Preparation process and application of ergothioneine, oyster, seaweed, selfheal, fritillaria, astragalus membranaceus and salvia miltiorrhiza compound preparation for dissipating nodules

Abstract

The invention belongs to the field of biological medicine, and particularly discloses a preparation process and application of an ergothioneine, oyster, seaweed, selfheal, thunberg fritillary bulb, astragalus membranaceus, salvia miltiorrhiza compound preparation for dissipating nodules, and the compound preparation is composed of ergothioneine, oyster, seaweed, selfheal, thunberg fritillary bulb, astragalus membranaceus, salvia miltiorrhiza, liquorice, edible tulip, curcuma zedoary and ganoderma lucidum spore powder. The core innovation lies in that a pseudobulbus cremastrae seu pleiones-curcuma zedoary medicine pair is introduced into a nodule treatment formula for the first time, the fibrosis process is blocked by inhibiting alpha-SMA expression, ganoderma lucidum spore powder-ergothioneine antioxidation synergy is achieved, and the bioavailability is improved by 6.6 times through embedding of ergothioneine-beta cyclodextrin. The preparation can be prepared into enteric capsules, pills and the like, the reduction rate of thyroid nodules can reach 83.3% when the dosage of levothyroxine is reduced by half in clinical combination, and the preparation has a good pharmaceutical prospect.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically discloses a preparation process and application of a compound preparation of ergothioneine, oyster, seaweed, prunella vulgaris, fritillaria cirrhosa, astragalus membranaceus, and salvia miltiorrhiza for dissipating nodules. Background Art

[0002] Nodular lesions (including thyroid nodules, breast nodules, and lung nodules, etc.) have become common clinical symptoms, and their pathogenesis is closely related to local tissue abnormal hyperplasia, continuous stimulation of the inflammatory microenvironment, and immune regulation imbalance. The current mainstream treatment options have significant limitations: Western medical therapies mainly rely on surgical resection or chemical drug intervention. Surgery is applicable to nodules with obvious malignant signs or compressive symptoms, but there are problems such as large trauma, postoperative functional defects (such as the permanent hypothyroidism incidence rate after thyroid surgery can reach 15%-30%), and it is difficult to eradicate multiple nodules. Drug treatment mostly uses hormone regulators (such as tamoxifen for breast nodules) or thyroid hormone suppression therapy (such as levothyroxine sodium for thyroid nodules). Although it can control nodule growth in the short term, long-term application is likely to cause endocrine disorders (manifested as hot flashes, menstrual disorders), abnormal liver function indicators (the ALT / AST elevation rate > 20%), and drug dependence, and the recurrence rate within 6 months after drug withdrawal is as high as 35%-40%. Although a large number of softening and resolving hard masses prescriptions (such as classic prescriptions like Haizao Yuhu Decoction and Xiaoluo Pills) have been accumulated in the field of traditional Chinese medicine treatment, two common defects have emerged in modern clinical applications: First, the traditional formula theory focuses on macroscopic "resolving phlegm and removing stasis" while ignoring the microscopic anti-hyperplasia mechanism. The compatibility of mineral herbs (such as oyster and pumice) and plant herbs (such as prunella vulgaris and fritillaria cirrhosa) lacks the support of a precise pharmacodynamic substance basis, resulting in low penetration efficiency for fibrotic nodules (especially collagen deposition type). Clinical studies show that the effective rate of existing Chinese patent medicines in treating TI-RADS class 3 thyroid nodules is only 42.7%-48.3%. Second, the preparation process is difficult to balance the stability of multiple active ingredients, showing three technical bottlenecks: (1) The ionized calcium dissolution rate of mineral drugs rich in calcium salts such as oyster after conventional water decoction is less than 18%, and the bioavailability is low; (2) The degradation rate of thermosensitive components (such as fritillaria cirrhosa alkaloids and tanshinones) during high-temperature extraction exceeds 35%; (3) The inactivation rate of trace highly active substances (such as ergothioneine) during oral delivery due to gastric acid damage exceeds 90%. More notably, the recently emerging "combination of traditional Chinese and Western medicines" model often leads to pharmacodynamic antagonism or superposition of toxic and side effects due to the lack of pharmacological synergistic design. A typical manifestation is that when tamoxifen is combined with sterol-containing traditional Chinese medicines, the targeting efficiency is reduced due to competitive binding to estrogen receptor β subtype. The above defects together make it difficult for the existing technology to meet the control requirements of progressive nodules, especially lacking effective means for postoperative recurrence prevention and drug-resistant nodules. In view of this industry technical dilemma, there is an urgent need to develop an innovative treatment plan with a scientific compatibility system and a targeted delivery process. Summary of the Invention

[0003] In view of the above problems, the present invention discloses a preparation process and application of a compound preparation of ergothioneine, oyster, seaweed, selfheal spike, fritillary bulb, astragalus root, and salvia root for dissipating nodules.

[0004] The object of the present invention is achieved through the following technical solutions.

[0005] A traditional Chinese medicine composition for dissipating nodules is made from the following raw materials in parts by weight:

[0006] Ergothioneine 0.01 - 0.05 parts, oyster 30 - 40 parts, seaweed 10 - 20 parts, selfheal spike 10 - 20 parts, fritillary bulb 8 - 15 parts, astragalus root 25 - 35 parts, salvia root 8 - 15 parts, liquorice root 2 - 5 parts, pseudobulb of appendiculate thunbergia 8 - 15 parts, zedoary 6 - 12 parts, ganoderma lucidum spore powder 3 - 10 parts.

[0007] Furthermore, the above traditional Chinese medicine composition is made from the following raw materials in parts by weight:

[0008] Ergothioneine 0.03 parts, oyster 35 parts, seaweed 15 parts, selfheal spike 15 parts, fritillary bulb 10 parts, astragalus root 30 parts, salvia root 10 parts, liquorice root 3 parts, pseudobulb of appendiculate thunbergia 12 parts, zedoary 10 parts, ganoderma lucidum spore powder 6 parts.

[0009] The present invention also discloses a preparation method of the above traditional Chinese medicine composition, including the following steps:

[0010] (1) Ultrafine pulverization of oyster: After calcining the oyster, it is ultrafinely pulverized at -40°C to D90 ≤ 15μm;

[0011] (2) Encapsulation of ergothioneine: Ergothioneine and β-cyclodextrin are subjected to molecular encapsulation in a ratio of 1:8;

[0012] (3) Stepwise extraction:

[0013] a. Seaweed, selfheal spike, and pseudobulb of appendiculate thunbergia are extracted twice with 8 times the amount of pH 8.5 alkaline water at 80°C;

[0014] b. Astragalus root, salvia root, and zedoary are refluxed and extracted three times with 70% ethanol;

[0015] c. Fritillary bulb and liquorice root are extracted by CO2 supercritical extraction;

[0016] (4) Combine the extracts with oyster powder, encapsulated ergothioneine, and ganoderma lucidum spore powder and mix evenly.

[0017] Furthermore, in the above preparation method, the alkaline water in step (3)a is 0.1% sodium bicarbonate solution, and the supercritical conditions in step (3)c are 45°C and 35 MPa.

[0018] The present invention also discloses a pharmaceutical preparation, which comprises the above-mentioned traditional Chinese medicine composition and pharmaceutical excipients, and the preparation forms include capsules, tablets, pills or granules.

[0019] Furthermore, for the above-mentioned preparation, the capsules adopt enteric-coated capsules, the tablets contain microcrystalline cellulose and croscarmellose sodium, and the granules adopt the fluidized bed granulation process.

[0020] The present invention also discloses the application of the above-mentioned medicine composition in the preparation of a medicine for dissipating human nodules, and it is characterized in that the nodules are thyroid nodules, breast nodules or lung nodules.

[0021] Furthermore, in the above-mentioned application, the traditional Chinese medicine composition is combined with a chemical medicine:

[0022] For thyroid nodules: combined with levothyroxine tablets, and the dosage is reduced to 50-70% of the conventional dosage

[0023] For breast nodules: combined with tamoxifen, and the medication cycle is shortened by 30-40%.

[0024] The present invention also discloses a combined traditional Chinese and western medicine set for treating nodules, and the set contains the above-mentioned traditional Chinese medicine composition and a chemical medicine, and the chemical medicine is levothyroxine tablets or tamoxifen.

[0025] Compared with the existing technology, the present invention has the following advantages and beneficial effects:

[0026] First, the compatibility of monarch, minister, assistant and envoy is scientific

[0027] In this compound prescription, oyster and seaweed are used as the monarch drugs, giving full play to the effect of softening hardness and dissipating nodules with salty taste. Among them, the oyster component can target and improve the calcified lesions of nodules, and the active substances of seaweed can effectively block the key signal pathways of fibrosis; Prunella vulgaris and Cremastra appendiculata are used as the minister drugs, playing an anti-hyperplasia role by inhibiting angiogenesis-related factors and interfering with the proliferation mechanism of fibroblasts; Astragalus membranaceus and Ganoderma lucidum spore powder are used as the assistant drugs, forming a two-way immunomodulatory effect and significantly enhancing the activity of immune cells; Salvia miltiorrhiza and Curcuma zedoaria are used as the envoy drugs, strengthening the overall curative effect through enhancing drug penetration and inducing apoptosis mechanism. The whole prescription constructs a multi-link treatment network of "softening lesions - inhibiting hyperplasia - regulating immunity".

[0028] Second, the innovation advantages of the core technology

[0029] Synergistic breakthrough of formula: Through the anti-fibrotic dual-target effects of Cremastra appendiculata and Curcuma zedoaria, synergistic effects are achieved from different mechanism levels, significantly reducing the expression of fibrosis markers; Ganoderma lucidum spore powder and antioxidant components form a composite protection system, significantly enhancing the body's antioxidant capacity. Process-directed synergistic enhancement: The segmented extraction technology is adopted to maximize the retention of active ingredients, the ultrafine comminution process improves the absorption efficiency of mineral drugs, and the special embedding technology solves the stability problem of core ingredients. Significant clinical value: The combined treatment plan adjusts the activity of key enzymes in thyroid hormone metabolism, achieving better control of hormone levels while reducing the dosage of western medicine. Advantages in commercial transformation: The preparation process effectively solves the stability problem of traditional Chinese medicine components, ensuring the quality stability of products during the shelf life and strong repeatability of clinical efficacy. Description of the Drawings

[0030] Figure 1 Comparison of the proliferation inhibition rate (%) in the in vitro anti-proliferative activity test;

[0031] Figure 2 Comparison of the down-regulation of α-SMA (%) in the in vitro anti-proliferative activity test. Detailed Description of the Invention

[0032] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below. However, it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the scope of the present invention. In addition, in the following description, the description of well-known structures and technologies is omitted to avoid unnecessarily confusing the concepts of the present invention. All raw materials in the embodiments of the present invention can be obtained through commercial channels.

[0033] It should be noted that, without conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present invention will be described in detail below in conjunction with the embodiments.

[0034] Example 1

[0035] Basic compound decoction extract

[0036] Raw material ratio (parts by weight):

[0037] L-ergothioneine 0.03 parts, oyster 35 parts, seaweed 15 parts, Prunella vulgaris 15 parts, Fritillaria thunbergii 10 parts, Astragalus membranaceus 30 parts, Salvia miltiorrhiza 10 parts, Glycyrrhiza uralensis 3 parts, Cremastra appendiculata 12 parts, Curcuma zedoaria 10 parts, Ganoderma lucidum spore powder 6 parts. (Except for L-ergothioneine and oyster, all are in the form of traditional Chinese medicine decoction pieces)

[0038] Preparation method:

[0039] (1) Pretreatment of oyster: The oyster is calcined (calcined with strong fire at 200°C until it turns bluish-gray), and ultrafinely comminuted at -40°C to D90 = 12 μm;

[0040] (2)Erythritol Thione Entrapment: Erythritol thione and β-cyclodextrin are mixed at a mass ratio of 1:8, magnetically stirred in an aqueous solution at 40 °C for 2 hours, and then freeze-dried;

[0041] (3)Stepwise Extraction:

[0042] a. Seaweed, Prunella vulgaris, and Cremastra appendiculata are added with 8 times the amount of 0.1% NaHCO3 solution (pH 8.5), and extracted at 80 °C for 2 times, 1.5 hours each time;

[0043] b. Astragalus membranaceus, Salvia miltiorrhiza, and Curcuma zedoaria are added with 10 times the amount of 70% ethanol and refluxed for extraction 3 times, 1 hour each time;

[0044] c. Fritillaria thunbergii and Glycyrrhiza uralensis are extracted by CO2 supercritical extraction (45 °C, 35 MPa, 2 hours);

[0045] (4)All the extracts are combined, concentrated under reduced pressure to a relative density of 1.25 (60 °C), and oyster powder, entrapped erythritol thione, and ganoderma spore powder are added and mixed evenly to obtain a decoction extract.

[0046] Example 2

[0047] Optimized Ratio Capsules

[0048] Raw Material Ratio:

[0049] Erythritol Thione 0.05 parts, Oyster 40 parts, Seaweed 20 parts, Prunella vulgaris 20 parts, Fritillaria thunbergii 15 parts, Astragalus membranaceus 35 parts, Salvia miltiorrhiza 15 parts, Glycyrrhiza uralensis 5 parts, Cremastra appendiculata 15 parts, Curcuma zedoaria 12 parts, Ganoderma Spore Powder 10 parts

[0050] Preparation Method:

[0051] Same as steps 1 - 3 of Example 1;

[0052] Concentrated to a relative density of 1.30, and microcrystalline cellulose (20%) and magnesium stearate (0.5%) are added;

[0053] Enteric coating with hydroxypropyl methylcellulose phthalate (HPMCP), and filled into capsules.

[0054] Example 3

[0055] Tablets

[0056] Raw Material Ratio:

[0057] Erythritol Thione 0.01 parts, Oyster 30 parts, Seaweed 10 parts, Prunella vulgaris 10 parts, Fritillaria thunbergii 8 parts, Astragalus membranaceus 25 parts, Salvia miltiorrhiza 8 parts, Glycyrrhiza uralensis 2 parts, Cremastra appendiculata 8 parts, Curcuma zedoaria 6 parts, Ganoderma Spore Powder 3 parts

[0058] Preparation Method:

[0059] Same as the fractional extraction process in Example 1;

[0060] After concentration and drying, mix with sodium carboxymethylcellulose cross-linked (5%) and microcrystalline cellulose (25%);

[0061] Dry granulation and tabletting, tablet weight 0.5 g.

[0062] Example 4

[0063] Dan agent

[0064] The raw material ratio is the same as that in Example 1

[0065] Preparation method:

[0066] Mix and pulverize all medicinal materials and pass through a 100-mesh sieve;

[0067] Adopt the method of ascending and descending Dan: Place the medicinal powder in a crucible, calcine at 250 °C for 3 hours, take out and grind, adjust the moisture to 10%, and make pills.

[0068] Example 5

[0069] Granules

[0070] The raw material ratio is the same as that in Example 2

[0071] Preparation method:

[0072] Same as the extraction process in Example 1;

[0073] Concentrate to a relative density of 1.20, add lactose (30%) and β-cyclodextrin (5%);

[0074] Fluidized bed granulation (inlet air temperature 60 °C) to obtain granules.

[0075] Comparative Examples 1-5

[0076] Comparative Examples 1-5 are configured as shown in Table 1 below.

[0077]

[0078] Test Example 1

[0079] In vitro anti-proliferative activity

[0080] Method:

[0081] Cell model: Human thyroid follicular epithelial cell (Nthy-ori 3-1) induced fibrosis nodule model (stimulated by TGF-β1)

[0082] Grouping: Example 1 group (100 μg / mL), D1 group (lacking Cremastra appendiculata / Rhizoma Zedoariae), D5 group (lacking Ganoderma lucidum spores)

[0083] Detection: CCK-8 was used to measure the proliferation rate, and Western blot was used to measure the expression of α-SMA.

[0084] Results (Table 2) and Figure 1-2 :

[0085]

[0086] *p < 0.01 vs Example 1

[0087] Conclusion: Colchicum (colchicine) and Curcuma zedoaria (β-elemene) synergistically inhibit the activation of myofibroblasts, and Ganoderma lucidum spore powder enhances immune regulation.

[0088] Test Example 2

[0089] In vivo nodule dissipation experiment

[0090] Method:

[0091] Model: SD rat thyroid nodule model (intragastric administration of propylthiouracil + local injection of TGF-β1 for 4 weeks)

[0092] Grouping (n = 10): Example 2 group (1 g / kg / d), D2 group (non-fractionated extraction), D3 group (ordinary oyster powder)

[0093] Evaluation: Ultrasonography was used to measure the nodule volume, ELISA was used to measure serum TgAb, and histological HE staining was performed.

[0094] The results are shown in Table 3

[0095]

[0096] Mechanism: Fractionated extraction retains the alkaloids of Fritillaria (the content measured by HPLC increased by 42%), and ultrafine grinding of oyster promotes calcium ions to inhibit nodule calcification.

[0097] Test Example 3

[0098] Synergistic effect with levothyroxine

[0099] Method:

[0100] Clinical samples: 60 patients with thyroid nodules (TI-RADS category 3)

[0101] Grouping:

[0102] Group A: Sodium levothyroxine (50 μg / day)

[0103] Group B: Tablets of Example 3 (4 tablets / day)

[0104] Group C: Combined group (levothyroxine 25 μg + tablets of Example 3, 4 tablets)

[0105] Treatment course: 12 weeks, evaluating nodule volume and TSH level

[0106] The results are shown in Table 4

[0107]

[0108] *p < 0.05 vs single-drug group

[0109] Conclusion: The compound preparation regulates the hypothalamic-pituitary axis through astragalus polysaccharide, reducing the dosage of levothyroxine by half and enhancing its efficacy.

[0110] Test Example 4

[0111] (Bioavailability evaluation)

[0112] Method:

[0113] Rats were gavaged with the preparation of Example 1 (containing 0.5 mg / kg of ergothioneine)

[0114] Control group: Group D4 (without embedded ergothioneine)

[0115] The plasma concentration of ergothioneine was determined by HPLC-MS (m / z 197 → 179)

[0116] Results:

[0117] For the embedded group, Cmax = 28.7 ng / mL, AUC0-24h = 312 h·ng / mL

[0118] For Group D4, Cmax = 5.2 ng / mL, AUC0-24h = 47 h·ng / mL

[0119] Key data: The embedding process increased the bioavailability by 6.6 times, and Tmax was shortened from 4 h to 1.5 h.

[0120] Summary of the examples

[0121] Innovation in formula: The combination of pleione bulbocodioides and zedoary turmeric can resist fibrosis (α-SMA decreased by 72.5%), and the combination of ganoderma lucidum spores and ergothioneine has antioxidant synergy.

[0122] Breakthrough in technology:

[0123] The yield of fritillaria alkaloids increased by 42% through segmented extraction

[0124] The dissolution rate of calcium ions in ultrafinely ground oyster increased by 67%

[0125] β-cyclodextrin embedding of ergothioneine increased the bioavailability by 6.6 times

[0126] Clinical value:

[0127] When the dosage of levothyroxine in combination is halved, the nodule shrinkage rate increases to 83.3%

[0128] Note: All experimental data were statistically verified by SPSS 26.0 (*p < 0.05, **p < 0.01), and the animal experiments followed the ARRIVE guidelines.

[0129] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Therefore, based on the innovative concept of the present invention, any changes and modifications made to the embodiments described herein, or equivalent structural or equivalent process transformations made using the content of the specification of the present invention, and directly or indirectly applying the above technical solutions to other related technical fields are all included in the protection scope of the present invention patent.

Claims

1. A traditional Chinese medicine composition for dissipating nodules, characterized in that, It is made from the following raw materials by weight parts: Ergothioneine 0.01 - 0.05 parts, oyster 30 - 40 parts, seaweed 10 - 20 parts, Prunella vulgaris 10 - 20 parts, Fritillaria thunbergii 8 - 15 parts, Astragalus membranaceus 25 - 35 parts, Salvia miltiorrhiza 8 - 15 parts, Glycyrrhiza glabra 2 - 5 parts, Cremastra appendiculata 8 - 15 parts, Curcuma zedoaria 6 - 12 parts, Ganoderma lucidum spore powder 3 - 10 parts.

2. The traditional Chinese medicine composition according to claim 1, wherein It is made from the following raw materials by weight parts: Ergothioneine 0.03 parts, oyster 35 parts, seaweed 15 parts, Prunella vulgaris 15 parts, Fritillaria thunbergii 10 parts, Astragalus membranaceus 30 parts, Salvia miltiorrhiza 10 parts, Glycyrrhiza glabra 3 parts, Cremastra appendiculata 12 parts, Curcuma zedoaria 10 parts, Ganoderma lucidum spore powder 6 parts.

3. The preparation method of the traditional Chinese medicine composition according to claim 1 or 2, characterized in that, It includes the following steps: (1) Ultrafine pulverization of oyster: After calcining the oyster, it is ultrafinely pulverized at -40°C to D90 ≤ 15μm. (2) Encapsulation of ergothioneine: Ergothioneine and β-cyclodextrin are subjected to molecular encapsulation in a ratio of 1:

8. (3) Fractional extraction: a. Seaweed, Prunella vulgaris, and Cremastra appendiculata are extracted twice with 8 times the amount of pH 8.5 alkaline water at 80°C. b. Astragalus membranaceus, Salvia miltiorrhiza, and Curcuma zedoaria are reflux-extracted three times with 70% ethanol. c. Fritillaria thunbergii and Glycyrrhiza glabra are extracted by CO2 supercritical extraction. (4) Combine the extracts with oyster powder, encapsulated ergothioneine, and Ganoderma lucidum spore powder and mix evenly.

4. The preparation method according to claim 3, characterized in that, In step (3)a, the alkaline water is 0.1% sodium bicarbonate solution, and the supercritical conditions in step (3)c are 45°C and 35 MPa.

5. A pharmaceutical preparation comprising the traditional Chinese medicine composition according to claim 1 or 2 and a pharmaceutical excipient, characterized in that The preparation forms include capsules, tablets, pills, or granules.

6. The preparation according to claim 5, wherein: The capsules use enteric-coated capsules, the tablets contain microcrystalline cellulose and cross-linked carboxymethylcellulose sodium, The granules use the fluidized bed granulation process.

7. A combined traditional Chinese and Western medicine set for treating nodules, characterized in that, The set contains the traditional Chinese medicine composition according to claim 1 or 2 and a chemical drug, and the chemical drug is levothyroxine sodium tablets or tamoxifen.

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