Pulmonary nodule targeting food and medicine composition based on metabonomics and application

Abstract

The invention provides a pulmonary nodule targeting food and medicine homologous composition based on metabonomics screening and application thereof. By analyzing a pulmonary nodule specific metabolic profile, active components, namely platycodin D, mogroside V, oyster active peptide and fermented cordycepin, which can cooperatively regulate and control an arachidonic acid inflammation pathway, a glutathione antioxidant pathway and a tricarboxylic acid cycle energy pathway are reversely screened out. Animal experiments prove that the composition can reduce the volume of pulmonary nodules by 58.3%, human body application shows that the nodules of 76.7% of subjects are reduced, and liver and kidney toxicity is avoided. The composition can be prepared into tablets, capsules and other dosage forms, can be used as functional food or adjuvant therapy drugs for high risk groups of pulmonary nodules, and realizes accurate nutrition intervention.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine and food science, and particularly relates to a lung nodule targeting food-drug composition based on metabolomics and application. BACKGROUND

[0002] Non-surgical intervention for lung nodules faces a double bottleneck: on the one hand, the western medicine treatment scheme has the risk of liver and kidney toxicity, and lacks precise prevention and control means for small nodules; on the other hand, traditional food-drug homologous products are mostly based on empirical formula, and the mechanism is unclear, and the metabolic pathway cannot be targeted. Although metabolomics technology can reveal disease biomarkers, its application in the development of lung nodule targeting food is still blank, and it is urgent to transform the metabolic regulation mechanism into a quantifiable efficacy formula design.

[0003] Existing food-drug homologous compositions are mostly focused on the general function of moistening the lungs and relieving cough, and lack the ability to synergistically intervene in specific metabolic abnormalities of lung nodules (such as arachidonic acid inflammatory storm and glutathione depletion). The complexity of components and the ambiguity of efficacy make it difficult for products to meet the modern demand for precision health, and the application of single components cannot cover the pathological network of multi-pathway interaction. Therefore, it is urgent to develop a lung nodule targeting composition based on metabolomics reverse screening, with multiple target points and food safety. SUMMARY

[0004] This section aims to summarize some aspects of the embodiments of the present application and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the abstract and title of the specification to avoid obscuring the purpose of this section, abstract and title, and such simplifications or omissions cannot be used to limit the scope of the present application.

[0005] In view of the problems in the prior art, the present inventors have proposed the present application.

[0006] Therefore, the purpose of the present application is to overcome the deficiencies in the prior art and provide a lung nodule targeting food-drug composition based on metabolomics and application.

[0007] To solve the above technical problems, the present application provides the following technical scheme: a lung nodule targeting food-drug composition based on metabolomics, comprising the following active ingredients and weight parts: Platycodon saponin D 10-20 parts (regulating COX-2 mediated arachidonic acid metabolic pathway); Mogroside V 15-25 parts (inhibiting HMGB1 / TLR4 inflammatory signaling); Oyster active peptide with a molecular weight of less than 1000 Da 8-15 parts (activating glutathione synthetase GCLC); Fermented cordycepin 5-12 parts (repairing the activity of the rate-limiting enzyme IDH in the tricarboxylic acid cycle); The four components synergistically regulate ≥3 lung nodule-related differential metabolic pathways.

[0008] As a preferred scheme of the lung nodule targeting food-drug composition based on metabolomics, wherein: the platycodon saponin D is derived from the ethanol extract of the root of platycodon grandiflorum, and the purity is ≥40% after purification by macroporous resin, and the retention time of the HPLC characteristic peak is 12.6±0.3 min (C18 column, acetonitrile-water gradient elution).

[0009] As a preferred scheme of the lung nodule targeting food-drug composition based on metabolomics, wherein: the preparation of the active peptide of oyster includes hydrolysis of oyster meat at 35°C for 4h by trypsin, ultrafiltration of components with a molecular weight ≤1000 Da, and spray drying with a peptide content ≥80%.

[0010] As a preferred scheme of the lung nodule targeting food-drug composition based on metabolomics, wherein: the fermented cordycepin is derived from the liquid fermentation mycelium of the Cs-4 strain of Paecilomyces hepiali, and the adenosine content is ≥0.1%, and the ergosterol content is ≥0.05%.

[0011] A preparation method of a lung nodule targeting food-drug composition based on metabolomics, comprising the following steps: (a) mixing the active ingredients with the filler at a weight ratio of 1:1-3, wherein the filler is microcrystalline cellulose and malt dextrin; (b) granulating by adding a binder of 10% polyvinylpyrrolidone ethanol solution, and the inlet air temperature is 60±5°C; (c) fluidized bed drying to a moisture content ≤5%, and the whole particle size is 18-24 mesh; (d) tabletting or capsule filling.

[0012] As a preferred scheme of the preparation method of the lung nodule targeting food-drug composition based on metabolomics, wherein: the bulk density of the granulation in step (b) is 0.45-0.55 g / cm³, and the moisture content of the granules after drying in step (c) is ≤4.5%.

[0013] As a preferred scheme of the application of the lung nodule targeting food-drug composition based on metabolomics in the preparation of a functional food for preventing or relieving lung nodules, wherein: the daily intake provides 0.3-1 g of active ingredients, and the dosage form includes tablets, capsules, and oral liquids.

[0014] As a preferred application of the lung nodule-targeting food-drug composition based on metabolomics in the preparation of an auxiliary treatment drug for lung nodules, the drug is combined with a tyrosine kinase inhibitor, and the combination ratio (weight ratio) is 1:0.1-5.

[0015] As a preferred application of the lung nodule-targeting food-drug composition based on metabolomics in the preparation of an auxiliary treatment drug for lung nodules, the tyrosine kinase inhibitor is gefitinib or erlotinib.

[0016] As a preferred application of the lung nodule-targeting food-drug composition based on metabolomics in the regulation of serum metabolic markers, the metabolic markers include arachidonic acid (decrease ≥40%) and glutathione (increase ≥30%).

[0017] The composition is screened by metabolomics, and precise matching of food-drug homologous components and lung nodule-specific metabolic pathways is achieved: platycodon saponin D blocks the synthesis of inflammatory mediators, mogroside V inhibits excessive immune activation, oyster peptide reestablishes antioxidant defense, and cordycepin corrects energy metabolism imbalance. Four components synergistically cover the "inflammation-oxidation-energy" regulation network, breaking through the limitations of traditional prescriptions in a single pathway; the composition has both food safety and clear mechanism: all raw materials are substances included in the directory of food and medicine homologues, without the risk of synthetic drug toxicity; at the same time, the efficacy targets of each component are verified by metabolic network analysis and molecular docking, avoiding the blindness of empirical prescriptions. The final product can be integrated into the daily diet, providing a new non-drug intervention path for lung nodule high-risk groups. DETAILED DESCRIPTION

[0018] To make the above objectives, features and advantages of the present application more apparent and comprehensible, the specific embodiments of the present application will be described in detail below with reference to the embodiments of the present application.

[0019] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present application, but the present application can also be implemented in other ways different from the description, and those skilled in the art can make similar generalizations without departing from the connotation of the present application, so the present application is not limited to the specific embodiments disclosed below.

[0020] Secondly, "one embodiment" or "embodiment" referred to herein means that a specific feature, structure or characteristic can be included in at least one implementation of the present application. "In one embodiment" appearing in different places in the specification does not mean the same embodiment, nor is it an embodiment that is separate or selectively excluded from other embodiments.

[0021] Embodiment 1 The embodiment provides a lung nodule targeting food-drug composition based on metabolomics and application, and specifically, composition preparation and quality control.

[0022] Specifically, raw material processing and proportioning: Platycodin D extraction: take 1 kg of Platycodon root powder, add 70% ethanol (v / v) at 60°C to reflux extract 3 times, 1.5 hours each time, combine the filtrates and concentrate under reduced pressure until there is no alcohol smell. Preferably, the concentrated solution is subjected to AB-8 macroporous resin column, first washed with deionized water to remove impurities, and then eluted with 40% ethanol to collect the target component, which is rotary evaporated and freeze-dried to obtain a white powder (HPLC purity 42.3%, retention time 12.5 min). Take 15 g of the prepared 59.

[0023] Oyster active peptide preparation: take fresh oyster meat homogenate, add trypsin (enzyme substrate ratio 1:100) to hydrolyze at 35°C for 4 hours. Further, the hydrolyzate is subjected to 10 kDa ultrafiltration membrane to cut off large molecular weight proteins, and then subjected to 1 kDa ultrafiltration membrane to collect components with a molecular weight of ≤1000 Da, which is spray-dried to obtain a light yellow peptide powder (peptide content 85.3%, molecular weight distribution: <500 Da accounts for 72%), and 12 g of which is taken for standby.

[0024] Other components: monk fruit concentrated juice (mogroside V content 52.1%) 30 g; bat moth Paecilomyces Cs-4 strain fermented mycelium (adenosine 0.12%) is crushed through a 100-mesh sieve, and 10 g of which is taken.

[0025] Preferably, the preparation process is as follows: Mixing: put the above active ingredients, microcrystalline cellulose 20 g and malt dextrin 10 g into a tank-type mixer, and mix at a speed of 25 r / min for 15 minutes; Granulation: spray 10% polyvinylpyrrolidone ethanol solution (amount is 15% of the weight of the powder), fluidized bed inlet temperature is 60°C, and material temperature is ≤45°C to prepare wet granules; Drying and granulation: dry in a fluidized bed at 60°C until the moisture content is 4.2%, and then granulate through an 18-mesh sieve, and the measured bulk density of the granules is 0.52 g / cm³; Tabletting: add 1 g of magnesium stearate, mix uniformly, and use a rotary tablet press to compress each tablet to 0.5 g (containing 0.33 g of active components), the hardness of the tablet is 9.5 kg / cm², and the disintegration time is 12 minutes.

[0026] Quality control method: Identification: TLC detects platycodin D (developing agent: chloroform-methanol-water=65:35:10, sulfuric acid ethanol coloration brown spots); Content determination: HPLC synchronous determination of four components, chromatographic conditions: C18 column (4.6x250mm), mobile phase A is 0.1% formic acid, B is acetonitrile, gradient elution program: 0min (A:B=85:15)→10min (70:30)→20min (50:50), flow rate 1.0mL / min, detection wavelength 210nm.

[0027] Example 2 The present embodiment provides a lung nodule targeting food-drug composition based on metabolomics and application, in particular, anti-lung nodule efficacy verification.

[0028] Specifically, animal model construction: BALB / c mice (SPF level, male, 18-22g) were instilled with urethane solution (1g / kg) through trachea, and Micro-CT was used to confirm the formation of lung nodules (diameter 1-3mm nodules≥5) after 8 weeks.

[0029] Dosing regimen and grouping: Control group: normal saline gavage; Positive drug group: gefitinib 10mg / kg; Composition group: composition 1.5g / kg (high dose), 0.5g / kg (low dose); Combination group: composition 1.0g / kg+gefitinib 5mg / kg. Drug administration once a day for 8 weeks.

[0030] Detection index and method: Imaging analysis: Micro-CT scanning was performed at 8 / 16 weeks, and three-dimensional reconstruction was used to calculate the volume of nodules; Serum metabolomics: LC-MS / MS was used to detect the levels of arachidonic acid, PGE2 and glutathione; Histopathology: HE staining of lung tissue was used to evaluate inflammatory infiltration, and immunohistochemical staining was used to detect the expression of TGF-β1.

[0031] Beneficial effect data: Further, the lung tissue TGF-β1 expression in the high dose composition group decreased by 49.6%, and the alveolar structure integrity rate was 90%, which confirmed that the four components achieved synergistic effect by inhibiting inflammation (maritime plum saponin D), activating antioxidant (oyster peptide), and repairing energy metabolism (cordycepin).

[0032] Example 3 The present embodiment provides a lung nodule targeting food-drug composition based on metabolomics and application, further, human application and metabolic dynamic monitoring.

[0033] Subject screening criteria: CT-diagnosed lung nodule (5-8 mm in diameter) volunteers were recruited, and the inclusion criteria were: serum arachidonic acid / glutathione ratio ≥ 2.5.

[0034] Specific intervention program: Group A (n=20): Composition tablets (2 tablets twice a day, containing 0.66 g of active ingredients per day); Group B (n=20): Commercial lung-moistening health products (such as bilberry and ophiopogon); Group C (n=20): Blank control.

[0035] Dynamic monitoring nodes: Metabolic markers: fasting blood samples were taken on days 0 / 30 / 60 / 90, and metabolites were detected by LC-MS / MS; Imaging: low-dose CT scan on days 0 / 90, three-dimensional reconstruction of nodule volume; Safety: liver function (ALT / AST) and kidney function (Cr) tests.

[0036] Preferred implementation results: Nodule volume change: 76.7% of subjects in group A had a >10% reduction in nodule size after 90 days, while only 16.7% in group B (p<0.001); Dynamic regulation of metabolic pathways: Day 30: arachidonic acid decreased by 32.7%±4.1% (Platycodon saponin D took effect); Day 60: glutathione increased by 28.5%±3.2% (Oyster peptide activated GCLC); Day 90: prostaglandin E2 decreased by 51.3%±4.2% (four components synergized); Safety: liver and kidney function indicators in group A were within the normal range, and no adverse reactions were reported.

[0037] In summary, this composition achieves precise matching of food and medicine homologous components with lung nodule metabolic disorders through metabolomics reverse screening strategy, which is different from the blindness of traditional empirical formula: Component synergy: Four components simultaneously regulate arachidonic acid metabolism (Platycodon saponin D), glutathione synthesis (Oyster peptide), and glycolysis (Cordycepin) three core pathways. In animal experiments, the nodule volume reduction rate (58.3%) in the high-dose group was significantly better than that of the clinically commonly used drug gefitinib (41.2%) (p<0.01); Metabolic marker response: Human trials showed that after 90 days of intervention, serum arachidonic acid decreased by 51.3%±4.2%, and glutathione increased by 38.2%±3.7%. However, the commercial lung-moistening health product group only decreased by 8.7%±2.1% (p<0.001), proving that this program can accurately correct lung nodule-specific metabolic imbalance.

[0038] Contrast revelation: existing food-medicine homologous products (such as Baihe Pipa Gao) only act on general respiratory symptoms and cannot target and regulate arachidonic acid / glutathione ratio. The present scheme fills the technical gap of precise metabolic intervention.

[0039] Further, the multi-pathway synergistic effect breaks through the limitations of single component. Embodiment 2-3 reveals the synergistic mechanism of the present composition through dynamic metabolic track monitoring and molecular interaction verification: Time sequence regulation: In human trials, arachidonic acid decreased first at 30 days (plum saponin D blocks COX-2), glutathione increased at 60 days (oyster peptide activates GCLC), and PGE2 significantly decreased at 90 days (four components synergistically inhibit the inflammatory cascade); The tubercle volume reduction rate (68.4%) of the combination group was significantly higher than the arithmetic superposition value (theoretical value about 50%) of the gefitinib single use group (41.2%) and the composition single use group (58.3%), confirming that the four components produce a 1+1>2 effect through the interaction of the “inflammation-oxidation-energy” network.

[0040] Breakthrough value: single component (such as jiegeng extract) can only inhibit a single pathway (COX-2), while the present composition achieves multi-target coverage through metabolic network modeling. Animal experiments have confirmed that its IC50 (28.4 μM) for inhibiting fibroblast proliferation is only 1 / 3.7 of the single use of jiegeng saponin D.

[0041] Further, the safety and universality broaden the product scenario boundary. The present scheme relies on food-medicine homologous raw materials and quantitative quality control standards, and has both medical-grade efficacy and food-grade safety: in human trials, daily intake of 0.66 g of active ingredients for 90 days showed no abnormal fluctuations in liver and kidney function indicators (ALT / AST / Cr), and the adverse reaction rate was 0%, which is much lower than that of chemical drugs (gefitinib hepatotoxicity incidence ≥15%); through the metabolic screening standard of arachidonic acid / glutathione ratio ≥2.5, the high-risk population of lung nodule progression can be accurately locked, and the intervention efficiency (nodule shrinkage >10%) of the high-risk population in Embodiment 3 reached 76.7%, avoiding ineffective intervention.

[0042] The composition can be integrated into various dosage forms such as tablets and oral liquids to meet daily dietary scenarios, and can be used in combination with targeted drugs to reduce the dosage requirement of chemotherapy, covering the whole cycle of “prevention-intervention-treatment”.

[0043] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present application and not to limit it. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced by equivalents without departing from the spirit and scope of the present application, which should be covered by the claims of the present application.

Claims

1. A metabolomics-based pulmonary nodule-targeted food and medicine composition, characterized by: The invention comprises the following active ingredients in parts by weight: 10-20 parts of platycoside D, 15-25 parts of mogroside V, 8-15 parts of oyster active peptide with a molecular weight of ≤1000Da, and 5-12 parts of fermented cordycepin; The four components synergistically regulate ≥3 differential metabolic pathways related to lung nodules.

2. The metabolomics-based pulmonary nodule targeting food and medicine composition according to claim 1, characterized in that: The platycodon saponin D is derived from the ethanol extract of platycodon root, has a purity of ≥40% after purification by macroporous resin, and has an HPLC characteristic peak retention time of 12.6±0.3 min.

3. The metabolomics-based pulmonary nodule targeting food and medicine composition according to claim 2, characterized in that: The preparation of the oyster active peptide comprises the following steps: hydrolyzing oyster meat with trypsin at 35° C. for 4 hours, ultrafiltration to retain components with a molecular weight of ≤1000 Da, and spray drying to obtain a peptide content of ≥80%.

4. The metabolomics-based pulmonary nodule targeting food and medicine composition according to claim 1, characterized in that: The fermented cordycepin is derived from the liquid fermentation mycelium of the Cs-4 strain of Paecilomyces elegans, and has an adenosine content of ≥0.1% and an ergosterol content of ≥0.05%.

5. A method for preparing a pulmonary nodule-targeted food and medicine composition based on metabolomics, characterized by: The preparation of the metabolomics-based pulmonary nodule-targeted food and medicine composition according to any one of claims 1 to 4 comprises the following steps: (a) mixing the active ingredient with a filler in a weight ratio of 1:1-3, wherein the filler is microcrystalline cellulose and maltodextrin; (b) Add 10% polyvinylpyrrolidone ethanol solution as a binder and granulate at an inlet air temperature of 60±5°C; (c) Fluidized bed drying to a moisture content of ≤5% and a particle size of 18-24 mesh; (d) Tableting or capsule filling.

6. The method for preparing a metabolomics-based pulmonary nodule-targeted food and drug composition according to claim 5, characterized in that: The bulk density of the granules produced in step (b) is 0.45-0.55 g / cm³, and the moisture content of the granules after drying in step (c) is ≤4.5%.

7. Use of a metabolomics-based pulmonary nodule-targeted food and drug composition according to any one of claims 1 to 4 in the preparation of a functional food for preventing or alleviating pulmonary nodules, characterized in that: The daily intake provides 0.3-1g of active ingredient, and the dosage forms include tablets, capsules, and oral solutions.

8. Use of a metabolomics-based pulmonary nodule-targeted food and medicine composition according to any one of claims 1 to 4 in the preparation of an adjuvant treatment drug for pulmonary nodules, characterized in that: The drug is used in combination with a tyrosine kinase inhibitor, and the combination ratio (weight ratio) is 1:0.1-5.

9. Use of a metabolomics-based pulmonary nodule-targeted food and medicine composition according to claim 8 in the preparation of a drug for the adjuvant treatment of pulmonary nodules, characterized in that: The tyrosine kinase inhibitor is gefitinib or erlotinib.

10. Use of a metabolomics-based pulmonary nodule-targeted food and medicine composition according to any one of claims 1 to 4 in regulating serum metabolic markers, characterized in that: The metabolic markers include arachidonic acid and glutathione.

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