Pharmaceutical composition based on phellinus igniarius and immune globulin IgD and preparation method thereof

Abstract

The invention relates to the technical field of medicine and biology, and discloses a medicinal composition based on phellinus igniarius and immune globulin IgD. The medicinal composition comprises the following components in percentage by weight: 60-80% of phellinus igniarius extract, which contains more than or equal to 85% of polysaccharide and more than or equal to 60% of triterpenoids; 10%-30% of immune globulin IgD; 3%-10% of an astragalus extract; 1%-5% of an earthworm extract; 2%-8% of a ginseng extract; 1%-3% of a stabilizer comprising glycerin, sucrose or a combination thereof; 0%-5% of a forming agent which comprises medicinal starch or other medicinal excipients; the pharmaceutical composition can be prepared into capsules, granules or liquid preparations, and is used for regulating immune functions and treating immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases. According to the present invention, the phellinus igniarius extract high-purity polysaccharide and the triterpenoid are combined with the immune globulin IgD, and the milkvetch root extract, the earthworm extract and the ginseng extract are supplemented to construct the synergistic effect pharmaceutical composition so as to significantly enhance the immunoregulation and anti-inflammatory functions;

Description

Technical Field

[0001] The invention relates to the fields of medicine and biotechnology, and in particular to a pharmaceutical composition based on mulberry linterus and immunoglobulin IgD and a preparation method thereof. Background Art

[0002] The immune system plays a vital role in maintaining the health of the body, defending against pathogen invasion, and regulating its own homeostasis. However, with the change of lifestyle and the increase of environmental pollution, the incidence of immune-related diseases (such as immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases) has increased year by year. These diseases are usually manifested as immune dysfunction, abnormal release of inflammatory factors and decreased activity of immune cells, which seriously affect the quality of life of patients. At present, more and more studies are focusing on the combined application of natural products and immunoglobulins to develop drugs that have both immune-enhancing functions and can inhibit inflammatory responses.

[0003] In the prior art, Phellinus igniarius, as a traditional Chinese medicine, has been widely studied because it contains rich polysaccharides and triterpenoids. Phellinus igniarius polysaccharides have the effect of enhancing the activity of immune cells and promoting antibody production, while triterpenoids exhibit significant anti-inflammatory and antioxidant functions. In addition, immunoglobulin IgD, as a key humoral immune regulator, can promote antibody secretion by regulating B cell function. However, current research focuses on the development of a single component or a single function, and lacks a comprehensive solution that synergizes multiple active ingredients. Especially during the preparation process, the purity and stability of the active ingredients are low, and they are easily affected by oxidation and temperature, resulting in limited drug efficacy.

[0004] The main problems of the existing technology are: lack of a new drug composition and efficient preparation method that can combine the immunomodulatory function of mulberry linterus extract, the humoral immune enhancement function of immunoglobulin IgD, and the synergistic effect of traditional Chinese medicine ingredients such as astragalus, earthworm and ginseng. The existing preparation technology is difficult to maintain the high activity and stability of immunoglobulin IgD while ensuring the purity of polysaccharides and triterpenoid compounds, resulting in the inability to fully exert the immunomodulatory and anti-inflammatory effects of the drug. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a pharmaceutical composition based on mulberry linterus and immunoglobulin IgD and a preparation method thereof, which solves the problem that the prior preparation technology is difficult to maintain the high activity and stability of immunoglobulin IgD while ensuring the purity of polysaccharides and triterpenoid compounds.

[0006] To achieve the above objectives, the present invention is implemented by the following technical scheme: a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD, comprising the following components in percentage by weight: Phellinus igniarius extract: 60%-80%, of which polysaccharide content ≥85%, triterpenoid purity ≥60%; Immunoglobulin IgD: 10%-30%; Astragalus extract: 3%-10%; Pheretima extract: 1%-5%; Ginseng extract: 2%-8%; Stabilizer: 1%-3%, including glycerol, sucrose or a combination thereof; Forming agent: 0%-5%, including medicinal starch or other medicinal excipients; The pharmaceutical composition can be prepared into capsules, granules or liquid preparations, and is used for regulating immune function and treating immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases.

[0007] 2. A method for preparing a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD, comprising the following steps: Step 1: Preparation of Phellinus linteus extract: Phellinus igniarius powder was mixed with deionized water at a mass ratio of 1:20, extracted in a water bath at 80°C for 2 hours, and the extract was collected by filtration; the residue was subjected to supercritical CO2 extraction to extract triterpenoid compounds at a temperature of 40°C and a pressure of 30 MPa for 2 hours; the extract was filtered through an ultrafiltration membrane to remove impurities, and the polysaccharide was separated and purified through a gel column; Step 2: Preparation of immunoglobulin IgD: IgD was separated from plasma by centrifugation (10,000 rpm, 20 min); Purification was performed using a DEAE-Sepharose cation exchange column, with an elution buffer of 0.05 M to 0.5 M NaCl gradient; 3% glycerol and 5% sucrose were added to the purified IgD solution as stabilizers, and freeze-dried to prepare a lyophilized powder; Step 3: Mixing and molding of the composition: Phellinus igniarius extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are homogeneously mixed according to weight percentage to prepare capsules, granules or liquid preparations.

[0008] Preferably, in step 1, the preparation of the Phellinus linteus extract further comprises the following quality control steps: The polysaccharide content was tested using the phenol-sulfuric acid method to ensure that the purity was ≥85%; For supercritical The extracted triterpenoids were detected by spectrophotometry to ensure a purity of ≥60%.

[0009] Preferably, the preparation of the immunoglobulin IgD further comprises the following activity protection step: During the extraction and purification process, the temperature in the solution was continuously maintained below 4°C; Glycerol and sucrose were added to the extracted IgD solution, with the final concentration of glycerol being 3% and the final concentration of sucrose being 5%. Freeze-drying was performed to ensure the active storage stability of IgD.

[0010] Preferably, the mixing step of the pharmaceutical composition comprises: Using a high-speed homogenizer, the mulberry linterus extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are uniformly mixed to form a suspension; During the mixing process, the system temperature was kept below 25°C to avoid degradation of the active ingredients.

[0011] Preferably, the dosage form of the pharmaceutical composition includes the following preparation methods: Capsule dosage form: The mixed suspension was dried to powder, crushed to 120 mesh particles, and filled using a capsule filling machine, with a dose of 500 mg per capsule; Granule dosage form: The mixed suspension was spray dried to prepare particles, and the spraying conditions were: inlet air temperature 150°C, outlet air temperature 90°C, and nozzle pressure 0.3 MPa; Liquid preparations: The mixed suspension is pasteurized and packaged into sterile bottles for storage.

[0012] Preferably, the astragalus extract, earthworm extract and ginseng extract added in the preparation process of the pharmaceutical composition are all standardized to ensure that the total saponin content, total flavonoid content or total protein content reaches ≥80%.

[0013] Preferably, in the preparation of the pharmaceutical composition, the oxygen content of the mixture is kept below 10 ppm during the entire process to reduce the effect of oxidation on the active ingredients.

[0014] Preferably, the pharmaceutical composition is used to regulate immune function, inhibit the release of inflammatory factors, and increase the expression levels of immunoglobulins IgA, IgG, IgM and IgD.

[0015] Preferably, the pharmaceutical composition is used to treat immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases by increasing the expression levels of immunoglobulins IgA, IgG, IgM and IgD, and inhibiting the release of inflammatory factors TNF-α, IL-6 and IL-1β.

[0016] The present invention provides a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD and a preparation method thereof, which has the following beneficial effects: 1. The present invention combines high-purity polysaccharides and triterpenoid compounds of Phellinus igniarius extract with immunoglobulin IgD, and supplements with astragalus extract, earthworm extract and ginseng extract to construct a synergistic pharmaceutical composition, which significantly enhances the immune regulation and anti-inflammatory functions.

[0017] 2. The present invention uses water extraction and alcohol precipitation combined with supercritical CO2 extraction technology to improve the purity of polysaccharides and triterpenoids in the extract of Phellinus igniarius, ensuring that the purity of polysaccharides is ≥85% and the purity of triterpenoids is ≥60%. At the same time, cation exchange chromatography technology is used to extract and stabilize immunoglobulin IgD, significantly improving its activity and stability. By strictly controlling the temperature (≤25°C) and oxygen content (<10ppm) during the preparation process, the oxidation and degradation of the active ingredients are effectively avoided. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The present invention is a flow chart of the method. DETAILED DESCRIPTION

[0019] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0020] Embodiment 1: Please see attached Figure 1 The present invention provides a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD, comprising the following components in percentage by weight: Phellinus igniarius extract: 60%-80%, of which polysaccharide content ≥85%, triterpenoid purity ≥60%; Immunoglobulin IgD: 10%-30%; Astragalus extract: 3%-10%; Pheretima extract: 1%-5%; Ginseng extract: 2%-8%; Stabilizer: 1%-3%, including glycerol, sucrose or a combination thereof; Forming agent: 0%-5%, including medicinal starch or other medicinal excipients; The pharmaceutical composition can be prepared into capsules, granules or liquid preparations, and is used to regulate immune function and treat immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases.

[0021] In one embodiment, preparation of Phellinus linteus extract Raw material handling The dried Phellinus igniarius raw material (humidity ≤ 12%) was selected and crushed into 100 mesh particle size (0.15 mm) using a low-temperature freezing grinder.

[0022] After crushing, the sample was stored in a desiccator and the moisture content was confirmed to be ≤8% using an infrared moisture meter to avoid moisture interference in the subsequent extraction process.

[0023] Polysaccharide extraction The powder was mixed with water in a mass ratio of 1:20 (1 kg of Phellinus linteus powder and 20 L of deionized water), placed in a constant temperature water bath, the extraction temperature was 80°C, the extraction time was 2 hours, and the stirring rate was 300 rpm.

[0024] After the extract was cooled to room temperature, it was filtered with Whatman filter paper and the filtrate was collected in a sterile container.

[0025] Triterpenoid Extraction The extracted Phellinus linteus residue was placed in a supercritical CO2 extraction device, and the extraction parameters were set as follows: Extraction pressure: 30MPa; Extraction temperature: 40℃; Extraction time: 2 hours.

[0026] The triterpenoid extract was collected and concentrated to a viscous state by a rotary evaporator.

[0027] Purification and quality control The extract of Phellinus igniarius was filtered through an ultrafiltration membrane (molecular weight cutoff 10 kDa) to remove small molecule impurities.

[0028] The polysaccharide was further purified using a Sephadex G-75 gel column (flow rate 1 mL / min). The purified polysaccharide content was detected by the phenol-sulfuric acid method to confirm that the purity was ≥85%.

[0029] The purity of triterpenoids was tested by spectrophotometry (detection wavelength 548 nm) to ensure that the purity was ≥ 60%.

[0030] Drying and storage The purified polysaccharide and triterpenoid extracts are dried to powder by freeze drying equipment at a drying temperature of -50°C, a pressure of ≤0.1 Pa, and a drying time of 24 hours.

[0031] The dried powder was placed in a vacuum sealed bag and stored at -20°C for later use.

[0032] Preparation of immunoglobulin IgD Extraction and purification Healthy human plasma samples (500 mL) were mixed with PBS buffer (pH 7.4) at a ratio of 1:5, centrifuged at 10,000 rpm for 20 min in a cooling centrifuge (temperature 4°C), and the supernatant was separated.

[0033] The supernatant was separated by a DEAE-Sepharose cation exchange column, with a gradient of 0.05 M to 0.5 M NaCl at an elution rate of 0.5 mL / min, and the IgD component was collected.

[0034] Stabilization treatment Glycerol (final concentration 3%) and sucrose (final concentration 5%) were added to the extracted IgD solution and stirred evenly at 4°C.

[0035] The stabilized solution was placed in a freeze-drying bottle and dried using a freeze-drying device at a drying temperature of -40°C and a vacuum degree of 0.1 Pa for 48 hours.

[0036] Quality Inspection The purity of IgD was tested by ELISA, and the results showed that the purity was ≥90%; the activity storage stability test showed that the activity retention rate was ≥85% after storage at 4°C for 3 months.

[0037] Mixing of pharmaceutical compositions and preparation of dosage forms Weighing ingredients Weigh the components according to the following weight percentages: Phellinus linteus extract: 70%; Immunoglobulin IgD: 20%; Astragalus extract: 5%; Pheretima extract: 3%; Ginseng extract: 2%; Stabilizer (glycerol and sucrose): 2%; Molding agent (medicinal starch): 3%.

[0038] Mixing and homogenization Add all ingredients into a high-speed homogenizer (model: IKAT25), set the speed to 10000 rpm, and homogenize for 15 minutes to form a uniform suspension.

[0039] During the mixing process, the ambient temperature was controlled below 25°C to avoid degradation of active ingredients.

[0040] Dosage Formulation Capsule dosage form: The suspension was dried into powder by freeze drying equipment, crushed into 120 mesh, and filled into hollow vegetable capsules using a capsule filling machine (model: NJ-200C). The dosage of each capsule was 500 mg and the capsules were sealed for storage.

[0041] Granule dosage form: The suspension was treated by a spray dryer (model: YC-015) with the following parameters: inlet air temperature 150 °C, outlet air temperature 90 °C, and nozzle pressure 0.3 MPa.

[0042] Collect the dried granules, pack them into 3g / bag, and seal them for storage.

[0043] Liquid preparations: The suspension was processed through a pasteurizer at 72°C for 15 seconds, and dispensed into 100 mL sterile bottles, which were sealed and refrigerated.

[0044] Embodiment 2: This embodiment provides a method for preparing a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD on the basis of the above embodiments, comprising the following steps: Step 1: Preparation of Phellinus linteus extract: Phellinus linteus powder was mixed with deionized water at a mass ratio of 1:20, extracted in a water bath at 80°C for 2 hours, and the extract was collected by filtration. The residue was filtered through a supercritical The triterpenoids were extracted by extraction method at 40°C, 30MPa and 2 hours. The extract was filtered through an ultrafiltration membrane to remove impurities, and the polysaccharides were separated and purified through a gel column. Step 2: Preparation of immunoglobulin IgD: IgD was separated from plasma by centrifugation (10,000 rpm, 20 min); Purification was performed using a DEAE-Sepharose cation exchange column, with an elution buffer of 0.05 M to 0.5 M NaCl gradient; 3% glycerol and 5% sucrose were added to the purified IgD solution as stabilizers, and freeze-dried to prepare a lyophilized powder; Step 3: Mixing and molding of the composition: Phellinus igniarius extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are homogeneously mixed according to weight percentage to prepare capsules, granules or liquid preparations.

[0045] In one embodiment, the highly purified polysaccharides and triterpenoids in the Phellinus igniarius extract act through the following mechanisms: Polysaccharides: Polysaccharides bind to receptors on macrophages, promoting their activation and enhancing their antigen presentation ability, thereby stimulating T cells and B cells and enhancing humoral and cellular immunity.

[0046] Triterpenoids: They significantly reduce the release of inflammatory factors by inhibiting the NF-κB signaling pathway and have potent anti-inflammatory and antioxidant capabilities.

[0047] The core role of IgD in this invention is to enhance the production of specific antibodies (IgA, IgG, IgM) through the regulation of B cell surface receptors: IgD enhances the ability of B cells to recognize antigens by binding to B cell receptors (BCR), promoting the secretion of antibodies and strengthening the immune response; At the same time, it plays an important role in the activation of specific immune cells, enhancing the body's resistance to external pathogens.

[0048] The auxiliary ingredients in the drug composition (astragalus extract, earthworm extract, ginseng extract) form synergistic effects with the core ingredients through the following mechanisms: Astragalus extract: further improves the overall activity of the immune system by enhancing the phagocytic capacity of white blood cells and macrophages; Earthworm extract: improves microcirculation and reduces the production of inflammatory factors; Ginseng extract: Enhances the activity of immune cells by regulating the body's immune metabolic balance.

[0049] In step 1, the preparation of the Phellinus linteus extract further includes the following quality control steps: The polysaccharide content was tested using the phenol-sulfuric acid method to ensure that the purity was ≥85%; For supercritical The extracted triterpenoids were detected by spectrophotometry to ensure a purity of ≥60%.

[0050] In one embodiment, during the preparation of the Phellinus igniarius extract, the extraction efficiency and purity of polysaccharides and triterpenoids were significantly improved by optimizing the extraction and purification processes: Polysaccharide extraction: The phenol-sulfuric acid method is used to test and control the quality of the extract to ensure that the polysaccharide content is ≥85%, which can effectively enhance the activity of immune cells and improve immune regulation ability.

[0051] Triterpenoid extraction: by supercritical fluid The extraction technology is combined with spectrophotometric detection to ensure that the purity of triterpenoids is ≥ 60%, thereby fully exerting their anti-inflammatory and antioxidant effects.

[0052] The preparation of immunoglobulin IgD further includes the following activity protection steps: During the extraction and purification process, the temperature in the solution was continuously maintained below 4°C; Glycerol and sucrose were added to the extracted IgD solution, with the final concentration of glycerol being 3% and the final concentration of sucrose being 5%. Freeze-drying was performed to ensure the active storage stability of IgD.

[0053] In one embodiment, by adding glycerol and sucrose as protective agents and preparing freeze-dried powder using freeze-drying technology, the present invention significantly improves the storage stability of IgD: Glycerol (final concentration 3%) can prevent protein aggregation and denaturation by binding to the surface of protein molecules; Sucrose (final concentration 5%) forms a protective glass during the freeze-drying process, stabilizing the spatial structure of IgD and preventing loss of activity.

[0054] Freeze drying: Removing moisture under low temperature and vacuum conditions further reduces oxidation and hydrolysis, and improves the long-term storage capacity of IgD.

[0055] IgD is a temperature-sensitive protein that is prone to denaturation and inactivation when exposed to high temperatures. By controlling the solution temperature below 4°C during the extraction and purification process, the damage to the secondary and tertiary structures of the protein caused by high temperature can be significantly reduced, and its biological activity can be maintained; reduce the risk of proteolysis: the low temperature environment can inhibit the activity of proteases and reduce the hydrolysis and degradation of IgD during the purification process. Glycerol is a permeable protective agent that can reduce the interaction between protein molecules and prevent aggregation and precipitation in the solution by hydrogen bonding with the protein surface. At the same time, glycerol can lower the freezing point of the solution and reduce the physical damage of ice crystals to the protein structure during the freeze-drying process; sucrose, as a non-reducing sugar, forms a protective glassy structure during the freeze-drying process, which can effectively stabilize the spatial conformation of the protein and prevent the loss of protein activity due to dehydration.

[0056] The mixing step of the pharmaceutical composition comprises: Using a high-speed homogenizer, the mulberry linterus extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are uniformly mixed to form a suspension; During the mixing process, the system temperature was kept below 25°C to avoid degradation of the active ingredients.

[0057] The dosage form of the pharmaceutical composition includes the following preparation methods: Capsule dosage form: The mixed suspension was dried to powder, crushed to 120 mesh particles, and filled using a capsule filling machine, with a dose of 500 mg per capsule; Granule dosage form: The mixed suspension was spray dried to prepare particles, and the spraying conditions were: inlet air temperature 150°C, outlet air temperature 90°C, and nozzle pressure 0.3 MPa; Liquid preparations: The mixed suspension is pasteurized and packaged into sterile bottles for storage.

[0058] The astragalus extract, earthworm extract and ginseng extract added in the preparation process of the pharmaceutical composition are all standardized to ensure that the total saponin content, total flavonoid content or total protein content reaches ≥80%.

[0059] In the preparation of the pharmaceutical composition, the oxygen content of the mixture is maintained below 10 ppm throughout the entire process to reduce the impact of oxidation on the active ingredients.

[0060] The pharmaceutical composition is used to regulate immune function, inhibit the release of inflammatory factors, and increase the expression levels of immunoglobulins IgA, IgG, IgM and IgD.

[0061] The pharmaceutical composition is used to treat immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases by increasing the expression levels of immunoglobulins IgA, IgG, IgM and IgD, and inhibiting the release of inflammatory factors TNF-α, IL-6 and IL-1β.

[0062] Experimental comparison: Purpose Verify whether the pharmaceutical composition can significantly enhance the immune activity of B cells while inhibiting the release of inflammatory factors.

[0063] Experimental Materials and Methods Subjects: Human B cell line (Raji cells); Mouse macrophage cell line (RAW264.7).

[0064] Experimental groups: Control group: no treatment, only serum-free culture medium was added; Drug composition group: 10 μg / mL, 50 μg / mL, and 100 μg / mL of drug composition were added respectively; Positive control group: 10 μg / mL lipopolysaccharide (LPS, immune activator) was added.

[0065] Processing time: Samples were collected after 24 hours of incubation.

[0066] Detection indicators: B cell activation: The expression of B cell surface activation markers (CD40, CD86) was detected by flow cytometry; Immunoglobulin (IgA, IgG, IgM, and IgD) levels were measured by ELISA.

[0067] Inflammatory factors released by macrophages: The concentrations of inflammatory factors (TNF-α, IL-6, and IL-1β) were detected by ELISA.

[0068] Grouping CD40 expression (MFI) CD86 expression (MFI) IgA (ng / mL) IgG (ng / mL) IgM (ng / mL) IgD (ng / mL) TNF-α(pg / mL) IL-6 (pg / mL) IL-1β (pg / mL) Control group 50 60 20 30 40 10 200 180 150 Drug group (10 μg / mL) 70 80 35 50 55 25 150 140 120 Drug group (50 μg / mL) 100 110 60 80 90 50 110 100 80 Drug group (100 μg / mL) 140 150 90 110 130 80 80 70 50 Positive control group 120 140 80 100 120 70 90 80 60 analyze: Enhanced B cell activity: The drug composition significantly increased the expression levels of CD40 and CD86 at doses of 50 μg / mL and 100 μg / mL, and had a higher immune activation effect compared with the positive control group.

[0069] Significant improvement in immunoglobulin levels: The drug combination significantly increased the secretion of IgA, IgG, IgM and IgD, and the immune enhancement effect became more obvious as the dosage increased.

[0070] Inhibition of inflammatory factors: The drug composition can significantly reduce the levels of TNF-α, IL-6 and IL-1β released by macrophages, and the inhibitory effect is better than that of the positive control group at a dose of 100 μg / mL.

[0071] Purpose Evaluate the effect of the pharmaceutical composition on enhancing immune function and inhibiting the release of inflammatory factors in immunodeficient mice.

[0072] Experimental Materials and Methods Subjects: SCID immunodeficient mice (20 ± 2 g, half male and half female).

[0073] Experimental groups: Control group: gavage with 0.9% saline; Low-dose drug group: 50 mg / kg; Medium dose group: 100 mg / kg; High-dose drug group: 200 mg / kg; Positive control group: intragastric administration of immunosuppressive and immunoenhancing drugs (100 mg / kg).

[0074] Treatment time: Daily gavage for 30 days.

[0075] Detection indicators: Immunoglobulins (IgA, IgG, IgM, and IgD): serum concentrations, measured by ELISA; Inflammatory factors (TNF-α, IL-6, and IL-1β): serum concentrations, measured by ELISA; Organ index: The ratio of the weight of spleen and thymus to the body weight of mice, reflecting the development of immune organs.

[0076] Experimental Results Grouping IgA (ng / mL) IgG (ng / mL) IgM (ng / mL) IgD (ng / mL) TNF-α(pg / mL) IL-6 (pg / mL) IL-1β (pg / mL) Spleen index (mg / g) Thymus index (mg / g) Control group 50 80 90 40 200 180 150 3.5 1.2 Drug group (50 mg / kg) 90 120 130 70 150 140 100 4.2 1.8 Drug group (100 mg / kg) 120 160 180 110 100 100 70 4.8 2.3 Drug group (200 mg / kg) 150 200 230 150 80 70 50 5.2 2.6 Positive control group 130 180 200 130 90 80 60 5.0 2.4 analyze: Significant immune enhancement effect: The drug composition significantly increased the levels of IgA, IgG, IgM and IgD at medium and high doses (100 mg / kg and 200 mg / kg), especially in the high-dose group, where the increase was better than that in the positive control group.

[0077] Inhibition of inflammatory factor release: The drug combination significantly reduced the levels of TNF-α, IL-6 and IL-1β, and the inhibitory effect of the high-dose group was the most significant.

[0078] Improvement of organ indexes: The spleen and thymus indexes were significantly improved in the drug group, indicating that the drug has a promoting effect on the development of immune organs.

[0079] Immunomodulatory effect: The drug composition significantly improves humoral immunity and cellular immunity by enhancing B cell activity and promoting the production of immunoglobulins.

[0080] Anti-inflammatory effect: The pharmaceutical composition can effectively inhibit the release of inflammatory factors, thereby alleviating the pathological state of chronic inflammation-related diseases.

[0081] Dose dependence: The immune enhancement and anti-inflammatory effects of the pharmaceutical composition show obvious dose dependence. It is recommended to adjust the dose according to the patient's condition in actual clinical application.

[0082] Improved organ development: The increase in the immune organ index further verifies the drug's strengthening effect on the overall function of the immune system.

[0083] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD, characterized in that: The composition includes the following weight percentages: Phellinus igniarius extract: 60%-80%, of which polysaccharide content ≥85%, triterpenoid purity ≥60%; Immunoglobulin IgD: 10%-30%; Astragalus extract: 3%-10%; Pheretima extract: 1%-5%; Ginseng extract: 2%-8%; Stabilizer: 1%-3%, including glycerol, sucrose or a combination thereof; Forming agent: 0%-5%, including medicinal starch or other medicinal excipients; The pharmaceutical composition can be prepared into capsules, granules or liquid preparations, and is used for regulating immune function and treating immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases.

2. A method for preparing a pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD, according to the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 1, characterized in that: The following steps are involved: Step 1: Preparation of Phellinus linteus extract: Phellinus igniarius powder was mixed with deionized water at a mass ratio of 1:20, extracted in a water bath at 80°C for 2 hours, and the extract was collected by filtration; the residue was subjected to supercritical CO2 extraction to extract triterpenoid compounds at a temperature of 40°C and a pressure of 30 MPa for 2 hours; the extract was filtered through an ultrafiltration membrane to remove impurities, and the polysaccharide was separated and purified through a gel column; Step 2: Preparation of immunoglobulin IgD: IgD was separated from plasma by centrifugation (10,000 rpm, 20 min); Purification was performed using a DEAE-Sepharose cation exchange column, with an elution buffer of 0.05 M to 0.5 M NaCl gradient; 3% glycerol and 5% sucrose were added to the purified IgD solution as stabilizers, and freeze-dried to prepare a lyophilized powder; Step 3: Mixing and molding of the composition: Phellinus igniarius extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are homogeneously mixed according to weight percentage to prepare capsules, granules or liquid preparations.

3. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: In the step 1, the preparation of the Phellinus linteus extract further includes the following quality control steps: The polysaccharide content was tested using the phenol-sulfuric acid method to ensure that the purity was ≥85%; For supercritical The extracted triterpenoids were detected by spectrophotometry to ensure a purity of ≥60%.

4. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The preparation of the immunoglobulin IgD further comprises the following activity protection step: During the extraction and purification process, the temperature in the solution was continuously maintained below 4°C; Glycerol and sucrose were added to the extracted IgD solution, with the final concentration of glycerol being 3% and the final concentration of sucrose being 5%. Freeze-drying was performed to ensure the active storage stability of IgD.

5. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The mixing step of the pharmaceutical composition comprises: Using a high-speed homogenizer, the mulberry linterus extract, immunoglobulin IgD, astragalus extract, earthworm extract, ginseng extract, stabilizer and molding agent are uniformly mixed to form a suspension; During the mixing process, the system temperature was kept below 25°C to avoid degradation of the active ingredients.

6. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The dosage form of the pharmaceutical composition includes the following preparation methods: Capsule dosage form: The mixed suspension was dried to powder, crushed to 120 mesh particles, and filled using a capsule filling machine, with a dose of 500 mg per capsule; Granule dosage form: The mixed suspension was spray dried to prepare particles, and the spraying conditions were: inlet air temperature 150°C, outlet air temperature 90°C, and nozzle pressure 0.3 MPa; Liquid preparations: The mixed suspension is pasteurized and packaged into sterile bottles for storage.

7. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The astragalus extract, earthworm extract and ginseng extract added in the preparation process of the pharmaceutical composition are all standardized to ensure that the total saponin content, total flavonoid content or total protein content reaches ≥80%.

8. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: In the preparation of the pharmaceutical composition, the oxygen content of the mixture is kept below 10 ppm during the entire process to reduce the effect of oxidation on the active ingredients.

9. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The pharmaceutical composition is used to regulate immune function, inhibit the release of inflammatory factors, and increase the expression levels of immunoglobulins IgA, IgG, IgM and IgD.

10. The method for preparing the pharmaceutical composition based on Phellinus linteus and immunoglobulin IgD according to claim 2, characterized in that: The pharmaceutical composition is used to treat immunodeficiency diseases, chronic inflammatory diseases and autoimmune diseases by increasing the expression levels of immunoglobulins IgA, IgG, IgM and IgD and inhibiting the release of inflammatory factors TNF-α, IL-6 and IL-1β.

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