Preparation method of fungus polysaccharide adjuvant drug composite component and fungus component compound

Abstract

The invention is suitable for the technical field of functional food preparation, and provides a preparation method of a fungus polysaccharide adjuvant drug composite component and a fungus component compound. The preparation method comprises the following steps: sequentially carrying out physical adsorption and functional integration extraction on four adjuvant raw materials such as shiitake mushrooms and pleurotus nebrodensis by adopting granular and dissolved pachymaran, applying an ultrasonic field in a low-temperature vacuum concentration process for dynamic co-coagulation, and finally forming to obtain the adjuvant composite gel particles with the particle size of 100-500 microns. Further, the adjuvant and conductant drug composite component is used as a wrapping layer, and solid-phase cores of the monarch and ministerial drugs are wrapped by a fluidized bed coating technology to form a fungus component compound with a core-shell space structure; through multiple physical structure design, controllable materialization of a traditional Chinese medicine monarch, minister, assistant and guide compatibility theory is realized, and the prepared product has the advantages of synergistic interaction of components, programmed release and remarkable improvement of immunomodulatory activity.

Description

Technical Field

[0001] This invention belongs to the field of functional food preparation technology, and particularly relates to a method for preparing a fungal polysaccharide adjuvant compound and a fungal component complex. Background Technology

[0002] In traditional Chinese medicine compound preparations, the theory of "principal, assistant, adjuvant, and guide" is the essence. Among them, "adjuvant drugs" are often used to assist the principal and assistant drugs, or to eliminate or reduce the toxicity or potency of the principal drug, or to treat concurrent symptoms. "Guide drugs" play a role in harmonizing the drugs and guiding the medicinal power directly to the disease site. However, in modern industrial production, how to transform this abstract theory into concrete, controllable, and repeatable formulation technology is a major challenge.

[0003] While existing technologies have attempted to combine multiple fungal polysaccharides, the processes are essentially simple additives, failing to achieve function-oriented structured integration. For example, prior art document CN101228951B discloses a method for preparing a compound polysaccharide nutrient solution, which involves fermenting and extracting multiple fungal strains such as Ganoderma lucidum, Poria cocos, Lentinus edodes, and Pleurotus ostreatus separately, and then physically mixing their culture solutions. This method has the following inherent defects: Functional role missing: In this method, Poria cocos is treated the same as other fungal species and is added only as one of the common active ingredients. Its core function as a harmonizing agent and guiding other drugs to their respective meridians is not designed or reflected in the process.

[0004] Lack of structural guidance: The process lacks targeted utilization of the physical forms (particulate and dissolved states) of Poria cocos polysaccharides, failing to enable them to act as structural cores or guiding agents to actively adsorb and integrate other polysaccharide molecules. After mixing, the components remain physically discrete, like a pile of loose sand.

[0005] Loose product structure: The final product is just a simple blend of various polysaccharide molecules. The components lack stable interactions and orderly spatial arrangement, which makes it impossible to improve its physicochemical properties (such as stability and release behavior) and biological functions (such as synergy and targeting).

[0006] Therefore, when such simple mixed adjuvant and guide drug components are combined with principal and assistant drugs, their unblocking effect is scattered and uncontrolled, and they cannot achieve orderly and synergistic efficacy. They may even affect the overall stability of the prescription due to mutual interference of their biases. How to materialize the theory of adjuvant and guide drugs in traditional Chinese medicine, especially the harmonizing and guiding function of the guide drug, through quantifiable and controllable modern pharmaceutical processes has become a long-standing technical problem in this field. Summary of the Invention

[0007] This invention provides a method for preparing a fungal polysaccharide adjuvant compound and a fungal component complex, aiming to solve the above-mentioned problems.

[0008] This invention is achieved by a method for preparing a fungal polysaccharide adjuvant compound component, comprising the following steps: S1. Step-by-step guided extraction: S1a. Physical adsorption guidance: The dried powders of four raw materials, namely shiitake mushroom, white lingzhi mushroom, white-backed wood ear fungus and button mushroom, are mixed together with water and granular Poria cocos polysaccharide to form an extraction system, and the first stage of extraction is carried out at 40-60℃. S1b. Functional integration guidance: Add dissolved Poria cocos polysaccharide solution to the S1a system, adjust the pH of the system, and carry out the second stage of extraction at 60-95℃. After the extraction is completed, the first complex polysaccharide extract is obtained by separation. First, tiny Poria cocos particles are used as "seeds" or "anchor points" to adsorb other polysaccharide molecules; then, dissolved Poria cocos polysaccharides are used as "functional media" to further guide and integrate the adsorbed polysaccharide active fragments in the solution.

[0009] S2. Dynamic co-condensation concentration: The first complex polysaccharide extract is concentrated under low temperature vacuum, and ultrasonic waves are applied to it during the concentration process to obtain a complex polysaccharide colloid; During vacuum concentration, low-intensity ultrasound with specific power is applied to interrupt the irregular thermal motion, which promotes the better extension of polysaccharide chains (especially long-chain Poria cocos polysaccharides) and the formation of a more uniform and denser network structure with other polysaccharides through hydrogen bonding and other interactions.

[0010] S3. Composite component molding: The composite polysaccharide colloid is formed into 100-500 micrometer gel particles to obtain the adjuvant-guide drug composite component.

[0011] Preferably, in step S1a, the mass ratio of the dried powders of shiitake mushrooms, white lingzhi mushrooms, white-backed wood ear mushrooms, and button mushrooms is 1-3:1-2:1-2:1-3; the particle size of the granular Poria cocos polysaccharide is 1-100 micrometers, and its addition amount is 10%-15% of the total mass of the mixed powder of the four raw materials; the first extraction time is 0.5-2 hours, and the material-to-liquid ratio is 1:10-20 (g / mL); in step S1b, the concentration of the dissolved Poria cocos polysaccharide is 1%-3% (w / v), and its addition amount, on a dry matter basis, is 5%-10% of the total mass of the mixed powder of the four raw materials; the second extraction time is 1-4 hours, and the pH is 4.0-6.0.

[0012] Preferably, in step S2, the conditions for low-temperature vacuum concentration are: initial temperature 60℃, maintaining vacuum degree -0.06 to -0.1MPa, using a programmed gradient cooling mode, uniformly cooling to 40℃ at a rate of 0.5-1℃ / min until the material is concentrated to 10-20% of its original volume; the applied ultrasonic frequency is 20-40kHz, and the power density is 50-200W / L.

[0013] Preferably, in step S3, the molding method includes one of spray granulation, drop molding, or extrusion spheroidization.

[0014] The present invention also provides a fungal polysaccharide adjuvant compound component, which is prepared by the above method.

[0015] The present invention also provides a fungal component complex, comprising a fungal polysaccharide adjuvant complex, a principal ingredient, and an assistant ingredient; the principal ingredient comprises Grifola frondosa polysaccharide and Cordyceps militaris polysaccharide; the assistant ingredient comprises Ganoderma lucidum extract (preferably Ganoderma lucidum fruiting body water extract (Ganoderma lucidum polysaccharide content ≥30%)), Hericium erinaceus polysaccharide, and Agaricus blazei polysaccharide.

[0016] Preferably, by weight, the fungal polysaccharide adjuvant compound component comprises 24-36 parts, Grifola frondosa polysaccharide 12-17 parts, Cordyceps militaris polysaccharide 12-17 parts, Ganoderma lucidum extract 15-20 parts, Hericium erinaceus polysaccharide 10-14 parts, and Agaricus blazei polysaccharide 8-12 parts.

[0017] The present invention also provides a method for preparing the above-mentioned fungal component complex, comprising the following steps: P1. Preparation of the solid core of the principal and assistant medicine: Take Grifola frondosa polysaccharide, Cordyceps militaris polysaccharide, Ganoderma lucidum extract, Hericium erinaceus polysaccharide, and Agaricus blazei polysaccharide; after thoroughly mixing the above materials, use a dry granulation process (dry granulation pressure is 5-10MPa) to make solid microspheres with a particle size of 100-1000 micrometers, which serve as the solid core of the principal and assistant medicine; P2. Construction of the core-shell structure: The solid core of the principal and assistant herbs is placed in a fluidized bed coating device and kept in a fluidized state; the fungal polysaccharide adjuvant compound component is dispersed or dissolved in an adhesive solution as a coating slurry; the coating slurry is uniformly coated on the surface of the fluidized solid core of the principal and assistant herbs using a fluidized bed spraying process to form a coating layer, which is then dried to obtain micro-units (drying conditions are 40-50℃, vacuum degree -0.05 to -0.08MPa, dried to the micro-unit moisture content of 10%-15%); wherein, the dry weight of the coating layer accounts for 20%-40% of the total weight of the micro-units; P3. Final formulation: The microunits are dried (preferably using vacuum drying process, drying at 45-55℃ and vacuum degree -0.07 to -0.1MPa for 1-2 hours) to make their water content less than 5%, thus obtaining the microbial component complex.

[0018] Preferably, in step P2, the fluidized bed spraying process is a bottom spraying process or a tangential spraying process; the adhesive solution is a 1%-5% (w / v) aqueous solution of hydroxypropyl methylcellulose or an aqueous solution of sodium alginate.

[0019] The present invention also provides the application of the above-mentioned fungal component complex in the preparation of health foods or medicines for immune regulation, warming and strengthening the body, or improving deficiency and cold syndrome.

[0020] Compared with the prior art, the embodiments of this application have the following main advantages: The preparation method of the fungal polysaccharide adjuvant drug complex provided by the present invention uses a step-by-step guided extraction process. Through the sequential interaction between granular and dissolved Poria cocos polysaccharides, the function of adjuvant drug first adsorbing and fixing and then integrating and guiding is simulated at the molecular level, so that the adjuvant drug component is transformed from a simple mixture into a composite gel microparticle with a stable cohesive structure.

[0021] The fungal component complex provided by this invention is constructed through a "core-shell structure," which physically achieves a spatial layout of "principal and assistant herbs inside, adjuvant and guide herbs outside." This allows for the preferential release of adjuvant and guide herbs to "unblock" the environment, while the sustained release of principal and assistant herbs to "strengthen the foundation," thereby enabling the overall efficacy of the formula to be released in a programmed and synergistic manner, which is significantly superior to physical mixtures. Attached Figure Description

[0022] Figure 1 This is a flowchart of the preparation method of the fungal polysaccharide adjuvant compound component provided by the present invention; Figure 2 This is a flowchart of the preparation method of the fungal component complex provided by the present invention. Detailed Implementation

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the terminology used herein in the specification of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application; the terms "comprising" and "having," and any variations thereof, in the specification, claims, and foregoing drawings of this application are intended to cover non-exclusive inclusion. The terms "first," "second," etc., in the specification, claims, or foregoing drawings of this application are used to distinguish different objects, not to describe a particular order.

[0024] In this document, the term "embodiment" means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The appearance of this phrase in various places throughout the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. It will be explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0025] This invention provides a method for preparing a fungal polysaccharide adjuvant compound component, such as... Figure 1 As shown, it includes the following steps: S1. Step-by-step guided extraction: a. Physical adsorption-guided extraction: Weigh 100g of dried powders of shiitake mushrooms, white lingzhi mushrooms, white-backed wood ear mushrooms, and button mushrooms (each passed through an 80-mesh sieve) at a mass ratio of 2:1.5:1.5:2; add 12.5g of granular Poria cocos polysaccharide with a particle size of approximately 50μm (accounting for 12.5% ​​of the total mass of the mycelium powder) and 1500mL of purified water (material-to-liquid ratio 1:15); and extract by stirring in a water bath at 50℃ for 1.25 hours.

[0026] b. Functional integration guidance: Add 375 mL of 2% (w / v) dissolved Poria cocos polysaccharide solution (equivalent to 7.5 g of dry matter, accounting for 7.5% of the total mass of the mycelium powder) to the above system; adjust the pH of the system to 5.0 with 1M HCl; heat to 80℃ and continue stirring for 2.5 hours; filter while hot through a 200-mesh filter cloth to obtain approximately 1900 mL of filtrate.

[0027] S2. Dynamic co-condensation concentration: Transfer the filtrate to a rotary evaporator equipped with an ultrasonic amplitude transformer or a dedicated ultrasonic concentration reactor; set the initial water bath temperature to 60℃ and the vacuum degree to -0.08MPa; turn on the concentration and ultrasonic generator, set the ultrasonic frequency to 30kHz and the power density to 120W / L, and perform ultrasonic treatment throughout the entire concentration process; simultaneously start the program to cool down at a rate of 0.75℃ / min; concentrate to a colloidal state with a volume of approximately 300mL (approximately 15% of the original volume), and stop concentration and ultrasonication.

[0028] S3. Composite component molding: The concentrated colloid is transported to the spray drying tower via a peristaltic pump (inlet air temperature 140℃, outlet air temperature 70℃, atomizer frequency 50Hz) and free-flowing light yellow gel particles are collected with an average particle size of about 280μm.

[0029] Example 2 The difference from Example 1 is as follows: In S1a, the mass ratio of the four raw materials is 1:1:1:1, the amount of granular Poria cocos polysaccharide added is 10% (10g), the material-to-liquid ratio is 1:10, the extraction temperature is 40℃, and the extraction time is 0.5 hours; In S1b, 500mL of 1% (w / v) dissolved Poria cocos polysaccharide solution (5g of dry matter, accounting for 5%) is added, the pH is adjusted to 4.0, the extraction temperature is 60℃, and the extraction time is 1 hour; In S2, the concentration program has a cooling rate of 0.5℃ / min, an ultrasonic frequency of 20kHz, and a power density of 50W / L; In S3, gel microspheres are obtained by the dripping method (dropped into cold vegetable oil to form), with an average particle size of about 100μm.

[0030] Example 3 The difference from Example 1 is as follows: In S1a, the mass ratio of the four raw materials is 3:2:2:3, the amount of granular Poria cocos polysaccharide added is 15% (15g), the material-to-liquid ratio is 1:20, the extraction temperature is 60℃, and the extraction time is 2 hours; In S1b, about 333mL of 3% (w / v) dissolved Poria cocos polysaccharide solution (10g of dry matter, accounting for 10%) is added, the pH is adjusted to 6.0, the extraction temperature is 95℃, and the extraction time is 4 hours; In S2, the concentration program has a cooling rate of 1℃ / min, an ultrasonic frequency of 40kHz, and a power density of 200W / L; In S3, granulation is carried out by extrusion spheronization, and the average particle size is about 480μm.

[0031] Example 4 The difference from Example 1 is as follows: In S1a, the amount of granular Poria cocos polysaccharide added is 11% (11g); in S1b, the amount of dissolved Poria cocos polysaccharide added is 6% (6g, using 400mL of 1.5% concentration solution), the pH is adjusted to 4.5, and the extraction temperature is 70℃; in S2, the concentration and cooling rate is 0.6℃ / min, the ultrasonic frequency is 25kHz, and the power density is 80W / L; after spray drying, the average particle size is about 180μm.

[0032] Example 5 The difference from Example 1 is as follows: In S1a, the amount of granular Poria cocos polysaccharide added is 14% (14g); in S1b, the amount of dissolved Poria cocos polysaccharide added is 8% (8g, using 320mL of 2.5% concentration solution), the pH is adjusted to 5.5, and the extraction temperature is 90℃; in S2, the concentration and cooling rate is 0.9℃ / min, the ultrasonic frequency is 35kHz, and the power density is 150W / L; after spray drying, the average particle size is about 400μm.

[0033] Comparative Example 1 The step-by-step guidance was cancelled; all four types of mushroom powder (100g), granular Poria cocos polysaccharide (12.5g), and dissolved Poria cocos polysaccharide (added in solid form) equivalent to 7.5g of dry matter were mixed at one time, and 2125mL of purified water was added (total liquid volume is the same as in Example 1). The pH was directly adjusted to 5.0, and the mixture was extracted at 80℃ for 3.75 hours at one time (total time is the same as in Example 1). In the subsequent concentration process, no ultrasound was applied, and only low-temperature vacuum concentration was carried out under the same conditions. The shaping steps were the same as in Example 1.

[0034] Comparative Example 2 Five single polysaccharide powders of shiitake mushroom, white lingzhi mushroom, white-backed wood ear fungus, button mushroom, and poria cocos were independently extracted and refined using conventional hot water extraction methods. After being accurately weighed according to the theoretical content ratio of each polysaccharide in the final product of Example 1, they were mixed in a three-dimensional mixer for 1 hour to obtain physically mixed adjuvant powder.

[0035] Comparative Example 3 The extraction steps are exactly the same as in Example 1, but no ultrasound is applied during the S2 concentration process. Instead, low-temperature vacuum concentration is performed at the same temperature, vacuum level, and programmed cooling.

[0036] Example 6 This invention provides a method for preparing a fungal component complex, such as... Figure 2 As shown, it includes the following steps: P1. Preparation of the solid core of the principal and assistant herbs: Weigh out 14.5 parts by weight of Grifola frondosa polysaccharide, 14.5 parts by weight of Cordyceps militaris polysaccharide, 17.5 parts by weight of Ganoderma lucidum extract (after crushing the fruiting body of Ganoderma lucidum, extract with water at a ratio of 1:20 and 80℃ for 2 hours, concentrate and dry to obtain the extract, with a polysaccharide content of 35%), 12 parts by weight of Hericium erinaceus polysaccharide, and 10 parts by weight of Agaricus blazei polysaccharide, and mix them evenly; use a dry granulator to compress and granulate, and sieve out solid microspheres of 150-300 micrometers (about 50-100 mesh) as the core.

[0037] P2. Core-shell structure construction: Take 30 parts (based on dry matter) of the adjuvant-guide drug composite component prepared in Example 1, disperse it in 600 mL of 3% (w / v) hydroxypropyl methylcellulose (HPMC) aqueous solution, homogenize it to obtain a coating slurry; put 100 parts of the principal-assistant drug core obtained in P1 into a fluidized bed coating machine (bottom spray process), with an inlet air temperature of 55℃; spray the coating slurry uniformly onto the surface of the fluidized core; after coating, the pellets continue to dry in the equipment for 10 minutes.

[0038] P3. Final formulation: The microspheres were dried in a vacuum drying oven at 40°C until the moisture content was <5%, thus obtaining the core-shell structured composite product; according to statistics, the dry weight of the coating layer accounted for about 30% of the total weight of the microspheres.

[0039] Example 7 The ratio of principal and assistant herbs was adjusted as follows: 12 parts of Grifola frondosa polysaccharide, 17 parts of Cordyceps militaris polysaccharide, 15 parts of Ganoderma lucidum extract, 14 parts of Hericium erinaceus polysaccharide, and 8 parts of Agaricus blazei polysaccharide; 24 parts of adjuvant and guide herbs were taken; the coating adhesive was changed to 2% sodium alginate solution; after coating, the dry weight of the coating layer accounted for about 20% of the total weight of the microcapsules; the remaining steps were the same as in Example 6.

[0040] Example 8 The ratio of principal and assistant herbs is as follows: 17 parts of Grifola frondosa polysaccharide, 12 parts of Cordyceps militaris polysaccharide, 20 parts of Ganoderma lucidum extract, 10 parts of Hericium erinaceus polysaccharide, and 12 parts of Agaricus blazei polysaccharide; 36 parts of adjuvant and guide herbs are taken; after coating, the dry weight of the coating layer accounts for about 40% of the total weight of the microcapsules; the remaining steps are the same as in Example 6.

[0041] Comparative Example 4 All the raw materials of the principal and assistant herbs (five kinds including Grifola frondosa polysaccharide) used in Example 6 were weighed together with the physically mixed adjuvant powder prepared in Comparative Example 2 in the same proportion as in Example 6 (principal and assistant herbs: adjuvant herbs = 100:30), and then thoroughly mixed in a three-dimensional mixer for 1 hour to obtain a fully mixed physical sample.

[0042] Comparative Example 5 The stepwise, non-ultrasound-free adjuvant drug composite component prepared in Comparative Example 3, along with the principal and adjuvant drug raw material dry powder in the same proportion as in Example 6, were placed in a three-dimensional mixer and thoroughly mixed for 1 hour at a ratio of principal and adjuvant drug: adjuvant drug dry matter = 100:30 to obtain the final product.

[0043] Performance testing and data analysis Test 1: Processing characteristics and physical properties of adjuvant and guide drug components The products of Examples 1-5 and Comparative Examples 1, 2, and 3 were subjected to the following tests: Total polysaccharide yield: determined by phenol-sulfuric acid method.

[0044] Gel microparticle characteristics: Particle size distribution was measured using a laser particle size analyzer, and flowability was measured using an angle of repose meter.

[0045] Colloidal stability: The sample was reconstituted in water (1% w / v), allowed to stand for 24 hours to observe the layering, and the zeta potential was measured.

[0046] The results are shown in Table 1: Table 1 Comparison of preparation processes and product properties of adjuvant and guide drug components Note: Comparative Example 2 is a physical mixture of five single polysaccharides in dry powder, with no fixed particle size, so the average particle size is marked as "not applicable (dry powder)"; the zeta potential measurement conditions are as follows: the dry powder is dispersed in purified water to prepare a 1% (w / v) dispersion, and the dispersion is ultrasonically dispersed for 5 minutes before measurement.

[0047] Conclusion: The adjuvant drug components prepared by the method of the present invention (Examples 1-5) are significantly superior to simple mixing extraction (Comparative Example 1) and physical mixing (Comparative Example 2) in terms of yield, particle uniformity, flowability and colloidal stability; in particular, the zeta potential is more negative, indicating that the colloidal dispersion system is more stable, and stepwise guidance and dynamic co-condensation promote intermolecular interactions and form a more stable composite structure.

[0048] Test 2: In vitro release behavior study Take an equivalent amount of the product from Example 6 (core-shell structure), the product from Comparative Example 4 (physical mixture of the whole formula), and an equal amount of the principal and assistant herbs core (uncoated), containing 100 mg of total polysaccharides. Dissolve the samples according to the Chinese Pharmacopoeia (slurry method). First, measure the dissolution in 900 mL of pH 1.2 hydrochloric acid solution (simulated gastric juice) for 2 hours. Then, quickly add an equal amount of pH 6.8 phosphate buffer and continue measuring in simulated intestinal juice for 12 hours. Take samples periodically to determine the cumulative release rate of polysaccharides. The cumulative release rate is calculated based on 100% of the total polysaccharide mass in the sample.

[0049] The results are shown in Table 2: Table 2. Key data points for in vitro release (cumulative release rate %) Conclusion: The core-shell structure product of the present invention (Example 6) exhibits significant sustained-release and programmed release characteristics; it releases slowly in gastric juice and smoothly after entering intestinal juice; while the uncoated core and physically mixed samples both showed a burst release of more than 65% within the first 2 hours; indicating that the core-shell structure of the present invention effectively achieves the design goal of the adjuvant drug clearing the obstruction first and the principal drug releasing slowly to consolidate the foundation.

[0050] Test 3: Evaluation of in vitro immunomodulatory activity The anti-inflammatory and immunomodulatory activities of the samples were evaluated using a lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophage inflammation model. The following sample groups were set up: a blank control group, an LPS model group, and the following sample groups: The concentration of the whole sample (Example 6, Comparative Example 4, Comparative Example 5) was tested at 100 μg / mL (calculated as total polysaccharides). The adjuvant and guide drug component samples (Examples 1, Comparative Examples 2 and 3): the test concentration was calculated based on its mass percentage (approximately 30%) in the whole formula of Example 6, and was 30 μg / mL; Samples of the principal and assistant medicinal components: The principal and assistant medicinal raw materials used in Example 6 were physically mixed in the same proportion (Grifola frondosa polysaccharide: Cordyceps militaris polysaccharide: Ganoderma lucidum extract: Hericium erinaceus polysaccharide: Agaricus blazei polysaccharide = 14.5:14.5:17.5:12:10). The test concentration was calculated based on its proportion in the whole formula (approximately 70%), and was 70 μg / mL. The ELISA method was used to determine the content of tumor necrosis factor-α (TNF-α) in cell supernatant and calculate the inhibition rate; the MTT assay was used to detect the effect of the sample on cell viability to ensure that the experimental concentration was non-toxic.

[0051] The results are shown in Table 3: Table 3. Inhibition rate of LPS-induced TNF-α secretion from RAW264.7 cells (%, x±s, n=3) Conclusion: The structured adjuvant component (Example 1) showed significantly higher anti-inflammatory activity than the physically mixed adjuvant (Comparative Example 2) and the control without ultrasonic treatment (Comparative Example 3), indicating that the stepwise guided extraction process produced a synergistic effect. The complete core-shell structure compound (Example 6) exhibited the highest immunomodulatory activity, which was significantly higher than that of the physical mixture of its components (adjuvant drug + principal drug) (Comparative Example 4) (P<0.01); indicating that the dual structuring of the present invention (internal structuring of adjuvant drug + whole core-shell structure) produced an unexpected synergistic effect.

[0052] Test 4: Validation of an animal model of deficiency-cold syndrome Experimental materials: ICR mice were used to establish a cold-induced hypothermia model by cold stimulation and reserpine injection. They were randomly divided into a model group, the drug administration group of Example 6 (gavage dose 200mg / kg·d), a physical mixed control group (comparative example 4, same dose), and a positive control group (Fuzi Lizhong Wan, 100mg / kg·d). Each group consisted of 10 mice, and the drugs were administered for 14 consecutive days.

[0053] Detection indicators: weight change, rectal temperature, serum IL-2 (immune-related cytokine), serum cortisol (indicator related to deficiency and cold).

[0054] The results are shown in Table 4: Conclusion: The drug administration group in Example 6 significantly improved the body weight, body temperature and serum indicators of the cold-deficiency model mice, with better results than the physical mixed control group and close to the positive control group.

[0055] It should be noted that, for the sake of simplicity, the foregoing embodiments are all described as a series of actions. However, those skilled in the art should understand that the present invention is not limited to the described order of actions, as some steps may be performed in other orders or simultaneously according to the present invention. Furthermore, those skilled in the art should also understand that the embodiments described in the specification are preferred embodiments, and the actions and modules involved are not necessarily essential to the present invention.

[0056] It should be understood that the disclosed apparatus can be implemented in other ways, as illustrated in the embodiments provided in this application. For example, the apparatus embodiments described above are merely illustrative; the division of units described above is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple units or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the coupling or communication connections shown or discussed may be through some interfaces; the indirect coupling or communication connections between devices or units may be telecommunications or other forms.

[0057] The units described above as separate components may or may not be physically separate. The components shown as units may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the units can be selected to achieve the purpose of this embodiment according to actual needs.

[0058] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the scope of protection of the invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on these embodiments, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art can still combine, add, delete, or otherwise adjust the features of the various embodiments of the present invention according to the circumstances without conflict or creative effort, thereby obtaining different technical solutions that do not fundamentally depart from the concept of the present invention. These technical solutions also fall within the scope of protection of the present invention.

Claims

1. A method for preparing a complex of fungal polysaccharide adjuvants, characterized in that, Includes the following steps: S1. Step-by-step guided extraction: S1a. Physical adsorption guidance: The dried powders of four raw materials, namely shiitake mushroom, white lingzhi mushroom, white-backed wood ear fungus and button mushroom, are mixed together with water and granular Poria cocos polysaccharide to form an extraction system, and the first stage of extraction is carried out at 40-60℃. S1b. Functional integration guidance: Add dissolved Poria cocos polysaccharide solution to the S1a system, adjust the pH of the system, and carry out the second stage of extraction at 60-95℃. After the extraction is completed, the first complex polysaccharide extract is obtained by separation. S2. Dynamic co-condensation concentration: The first complex polysaccharide extract is concentrated under low temperature vacuum, and ultrasonic waves are applied to it during the concentration process to obtain complex polysaccharide colloid; S3. Composite component molding: The composite polysaccharide colloid is formed into 100-500 micrometer gel particles to obtain the adjuvant-guide drug composite component.

2. The method for preparing the fungal polysaccharide adjuvant compound component as described in claim 1, characterized in that, In step S1a, the mass ratio of the dried powders of shiitake mushrooms, white lingzhi mushrooms, white-backed wood ear mushrooms, and button mushrooms is 1-3:1-2:1-2:1-3; the particle size of the granular Poria cocos polysaccharide is 1-100 micrometers, and its addition amount is 10%-15% of the total mass of the mixed powder of the four raw materials; the first extraction time is 0.5-2 hours, and the material-to-liquid ratio is 1:10-20 (g / mL); in step S1b, the concentration of the dissolved Poria cocos polysaccharide is 1%-3% (w / v), and its addition amount, on a dry matter basis, is 5%-10% of the total mass of the mixed powder of the four raw materials; the second extraction time is 1-4 hours, and the pH is 4.0-6.

0.

3. The method for preparing the fungal polysaccharide adjuvant compound component as described in claim 1, characterized in that, In step S2, the conditions for low-temperature vacuum concentration are as follows: initial temperature 60℃, maintaining vacuum degree -0.06 to -0.1MPa, using a programmed gradient cooling mode, uniformly cooling to 40℃ at a rate of 0.5-1℃ / min until the material is concentrated to 10-20% of its original volume; the applied ultrasonic frequency is 20-40kHz, and the power density is 50-200W / L.

4. The method for preparing the fungal polysaccharide adjuvant compound component as described in claim 1, characterized in that, In step S3, the molding method includes one of spray granulation, drop molding, or extrusion spheroidization.

5. A fungal polysaccharide adjuvant compound component, characterized in that, Prepared by the method described in any one of claims 1-4.

6. A fungal component complex, characterized in that, It includes the fungal polysaccharide adjuvant compound component as described in claim 5, the principal drug and the assistant drug; the principal drug includes Grifola frondosa polysaccharide and Cordyceps militaris polysaccharide; the assistant drug includes Ganoderma lucidum extract, Hericium erinaceus polysaccharide and Agaricus blazei polysaccharide.

7. The fungal component complex as described in claim 6, characterized in that, By weight, the fungal polysaccharide adjuvant compound consists of 24-36 parts, Grifola frondosa polysaccharide 12-17 parts, Cordyceps militaris polysaccharide 12-17 parts, Ganoderma lucidum extract 15-20 parts, Hericium erinaceus polysaccharide 10-14 parts, and Agaricus blazei polysaccharide 8-12 parts.

8. The method for preparing the fungal component complex as described in claim 6, characterized in that, Includes the following steps: P1. Preparation of the solid core of the principal and assistant medicine: Take Grifola frondosa polysaccharide, Cordyceps militaris polysaccharide, Ganoderma lucidum extract, Hericium erinaceus polysaccharide, and Agaricus blazei polysaccharide; after thoroughly mixing the above materials, use a dry granulation process to make solid microspheres with a particle size of 100-1000 micrometers, which serve as the solid core of the principal and assistant medicine. P2. Construction of the core-shell structure: The solid core of the principal and adjuvant drugs is placed in a fluidized bed coating device and kept in a fluidized state; the fungal polysaccharide adjuvant drug composite components are dispersed or dissolved in an adhesive solution as a coating slurry; The coating slurry is uniformly coated onto the surface of the fluidized solid core of the medicinal material using a fluidized bed spraying process to form a coating layer. After drying, micro-units are obtained. The dry weight of the coating layer accounts for 20%-40% of the total weight of the micro-units. P3. Final formulation: The micro-units are dried to a moisture content of less than 5%, thus obtaining the microbial component complex.

9. The method for preparing the fungal component complex as described in claim 8, characterized in that, In step P2, the fluidized bed spraying process is either a bottom spraying process or a tangential spraying process; the adhesive solution is a 1%-5% (w / v) aqueous solution of hydroxypropyl methylcellulose or an aqueous solution of sodium alginate.

10. The use of the fungal component complex as described in claim 6 in the preparation of health foods or medicines for immune regulation, warming and strengthening the body, or improving deficiency-cold syndrome.

Images