Artemisia soup composition with the effect of protecting liver and its application
By combining amorphous micronized powder composition with twin-screw extrusion technology, the problems of solubility and storage stability of traditional Chinese medicine composition were solved, and the highly effective hangover relief and liver protection effect of Yin Chen Hao Tang composition was achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HUNAN NORMAL UNIVERSITY
- Filing Date
- 2026-05-06
- Publication Date
- 2026-06-12
Smart Images
Figure CN122182584A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of traditional Chinese medicine preparation technology, specifically to a Yin Chen Hao Tang composition with hangover relief and liver protection effects and its application. Background Technology
[0002] Yin Chen Hao Tang, a classic traditional formula for liver protection, contains flavonoids such as quercetin and kaempferol, as well as anthraquinone components such as aloe-emodin. These active substances have pharmacological effects of downregulating the release of inflammatory cytokines and repairing liver damage. However, when these components are extracted and purified from their original medicinal environment, they often spontaneously form a stable crystalline state with high lattice energy. This dense crystal structure directly leads to low solubility in water and gastrointestinal physiological fluids, slow dissolution of drug molecules, and difficulty in penetrating intestinal cell membranes to enter the bloodstream. Insufficient bioavailability constitutes the main technical bottleneck restricting the actual liver-protecting and hangover-relieving effects of these components.
[0003] To address the aforementioned solubility bottleneck, pharmaceutical process engineering typically favors fusing crystalline drugs with polymer carriers, attempting to transform them into high-energy amorphous solid dispersions. Traditional direct hot-melt extrusion processes are widely used due to their continuous production advantages, but this comes with the problem of processing heat damage. The flavonoids and anthraquinones in traditional Chinese medicine generally have high melting points; to ensure sufficient plasticization and melting of the mixture within the extruder chamber, the equipment operating temperature must be set within a high range. Under this continuous high-temperature processing environment accompanied by high-intensity mechanical shearing, the heat-sensitive active ingredients of traditional Chinese medicine inevitably undergo thermal degradation or spatial structure destruction, directly weakening the original pharmacodynamic basis during the formulation stage.
[0004] Even if the risk of thermal degradation during processing is overcome, the inherent physical instability of amorphous systems remains another challenge. Amorphous solid dispersions are inherently thermodynamically unstable. During long shelf-life storage or when the formulation is released into the gastrointestinal tract, drug molecules in a high-energy state tend to undergo microscopic thermal motion within the matrix. If the formulation lacks sufficient intermolecular anchoring to limit this physical migration, drug molecules will aggregate and spontaneously rearrange themselves towards a more thermodynamically stable crystalline structure, a phenomenon known as crystallization re-aggregation. Once crystallization re-aggregation occurs, the solubility advantage gained through early processing will be lost, and the drug release profile will fluctuate, ultimately preventing the drug from maintaining a uniform and long-lasting effect in vivo. Summary of the Invention
[0005] To address the shortcomings of existing technologies, this invention provides a Yin Chen Hao Tang composition with hangover relief and liver protection effects and its application, solving the problems of poor water solubility, low bioavailability, and easy crystal lattice recombination and re-aggregation of poorly soluble active ingredients in traditional Chinese medicine during storage or dissolution.
[0006] To achieve the above objectives, the present invention provides the following technical solution: Firstly, the following technical solution is adopted: A composition of Artemisia capillaris decoction with hangover-relieving and liver-protecting effects, wherein the composition is in an amorphous and micronized powder state, and the composition is made from raw materials comprising the following parts by weight: Quercetin: 10.0 parts; Kaempferol: 10.0 parts; Aloe-emodin: 10.0 parts; L-arginine: 1.5–6.0 parts; Glycyrrhizic acid: 15.0–60.0 parts; Polyvinylpyrrolidone K30: 60.0-180.0 parts.
[0007] By adopting the above technical solution, the physicochemical state of the active ingredients is changed at the thermodynamic and kinetic levels by combining the basic amino acid L-arginine with the polymer polyvinylpyrrolidone K30 and the surface active ingredient glycyrrhizic acid. Therefore, the original crystal lattice of the insoluble components is deconstructed, the in vitro dissolution rate is increased, and the re-aggregation of crystals is prevented.
[0008] To achieve the aforementioned changes in physicochemical state, the underlying reaction mechanism lies in the following: the phenolic hydroxyl groups in the molecular structures of quercetin, kaempferol, and aloe-emodin in the system dissociate and release protons under the heat of processing. Simultaneously, the free guanidin and amino groups in the L-arginine molecular chain are basic and can act as proton acceptors, binding with the aforementioned dissociated protons. This microscopic matching between functional groups promotes spontaneous acid-base proton transfer reactions among the components in the system, leading to the formation of ion-pair complexes. With the establishment of the complexation reaction, the lactam carbonyl functional group present in the polyvinylpyrrolidone K30 molecular structure acts as a hydrogen bond acceptor, further interacting with the undissociated hydroxyl or amino groups in the complex through hydrogen bonding. This series of physicochemical interactions causes the original lattice of the poorly soluble active ingredients to decompose, allowing them to be fixed in a monomolecular dispersed state within the three-dimensional polymer network framework of polyvinylpyrrolidone K30, thus exhibiting a rheologically uniform amorphous physical state.
[0009] Preferably, the composition is made from raw materials comprising the following parts by weight: 10.0 parts quercetin; 10.0 parts kaempferol; 10.0 parts aloe-emodin; 3.75 parts L-arginine; 37.5 parts glycyrrhizic acid; and 0.0 parts polyvinylpyrrolidone K3012.
[0010] By adopting the above technical solution, the raw material ratio is matched to achieve stoichiometric equilibrium, meeting the molar ratio requirements of groups for acid-base proton transfer reactions. Furthermore, the proportion of polymeric carrier is within a moderate range, which can both inhibit the thermal motion of drug molecules and avoid the increase in viscosity of the dissolution medium caused by excessive carrier, thus balancing the system's dissolution rate and drug release rate.
[0011] Preferably, the method for preparing the composition includes the following steps: The raw materials are added together into a mixer according to the weight proportions and mixed at room temperature to obtain a premixed dry powder with no color difference and uniform dispersion. The premixed dry powder is continuously fed into the first zone of a co-rotating twin-screw extruder. The barrel temperature of the first zone is controlled. As the screw conveyor advances to the second zone, purified water is continuously injected into the barrel of the second zone to obtain water-plasticized material. The water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements to induce the material to melt, thereby obtaining a ternary homogeneous melt with uniform rheological state; The ternary homogeneous melt is pushed into the fifth zone equipped with a large-lead delivery thread, and the vacuum pump system is turned on to maintain the pressure at the exhaust port, thus obtaining a dehydrated and quenched solid precursor. The dehydrated and quenched solid precursor is further advanced to the sixth zone and the extrusion die, which are sealed with reverse thread, and extruded to obtain an amorphous, semi-transparent, brittle shaped strand. The shaped strip is fed onto a conveyor belt, cooled, solidified, pulverized, and collected and sieved to obtain the composition in a micro powder state.
[0012] By adopting the above technical solution, the technical defects of high melting point drugs being easily thermally degraded by direct hot-melt extrusion are overcome by using co-rotating twin-screw extrusion combined with continuous water injection and flash evaporation dehydration processes. Therefore, the effects of low processing temperature, high amorphous conversion rate and continuous production are achieved.
[0013] In the specific equipment processing stage, the driving principle of the amorphous transformation of the system is as follows: Initially, water molecules, acting as a plasticizing medium, penetrate between the molecular chain segments of polyvinylpyrrolidone K30. This penetration increases the free volume of the polymer and lowers the glass transition temperature of the system, allowing the material to exhibit a plastic state even under heating conditions below the melting point of the drug. When the material is conveyed by the screw to the area with meshing disc elements, mechanical shear forces continuously renew the material surface. More importantly, purified water exhibits a high dielectric constant under local high temperature and high pressure conditions, constructing a polar microenvironment for the system. This environment weakens the resistance to proton transfer, thereby inducing the acid-base proton transfer complexation reaction mentioned above. As processing progresses, the reacted melt enters the large-lead threaded space of the fifth zone. Due to the sudden expansion of the system volume and the vacuum negative pressure state, the water molecules trapped inside the system vaporize, boil, and flash evaporate. The vaporization of water inevitably absorbs the latent heat from the surroundings, causing the system temperature to drop sharply, producing a phase change rapid cooling effect. After rapid cooling, the viscosity of the system increases immediately. At this point, the drug molecules are frozen and solidified, losing their ability to rearrange into a crystal lattice, thus establishing and maintaining their amorphous structure.
[0014] Preferably, the mixer is a three-dimensional motion mixer; the mixing at room temperature is carried out at a speed of 15-35 rpm for 15-30 minutes; the premixed dry powder is continuously fed in using a loss-in-weight twin-screw feeder at a rate of 1.0-3.0 kg / h; the temperature of the first zone barrel is controlled to be maintained at 70-85°C; the continuous injection of purified water is carried out using a plunger-type industrial metering pump, and the water injection rate is controlled to be constant at 2.0%-6.0% of the premixed dry powder feed rate.
[0015] By adopting the above technical solution, the constant proportion of water injection ensures that the polymer chain segments obtain sufficient plasticizing effect to reduce the plasticizing temperature, while avoiding the problems of insufficient melt strength of the system and excessive load in the subsequent dehydration step caused by excessive water.
[0016] Preferably, after the water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements, the barrel temperature is set to 105-115°C and the spindle speed is set to 150-250 rpm; the induced melting of the material is carried out under strong shear and the dielectric action of water molecules.
[0017] By adopting the above technical solution, the set thermal field temperature and screw spindle speed generate sufficient mechanical and thermal energy to ensure that the acid-base proton transfer complexation reaction reaches the reaction endpoint within the limited residence time in the extruder chamber.
[0018] Preferably, after the ternary homogeneous melt is pushed into the fifth zone equipped with a large-lead delivery thread, the barrel temperature is set to 110-115°C; the exhaust port gauge pressure is maintained at -0.095MPa to -0.07MPa.
[0019] By adopting the above technical solution, the negative pressure environment changes the boiling point threshold of water, forming a significant pressure difference driving force, which drives the water molecules encapsulated inside the polymer to be removed, thereby achieving water stripping and system cooling and solidification.
[0020] Preferably, after the dehydrated and quenched solid precursor is further advanced to the sixth zone with reverse thread seal and the extrusion die, the temperature of the sixth zone and the die is independently compensated and raised to 120-125°C, and the die pressure is maintained at 2.0-5.0 MPa by using secondary shearing and pressure building; after the formed strip is introduced onto the conveyor belt, the cooling and solidification is carried out on the conveyor belt with a forced air cooling system at 20-25°C for 2-5 minutes; the crushing and collection sieving is carried out by crushing the powder with a universal crusher and collecting it through an 80-120 mesh standard sieve.
[0021] By adopting the above technical solution, the reverse threaded seal combined with independent compensation heating eliminates residual micropores inside the extrudate, establishes sufficient die back pressure, and extrudes to form a uniform and dense solid strand structure; ultimately controlling particle size distribution and improving the flow properties of the powder and its adaptability to formulation processing.
[0022] Secondly, the following technical solution is adopted: Application of Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects in the preparation of hangover-relieving and liver-protecting drugs or health foods.
[0023] By adopting the above technical solution, based on the physical structural characteristics of the single-molecule dispersed state of the composition, the transmembrane absorption efficiency and blood drug concentration of the active ingredients of Chinese herbal flavonoids and anthraquinones are improved. It can block or downregulate the release level of inflammatory cytokines in the body, and achieve the physiological effects of relieving hangover and repairing liver damage.
[0024] This invention provides an Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects, and its application. It possesses the following beneficial effects: 1. This invention combines the active ingredients of traditional Chinese medicine with L-arginine and polyvinylpyrrolidone K30. The basic groups provided by L-arginine react with the phenolic hydroxyl groups of the poorly soluble components via an acid-base proton transfer reaction, forming ion-pair complexes that are then immobilized within the polymer backbone. This alteration of the microstructure disrupts the original crystal structure of the flavonoids and anthraquinones, transforming them into a monomolecularly dispersed amorphous state. This improves the water solubility and transmembrane absorption efficiency of the poorly soluble components, overcoming the low bioavailability of traditional Chinese medicine compositions.
[0025] 2. This invention employs a twin-screw extrusion combined with a negative pressure flash dehydration process during preparation. This allows the molten system to instantly lose moisture and undergo a phase change rapid cooling at the tail end of the extruder. The sudden drop in system temperature leads to a rapid increase in viscosity, directly freezing the drug molecules within a solid framework. Combined with the hydrogen bonding interactions formed between polyvinylpyrrolidone K30 and the drug molecules, this prevents them from rearranging their molecular mobility. This mechanism effectively prevents lattice recombination and re-aggregation during long-term storage and gastrointestinal dissolution, ensuring the physical stability of the system.
[0026] 3. The preparation method of this invention involves continuously injecting purified water into the extruder barrel. The plasticizing effect of water molecules lowers the glass transition temperature of the polymer, allowing the mixture to plasticize and melt at a temperature below the drug's melting point. This low-temperature extrusion environment avoids the thermal degradation of active ingredients that easily occurs in conventional high-temperature hot-melt extrusion, preserving the complete pharmacodynamic material basis of the composition. This ensures that the composition can effectively exert its hepatoprotective and alcohol-detoxifying effects in practical applications, such as downregulating the release of inflammatory factors and repairing liver damage. Attached Figure Description
[0027] Figure 1 This is a three-dimensional simulation diagram of molecular docking between the active ingredient released in Example 2 of the present invention and the target proteins TNF-α, IL-1β and IL-6; Figure 2 This is a cell thermal transfer analysis curve of the active ingredient released in Example 2 of the present invention to the TNF-α target, showing its thermal stability. Figure 2 The subplot of figure 'a' is a heat transfer analysis curve in the RAW264.7 cell system. Figure 2 The subplot in b is a heat transfer analysis curve in the recombinant protein system; Figure 3 This is a scatter plot showing the quantitative scatter distribution of the effect of the active ingredient released in Example 2 of the present invention on lipopolysaccharide-induced cells, wherein... Figure 3 The subplot of 'a' is a scatter plot showing the quantitative distribution of the relative fluorescence intensity of p65 in the cell nucleus. Figure 3 Subplot b is a scatter plot showing the quantitative distribution of the relative transcriptional level of TNF-α mRNA; Figure 4 This is a time-line graph showing the effect of the monomer released in Example 2 of the present invention on cell viability, wherein... Figure 4 The subplot of figure 'a' is a line graph showing the time of quercetin release at different concentrations. Figure 4 The subplot in section b is a line graph showing the time of kaempferol release at different concentrations; Figure 5 This is a line graph showing the dynamic changes in the retention rate of quercetin at different processing stages under different process treatments according to the present invention. Figure 6 Line graph showing the change in cumulative quercetin dissolution rate of different groups of samples in this invention during storage under accelerated conditions; Figure 7 This is a line graph showing the changes in the cumulative dissolution rate in vitro for different groups of samples according to the present invention. Detailed Implementation
[0028] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0029] Examples 1-3: Example 1: This embodiment provides a composition of Artemisia capillaris decoction with hangover-relieving and liver-protecting effects, comprising the following process: 10.0g quercetin, 10.0g kaempferol, 10.0g aloe-emodin, 1.5g L-arginine, 15.0g glycyrrhizic acid and 60.0g polyvinylpyrrolidone K30 were put into a three-dimensional motion mixer and mixed at 15 rpm for 15 minutes at room temperature to obtain a premixed dry powder with no color difference and uniform dispersion. The premixed dry powder is continuously fed into the first zone of the co-rotating twin-screw extruder at a rate of 1.0 kg / h via a loss-in-weight twin-screw feeder. The barrel temperature in the first zone is maintained at 70°C. As the screw conveyor advances to the second zone, purified water is continuously injected into the barrel via a plunger-type industrial metering pump. The water injection rate is kept constant at 2.0% of the premixed dry powder feed rate, which promotes the increase of the free volume of polymer chain segments and obtains water-plasticized material. The water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements. The barrel temperature is set to 105℃ and the spindle speed is 150rpm. Under strong shear and the dielectric effect of water molecules, the material is induced to melt and spontaneous acid-base proton transfer complexation reaction occurs between the components, resulting in a ternary homogeneous melt with uniform rheological state. The ternary homogeneous melt is pushed into the fifth zone equipped with a large lead feed thread. The barrel temperature is set to 110℃, and the vacuum pump system is turned on to maintain the exhaust port gauge pressure at -0.07MPa. This induces the plasticized water inside the system to boil and flash evaporate instantly, and undergo phase transformation quenching and rapid cooling to obtain a dehydrated and quenched solid precursor with frozen molecules in an amorphous state. The dehydrated and quenched solid precursor is further pushed to the sixth zone and the extrusion die head with reverse thread seal. The temperature of the sixth zone and the die head is set to be independently compensated and heated to 120°C. The die head pressure is maintained at 2.0MPa by using secondary shear pressure building. The densified extrusion yields amorphous, semi-transparent, brittle shaped strands. The shaped strips are fed onto a conveyor belt with a forced air cooling system and cooled and solidified at 20°C for 2 minutes. The powder is then pulverized by a universal pulverizer and collected through an 80-mesh standard sieve to obtain the final micronized Artemisia capillaris decoction composition.
[0030] Example 2: This embodiment provides a composition of Artemisia capillaris decoction with hangover-relieving and liver-protecting effects, comprising the following process: 10.0g quercetin, 10.0g kaempferol, 10.0g aloe-emodin, 3.75g L-arginine, 37.5g glycyrrhizic acid and 120.0g polyvinylpyrrolidone K30 were put into a three-dimensional motion mixer and mixed at 25 rpm for 22 minutes at room temperature to obtain a premixed dry powder with no color difference and uniform dispersion. The premixed dry powder is continuously fed into the first zone of the co-rotating twin-screw extruder at a rate of 2.0 kg / h via a loss-in-weight twin-screw feeder. The barrel temperature in the first zone is maintained at 78°C. As the screw conveyor advances to the second zone, purified water is continuously injected into the barrel via a plunger-type industrial metering pump. The water injection rate is controlled to be constant at 4.0% of the premixed dry powder feed rate, which promotes the increase of the free volume of polymer chain segments and obtains water-plasticized material. The water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements. The barrel temperature is set to 110℃ and the spindle speed is 200rpm. Under strong shear and the dielectric effect of water molecules, the material is induced to melt and spontaneous acid-base proton transfer complexation reaction occurs between the components, resulting in a ternary homogeneous melt with uniform rheological state. The ternary homogeneous melt is pushed into the fifth zone equipped with a large lead feed thread. The barrel temperature is set to 112℃, and the vacuum pump system is turned on to maintain the exhaust port gauge pressure at -0.08MPa. This induces the plasticized water inside the system to boil and flash evaporate instantly, and undergo phase transformation quenching and rapid cooling to obtain a dehydrated and quenched solid precursor with frozen molecules in an amorphous state. The dehydrated and quenched solid precursor is further pushed to the sixth zone and the extrusion die with reverse thread seal. The temperature of the sixth zone and the die is set to be independently compensated and heated to 123°C. The die pressure is maintained at 3.5MPa by using secondary shear pressure building. The densified extrusion yields amorphous, semi-transparent, brittle shaped strands. The shaped strips are fed onto a conveyor belt with a forced air cooling system and cooled and solidified at 22°C for 3 minutes. Then, they are pulverized by a universal pulverizer and the powder is collected and passed through a 100-mesh standard sieve to obtain the final micronized Artemisia capillaris decoction composition.
[0031] Example 3: This embodiment provides a composition of Artemisia capillaris decoction with hangover-relieving and liver-protecting effects, comprising the following process: 10.0g quercetin, 10.0g kaempferol, 10.0g aloe-emodin, 6.0g L-arginine, 60.0g glycyrrhizic acid and 180.0g polyvinylpyrrolidone K30 were put into a three-dimensional motion mixer and mixed at 35 rpm for 30 minutes at room temperature to obtain a premixed dry powder with no color difference and uniform dispersion. The premixed dry powder is continuously fed into the first zone of the co-rotating twin-screw extruder at a rate of 3.0 kg / h via a loss-in-weight twin-screw feeder. The barrel temperature in the first zone is maintained at 85°C. As the screw conveyor advances to the second zone, purified water is continuously injected into the barrel via a plunger-type industrial metering pump. The water injection rate is kept constant at 6.0% of the premixed dry powder feed rate, which promotes the increase of the free volume of polymer chain segments and yields water-plasticized material. The water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements. The barrel temperature is set to 115℃ and the spindle speed is 250rpm. Under strong shear and the dielectric effect of water molecules, the material is induced to melt and spontaneous acid-base proton transfer complexation reaction occurs between the components, resulting in a ternary homogeneous melt with uniform rheological state. The ternary homogeneous melt is pushed into the fifth zone equipped with a large lead feed thread. The barrel temperature is set to 115℃, and the vacuum pump system is turned on to maintain the exhaust port gauge pressure at -0.095MPa. This induces the plasticized water inside the system to boil and flash evaporate instantly, and undergo phase transformation quenching and rapid cooling to obtain a dehydrated and quenched solid precursor with frozen molecules in an amorphous state. The dehydrated and quenched solid precursor is further pushed to the sixth zone and the extrusion die head with reverse thread seal. The temperature of the sixth zone and the die head is set to be independently compensated and heated to 125°C. The die head pressure is maintained at 5.0MPa by using secondary shear pressure building. The densified extrusion yields amorphous, semi-transparent, brittle shaped strands. The shaped strips are fed onto a conveyor belt with a forced air cooling system and cooled and solidified at 25°C for 5 minutes. Then, they are pulverized by a universal pulverizer and the powder is collected through a 120-mesh standard sieve to obtain the final micronized Artemisia capillaris decoction composition.
[0032] Comparative Examples 1-4: Comparative Example 1: Compared with Example 2, the difference is that L-arginine is not added to the formula, while the proportions of other materials and the preparation process parameters are the same.
[0033] Comparative Example 2: Compared with Example 2, the difference is that purified water is not injected in the preparation process, and the vacuum pump is not turned on in the fifth zone. Due to the lack of water plasticization, in order to ensure the fluidity of the polyvinylpyrrolidone matrix, the barrel temperature of the third to sixth zones is set to 155°C. After demolding, air cooling is not used for rapid cooling, but rather the material is allowed to cool slowly at room temperature. The other material ratios and preparation process parameters are the same.
[0034] Comparative Example 3: Compared with Example 2, the difference is that glycyrrhizic acid is not added to the formula, while the proportions of other materials and the preparation process parameters are the same.
[0035] Comparative Example 4: Compared with Example 2, the difference is that all raw materials are simply put into a three-dimensional motion mixer according to the formula, mixed evenly, and then directly crushed and sieved. No twin-screw water injection hot melt extrusion and flash evaporation dehydration process is performed. All other processes are the same.
[0036] Test Examples 1-5: Test Example 1: Crystal structure files of TNF-α, IL-1β, and IL-6 were extracted from a general protein database and then dehydrated and hydrogenated.
[0037] The composition prepared in Example 2 was placed in a simulated physiological medium for isothermal dissolution, and quercetin, kaempferol and aloe-emodin monomers were separated and purified as docking ligands.
[0038] The interaction between ligand molecules and the binding pockets of various receptor proteins was simulated using a molecular docking program. The docking conformation was recorded, binding affinity data was extracted, and three-dimensional docking conformation diagrams of each component were output. The interaction sites of amino acid residues were observed.
[0039] RAW264.7 macrophages in the logarithmic growth phase were cultured, and the corresponding human recombinant target protein solution was prepared simultaneously.
[0040] RAW264.7 macrophage samples and human recombinant target protein samples were exposed to the dissolution media of Example 2 and Comparative Example 1, respectively. An equal amount of blank solvent was added to the control group, and incubation was maintained until drug molecules penetrated and established binding equilibrium.
[0041] After incubation, each group of samples was aliquoted into microcentrifuge tubes.
[0042] Multiple temperature points were set within the range of 40℃ to 80℃ using a gradient PCR instrument, and the samples were heated for three minutes.
[0043] After heating, the material is cooled to room temperature to terminate the thermal denaturation process.
[0044] The heated RAW264.7 macrophage samples were subjected to cell membrane disruption by adding lysis buffer and then precipitated in a low-temperature centrifuge to form denatured protein complexes.
[0045] Soluble target protein fragments were extracted from the supernatant, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to a membrane for sequential incubation with primary and secondary antibodies.
[0046] The luminescent band signal was captured and the gray value was measured. The residual ratio of the target protein to the baseline temperature at each temperature point was calculated, and the thermal melting curve of the target protein was plotted.
[0047] Table 1. Affinity assessment of the binding affinity of the active monomers released by the composition to various inflammatory cytokine target proteins
[0048] Note: "-" indicates that the binding free energy is negative.
[0049] exist Figure 2 In the diagram, the solid black line represents the control group, the dark gray dashed line represents the quercetin-releasing treatment group in Example 2, the medium gray dotted line represents the aloe-emodin-releasing treatment group in Example 2, and the light gray dotted line represents the kaempferol-releasing treatment group in Example 2. (Appendix) Figure 1 The meanings of the English terms mentioned are explained below: Quercetin act on TNF: Quercetin acts on TNF. Quercetin act on IL-1β: Quercetin acts on IL-1β. Quercetin acts on IL-6. Meaning of Aloc-emodin act on TNF: Aloe-emodin acts on TNF; Meaning: Aloc-emodin acts on IL-1β; Meaning: Aloc-emodin acts on IL-6; Kaempferol acts on TNF; Meaning: Kaempferol acts on IL-1β; Kaempferol acts on IL-6.
[0050] Test conclusion: according to Figure 1 Table 1 and Figure 2 The data were used to evaluate the transformation effect of the composition in Example 2 at the molecular pharmacological level. The release of free active monomers provides a material basis for multi-target intervention of the inflammatory network. When the unextruded flavonoid mixture crystals were tested alone in the laboratory, the low water solubility of the material led to easy aggregation and sedimentation in the cell culture medium, which limited the amount of drug components that could permeate the cell membrane.
[0051] The composition, after being treated with water plasticization and vacuum flash evaporation, undergoes a transformation of its original crystal structure into an amorphous state due to the influence of L-arginine in the formulation. This change in microscopic physical morphology is reflected in the binding affinity data. This is illustrated by the -9.1 eV binding affinity data in Table 1. Figure 1 The docking conformation in the study shows that quercetin molecules enter the hydrophobic binding pocket of TNF-α and form a hydrogen bond network with amino acid residues S99, Q102, and P100. This single-molecule form allows quercetin to act on the E64 and K63 sites when binding to IL-1β, completing receptor occupancy.
[0052] The interaction at the binding sites causes changes in thermodynamic properties. Figure 2 Subgraph of a in the middle and Figure 2 This is reflected in the thermal melting curve of subplot b. In the range where the ambient temperature rises above 50°C, whether in the intact macrophage environment or the recombinant protein medium, the control group represented by the black solid line shows that the target protein undergoes unfolding denaturation due to internal molecular thermal motion, and the relatively retained protein content shows a decreasing trend.
[0053] The test samples that underwent the leachate intervention of Example 2 exhibited resistance to thermal stress. Figure 2 Subgraph of a in the middle and Figure 2 In the subplot of b, the dark gray dashed line and the two other broken lines representing different active ingredient treatment groups shift to the right towards the high-temperature region. The increased structural rigidity of the target protein reflects the stable binding of drug molecules in the physiological environment, verifying the influence of formulation components and thermodynamic control during the preparation process on improving the dissolution of poorly soluble traditional Chinese medicine components and their ability to intervene in inflammatory signaling pathways.
[0054] Test Example 2: RAW264.7 macrophages were cultured, seeded in cell culture dishes, and maintained at the temperature and carbon dioxide conditions of a cell culture incubator.
[0055] Once the cells have adhered to the culture medium and entered the logarithmic growth phase, discard the original culture medium.
[0056] A culture medium containing lipopolysaccharide was prepared and added to the corresponding cell culture dish to construct an in vitro inflammatory cell model; the control group was given an equal volume of culture medium without lipopolysaccharide.
[0057] The composition prepared in Example 2 was used to extract quercetin, kaempferol and aloe-emodin monomers by isothermal dissolution and purification.
[0058] The extracted active monomers were added to cell culture dishes that had been stimulated with lipopolysaccharide at gradient concentrations, and incubation was continued.
[0059] Total protein was extracted from cells, and the proteins were separated by polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane.
[0060] The target protein expression bands were detected by chemiluminescence immunoassay and grayscale data after incubation with antibodies against p65, phosphorylated p65, and internal reference protein.
[0061] Another portion of the treated cells were fixed and permeabilized, and then incubated with p65 primary antibody and fluorescently labeled secondary antibody.
[0062] The cell nucleus was counterstained with nuclear dye, and the distribution of p65 protein in the cytoplasm and nucleus was observed under a fluorescence microscope. Images were acquired and the relative fluorescence intensity in the cell nucleus was quantified.
[0063] Cells from each treatment group were collected, and total RNA was extracted from the cells using an RNA extraction reagent. cDNA was then synthesized by reverse transcription.
[0064] Using cDNA as a template, primers were added, and the transcriptional levels of tumor necrosis factor (TNF-α), interleukins (IL-1β, IL-6), and inducible nitric oxide synthase (iNOS) genes were detected in a real-time quantitative PCR instrument.
[0065] RAW264.7 macrophages were seeded in 96-well plates and cultured with different concentrations of active monomers.
[0066] CCK-8 detection solution was added to each well at the set time points. After incubation, the absorbance value at the specified wavelength was measured using an ELISA reader, and cell proliferation data at different drug concentrations were recorded.
[0067] Table 2. Relative transcription levels of inflammation-related gene mRNAs in cells of different intervention groups
[0068] Note: "±" represents the standard deviation of independent experimental samples.
[0069] exist Figure 3 and Figure 4In the diagram, a solid black line with a solid circle represents the control group, a black dotted line with a solid square represents the solvent control group containing dimethyl sulfoxide, a dark gray dashed line with an upper triangle represents the quercetin (1 μM) treatment group of Example 2, a dark gray dashed line with a lower triangle represents the quercetin (5 μM) treatment group of Example 2, a dark gray dashed line with a rhombus represents the quercetin (10 μM) treatment group of Example 2, and a dark gray dashed line with a hollow circle represents the quercetin (20 μM) treatment group of Example 2.
[0070] Test conclusion: According to Table 2 and Figure 3 , Figure 4 The data can be used to evaluate the mechanism of the composition's intervention in inflammatory signaling pathways and its safety after administration. When cultured macrophages are stimulated by lipopolysaccharide, the p65 protein in the cytoplasm translocates to the nucleus, inducing... Figure 3 In the subplot of 'a', the intranuclear fluorescence intensity corresponding to the model group increases.
[0071] The accumulation of p65 in the cell nucleus promotes the transcription of downstream inflammation-related genes, as evidenced by the high expression of TNF-α and IL-1β in the model group shown in Table 2. After intervention with the active monomer from Example 2, the scatter plot distribution of nuclear fluorescence intensity shifted downwards. The active ingredient interacts with the receptor protein, reducing the nuclear translocation activity of p65. The reduction in nuclear transcription factors affects the expression of downstream genes, as shown in Table 2 and... Figure 3 In the subplot of b, the data on the mRNA transcription levels of various inflammatory factors in the drug intervention group showed a decline.
[0072] The safety of drugs when they exert their regulatory effects on cellular pathways is a key consideration in evaluating the application environment of drug formulations. Conventional physical mixtures are limited by solubility conditions, requiring higher dosage concentrations to achieve the desired intervention, while the deposition of drug crystals on cell surfaces can cause physical pressure damage.
[0073] Figure 4 Subgraph of a in the middle and Figure 4 Subplot b records the cell proliferation status of different components over four consecutive days. Within the concentration range of 10 μM or below, the light gray and dark gray trajectories representing the drug treatment group follow the same trend as the black control group, reflecting that the released monomers did not produce additional inhibition on the cell's own cell cycle. The amorphous formulation improves the water solubility of the monomers, allowing the drug to reach the desired concentration within a lower dose window, thus controlling the cytotoxicity issues associated with increased doses.
[0074] Test Example 3: Process intermediates and final extrusions were taken from the processing equipment settings (feeding section, melting section, mixing section, venting section, and die extrusion section) of Examples 1 to 3 and Comparative Example 2 as test samples.
[0075] The collected solid sample was crushed in a mortar, the powder was weighed and placed in a centrifuge tube, and a methanol-water mixed solvent was added for ultrasonic extraction to release the active ingredients in the matrix.
[0076] The extracted suspension was centrifuged and precipitated. The supernatant was collected and filtered through a microporous membrane. The filtrate was collected as the test solution for high performance liquid chromatography (HPLC).
[0077] Weigh out the reference standards of quercetin, kaempferol and aloe-emodin, and prepare a mixed reference solution with a known gradient using a solvent.
[0078] The test solution and the reference solution were injected sequentially into the high-performance liquid chromatograph. A C18 reversed-phase column and an isocratic elution program were used. The peak areas of each component were recorded at the detection wavelength. The mass fractions were calculated based on the standard curve, and the retention rates relative to the initial materials were estimated.
[0079] Table 3. Retention rate of active ingredients in the final extrudates of different preparation processes
[0080] Note: "±" represents the standard deviation of the independent test samples.
[0081] exist Figure 5 In the diagram, a dark gray solid line with a solid square represents the treatment group of Example 1, a medium gray solid line with a solid triangle represents the treatment group of Example 2, a light gray solid line with a solid rhombus represents the treatment group of Example 3, and a light gray dashed line with a solid circle represents the treatment group of Comparative Example 2.
[0082] Test conclusion: According to Table 3 and Figure 5 The data can be used to assess the impact of differences in thermodynamic conditions during the preparation process on the chemical stability of polyphenolic active ingredients. Flavonoids and anthraquinones contain multiple phenolic hydroxyl groups in their molecular structure, which are prone to oxidation reactions under elevated temperatures and aerobic conditions to generate corresponding quinone derivatives. Under conventional hot-melt extrusion processes, the polymer matrix requires higher temperatures to weaken intermolecular forces and achieve melt flow dynamics. Comparative Example 2 used an anhydrous hot extrusion mode, with processing temperatures exceeding the conventional heat resistance range of the active monomers. The shear friction heat of the equipment, along with residual oxygen in the material gaps, triggered a degradation reaction, leading to a decrease in the final test retention rates of each component in Comparative Example 2 as shown in Table 3. Figure 5 The light gray dashed line in the middle shows a downward trend as the processing section progresses.
[0083] Water plasticization and vacuum flash evaporation phase change technology alter the phase transition path of materials within the equipment. Water molecules enter the hydrogen bond network of polymer chains, increasing the mobility of the molecular chains. The plasticizing effect lowers the glass transition temperature of the material system, leading to a reduction in the set parameters of each temperature zone of the extruder. Under reduced heat exposure, the dark gray, medium gray, and light gray solid lines corresponding to Examples 1, 2, and 3 maintain a relatively gentle trend in the melt mixing section, and the reduced processing temperature alleviates structural damage to heat-sensitive components. The latter half of the vacuum flash evaporation stage removes some oxygen and excess moisture from the system through negative pressure conditions, controlling the probability of thermo-oxidative aging and providing the process conditions for the retention rates of Examples 1 to 3 in Table 3. Adjusting the high-temperature melting logic relied upon by traditional processes helps maintain the original chemical form of the formulation components during drug molding.
[0084] Test Example 4: Solid dispersion samples prepared in Examples 1, 2, and 3, as well as Comparative Examples 2 and 3, were used as test subjects.
[0085] Each group of test samples was laid flat in a petri dish.
[0086] Place the petri dish containing the sample inside the test chamber and adjust the environmental parameters to maintain an accelerated placement at a temperature of 40℃ and a relative humidity of 75%.
[0087] At the initial placement point (0 months), the end of the 3rd month, and the end of the 6th month, equal amounts of each group of test samples were taken out of the test chamber.
[0088] The paddle method, a dissolution test method, was used to conduct an in vitro dissolution test on the extracted samples under set rotation speed and water bath temperature conditions, using simulated physiological fluid as the dissolution medium.
[0089] The dissolution medium was aspirated at the set dissolution time point and filtered using a microporous membrane.
[0090] Collect the filtrate and dilute it according to the requirements of chromatographic analysis to prepare the test solution.
[0091] The concentration of the target component in the filtrate was determined using high performance liquid chromatography (HPLC), the cumulative dissolution rate was calculated, and the dissolution rate data at different storage time points were recorded.
[0092] Table 4. Cumulative dissolution of quercetin in each group of samples after different storage times under accelerated environment (40℃, RH75%)
[0093] Note: "±" represents the standard deviation of the independent test samples.
[0094] exist Figure 6In the diagram, the dark gray solid line with a solid square represents the treatment group of Example 1, the medium gray solid line with a solid triangle represents the treatment group of Example 2, the light gray solid line with a solid rhombus represents the treatment group of Example 3, the light gray dashed line with a solid circle represents the treatment group of Comparative Example 2, and the medium gray dashed line with a hollow square represents the treatment group of Comparative Example 3.
[0095] Test conclusion: According to Table 4 and Figure 6 The data were analyzed to assess the miscibility and resistance to crystallization and repolymerization of the drug composition under increased ambient temperature and humidity. Amorphous solid dispersions are thermodynamically metastable. The infiltration of external heat and moisture increases the activity of molecular chains within the polymer matrix, providing kinetic energy for drug molecule migration and rearrangement. In Comparative Example 2, natural cooling was used during preparation, resulting in localized enrichment of drug molecules within the cooling range, forming crystal nuclei. After accelerated storage in Comparative Example 2, the test data in Table 4 showed a decrease in values. Figure 6 The light gray dashed line with a solid circle trending downwards reflects the physical process of amorphous drug transforming into crystalline form due to residual crystal nuclei-induced phase separation. The formulation system of Comparative Example 3 lacks the surfactant glycyrrhizic acid, making it difficult to construct effective wetting, dispersion, and auxiliary anchoring structures within the matrix. With prolonged storage, intermolecular forces become insufficient to restrict the proximity of monomeric components, resulting in a decreasing dissolution rate.
[0096] The synergy between process conditions and formulation structure plays a role in maintaining the high-energy state of the system. The thermodynamic phase change effect brought about by water plasticization combined with vacuum flash evaporation alters the temperature gradient when the material leaves the die. Negative pressure accelerates the endothermic vaporization of residual moisture, causing the system temperature to drop and fixing the active monomers in a dispersed state within the polymer backbone, controlling the nucleation stage. The rapid cooling phase change process complements the interaction network established within the formulation system, increasing the molecular diffusion resistance at the microscopic level. After six months of accelerated storage under high temperature and humidity, Examples 1, 2, and 3 retained their dissolution and release properties, and the test deviations at each time point in Table 4 remained within a small range. Figure 6 The solid squares (medium-dark gray), triangles (medium gray), and rhombuses (light gray) exhibited smooth trajectories without abrupt numerical changes. The test results validate that the preparation logic can reduce the dissolution decrease of polyphenolic drugs due to phase separation during long-term storage.
[0097] Test Example 5: The products prepared in Examples 1, 2, and 3, as well as Comparative Examples 1 and 4, were used as test objects, and samples containing equal amounts of active ingredients were weighed.
[0098] According to the requirements of the paddle apparatus in the dissolution test method, the degassed medium was measured and added to the dissolution cup, and the water bath circulation system was turned on to maintain constant temperature conditions.
[0099] Place the weighed samples into the dissolution cups, lower the stirring paddle, set the speed, and start the test.
[0100] At 5, 15, 30, 45 and 60 minutes, the dissolution solution was drawn from the dissolution vessel using a syringe with a microporous filter membrane, while an equal volume of blank medium at the same temperature was added to the dissolution vessel.
[0101] The collected filtrate was used as the test solution, and the concentration of polyphenolic active ingredients was determined by high performance liquid chromatography.
[0102] Based on the concentration data and dilution factor, calculate the cumulative dissolution percentage of the corresponding component at each sampling time point.
[0103] Table 5. Cumulative dissolution rates of different sample groups at various time points
[0104] Note: "±" represents the standard deviation of the independent test samples.
[0105] exist Figure 7 In the diagram, a dark gray solid line with a solid square represents Example 1, a medium gray solid line with a solid triangle represents Example 2, a light gray solid line with a solid rhombus represents Example 3, a dark gray dashed line with a hollow upper triangle represents Comparative Example 1, and a black dotted line with a cross represents Comparative Example 4.
[0106] Test conclusion: According to Table 5 and Figure 7 Data was recorded and the effects of preparation logic and formulation components on the dissolution behavior of polyphenolic drugs were analyzed. Polyphenolic compounds such as quercetin and kaempferol possess lattice energy and hydrophobic structures, limiting their release capacity in aqueous media. Comparative Example 4 used physical pulverization and mixing to reduce drug particle size on a macroscopic scale without altering the internal crystal structure properties of the drug. The dissolution rate of Comparative Example 4 within 60 minutes fell within the low range shown in Table 5. Figure 7 The straight trend of the black dotted line with the cross indicates that the untreated drug crystals have difficulty overcoming the thermodynamic barrier to enter the dissolved state.
[0107] Introducing excipient molecules into the extrusion process intervenes in the drug lattice deconstruction process. Comparative Example 1, by removing L-arginine during preparation, lacks molecules with charged groups to act as steric breaker. In the absence of ionic recombination, the degree of lattice deconstruction by the polymer matrix is affected, and some drug molecules maintain an aggregated structure.
[0108] The dissolution trajectory of Comparative Example 1 is represented by a dark gray dashed line with a hollow upper triangle, higher than that of the physical mixing group, with a release level in the 50% range at 45 minutes. The formulations of Examples 1, 2, and 3 contain L-arginine. The basic groups of L-arginine interact with the polyphenolic hydroxyl groups in the thermally sheared microenvironment, breaking the original hydrogen bond arrangement at the molecular level. The disintegration of the crystal structure, combined with the dispersion state of hot-melt extrusion, alters the hydration kinetics of the drug in the medium. Table 5 shows the increased dissolution values at 45 minutes for Examples 1, 2, and 3. Figure 7 The solid squares with medium-dark gray solid lines, the solid triangles with medium gray solid lines, and the solid rhombuses with light gray solid lines present an ascending dissolution profile, reflecting the test results of how the combination of formulation mechanism and processing technology alters the in vitro release rate of the drug.
Claims
1. A composition of Artemisia capillaris decoction with hangover-relieving and liver-protecting effects, characterized in that, The composition is an amorphous and micronized powder composition, and the composition is made from raw materials comprising the following parts by weight: Quercetin: 10.0 parts; Kaempferol: 10.0 parts; Aloe-emodin: 10.0 parts; L-arginine: 1.5–6.0 parts; Glycyrrhizic acid: 15.0–60.0 parts; Polyvinylpyrrolidone K30: 60.0~180.0 parts.
2. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 1, characterized in that, The composition is made from raw materials comprising the following parts by weight: Quercetin: 10.0 parts; Kaempferol: 10.0 parts; Aloe-emodin: 10.0 parts; L-arginine: 3.75 parts; Glycyrrhizic acid: 37.5 parts; Polyvinylpyrrolidone K30: 120.0 parts.
3. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 1, characterized in that, The composition undergoes spontaneous acid-base proton transfer complexation reactions among its raw material components, resulting in an amorphous state of frozen molecules.
4. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 3, characterized in that, The composition is prepared by a process comprising co-rotating twin-screw extrusion, continuous injection of purified water, flash evaporation to remove impurities, and phase change quenching.
5. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 4, characterized in that, The method for preparing the composition includes the following steps: The raw materials are added together into a mixer according to the weight proportions and mixed at room temperature to obtain a premixed dry powder with no color difference and uniform dispersion. The premixed dry powder is continuously fed into the first zone of a co-rotating twin-screw extruder. The barrel temperature of the first zone is controlled. As the screw conveyor advances to the second zone, purified water is continuously injected into the barrel of the second zone to obtain water-plasticized material. The water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements to induce the material to melt, thereby obtaining a ternary homogeneous melt with uniform rheological state; The ternary homogeneous melt is pushed into the fifth zone equipped with a large-lead delivery thread, and the vacuum pump system is turned on to maintain the pressure at the exhaust port, thus obtaining a dehydrated and quenched solid precursor. The dehydrated and quenched solid precursor is further advanced to the sixth zone and the extrusion die, which are sealed with reverse thread, and extruded to obtain an amorphous, semi-transparent, brittle shaped strand. The shaped strip is fed onto a conveyor belt, cooled, solidified, pulverized, and collected and sieved to obtain the composition in a micro powder state.
6. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 5, characterized in that, The mixer is a three-dimensional motion mixer; The mixing process is carried out at room temperature using a speed of 15-35 rpm for 15-30 minutes. The premixed dry powder is continuously fed in using a loss-in-weight twin-screw feeder at a rate of 1.0 to 3.0 kg / h. The temperature of the barrel in the first zone is maintained at 70-85°C. The continuous injection of purified water is carried out by a plunger-type industrial metering pump, and the water injection rate is controlled to be constant at 2.0% to 6.0% of the feed rate of the premixed dry powder.
7. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 5, characterized in that, After the water-plasticized material is pushed into the third to fourth zones equipped with meshing disc elements, the barrel temperature is set to 105-115°C and the spindle speed is set to 150-250 rpm. The induced melting of the material is carried out under strong shear and the dielectric effect of water molecules.
8. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 5, characterized in that, After the ternary homogeneous melt is pushed into the fifth zone equipped with a large-lead delivery thread, the barrel temperature is set to 110-115°C. The pressure at the exhaust port is maintained at -0.095 MPa to -0.07 MPa.
9. The Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects according to claim 5, characterized in that, After the dehydrated and quenched solid precursor is further pushed into the sixth zone with reverse thread seal and the extrusion die, the temperature of the sixth zone and the die is set to be independently compensated and heated to 120-125°C, and the die pressure is maintained at 2.0-5.0 MPa by using secondary shear pressure building. After the shaped strip is introduced onto the track conveyor belt, the cooling and curing is carried out on the track conveyor belt with a forced air cooling system at an environment of 20-25°C for 2-5 minutes. The pulverization and collection sieving refers to the pulverization and collection of powder by a universal pulverizer and passing it through a standard sieve of 80-120 mesh.
10. The use of the Artemisia capillaris decoction composition with hangover-relieving and liver-protecting effects as described in any one of claims 1-9 in the preparation of hangover-relieving and liver-protecting drugs or health foods.