A natural small molecule inhibits the expression of liver inflammatory factors and a composition for protecting liver from alcohol

Abstract

The application relates to the fields of medicine and health care products, and discloses a natural small-molecule alcoholism-relieving and liver-protecting composition for inhibiting expression of liver inflammation factors, which is prepared from a core group, polyvinylpyrrolidone, polysorbate-80 and micro-powder silica gel, and is composed of quercetin, kaempferol and aloe-emodin. During preparation, the core group and the polyvinylpyrrolidone are dissolved to form a homogeneous dispersion liquid, the anti-solvent phase containing the micro-powder silica gel and the polysorbate-80 is synchronously pumped into a pipeline high-shear emulsifier and collides against each other, phase change precipitation occurs, ceramic membrane cross-flow filtration washing and vacuum drying in stages are carried out, and finally the finished product is obtained. Through the synergistic effect of polymer hydrogen bond and micro-powder silica gel steric hindrance, the natural molecules are fixed in a stable co-amorphous state, the water solubility and oral bioavailability of the insoluble polyphenols and anthraquinone components are improved, and crystallization aging of the composition during the storage period is effectively prevented.

Description

Technical Field

[0001] This invention relates to the fields of pharmaceuticals and health products, specifically to a natural small molecule composition for inhibiting the expression of liver inflammatory factors and protecting the liver from alcohol damage. Background Technology

[0002] Quercetin, kaempferol, aloe-emodin, and other polyphenols and anthraquinones, as well as other natural small molecules, have shown potential in inhibiting liver inflammatory factors and have been used in the development of hangover relief and liver protection products. However, these natural components have poor water solubility, resulting in low oral bioavailability. To improve their solubility, existing technologies often employ solid dispersion techniques. However, conventional solid dispersion systems are prone to crystallization and aging during storage due to the thermal motion of internal free molecules, leading to decreased physical stability of the product and a reversal of the solubility of the active ingredients.

[0003] Liquid-phase antisolvent precipitation is a common process for preparing such amorphous compositions. Existing precipitation operations are mostly carried out in conventional stirred tanks, which easily leads to localized concentration differences and mixing delays during fluid mixing. Uneven supersaturation distribution within the system directly triggers abnormal crystal growth, resulting in a non-uniform phase state of the final precipitate, making it difficult to achieve the continuous and stable preparation of high-quality compositions.

[0004] Furthermore, the post-processing of precipitated products also has shortcomings. Conventional dead-end filtration separation methods tend to form a dense filter cake layer on the surface of the filter media, hindering fluid passage and making it difficult to completely remove residual organic solvents and soluble salts from the system, affecting product purity. In the subsequent drying and dehydration stage, conventional processes often directly use high-temperature drying. This drastic heating process can easily trigger a plasticizing effect of residual moisture inside the material, thereby inducing a crystallization phase transition in the amorphous matrix, destroying the stability of the product's physical structure, and making it difficult to control the quality of the finished product. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a natural small molecule composition for inhibiting the expression of liver inflammatory factors, which solves the problems of poor water solubility, low oral bioavailability, and easy crystallization aging during the preparation of conventional solid dispersions of existing natural small molecule hangover-relieving and liver-protecting ingredients.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a natural small molecule composition for inhibiting the expression of liver inflammatory factors, comprising the following raw materials by weight percentage: The core group of active ingredients is 15.0%–25.0%; Polyvinylpyrrolidone (PVP) 69.0%–80.0%; Polysorbate-80 1.0%~3.0%; Micronized silica gel 2.0%–5.0%; The core group of active ingredients consists of quercetin, kaempferol, and aloe-emodin.

[0007] By employing the above technical solution, and utilizing a composite carrier system composed of polyvinylpyrrolidone, polysorbate-80, and micronized silica powder to co-amorphize quercetin, kaempferol, and aloe-emodin, a significantly increased water solubility and high physical stability of the composition are achieved. Quercetin, kaempferol, and aloe-emodin are polyphenols and anthraquinones, natural small molecules containing multiple phenolic hydroxyl groups in their molecular structure, resulting in poor water solubility.

[0008] The formation mechanism of this composition involves a physical phase transition and intermolecular hydrogen bonding: Due to the long-chain backbone and amide groups of polyvinylpyrrolidone (PVP), the PPVP segments and the structure of the active molecule are spatially intertwined. Intermolecular hydrogen bonds are formed between the amide carbonyl group of PPVP and the phenolic hydroxyl group of the natural small molecule. These hydrogen bonds restrict the thermal motion of the natural small molecule, altering its original ordered arrangement and inhibiting crystal nucleation. Polysorbate-80 enters the phase interface of the amorphous system, reducing the solid-liquid interfacial tension and increasing the hydrophilicity of the particles. Micronized silica gel provides a high specific surface area, adsorbing and dispersing amorphous particles, preventing particle aggregation, and creating a steric hindrance effect within the system, cutting off the mass transfer pathway for crystal growth and maintaining the amorphous state. When the composition enters an aqueous medium, PPVP rapidly hydrates and dissolves, releasing the natural small molecules in an amorphous state, forming a supersaturated solution and improving bioavailability.

[0009] Preferably, the core group of active components consists of the following components in parts by mass: quercetin 30.0-30.5 parts; kaempferol 28.5-29.0 parts; aloe-emodin 13.0-13.5 parts.

[0010] By adopting the above technical solution, the combination of quercetin, kaempferol and aloe-emodin in the above ratio can synergistically regulate liver metabolic pathways, inhibit the expression of inflammatory factors, and enhance the overall effect of alcohol detoxification and liver protection.

[0011] Preferably, the K value of the polyvinylpyrrolidone is 25 to 35; and the average original particle size of the micronized silica is 7 nm to 16 nm.

[0012] By adopting the above technical solutions, polyvinylpyrrolidone with a K value of 25 to 35 has a moderate molecular weight, which can provide sufficient hydrogen bonding sites and maintain a suitable viscosity of the system, avoiding excessive viscosity that hinders molecular diffusion or excessive viscosity that leads to insufficient inhibition of crystallization; micro-powdered silica with a K value of 7 nm to 16 nm has a well-developed surface structure, which can be inserted into the interior of the amorphous matrix to improve the anti-aggregation effect.

[0013] Preferably, the preparation method of the hangover relief and liver protection composition includes the following steps: Anhydrous ethanol and ethyl acetate were mixed and placed in a reaction vessel. The fluid temperature inside the reaction vessel was kept constant. Aloe vera emodin was added and stirred until completely dissolved. The reaction vessel is heated and kept at a constant temperature. Quercetin and kaempferol are added to the reaction vessel in sequence. After the aloe-emodin, quercetin and kaempferol are completely dissolved, polyvinylpyrrolidone is added. The stirring speed is increased and the mixture is continuously heated and stirred under high shear to obtain a homogeneous dispersion for later use. Purified water was added to the mixing tank, and citric acid solution and sodium citrate solution were added separately to obtain a mixture. The pH value of the mixture was adjusted to obtain a buffer solution. Polysorbate-80 was added to the buffer solution and stirred evenly. Micronized silica gel was added and the micronized silica gel was evenly dispersed using a high-shear disperser to form a milky white suspension as an antisolvent phase. The temperature of the antisolvent phase was continuously cooled and kept constant. A pipeline high-shear emulsifier is used to simultaneously pump the homogeneous dispersion and the antisolvent into the working chamber of the pipeline high-shear emulsifier. The homogeneous dispersion and the antisolvent collide to form a co-amorphous precipitate suspension, which is continuously discharged into a temporary storage tank and the temperature inside the temporary storage tank is maintained constant. The amorphous precipitate suspension is pumped into an inorganic ceramic membrane cross-flow filtration system for concentration and dialysis washing to obtain a concentrated slurry. The concentrated slurry is spread on a drying tray and placed in a vacuum drying oven for staged drying. After dry pulverization and sieving, the hangover relief and liver protection composition is obtained.

[0014] By employing the above technical solution, continuous preparation of co-amorphous particles is achieved using a liquid-phase antisolvent precipitation method combined with high-shear collision technology. The preparation and reaction mechanism can be divided into the following processes: The first stage is the solvent phase preparation process. Anhydrous ethanol and ethyl acetate form a mixed solvent system. Ethyl acetate acts as a co-solvent to reduce the polarity of the system and increase the dissolution rate of aloe-emodin. The stepwise heating dissolution process avoids high temperatures damaging the natural components. High-shear stirring allows the polyvinylpyrrolidone macromolecular chains to fully extend, increasing the contact area with smaller molecules and providing space for subsequent intermolecular hydrogen bond formation.

[0015] The second stage involves the preparation of the antisolvent phase. A buffer solution composed of citric acid and sodium citrate stabilizes the pH of the aqueous phase within the acidic range, inhibiting the dissociation of polyphenols and anthraquinone molecules, thus maintaining their molecular state and facilitating subsequent hydrogen bonding. Micronized silica gel overcomes van der Waals forces under high shear stress to achieve nanoscale dispersion, forming a suspension.

[0016] The third stage is the micro-mixing and phase change precipitation process. The homogeneous dispersion and the antisolvent phase are simultaneously pumped into the working chamber of the inline emulsifier. The high-speed rotation of the rotor generates shear rates and creates a turbulent flow field. Through the collision and mixing of the two fluids, the organic solvent diffuses into the aqueous phase, and water diffuses into the organic solvent phase. The solvent composition within the micro-region changes rapidly, causing a sharp decrease in the solubility of natural small molecules and polymers. The system becomes supersaturated, driving the co-precipitation of natural small molecules and polymers. Within a short precipitation time, the molecular chain segments are fixed in a disordered state, and the micro-powdered silica particles coat the surface of the precipitate or are embedded within the particles, terminating the growth of amorphous particles. The low-temperature environment reduces the kinetic energy of the molecules, slowing down the tendency to transform into crystals.

[0017] The fourth stage is the separation and drying process. An inorganic ceramic membrane cross-flow filtration system separates the amorphous particles from the solution through surface retention. A constant-volume dialysis washing process utilizes the concentration gradient to displace and elute residual organic solvents and soluble salts from the particle surface and pores, purifying the product. Staged vacuum drying removes surface free water at low temperatures and then removes internal bound water by heating, preventing water-induced plasticizing effects and crystallization phase transitions.

[0018] Preferably, the volume of the added ethyl acetate accounts for 3.0% to 5.0% of the volume of the anhydrous ethanol, and the fluid temperature in the reactor is controlled to be constant at 20°C to 25°C; the reactor is heated and kept at 70°C to 75°C, the stirring speed is increased to 400 rpm to 600 rpm, and high-shear stirring is carried out continuously for 30 to 45 minutes.

[0019] By adopting the above technical solution, the limited organic solvent ratio and temperature parameters ensure that the three natural small molecule components are completely dissolved and form a homogeneous fluid; the shear time ensures that polyvinylpyrrolidone expands and complexes, avoiding the presence of undissolved seed crystals.

[0020] Preferably, the pH of the mixture is adjusted to 3.0–3.5; the mass-volume ratio of polysorbate-80 added to the buffer solution is 0.5%–1.0%; the mass-volume ratio of the micronized silica powder added is 1.5%–2.0%; and the antisolvent phase is cooled and maintained at 2°C–5°C.

[0021] By adopting the above technical solutions, the pH value limits the ionization of natural small molecules; the temperature of the antisolvent phase lowers the saturated vapor pressure of the system, delaying aggregation; and a specified concentration of polysorbate-80 forms a monolayer adsorption at the solid-liquid interface, avoiding excessive micelle solubilization that would lead to a decrease in yield.

[0022] Preferably, the stator-rotor gap of the inline high-shear emulsifier is set to 0.2 mm to 0.5 mm; the volumetric flow rate ratio of the homogeneous dispersion to the antisolvent phase is 1:4 to 1:5; the rotational speed of the inline high-shear emulsifier is set to 8000 rpm to 10000 rpm; and the maintenance temperature of the temporary storage tank is 2°C to 5°C.

[0023] By adopting the above technical solutions, the stator-rotor gap is matched with the high speed, forming shear stress and micro-mixing efficiency, ensuring that the mass transfer rate during phase change is greater than the crystal nucleation growth rate; the volumetric flow rate ratio ensures that there is no excess solvent and maintains supersaturation; low-temperature storage prevents secondary crystallization of precipitates during liquid-phase ripening.

[0024] Preferably, the operating pressure of the inorganic ceramic membrane cross-flow filtration system is maintained at 0.15 MPa to 0.25 MPa; when the volume of the co-amorphous precipitate suspension is concentrated to 20% to 30% of the initial volume, purified water at 2°C to 5°C is continuously added for dialysis washing, and the washing water volume is 3 to 5 times the volume of the concentrated co-amorphous precipitate suspension; the staged drying is as follows: the first stage temperature is set at 20°C to 25°C for drying; the second stage temperature is raised to 40°C to 45°C for continued vacuum drying until the moisture content is ≤3.0%.

[0025] By employing the above technical solutions, the shear stress of cross-flow filtration prevents particles from forming a dense filter cake layer on the membrane surface, maintaining membrane flux; low-temperature purified water washing avoids local redissolution of particles caused by changes in organic solvent concentration; and the stepped-heating vacuum dehydration process balances drying efficiency with the thermal stability of the amorphous matrix, controlling the moisture content to ≤3.0% to eliminate the drop in glass transition temperature caused by residual moisture.

[0026] This invention provides a natural small-molecule composition for inhibiting the expression of inflammatory factors in the liver, thus relieving hangovers and protecting the liver. It possesses the following beneficial effects: 1. This invention utilizes a composite carrier system composed of polyvinylpyrrolidone, polysorbate-80, and micronized silica gel to co-amorphize quercetin, kaempferol, and aloe-emodin, thereby improving the water solubility and oral bioavailability of these natural small-molecule components. The amide carbonyl group of polyvinylpyrrolidone forms intermolecular hydrogen bonds with the phenolic hydroxyl groups of the natural small molecules, restricting the ordered arrangement of molecules; micronized silica gel provides steric hindrance to prevent particle aggregation; and polysorbate-80 increases the hydrophilicity of the particle interface. The synergistic effect of these excipients allows the poorly soluble natural components to dissolve rapidly in an aqueous medium, forming a supersaturated solution, while effectively avoiding the crystallization and aging problems caused by molecular thermal motion during storage of the solid dispersion.

[0027] 2. This invention combines liquid-phase antisolvent precipitation with inline high-shear collision technology to achieve continuous and stable preparation of amorphous compositions and ensure the phase uniformity of the product. The homogeneous dispersion and the antisolvent phase maintained at acidic and low temperature are simultaneously pumped into the working chamber for high-speed collision, completing micro-mixing and phase transition in a turbulent field. This process promotes rapid and uniform supersaturation within the system, driving the rapid co-precipitation of natural small molecules and polymeric carriers, avoiding abnormal crystal growth caused by localized concentration differences or mixing delays in conventional stirred tanks.

[0028] 3. This invention employs an inorganic ceramic membrane cross-flow filtration system for concentration, dialysis, and washing, combined with a staged vacuum drying process, ensuring the purity and physical structural stability of the product. The tangential fluid shear force of the cross-flow filtration prevents particles from forming a dense filter cake layer on the membrane surface. Combined with low-temperature constant-volume dialysis washing, it effectively removes residual organic solvents and soluble salts from the system. The subsequent drying process uses a staged vacuum operation, starting at a low temperature and then gradually increasing the temperature, to remove free water and internally bound water. This avoids the water plasticization effect and crystallization phase transition caused by direct high-temperature drying, ensuring that the final moisture content of the product is stably controlled at a low level. Attached Figure Description

[0029] Figure 1 This is a comparison diagram of the physicochemical phase transition characterization of the present invention; Figure 2 This is a comparison diagram of the dissolution kinetics characteristics of the present invention; Figure 3 This is a graph showing the fitting and evaluation of the thermal displacement curve of the target protein in this invention; Figure 4 This is a scatter plot of IL-6 expression levels according to the present invention; Figure 5 This is a bar chart showing the relative mRNA expression level of TNF-α according to the present invention. Figure 6 This is a bar chart showing the relative mRNA expression level of IL-1β according to the present invention. Figure 7 This is a bar chart showing the relative mRNA expression level detection of iNOS according to the present invention; Figure 8 This is a bar chart showing the relative mRNA expression level of IL-6 as described in this invention. Detailed Implementation

[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings, examples, comparative examples, and test examples. 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.

[0031] Examples 1-3: Example 1: This embodiment provides a natural small molecule composition for inhibiting the expression of liver inflammatory factors, comprising the following steps: Weigh out the natural small molecules according to the mass fractions. The natural small molecules include 30.0 parts of quercetin, 28.5 parts of kaempferol, and 13.0 parts of aloe-emodin, and mix them as the core group of active ingredients.

[0032] Weigh out the following components according to the mass percentage of the total solid products: 15.0% of the active core group, 80.0% of polyvinylpyrrolidone (K value 25), 3.0% of polysorbate-80, and 2.0% of micronized silica (average original particle size of 7nm).

[0033] In a reaction vessel, add anhydrous ethanol and ethyl acetate (3.0% of the volume of anhydrous ethanol). Start mechanical stirring and maintain the fluid temperature at a constant 20°C. Add the weighed aloe-emodin and stir until completely dissolved. Heat and maintain the temperature to 70°C, then slowly add the weighed quercetin and kaempferol sequentially. After the natural small molecules are completely dissolved, slowly sieve in the prescribed ratio of polyvinylpyrrolidone, increase the stirring speed to 400 rpm, and maintain high-shear stirring at 70°C for 30 minutes to obtain a homogeneous dispersion for later use.

[0034] Purified water was added to a mixing tank, followed by citric acid solution and sodium citrate solution, to adjust the pH to 3.0, yielding a buffer solution. 0.5% (w / v) of polysorbate-80 was added to the buffer solution, and after thorough mixing, 1.5% (w / v) of micronized silica gel was uniformly dispersed in the solution using a high-shear disperser to form a milky white suspension, which served as the antisolvent phase. The solution was continuously cooled and maintained at 2°C.

[0035] A pipeline-type high-shear emulsifier was used, with the stator-rotor gap set to 0.2 mm. A homogeneous dispersion at 70°C and an antisolvent phase at 2°C were simultaneously pumped into the working chamber of the emulsifier, controlling the volumetric flow rate ratio of the two fluids to be 1:4, and the emulsifier speed was set to 8000 rpm. The two fluids collided to form a co-amorphous precipitate suspension, which was continuously discharged into a chilled water jacketed storage tank and maintained at 2°C.

[0036] The amorphous precipitate suspension was pumped into a cross-flow filtration system using an inorganic ceramic membrane with a pore size of 50 nm, and the operating pressure was maintained at 0.15 MPa. When the volume of the amorphous precipitate suspension was concentrated to 20% of its initial volume, purified water at 2°C was continuously added for constant-volume dialysis washing. The washing water volume was three times the volume of the concentrated amorphous precipitate suspension, resulting in a concentrated slurry.

[0037] The concentrated slurry is spread evenly on a drying tray and placed in a vacuum drying oven. The first stage of drying is carried out at a temperature of 20°C; the second stage involves raising the temperature to 40°C and continuing vacuum drying until the moisture content is ≤2.0%. After dry pulverization using an air jet mill, the powder that passes through an 80-mesh standard sieve is collected to obtain the finished product of the hangover relief and liver protection composition.

[0038] Example 2: This embodiment provides a natural small molecule composition for inhibiting the expression of liver inflammatory factors, comprising the following steps: Weigh out the natural small molecules according to the mass fractions. The natural small molecules include quercetin (30.2 parts), kaempferol (28.7 parts), and aloe-emodin (13.2 parts), and mix them as the core group of active components.

[0039] Weigh out the following components according to the mass percentage of the total solid products: 20.0% of the active core group, 74.5% of polyvinylpyrrolidone (K value 30), 2.0% of polysorbate-80, and 3.5% of micronized silica (average original particle size of 12nm).

[0040] In a reaction vessel, add anhydrous ethanol and ethyl acetate (4.0% of the volume of anhydrous ethanol). Start mechanical stirring and maintain the fluid temperature at a constant 22°C. Add the weighed aloe-emodin and stir until completely dissolved. Heat to 72°C and maintain the temperature thereafter, slowly adding the weighed quercetin and kaempferol sequentially. After the natural small molecules are completely dissolved, slowly sieve in the prescribed ratio of polyvinylpyrrolidone. Increase the stirring speed to 500 rpm and maintain high-shear stirring at 72°C for 37 minutes to obtain a homogeneous dispersion for later use.

[0041] Purified water was added to a mixing tank, followed by citric acid solution and sodium citrate solution, to adjust the pH to 3.2, yielding a buffer solution. 0.75% (w / v) of polysorbate-80 was added to the buffer solution, and after thorough mixing, 1.75% (w / v) of micronized silica gel was uniformly dispersed in the solution using a high-shear disperser to form a milky white suspension, which served as the antisolvent phase. The solution was continuously cooled and maintained at 3°C.

[0042] A pipeline-type high-shear emulsifier was used, with the stator-rotor gap set at 0.35 mm. A homogeneous dispersion at 72°C and an antisolvent phase at 3°C ​​were simultaneously pumped into the working chamber of the emulsifier, controlling the volumetric flow rate ratio of the two fluids at 1:4.5, and the emulsifier speed was set at 9000 rpm. The two fluids collided to form a co-amorphous precipitate suspension, which was continuously discharged into a chilled water jacketed storage tank maintained at 3°C.

[0043] The amorphous precipitate suspension was pumped into a cross-flow filtration system using an inorganic ceramic membrane with a pore size of 75 nm, and the operating pressure was maintained at 0.20 MPa. When the volume of the amorphous precipitate suspension was concentrated to 25% of its initial volume, purified water at 3°C ​​was continuously added for constant-volume dialysis washing. The washing water volume was four times the volume of the concentrated amorphous precipitate suspension, resulting in a concentrated slurry.

[0044] The concentrated slurry is spread evenly on a drying tray and placed in a vacuum drying oven. The first stage of drying is carried out at a temperature of 22℃; the second stage involves raising the temperature to 42℃ and continuing vacuum drying until the moisture content is ≤2.5%. After dry pulverization using an air jet mill, the powder that passes through a 90-mesh standard sieve is collected to obtain the finished product of the hangover relief and liver protection composition.

[0045] Example 3: This embodiment provides a natural small molecule composition for inhibiting the expression of liver inflammatory factors, comprising the following steps: Weigh out the natural small molecules according to the mass fractions. The natural small molecules include quercetin 30.5 parts, kaempferol 29.0 parts, and aloe-emodin 13.5 parts, and mix them as the core group of active ingredients.

[0046] Weigh out the following components according to the mass percentage of the total solid products: 25.0% of the active core group, 69.0% of polyvinylpyrrolidone (K value 35), 1.0% of polysorbate-80, and 5.0% of micronized silica (average original particle size of 16nm).

[0047] In a reaction vessel, add anhydrous ethanol and ethyl acetate (5.0% of the volume of anhydrous ethanol). Start mechanical stirring and maintain the fluid temperature at a constant 25°C. Add the weighed aloe-emodin and stir until completely dissolved. Heat to 75°C and maintain the temperature thereafter, slowly adding the weighed quercetin and kaempferol sequentially. After the natural small molecules are completely dissolved, slowly sieve in the prescribed ratio of polyvinylpyrrolidone. Increase the stirring speed to 600 rpm and maintain high-shear stirring at 75°C for 45 minutes to obtain a homogeneous dispersion for later use.

[0048] Purified water was added to a mixing tank, followed by citric acid solution and sodium citrate solution, to adjust the pH to 3.5, yielding a buffer solution. 1.0% (w / v) of polysorbate-80 was added to the buffer solution, and after thorough mixing, 2.0% (w / v) of micronized silica gel was uniformly dispersed in the solution using a high-shear disperser to form a milky white suspension, which served as the antisolvent phase. The solution was continuously cooled and maintained at 5°C.

[0049] A pipeline-type high-shear emulsifier was used, with the stator-rotor gap set to 0.5 mm. A homogeneous dispersion at 75°C and an antisolvent at 5°C were simultaneously pumped into the working chamber of the emulsifier, controlling the volumetric flow rate ratio of the two fluids to be 1:5, and the emulsifier speed was set to 10,000 rpm. The two fluids collided to form a co-amorphous precipitate suspension, which was continuously discharged into a chilled water jacketed storage tank and maintained at 5°C.

[0050] The amorphous precipitate suspension was pumped into a cross-flow filtration system using an inorganic ceramic membrane with a pore size of 100 nm, and the operating pressure was maintained at 0.25 MPa. When the volume of the amorphous precipitate suspension was concentrated to 30% of its initial volume, purified water at 5°C was continuously added for constant-volume dialysis washing. The washing water volume was 5 times the volume of the concentrated amorphous precipitate suspension, resulting in a concentrated slurry.

[0051] The concentrated slurry is spread evenly on a drying tray and placed in a vacuum drying oven. The first stage of drying is carried out at a temperature of 25°C; the second stage involves raising the temperature to 45°C and continuing vacuum drying until the moisture content is ≤3.0%. After dry pulverization using an air jet mill, the powder that passes through a 100-mesh standard sieve is collected to obtain the finished product of the hangover relief and liver protection composition.

[0052] Comparative Examples 1-5: Comparative Example 1: Compared with Example 1, the difference is that only purified water is added to the solution preparation tank, and citric acid solution and sodium citrate solution are not added for pH adjustment. That is, the antisolvent phase is neutral pure water, and everything else is the same.

[0053] Comparative Example 2: Compared with Example 1, the difference is that when weighing the materials according to the mass percentage of the total solid product, polyvinylpyrrolidone was not added, and the mass percentage of this part was replaced by an equal amount of purified water. In addition, the screening operation of polymeric excipients was no longer performed in the reactor preparation step. All other aspects are the same.

[0054] Comparative Example 3: Compared with Example 1, the difference is that when weighing the materials according to the mass percentage of the total solid product, no micronized silica gel was added, and the high-shear dispersion operation of micronized silica gel was not performed in the step of preparing the antisolvent phase in the mixing tank; all other aspects are the same.

[0055] Comparative Example 4: Compared with Example 1, the difference is that when weighing the natural small molecules by mass fraction, the mass fractions of quercetin, kaempferol, and aloe-emodin were modified to 23.8 parts, 23.8 parts, and 23.8 parts, respectively. That is, the original specific molar ratio framework was replaced by mixing in equal mass ratios. All other aspects are the same.

[0056] Comparative Example 5: Compared with Example 1, the difference is that the two-fluid collision process is replaced with a conventional drop-and-precipitate process, that is, the homogeneous dispersion at 70°C is slowly added dropwise at a constant flow rate to the antisolvent phase at 2°C with conventional mechanical stirring for crystallization and precipitation. The inline high-shear emulsifier is not used for millisecond-level rapid collision. All other aspects are the same.

[0057] Test Example 1-2: Test Example 1: Test steps: Take appropriate amounts of the hangover-relieving and liver-protecting compositions prepared in Examples 1 to 3, and Comparative Examples 1, 2, 3, and 5, and place them in an aluminum crucible, sealing it with a lid. Prepare an empty aluminum crucible as a reference. Under a nitrogen atmosphere, set the protective gas flow rate to 50 mL / min. Use a differential scanning calorimeter to heat the samples from 30°C to 250°C at a heating rate of 10°C / min. Record the heat flow curves and read the glass transition temperature and the relative enthalpy corresponding to the characteristic crystallization melting peak.

[0058] Each group of powder samples was laid flat in a stability test chamber, and the ambient temperature was set at 40℃ and the relative humidity at 75% for an accelerated placement period of 30 days. After the period, the samples were removed, and an equal amount of powder was weighed and added to a dissolution vessel containing 900 mL of pH 6.8 simulated artificial intestinal fluid. The paddle speed was set at 75 rpm, and the temperature was maintained at 37℃. At the 45th minute, an appropriate amount of dissolution medium was aspirated and filtered through a 0.45-micron filter membrane. The concentration of the active ingredient in the filtrate was determined by high-performance liquid chromatography (HPLC) and converted into the cumulative dissolution rate.

[0059] Take an initial powder sample that has not been placed and determine the powder mechanical parameters using a powder comprehensive tester. Measure the angle between the inclined plane of the cone formed by the powder after free fall and the horizontal plane using the fixed funnel method, and record it as the angle of repose. Weigh a fixed mass of powder and slowly pour it into a graduated cylinder to read the loose volume. Use a tapper to vibrate at a fixed frequency until the volume no longer changes, and then read the tapped volume. Calculate the percentage difference between the loose density and the tapped density relative to the tapped density, and record it as the Karl ellipse index.

[0060] Test data: Table 1. Comprehensive Test Data of Physicochemical Phase Transition Characterization and Macroscopic Process Efficiency

[0061] Test conclusion: According to Table 1 and Figure 1 and Figure 2 The data shows that the glass transition temperatures of Examples 1 to 3 were maintained within the range of 105°C to 108°C, and the relative enthalpy of the characteristic crystallization peak was zero. Figure 1 The heat flow curves of the intermediate example group remained smooth and showed no endothermic peaks, indicating that the molecules in the composition overcame the lattice energy barrier and were fixed in a co-amorphous state. Normally, conventionally physically mixed plant extract powders will agglomerate and crystallize under humid and hot conditions. The example group underwent 30 days of accelerated environmental placement, and its 45-minute dissolution rate remained above 86%. Figure 2 The dissolution curves of the intermediate example group rose steadily and remained within a stable range, proving that this state possesses structural stability.

[0062] In contrast, when the acid conditioning step was omitted in the preparation of Comparative Example 1, a crystallization enthalpy of 12.6 J / g appeared in the powder system, and the solubility decreased to 45.1%. This is because the uninhibited polyhydroxy structure retained high reactivity, altering the intermolecular dipole interactions within the microenvironment, causing some aloe-emodin to detach from the co-amorphous system and crystallize in situ. A similar phenomenon occurred in Comparative Example 2; due to the lack of a polyvinylpyrrolidone chain network for encapsulation, the system reverted to a crystalline mixture morphology. Figure 1 It can be observed that Comparative Example 1 and Comparative Example 2 exhibit crystallization melting peaks in their respective temperature ranges, accompanied by the thermal energy absorption process. Figure 2 The dissolution curve was at a low level, dropping to 32.8% at 45 minutes. The powder's hygroscopic clumping affected the fluid's penetration of the drug inside.

[0063] The data comparison demonstrates the impact of process methods on macroscopic properties. Comparative Example 5, which replaced the two-fluid collision process with a conventional dropwise precipitation process, showed similar data on the crystalline phase as Comparative Example 2. Figure 1 The same clear crystallization characteristics were also recorded. Figure 2This reflects the obstruction of its overall dissolution process. Slow fluid convergence prevents the formation of the temperature drop required to overcome the nucleation activation energy, allowing molecules time to arrange into an ordered lattice. Regarding the physical barrier effect of micronized silica, data from Comparative Example 3 show that although this formulation preserves the amorphous characteristics in thermodynamic tests, the lack of skeletal support from the micronized silica leads to physical collapse of the polymer matrix during the drying and dehydration process, with the angle of repose rising to 48.2 degrees and the Karl Fischer index reaching 36.5%. In pharmaceutical manufacturing, this lack of flowability makes it difficult to adapt to the operating rhythm of high-speed tableting or capsule filling equipment, indirectly confirming the role of silica particles in intervening in the secondary agglomeration of amorphous particles and maintaining the aerodynamic properties of the powder. The test results of the microstructure and macroscopic physicochemical properties verify the rationality of the formulation and process design of the composition of this invention.

[0064] Test Example 2: Test steps: RAW 264.7 macrophages were seeded at an appropriate density in culture plates and cultured in a CO2 incubator until the cells adhered. The experiment included a blank control group, a lipopolysaccharide (LPS) model group, monomeric intervention groups (containing quercetin, kaempferol, and aloe-emodin, respectively), a comparative example 4 intervention group, and intervention groups from Examples 1 to 3. Except for the blank control group, LPS was added to each well to a final concentration of 1 μg / mL to stimulate the cells to form an inflammatory environment. Subsequently, the corresponding test substances at predetermined concentrations were added, and the wells were continuously co-incubated in an incubator.

[0065] Cell suspensions from each group were collected for thermal displacement analysis to assess the rigidity of the target protein. The cell suspensions were aliquoted into PCR tubes and heated for 3 minutes at multiple temperature gradients within the range of 40°C to 80°C. After heating, the samples were cooled to room temperature, and the cells were lysed using a liquid nitrogen freeze-thaw cycle to obtain the protein extract. The extract was centrifuged at high speed to separate the supernatant. The residual soluble tumor necrosis factor-α (TNF-α) level in the supernatant was quantitatively detected using an enzyme-linked immunosorbent assay (ELISA) kit. A melting curve was fitted, and the denaturation temperature at which the target protein undergoes half-disintegration was calculated.

[0066] Another batch of cells treated under the same conditions was collected for culture. The culture supernatant and cell pellet were separated by centrifugation. Total RNA was extracted from the cells using TRIzol reagent, and its concentration and purity were determined. The RNA was then reverse transcribed into cDNA templates. The relative expression levels of mRNA of TNF-α, interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS), and interleukin-6 (IL-6) were detected by real-time quantitative PCR to investigate the blocking effects of specific downregulation folds and ratios of various transcription factors.

[0067] Test data: Table 2. Comprehensive Test Data on Synergistic Effects of Molecular Biology Targeting Mechanisms and Core Combinations

[0068] Test conclusion: Based on Table 2 and the results of the fitting evaluation of the target protein thermal displacement curve, Figure 3 The data showed that the thermodynamic stability of the target protein varied among groups when subjected to different drug interventions. Figure 3 The soluble retention curves of the target protein at different temperature gradients are shown for the lipopolysaccharide group, the monomer intervention group (quercetin), Comparative Example 4, and Example 1, reflecting the delaying effect of each treatment group on the protein denaturation temperature. Figure 3 As shown in the melting curve trend, the denaturation temperature of the uninterrupted lipopolysaccharide model group remained at a low level of 46.5℃, and the related proteins free in the cytoplasm underwent conformational disintegration after heating.

[0069] In contrast, the introduction of monomeric compounds can delay the pyrolysis and folding process to some extent, demonstrating that single molecules possess the basic affinity to intercalate into the target active pocket. After treating cells using the same mass mixing strategy as in Comparative Example 4, the protein denaturation limit increased to 52.8℃. The example groups are shown in Table 2 and... Figure 3 As shown, the disintegration temperature limit was raised to above 56°C, and the thermodynamic solidification phenomenon verified the mechanism by which the core group of the formulation anchors protein regions through a multi-site network. The molecular clusters constructed according to a specific molar ratio restrict the degree of freedom of the target protein peptide chain through steric hindrance, resulting in overall rigidity and interrupting the signaling cascade pathway of inflammatory factors.

[0070] Multi-point anchoring efficacy at the molecular level is mapped onto transcriptional regulation data of inflammatory genes. Table 2 shows a scatter plot of IL-6 expression levels. Figure 4 and Figures 5 to 8The detection results showed that the intervention of lipopolysaccharide led to high expression of various pro-inflammatory factor mRNAs in cells. The relative mRNA expression level of TNF-α was also detected. Figure 5 (The vertical axis represents TNF-α relative mRNA levels, i.e., the relative mRNA expression level of TNF-α, with a scale of 0 to 5.) Detecting the relative mRNA expression level of IL-1β. Figure 6 (The vertical axis represents IL-1β Relative mRNA levels, i.e., the relative mRNA expression level of IL-1β, with a scale from 0 to 40) and the detection of iNOS relative mRNA expression levels. Figure 7 (The vertical axis represents iNOS Relative mRNA levels, with a scale of 0 to 15.) The horizontal axis represents the experimental groups, arranged from left to right as follows: Control (dark blue bar), LPS (Lipopolysaccharide treatment group, dark purple bar), LPS+AE (Lipopolysaccharide + Aloe-emodin treatment group, light purple bar), LPS+Kae (Lipopolysaccharide + Kaempferol treatment group, light red bar), and LPS+QR (Lipopolysaccharide + Quercetin treatment group, orange bar). Each group corresponds to one bar, and the scatter points on the bars represent the individual values ​​of each group's samples.

[0071] Among the salient symbols marked above the figure, Figure 5 and Figure 6 The **** in the diagram represents highly significant differences between the control group and the LPS group, the LPS group and the LPS+AE group, the LPS group and the LPS+Kae group, and the LPS group and the LPS+QR group. The results showed that the relative mRNA expression levels of TNF-α and IL-1β in the LPS group were significantly higher than those in the control group, and the LPS+AE, LPS+Kae, and LPS+QR groups were all significantly lower than those in the LPS group. Figure 7 In the diagram, * indicates a significant difference between the LPS group and the LPS+Kae group, and **** indicates highly significant differences between the control group and the LPS group, the LPS group and the LPS+AE group, and the LPS group and the LPS+QR group. The results show that the relative iNOS mRNA expression level in the LPS group was significantly higher than that in the control group, and significantly lower in the LPS+AE, LPS+Kae, and LPS+QR groups compared to the LPS group. These results indicate that monomeric intervention exhibits anti-inflammatory and inhibitory effects, reducing the transcriptional activity of TNF-α, IL-1β, and iNOS compared to the LPS group.

[0072] Unlike the transcriptional repression of the aforementioned conventional factors, in Table 2, Figure 4and detection of relative mRNA expression levels of IL-6 Figure 8 (The vertical axis represents IL-6 Relative mRNA levels, i.e., the relative mRNA expression level of IL-6, with a scale of 0 to 15; the horizontal axis represents experimental groups, and the group and scatter plot bar color rules are consistent with...) Figure 5 In the study (consistent), the transcriptional level of IL-6 exhibited an intracellular pathway compensatory mechanism. Results showed that the relative mRNA expression level of IL-6 in the LPS group was significantly higher than that in the control group. During this phase, the laboratory observed that the monomeric intervention of aloe-emodin, kaempferol, and quercetin failed to inhibit IL-6, inducing a gradient increase in IL-6 transcription levels, such as... Figure 4 The graph shows the line graph and data point annotations illustrating the expression increase induced by monomeric intervention and the suppression effect of the examples. Figure 8 The relative mRNA expression levels of IL-6 in the LPS+AE, LPS+Kae, and LPS+QR groups all showed a gradient increase compared to the LPS group, with the LPS+QR group exhibiting the highest relative mRNA expression level of IL-6. (Table 2 and...) Figure 8 The value reached 16.42 under the action of LPS+QR. In the application of single-target blockers, the cellular inflammatory network will compensate by activating paralateral kinase pathways, leading to the burst of certain inflammatory mediators, which is a limiting factor for the efficacy of single anti-inflammatory drugs.

[0073] To overcome this limiting factor, the experiment further verified the intervention effect of the combined formulation. The equal-mass mixing in Comparative Example 4 only pulled back IL-6 expression to 10.36, failing to eliminate the compensatory response. The formulation of the example, calculated using specific molar ratios, reduced factor transcriptional activity, blocked the compensatory feedback loop of IL-6, and suppressed the expression of various inflammatory markers within the physiological metabolic baseline range. The complementary anchoring array formed by multiple active ingredients covered the intracellular inflammatory signal transduction pathways, demonstrating that biological regulation triggered by a specific proportional framework is superior to the linear superposition of substance activities.

Claims

1. A natural small molecule composition for inhibiting the expression of inflammatory factors in the liver, characterized in that, It contains the following raw materials by weight percentage: The core group of active ingredients is 15.0%–25.0%; Polyvinylpyrrolidone (PVP) 69.0%–80.0%; Polysorbate-80 1.0%~3.0%; Micronized silica gel 2.0%–5.0%; The core group of active ingredients consists of quercetin, kaempferol, and aloe-emodin.

2. The hangover-relieving and liver-protecting composition according to claim 1, characterized in that, The core group of active components consists of the following components in parts by mass: Quercetin 30.0–30.5 parts; Kaempferol 28.5–29.0 parts; Aloe-emodin 13.0–13.5 parts.

3. The hangover-relieving and liver-protecting composition according to claim 1, characterized in that, The K value of the polyvinylpyrrolidone is 25-35, and the average original particle size of the micronized silica is 7nm-16nm.

4. The hangover-relieving and liver-protecting composition according to claim 1, characterized in that, The preparation method of the hangover relief and liver protection composition includes the following steps: Anhydrous ethanol and ethyl acetate were mixed and placed in a reaction vessel. The fluid temperature inside the reaction vessel was kept constant. Aloe vera emodin was added and stirred until completely dissolved. The reaction vessel is heated and kept at a constant temperature. Quercetin and kaempferol are added to the reaction vessel in sequence. After the aloe-emodin, quercetin and kaempferol are completely dissolved, polyvinylpyrrolidone is added. The stirring speed is increased and the mixture is continuously heated and stirred under high shear to obtain a homogeneous dispersion for later use. Purified water was added to the mixing tank, and citric acid solution and sodium citrate solution were added separately to obtain a mixture. The pH value of the mixture was adjusted to obtain a buffer solution. Polysorbate-80 was added to the buffer solution and stirred evenly. Micronized silica gel was added and the micronized silica gel was evenly dispersed using a high-shear disperser to form a milky white suspension as an antisolvent phase. The temperature of the antisolvent phase was continuously cooled and kept constant. A pipeline high-shear emulsifier is used to simultaneously pump the homogeneous dispersion and the antisolvent into the working chamber of the pipeline high-shear emulsifier. The homogeneous dispersion and the antisolvent collide to form a co-amorphous precipitate suspension, which is continuously discharged into a temporary storage tank and the temperature inside the temporary storage tank is maintained constant. The amorphous precipitate suspension is pumped into an inorganic ceramic membrane cross-flow filtration system for concentration and dialysis washing to obtain a concentrated slurry. The concentrated slurry is spread on a drying tray and placed in a vacuum drying oven for staged drying. After dry pulverization and sieving, the hangover relief and liver protection composition is obtained.

5. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The volume of the added ethyl acetate is 3.0% to 5.0% of the volume of the anhydrous ethanol, and the temperature of the fluid in the reactor is controlled to be constant at 20°C to 25°C.

6. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The reactor is heated and kept at a constant temperature of 70°C to 75°C. The stirring speed is increased to 400 rpm to 600 rpm, and the reactor is continuously heated and stirred under high shear for 30 to 45 minutes.

7. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The pH of the mixture is adjusted to 3.0-3.5, the mass-volume ratio of polysorbate-80 added to the buffer solution is 0.5%-1.0%, the mass-volume ratio of micronized silica powder is 1.5%-2.0%, and the antisolvent phase is cooled and maintained at 2℃-5℃.

8. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The stator-rotor gap of the inline high-shear emulsifier is set to 0.2mm to 0.5mm, the volumetric flow rate ratio of the homogeneous dispersion to the antisolvent phase is 1:4 to 1:5, the rotational speed of the inline high-shear emulsifier is set to 8000rpm to 10000rpm, and the maintenance temperature of the temporary storage tank is 2℃ to 5℃.

9. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The operating pressure of the inorganic ceramic membrane cross-flow filtration system is maintained at 0.15 MPa to 0.25 MPa; When the volume of the co-amorphous precipitate suspension is concentrated to 20% to 30% of the initial volume, purified water at 2°C to 5°C is continuously added for dialysis washing. The washing water volume is 3 to 5 times the volume of the concentrated co-amorphous precipitate suspension.

10. The hangover-relieving and liver-protecting composition according to claim 4, characterized in that, The staged drying process is as follows: The first stage of drying is carried out at a temperature of 20℃~25℃. The second stage involves raising the temperature to 40℃~45℃ and continuing vacuum drying until the moisture content is ≤3.0%.

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