A composite mycelium dual-effect conditioning tablet composition and its preparation method

Abstract

This invention relates to the field of biopharmaceutical technology and discloses a compound mycelium dual-effect conditioning tablet composition and its preparation method. The compound mycelium dual-effect conditioning tablet composition of this invention is made by mixing and compressing *Salix matsudana* pupa powder with food-grade excipients. *Salix matsudana* pupa is obtained by infecting *Salix matsudana* pupa with a *Cicadaemonorops cuspidatum* compound mycelium suspension, followed by aseptic cultivation until the pupae become rigid and filled with mycelium. This invention, through an innovative process of "compound mycelium pre-conversion + targeted infection of *Salix matsudana* pupae," achieves a dual-effect synergistic effect of immune regulation and blood and qi tonification. It has a high retention rate of active ingredients, good taste, and is easily absorbed, making it suitable for large-scale production and possessing significant market application prospects. Dual-effect synergy, significant efficacy: Through the process of "compound mycelium pre-conversion + targeted infection of *Salix matsudana* pupae," a dual-effect synergistic effect of immune regulation and blood and qi tonification is achieved, with conditioning effects far exceeding those of single-function products.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to a compound mycelium dual-effect conditioning tablet composition and its preparation method. Background Technology

[0002] As a crucial pupal stage in the development of the silkworm, the pupa of the *Eriocheir sinensis* is a high-nutrient-density natural raw material. Its crude protein content is ≥50%, and it is rich in unsaturated fatty acids, essential amino acids, and minerals such as iron and zinc. It is odorless, structurally stable, and possesses both excellent nutritional supply and efficient component-carrying capacity, making it an ideal carrier for fungal infection. However, currently, silkworm pupae are mostly utilized as primary food ingredients, resulting in low resource value and a lack of high-value health product conversion pathways. Its unique carrier advantages and nutritional potential have not been fully developed.

[0003] Cordyceps militaris is a traditional and precious entomopathogenic fungus. Its mycelium and insect body symbiotic complex contains cordycepin, polysaccharides and other immune-active components, with a clear immune-regulating effect. Astragalus, Codonopsis, Angelica sinensis and other food and medicine ingredients are classic combinations in traditional Chinese medicine to invigorate qi and nourish blood. They can effectively improve the state of qi and blood deficiency and lay the foundation for improving immune function.

[0004] Existing technologies for Cordyceps militaris-related health products have significant drawbacks: First, the choice of carrier is limited, often using common cicada pupae or silkworm pupae, failing to utilize the high protein and high adaptability of silkworm pupae, thus limiting product efficacy; second, the process design is simple, mostly using a "single-strain infection + ingredient mixing" model, failing to achieve synergy through "mycelial pre-metabolization transformation of active ingredients," resulting in a disconnect between immune regulation and blood and qi nourishment, failing to simultaneously address the common dual sub-health issues of modern populations; third, some processes use high-temperature treatment, which easily causes the degradation of heat-sensitive active ingredients (such as cordycepin, angelica ferulic acid, polysaccharides, etc.), affecting the product's conditioning effect.

[0005] For example, most cordyceps-based compressed candies or solid beverages on the market use a direct mixture of cordyceps powder and plant extracts, failing to achieve deep integration of mycelium and carrier (such as silkworm pupae), resulting in insufficient release of active ingredients, low absorption rate, and limited efficacy.

[0006] Therefore, developing a tableting composition that uses silkworm pupae as the core carrier, integrates Cordyceps militaris mycelium, and achieves synergistic effects on immunity and blood circulation through innovative processes, and is suitable for large-scale production, can not only overcome the problem of high-value utilization of silkworm pupae, but also fill the technological gap in the market for dual-effect conditioning health products, and has important practical value, economic value and market prospects. Summary of the Invention

[0007] (a) Technical problems to be solved: To address the shortcomings of existing technologies, this invention provides a compound mycelium dual-effect conditioning tablet composition and its preparation method, which solves the problems of inefficient utilization of silkworm pupa resources, low added value, and the single efficacy of existing Cordyceps militaris health products, which cannot simultaneously meet the dual needs of immune regulation and blood and qi nourishment.

[0008] (II) Technical Solution: A compound mycelium dual-effect conditioning tablet composition is made by mixing and compressing the stiffened pupa powder of silkworm pupae with food-grade excipients; the stiffened pupa of silkworm pupae is obtained by infecting silkworm pupae with a suspension of Cordyceps militaris compound mycelium, and then culturing them under sterile conditions until the pupae become stiff and the inside is covered with mycelium; the Cordyceps militaris compound mycelium is obtained by inoculating Cordyceps militaris strains into a compound culture medium containing medicinal and edible functional groups and miscellaneous grain substrate.

[0009] Preferably, the tableting composition consists of the following raw materials in parts by weight: 100 parts of silkworm pupa powder, and excipients consisting of 6-11 parts of maltodextrin, 1.5-3.5 parts of magnesium stearate, and 3-6 parts of erythritol.

[0010] Preferably, the medicinal and edible functional group consists of the following ingredients in parts by weight: 10-16 parts Astragalus membranaceus, 7-13 parts Codonopsis pilosula, 12-20 parts red wolfberry, 4-9 parts Angelica sinensis, 8-14 parts Poria cocos, 6-11 parts Ophiopogon japonicus, 4-7 parts American ginseng, and 9-16 parts jujube; the mixed grain matrix consists of the following ingredients in parts by weight: 11-16 parts oats, 9-13 parts millet, 7-11 parts quinoa, and 5-9 parts yam powder.

[0011] Preferably, the preparation method of the compound mycelium dual-effect conditioning tablet composition is as follows: (1) Sterilize the compound culture medium, inoculate it with Cordyceps militaris, and culture it for 16-21 days at 23-27℃, 68-78% humidity and in the dark to obtain Cordyceps militaris compound mycelium suspension. The suspension contains cordycepin ≥0.12%, total polysaccharide ≥11%, and total saponin ≥2.2%.

[0012] (2) Sterilize the silkworm pupae, spray them with a compound mycelium suspension of Cordyceps militaris, and inoculate them with 9-13% of the weight of the silkworm pupae. Cultivate them for 21-26 days at 21-25℃, 62-72% humidity, and 0.35-0.55m³ / (h•m²) ventilation. The mycelium coverage of the stiff silkworm pupae is ≥98%, the crude protein content is ≥50%, the cordycepin content is ≥0.09%, and the total polysaccharide content is ≥8.5%. Freeze-dry the stiff silkworm pupae, crush and sieve them to obtain stiff silkworm pupae powder.

[0013] (3) Mix the silkworm pupa powder and excipients evenly, compress into tablets, sterilize and package to obtain a composite mycelium dual-effect conditioning tablet composition.

[0014] Preferably, the freeze-drying conditions in (3) are: pre-freezing at -40 to -35℃ for 3-4 hours, sublimation drying at 35-45℃, and desorption drying at 45-50℃, until the moisture content of the silkworm pupa powder is ≤7%, and then pulverized through a 100-120 mesh sieve.

[0015] Preferably, the molding pressure in (3) is 22-32 MPa, and the weight of the composite mycelium dual-effect conditioning tablet composition is 0.6-1 g.

[0016] Preferably, the application of the compound mycelium dual-effect conditioning tablet composition in food and medicine homology health products for the dual-effect synergistic use of immune regulation and qi and blood tonification.

[0017] (III) Beneficial Technical Effects: This invention provides a compound mycelium dual-effect conditioning tablet composition and its preparation method. Through the innovative process of "compound mycelium pre-conversion + directional infection with silkworm pupae," it achieves a dual-effect synergistic effect of immune regulation and qi and blood nourishment. It boasts high retention of active ingredients, good taste, and easy absorption, making it suitable for large-scale production and possessing significant market application prospects. Dual-effect synergy, significant efficacy: Through the process of "compound mycelium pre-conversion + directional infection with silkworm pupae," a dual-effect synergistic effect of immune regulation and qi and blood nourishment is achieved, with conditioning effects far exceeding those of single-function products.

[0018] High retention rate of active ingredients: Low-temperature culture and freeze-drying processes are used to avoid high temperature damage to heat-sensitive ingredients, and the retention rate of active ingredients is ≥90%.

[0019] Good taste and easy absorption: The silkworm pupa carrier has no odor, and the taste is adjusted with maltodextrin and erythritol. The powder is fine and the human body's absorption efficiency is increased by more than 20%.

[0020] Process adaptable to large-scale production: Parameters for each step are clearly defined and quantified, enabling mass production using existing equipment and ensuring stable quality between batches.

[0021] High resource value: Transforming low-value-added silkworm pupae into high-value-added health products improves resource utilization efficiency. Detailed Implementation

[0022] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. 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 of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0023] Example 1: (1) Mix 13 parts by weight of Astragalus membranaceus, 9 parts by weight of Codonopsis pilosula, 16 parts by weight of red wolfberry, 6 parts by weight of Angelica sinensis, 10 parts by weight of Poria cocos, 8 parts by weight of Ophiopogon japonicus, 5 parts by weight of American ginseng, 13 parts by weight of jujube, 14 parts by weight of oats, 11 parts by weight of millet, and 9 parts by weight of quinoa, grind them into powder, sieve them, add distilled water to adjust the moisture content to 63%, stir and mix, sterilize, cool, and obtain a sterilized composite culture medium.

[0024] (2) Sterilize the compound culture medium and inoculate it with Cordyceps militaris. Cultivate it at 25°C, 73% humidity and in the dark for 19 days to obtain Cordyceps militaris compound mycelial suspension.

[0025] (3) Sterilize the silkworm pupae, spray them with a suspension of Cordyceps militaris mycelium, and inoculate them with 11% of the weight of the silkworm pupae. Cultivate them for 23 days at 24℃, 68% humidity and 0.345m³ / (h•m²) ventilation. Freeze-dry the stiff silkworm pupae, pre-freeze at -38℃ for 3.5h, sublime dry at 40℃ and desorption dry at 48℃ until the moisture content of the stiff silkworm pupae powder is ≤7%. Crush the powder through a 120-mesh sieve to obtain stiff silkworm pupae powder.

[0026] (4) Mix 100 parts by weight of silkworm pupa powder, 9 parts by weight of maltodextrin, 2.5 parts by weight of magnesium stearate, and 4 parts by weight of erythritol at high speed for 20 minutes, press and mold under 28 MPa pressure, sterilize and package to obtain a composite mycelium dual-effect conditioning tablet composition.

[0027] The immunomodulatory efficacy was verified as follows: Objective: To verify the product's effect on enhancing the activity of immune cells.

[0028] Cells: Human peripheral blood mononuclear cells (PBMCs) and RAW264.7 macrophages.

[0029] Groups: control group, positive control group (LPS), and experimental group (product extract 0.1, 0.5, 1.0 mg / mL).

[0030] Detection indicators: NK cell killing activity (flow cytometry), macrophage phagocytic ability of fluorescent microspheres, and lymphocyte proliferation (MTT assay).

[0031] Results: NK cell activity increased 2.4-fold (1.0 mg / mL). Macrophage phagocytosis rate increased by 42%. T lymphocyte proliferation rate increased by 38%.

[0032] Conclusion: The product has a significant immunomodulatory effect.

[0033] The efficacy of this product in nourishing Qi and blood is verified as follows: Objective: To verify the product's effect on improving a model of Qi and blood deficiency.

[0034] Model: Cyclophosphamide-induced deficiency of qi and blood in rats.

[0035] Grouping: normal group, model group, positive control group (compound donkey-hide gelatin oral liquid), and experimental group (low, medium, and high doses).

[0036] Test indicators: hemoglobin, serum ferritin, and exhaustion swimming time.

[0037] Results: Hemoglobin increased by 18%, serum ferritin increased by 22%, and the time to exhaustion increased by 32%.

[0038] Conclusion: The product has a significant effect on nourishing qi and blood.

[0039] The consistency verification of active ingredient content between batches is as follows: Objective: To verify the content of key active ingredients in the product and batch-to-batch consistency.

[0040] Detection indicators: Cordycepin (HPLC), total polysaccharides (phenol-sulfuric acid method), total saponins (colorimetric method).

[0041] Sampling: 3 independent production batches, each batch was tested 3 times.

[0042] Results (mean ± RSD): Cordycepin: 0.112% ± 3.2%. Total polysaccharides: 9.05% ± 2.8%. Total saponins: 2.48% ± 3.5%.

[0043] Conclusion: The content of active ingredients in the product is stable, with batch-to-batch variation ≤5%.

[0044] The optimization and verification of key parameters in the preparation process are as follows: Objective: To optimize the inoculation process parameters.

[0045] Orthogonal experimental design: inoculum size (9%, 11%, 13%), temperature (21℃, 23℃, 25℃), humidity (62%, 67%, 72%).

[0046] Indicators: mycelial coverage and infection cycle.

[0047] Results: Optimal combination: inoculum 11%, temperature 24℃, humidity 68%.

[0048] Mycelial coverage: 99.2%. Infection cycle: 23 days.

[0049] Conclusion: The infection efficiency was significantly improved after the process parameters were optimized.

[0050] Stability and shelf life verification are as follows: Objective: To evaluate the stability of the product under accelerated storage conditions.

[0051] Accelerated testing conditions: 40℃, 75%RH, for 6 months.

[0052] Testing indicators: cordycepin, total polysaccharide content, tablet hardness, and disintegration time.

[0053] Results: Cordycepin retention rate: 92.5%. Total polysaccharide retention rate: 91.8%. Moisture content: 6.2%.

[0054] Disintegration time: ≤15 minutes.

[0055] Conclusion: The product has good stability, and the predicted shelf life is ≥24 months.

[0056] Safety assessment (acute and subacute toxicity tests) is as follows: Objective: To verify the food safety of the product.

[0057] Acute toxicity: Rats were administered a single dose of 5000 mg / kg by gavage and observed for 14 days.

[0058] Subacute toxicity: Rats were administered 10, 50, or 100 times the recommended daily dose by gavage for 28 days.

[0059] Results: LD > 5000 mg / kg, with no death or symptoms of poisoning. No significant abnormalities were found in blood routine tests, liver and kidney function tests, or organ indices.

[0060] Conclusion: The product has good safety.

[0061] The bioavailability comparison study is as follows: Objective: To verify the effect of silkworm pupa carrier on the bioavailability of active ingredients.

[0062] Grouping: Experimental group (this product), control group (ordinary silkworm pupa-based product).

[0063] Detection indicators: area under the cordycepin plasma concentration-time curve (AUC), peak concentration (Cmax), and time to peak concentration (Tmax).

[0064] Results: AUC (experimental group vs. control group): increased by 25%. Cmax: increased by 22%. Tmax: advanced by 1.2 hours.

[0065] Conclusion: The silkworm pupa carrier significantly improves the absorption efficiency of active ingredients.

[0066] Synergistic efficacy verification (comparison of dual-effect effects) is as follows: Objective: To verify the synergistic effect of immune regulation and qi and blood tonification.

[0067] Grouping: Blank group, single immune group (only Cordyceps militaris mycelium), single blood and qi group (only food and medicine homology ingredients), experimental group (all products).

[0068] Detection indicators: serum antibody (IgG), hemoglobin.

[0069] Results: Improvement rate of immune indicators: 28% higher in the experimental group compared to the single immune group. Improvement rate of Qi and blood indicators: 25% higher in the experimental group compared to the single Qi and blood group. Synergy index: 1.31 (>1).

[0070] Conclusion: The product has a significant synergistic effect.

[0071] Consumer acceptance and taste evaluation are as follows: Objective: To assess the market acceptance of the product.

[0072] Sample: 100 sub-healthy adults.

[0073] Evaluation method: blind taste test + questionnaire survey (5-point scale).

[0074] Results: Taste score: 4.3. Aroma score: 4.1. Dietability score: 4.5. Overall acceptability: 86%.

[0075] Conclusion: The product tastes good and has high market acceptance.

[0076] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A compound mycelium dual-effect conditioning tablet composition, characterized in that, The composite mycelium dual-effect conditioning tablet composition is made by mixing and compressing silkworm pupa pupa powder with food-grade excipients; The stiffened pupae of the silkworm were obtained by infecting silkworm pupae with a suspension of Cordyceps militaris mycelium and then culturing them under aseptic conditions until the pupae became stiff and their interiors were covered with mycelium. The Cordyceps militaris composite mycelium is cultured by inoculating Cordyceps militaris strains into a composite culture medium containing medicinal and edible functional groups and miscellaneous grain substrate.

2. The composite mycelium dual-effect conditioning tablet composition according to claim 1, characterized in that, The tableting composition consists of the following raw materials in parts by weight: 100 parts of silkworm pupa powder, and excipients consisting of 6-11 parts of maltodextrin, 1.5-3.5 parts of magnesium stearate, and 3-6 parts of erythritol.

3. The composite mycelium dual-effect conditioning tablet composition according to claim 1, characterized in that, The medicinal and edible functional group consists of the following ingredients in parts by weight: 10-16 parts Astragalus membranaceus, 7-13 parts Codonopsis pilosula, 12-20 parts red wolfberry, 4-9 parts Angelica sinensis, 8-14 parts Poria cocos, 6-11 parts Ophiopogon japonicus, 4-7 parts American ginseng, and 9-16 parts jujube; the mixed grain matrix consists of the following ingredients in parts by weight: 11-16 parts oats, 9-13 parts millet, 7-11 parts quinoa, and 5-9 parts yam powder.

4. The composite mycelium dual-effect conditioning tablet composition according to claim 1, characterized in that, The preparation conditions for the Cordyceps militaris compound mycelial suspension are as follows: sterilize the compound culture medium, inoculate with Cordyceps militaris strain, and culture for 16-21 days at 23-27℃, 68-78% humidity, and in the dark to obtain the Cordyceps militaris compound mycelial suspension.

5. The composite mycelium dual-effect conditioning tablet composition according to claim 1, characterized in that, The inoculation conditions for the silkworm pupae are as follows: sterilize the silkworm pupae, spray with a suspension of Cordyceps militaris mycelium, and inoculate with 9-13% of the weight of the silkworm pupae. Cultivate for 21-26 days at 21-25℃, humidity 62-72%, and ventilation rate 0.35-0.55 m³ / (h•m²). Freeze-dry the stiffened silkworm pupae, pulverize and sieve to obtain stiffened silkworm pupae powder.

6. The composite mycelium dual-effect conditioning tablet composition according to claim 1, characterized in that, The freeze-drying conditions are as follows: pre-freezing at -40 to -35℃ for 3-4 hours, sublimation drying at 35-45℃, and desorption drying at 45-50℃ until the moisture content of the silkworm pupa powder is ≤7%, and then pulverized through a 100-120 mesh sieve.

7. A method for preparing a compound mycelium dual-effect conditioning tablet composition as described in any one of claims 1-6, characterized in that, The preparation method is as follows: mix the silkworm pupa powder and excipients evenly, compress into tablets, sterilize and package to obtain a composite mycelium dual-effect conditioning tablet composition.

8. The method for preparing the composite mycelium dual-effect conditioning tablet composition according to claim 7, characterized in that, The tableting pressure is 22-32 MPa, and the weight of the compound mycelium dual-effect conditioning tableting composition is 0.6-1 g.

9. The application of a compound mycelium dual-effect conditioning tablet composition obtained by the preparation method according to any one of claims 7 in a food-medicine homology health product for the dual-effect synergistic use of immune regulation and qi and blood tonification.