleen, lungs, and kidneys were removed to determine the concentration of GA in each tissue. Figure 10 As shown, after gavage administration of GA raw material suspension, the overall GA deposition was less than 0.5 ng / mg after 10 minutes, and the deposition in the lungs was not significantly higher than in other organs. After administration of GA@CD-MOF dry powder inhaler to the lungs, 72 ng / mg of GA was deposited in the lungs, while other sites were within the range of 1.0-1.7 ng / mg. 92% of the GA was deposited in the lungs, indicating that GA delivered via the lungs can effectively deposit in the lungs, and the absorption rate is significantly improved compared to gavage administration of GA raw material suspension. Figure 7 Corresponding to the conclusion, it achieves a therapeutic effect on the lungs.

[0189] (9) Evaluation of efficacy

[0190] 36 healthy male rats, weighing 250±50g, 6 rats in each group, were divided into A, B, C, D, E, F, a total of 6 groups. Group A (gavaged with 5 mg / kg of normal saline for 14 days, blank (Blank) group), group B (after intratracheal instillation of 5 mg / kg bleomycin to make model, gavaged with 5 mg / kg of normal saline for 14 days after 7 days, model (Model) group), group C (after intratracheal instillation of 5 mg / kg bleomycin to make model, gavaged with 200 mg / kg of positive control drug pirfenidone for 14 days after 7 days, PFD group), group D (after intratracheal instillation of 5 mg / kg bleomycin to make model, 10 mg GA@CD-MOF was given to the lung for 14 days after 7 days, inhal GA@CD-MOF group), group E (after intratracheal instillation of 5 mg / kg bleomycin to make model, gavaged with 50 mg / kg of GA@CD-MOF for 14 days after 7 days, i.g. GA@CD-MOF group), group F (after intratracheal instillation of 5 mg / kg bleomycin to make model, 10 mg / kg GA was given to the lung for 14 days after 7 days, GA group). The rats were anesthetized, and blood was taken from the abdominal aorta, and lung tissue was taken. The lung coefficient (lung wet weight / body weight x 100%) was calculated, and the lung tissue was subjected to HE, Masson staining, hydroxyproline (HYP), transforming growth factor β1 (TGF-β1), and Smad3 content determination.

[0191] The results showed that bleomycin can promote the release of lung cell Smad3 and TGF-β1, and GA has different degrees of inhibition on this process. GA effectively delays the progression of pulmonary fibrosis by reducing collagen deposition, reducing inflammatory response, and inhibiting lung fibroblast proliferation. After drug loading, the treatment effect of GA is improved, especially the lung administration further improves the drug treatment effect Figure 11 , and the lung coefficient, HYP, TGF-β1, and Smad3 content results are shown in Table 2).

[0192] Table 2 Pharmacodynamic results of each group (n=6)

[0193]

[0194] Example 2

[0195] (1) Preparation of CD-MOF

[0196] Preparation of cyclodextrin-metal organic framework (CD-MOF): 100.0 g of γ-CD and 15.1 g of potassium acetate were weighed into 100 mL of pure water (n γ-CD : n CH3COOK= 1 : 2), 90°C water bath heating to dissolve, heating reaction 20 min, take out room temperature cooling to solution about 70°C, add 200 mL anhydrous ethanol, stirring uniform, 4000 rpm centrifugal 3 min, discard supernatant. Again add 100 mL anhydrous ethanol wash purification, stirring uniform, 4000 rpm centrifugal 3 min, discard supernatant, the precipitate spread in iron tray, 60°C drying to completely dry, namely get, particle size in 2~7μm.

[0197] Preparation of neutralized micro-CD-MOF: according to n γ-CD : n KOH = 1 : 8 feed molar ratio preparation, respectively weighing 64.8g γ-CD and 22.4g potassium hydroxide dissolved in 2000 mL pure water, ultrasonic 10 min, get reaction mother liquor. Add 1200 mL methanol to the mother liquor, solution system turbid, place in 60°C water bath heating to clear, continue heating reaction 20 min, add 12.8g PEG 20000, mix uniform. Cold water bath standing overnight, make crystal analysis precipitate. 4000 rpm centrifugal 3 min, discard supernatant, precipitate with anhydrous ethanol (containing 4% acetic acid) wash, ultrasonic dispersion, centrifugal discard supernatant, lower layer precipitate with anhydrous ethanol wash 1 time. Finally with methanol wash to pH 6.5-7.5, in 60°C oven drying 5h, namely get neutralized micro KOH-CD-MOF, particle size in 1~5μm.

[0198] Preparation of neutralized nano-CD-MOF: according to n γ-CD : n KOH = 1 : 8 feed molar ratio preparation, respectively weighing 64.8g γ-CD and 22.4g potassium hydroxide dissolved in 2000 mL pure water, ultrasonic 10 min, get reaction mother liquor. Add 1200 mL methanol to the mother liquor, solution system turbid, place in 60°C water bath heating to clear, continue heating reaction 20 min, add 12.8g PEG 20000, mix uniform. Cold water bath standing overnight, make crystal analysis precipitate. 4000 rpm centrifugal 3 min, discard supernatant, precipitate with anhydrous ethanol (containing 4% acetic acid) wash, ultrasonic dispersion, centrifugal discard supernatant, lower layer precipitate with anhydrous ethanol wash 1 time. Finally with methanol wash to pH 6.5-7.5, in 60°C oven drying 5h, namely get neutralized micro KOH-CD-MOF, particle size in 1~5μm.

[0199] (2) Preparation of GA@CD-MOF

[0200] 1.18 g of GA was added to a 10 mL vial, 10 mL of ethanol was added, and the mixture was sonicated until the drug was completely dissolved. GA and CD-MOF were added to the CD-MOF prepared in Example 2 at different molar ratios. The suspension was stirred at 400 rpm in a water bath using a magnetic stirrer for different times, and then filtered using a Buchner funnel. The filter cake was dried in a vacuum oven at 60°C overnight to obtain GA@CD-MOF. The molar ratio of GA to CD-MOF in GA@CD-MOF, the drug loading, and the solubility of GA@CD-MOF in water were determined by high-performance liquid chromatography. The resulting GA@CD-MOF was a regular cubic crystal with uniform particle size. The particle size, drug loading, drug loading molar ratio, and solubility are shown in Table 3.

[0201] Table 3 Process conditions and drug loading results for GA-loaded CD-MOF

[0202]

[0203] Example 3

[0204] Preparation of GA@micro-CD-MOF dry powder inhalation: GA@micro-CD-MOF was filled into HPMC second-generation capsules to form the final dry powder inhalation with a single-dose capsule inhalation device. Results: The fine particle fraction (FPF%) of GA@micro-CD-MOF dry powder inhalation was 27.79%, indicating that the micron-sized drug powder can effectively deposit in the deep lung to exert its efficacy. The emptying rate was 99.73%, indicating that the inhalation powder capsule has a good emptying rate.

[0205] Example 4

[0206] Preparation of GA@micro-CD-MOF dry powder inhalation: GA@micro-CD-MOF and lactose were weighed according to a mass ratio of 1:1, i.e., 1 g of GA@micro-CD-MOF and 1 g of lactose (fine lactose LH300(D 50 45% coarse lactose SV010(D 50The lactose / GA@micro-CD-MOF powder (95-125 μm, 1:9 ratio) was premixed in equal increments. The premixed powder was then added to a V-type mixer and mixed at 40 rpm for 15 minutes, followed by another 15 minutes at the same speed to obtain lactose / GA@micro-CD-MOF. This lactose / GA@micro-CD-MOF powder was then filled into HPMC second-generation capsules and combined with a single-dose capsule-type inhaler to form the final dry powder inhaler. Results: The fine particle fraction (FPF%) of the lactose / GA@micro-CD-MOF dry powder inhaler was 29.52%, and the emptying rate was 99.23%, demonstrating that increasing the amount of lactose allows more drug powder to deposit deep into the lungs and exert its therapeutic effect.

[0207] Example 5

[0208] Preparation of GA@micro-CD-MOF dry powder inhaler: Weigh 1g of GA@micro-CD-MOF and 1g of lactose (fine lactose LH300 (D) according to a GA@micro-CD-MOF to lactose mass ratio of 1:1. 50 (0-5μm): 45% crude lactose SV003 (D 50 The fine particle size (53-66 μm) was premixed at a ratio of 1:9 in equal increments. The premixed powder was then added to a V-type mixer and mixed at 40 rpm for 15 minutes, followed by another 15 minutes at the same speed to obtain lactose / GA@micro-CD-MOF. This lactose / GA@micro-CD-MOF was then filled into HPMC second-generation capsules and combined with a single-dose capsule-type inhaler to form the final dry powder inhaler. Results: The fine particle fraction (FPF%) of the lactose / GA@micro-CD-MOF dry powder inhaler was 30.78%, and the emptying rate was 99.44%, demonstrating that increasing the amount of lactose allows more drug powder to deposit deep into the lungs and exert its therapeutic effect.

[0209] Example 6

[0210] Preparation of GA@micro-CD-MOF dry powder inhalant: 1 g of GA@micro-CD-MOF and 30 mg of cholesterol were weighed separately, mixed in an equal incremental manner, and then mixed at 40 r / min for 15 min, and then mixed at the same speed for another 15 min to obtain cholesterol / GA@micro-CD-MOF. The obtained cholesterol / GA@micro-CD-MOF was filled into HPMC second-generation capsules to form a final dry powder inhalant together with a single-dose capsule-type inhalation administration device. Results: The fine particle fraction (FPF%) of the cholesterol / GA@micro-CD-MOF dry powder inhalant was 32.54%, and the emptying rate was 98.79%, proving that increasing the amount of cholesterol can enable more drug powder to be deposited in the deep lung to exert a pharmaceutical effect.

[0211] Example 7

[0212] Preparation of GA@micro-CD-MOF dry powder inhalant: 1 g of GA@micro-CD-MOF and 30 mg of cholesterol were weighed separately, mixed in an equal incremental manner, and then mixed at 40 r / min for 15 min, and then mixed at the same speed for another 15 min to obtain cholesterol / GA@micro-CD-MOF. The obtained cholesterol / GA@micro-CD-MOF was filled into HPMC second-generation capsules to form a final dry powder inhalant together with a single-dose capsule-type inhalation administration device. Results: The fine particle fraction (FPF%) of the cholesterol / GA@micro-CD-MOF dry powder inhalant was 32.54%, and the emptying rate was 98.79%, proving that increasing the amount of cholesterol can enable more drug powder to be deposited in the deep lung to exert a pharmaceutical effect.

[0213] All documents mentioned in this application are incorporated herein by reference as if each individual document were specifically and individually incorporated by reference. In addition, it is to be understood that the application may be carried out by specifically different embodiments and that each described embodiment can be implemented in either hardware or software or a combination of both, but that the application is not limited to any particular such embodiment.

Claims

1. A complex, characterized in that, The complex comprises (a) glycyrrhetinic acid; and (b) a cyclodextrin-metal-organic framework, wherein the complex is prepared by the following method. (s1) provides a cyclodextrin-metal-organic framework; (s2) Glycyrrhetinic acid was mixed with cyclodextrin-metal-organic framework and incubated at temperature T1 for time t1 to obtain the complex; The reaction conditions in step (s2) have the following characteristics: (1) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework is 1-15:1; (2) The temperature T1 is 40-70℃; (3) The time t1 is 1-12h; (4) The incubation is carried out in the presence of a solvent, the solvent being selected from methanol, ethanol, or a combination thereof.

2. The complex according to claim 1, characterized in that, The complex is a biphasic release complex of glycyrrhetinic acid.

3. The complex according to claim 2, characterized in that, The biphasic release refers to the fact that in water, glycyrrhetinic acid is rapidly released at a rate of over 40% within 0.5 hours and remains stable for 10-16 hours; then, after 10-16 hours, glycyrrhetinic acid is slowly released, reaching 100% in about 96 hours.

4. The complex according to claim 1, characterized in that, The complex has one or more features selected from the group consisting of: (1) The complex is a crystal; (2) The particle size of the composite is 0.3-10 μm; (3) In the complex, (a) glycyrrhetinic acid and (b) cyclodextrin-metal-organic framework are combined in such a way that glycyrrhetinic acid enters the cavity of the cyclodextrin-metal-organic framework and / or glycyrrhetinic acid is partially encapsulated by cyclodextrin. (4) The glycyrrhetinic acid loading in the complex is 5%-30% by weight; (5) The solubility of the complex in water at 25°C is 0.5-5 mg / mL; (6) The solubility of the complex in water at 25°C is increased by 3000-10000 times compared to the raw material; (7) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework in the complex is 0.02-0.8:1; (8) The particle size of the cyclodextrin-metal-organic framework in the complex is 0.3-10 μm.

5. The complex according to claim 4, characterized in that, The complex has one or more features selected from the group consisting of: (1) The composite is a cubic crystal; (2) The particle size of the composite is 0.5-5 μm; (3) The glycyrrhetinic acid loading in the complex is 8%-20% by weight; (4) The solubility of the complex in water at 25°C is 1-4 mg / mL; (5) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework in the complex is 0.3-0.8:1; (6) The particle size of the cyclodextrin-metal-organic framework in the complex is 0.5-5 μm.

6. The complex according to claim 1, characterized in that, The cyclodextrin is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or combinations thereof.

7. The complex according to claim 1, characterized in that, The cyclodextrin is γ-cyclodextrin.

8. The complex according to claim 1, characterized in that, The reaction conditions in step (s2) have the following characteristics: (1) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework is 1-15:1; (2) The temperature T1 is 40-70℃; (3) The incubation is carried out under water bath conditions; (4) The time t1 is 1-12h; (5) The incubation is carried out in the presence of a solvent, the solvent being selected from methanol, ethanol, or a combination thereof.

9. The complex according to claim 1, characterized in that, The reaction conditions of step (s2) have one or more characteristics selected from the group consisting of: (1) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework is 5-15:1; (2) The temperature T1 is 50-60℃; (3) The time t1 is 1-6h.

10. A method for preparing the complex as described in claim 1, characterized in that, The complex was prepared by the following method: (s1) provides a cyclodextrin-metal-organic framework; (s2) Glycyrrhetinic acid was mixed with cyclodextrin-metal-organic framework and incubated at temperature T1 for time t1 to obtain the complex.

11. The preparation method according to claim 10, characterized in that, The reaction conditions in step (s2) have the following characteristics: (1) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework is 1-15:1; (2) The temperature T1 is 40-70℃; (3) The incubation is carried out under water bath conditions; (4) The time t1 is 1-12h; (5) The incubation is carried out in the presence of a solvent, the solvent being selected from methanol, ethanol, or a combination thereof.

12. The preparation method according to claim 11, characterized in that, The reaction conditions of step (s2) have one or more characteristics selected from the group consisting of: (1) The molar ratio of glycyrrhetinic acid to cyclodextrin-metal-organic framework is 5-15:1; (2) The temperature T1 is 50-60℃; (3) The time t1 is 1-6h.

13. The preparation method according to claim 10, characterized in that, The cyclodextrin-metal-organic framework was prepared by the following method: (a) In a first solvent, cyclodextrin is mixed with a base to obtain a first mixed solution; (b) Add a second solvent to the first mixed solution, mix and react to obtain a second mixture; (c) Adding a size regulator to the second mixture to crystallize and form a third mixture containing the cyclodextrin-metal-organic framework material; and (d) The third mixture is separated to obtain the cyclodextrin-metal-organic framework material.

14. The preparation method according to claim 13, characterized in that, The method for preparing cyclodextrin-metal-organic frameworks has one or more features selected from the group consisting of: (1) In step (a), the first solvent is water; (2) In step (a), the base contains (a) a metal ion and (b) an acid radical, wherein the metal ion is selected from K. + Li + Na + Mg 2+ Ca 2+ or a combination thereof; the acid radical is selected from OH - CH3COO - Halogen ions, or combinations thereof; (3) In step (b), the second solvent is selected from ethanol, methanol, or a combination thereof; (4) In step (c), the size regulator is polyethylene glycol.

15. The preparation method according to claim 14, characterized in that, The method for preparing cyclodextrin-metal-organic frameworks has one or more features selected from the group consisting of: (1) In step (a), the first solvent is pure water; (2) In step (a), the base is KOH or CH3COOK.

16. A formulation, characterized in that, The formulation comprises the complex of claim 1.

17. The formulation of claim 16, characterized in that, The formulation is administered by inhalation or oral administration.

18. The formulation of claim 17, characterized in that, The formulation is administered by inhalation.

19. The formulation of claim 16, characterized in that, The formulation is an inhaled formulation or an oral formulation, and the inhaled formulation includes a spray, powder, or aerosol.

20. An oral inhalation drug delivery device, characterized in that, The device includes an inhalation device and the formulation of claim 16.

21. Use of a complex as claimed in claim 1, or an formulation as claimed in claim 16, characterized in that, Used for biphasic release of glycyrrhetinic acid in solution.

22. The use as described in claim 21, characterized in that, The biphasic release refers to the fact that in water, glycyrrhetinic acid is rapidly released at a rate of over 40% within 0.5 hours and remains stable for 10-16 hours; then, after 10-16 hours, glycyrrhetinic acid is slowly released, reaching 100% in about 96 hours.

23. Use of a complex as claimed in claim 1, or an formulation as claimed in claim 16, characterized in that, This is used to prepare a medicine for treating and / or preventing lung diseases selected from the group consisting of pulmonary fibrosis, lung injury, and lung tumors.

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