A traditional Chinese medicine composition for treating lung cancer and its preparation method

Abstract

This invention relates to the field of traditional Chinese medicine composition technology, and in particular to a traditional Chinese medicine composition for treating lung cancer and its preparation method, comprising composition one, composition two, and composition three; composition one, by weight parts, comprises: Astragalus membranaceus 20-45 parts, Polygonatum odoratum 15-30 parts, Ophiopogon japonicus 8-18 parts, Coix lacryma-jobi 20-45 parts, Phragmites communis 20-45 parts, and Glycyrrhiza uralensis 3-6 parts; composition two, by weight parts, comprises: Clematis armandii 9-15 parts, Actinidia chinensis root 15-30 parts, Scutellaria barbata 10-22 parts, Pinellia ternata 6-13 parts, Fritillaria thunbergii 6-13 parts, and Citrus reticulata peel 6-13 parts; composition three, by weight parts, comprises: Buthus martensii 3-8 parts. The traditional Chinese medicine composition prepared by this invention has the effects of invigorating qi and nourishing yin, clearing heat and detoxifying, resolving phlegm and dissipating nodules, and promoting blood circulation and unblocking collaterals. It overcomes the technical problems of existing anti-lung cancer chemotherapy drugs having large toxic side effects and easily developing drug resistance, as well as the limited anti-lung cancer effects of existing single or simple compound traditional Chinese medicines, and the lack of optimization in compatibility and process.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine composition technology, and in particular to a traditional Chinese medicine composition for treating lung cancer and its preparation method. Background Technology

[0002] Lung cancer is the leading cause of both morbidity and mortality worldwide, with non-small cell lung cancer accounting for over 80% of cases. Clinical treatment primarily involves surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. However, chemotherapy drugs commonly cause toxic side effects such as bone marrow suppression, gastrointestinal reactions, and liver and kidney damage, and are prone to multidrug resistance, leading to decreased efficacy. Traditional Chinese medicine (TCM) treatment for lung cancer is characterized by its ability to strengthen the body's resistance and eliminate pathogenic factors, address both the root cause and symptoms, target multiple areas, and reduce toxicity. It possesses unique advantages in improving patients' quality of life, prolonging survival, and mitigating the adverse reactions of radiotherapy and chemotherapy.

[0003] Currently, most traditional Chinese medicine (TCM) compound formulas or single-component formulas used for lung cancer focus on a single efficacy, or their formulations are not optimized enough, failing to fully leverage the synergistic effects of invigorating qi and nourishing yin, clearing heat and detoxifying, resolving phlegm and dissipating nodules, and unblocking collaterals to attack toxins. Consequently, their effectiveness in inhibiting key aspects such as tumor cell proliferation, invasion, and metastasis is limited. Therefore, developing a TCM composition with rigorous formulation, stable processing, and significantly superior anti-lung cancer efficacy compared to single-component formulas has significant clinical value and market potential. Summary of the Invention

[0004] The purpose of this invention is to provide a traditional Chinese medicine composition for lung cancer and its preparation method. The prepared traditional Chinese medicine composition has the effects of invigorating qi and nourishing yin, clearing heat and detoxifying, resolving phlegm and dissipating nodules, and promoting blood circulation and unblocking collaterals. It overcomes the technical problems of existing anti-lung cancer chemotherapy drugs having large toxic side effects and easy to produce drug resistance, as well as the limited anti-lung cancer effects of existing single or simple compound traditional Chinese medicines, and the lack of optimization in compatibility and process.

[0005] To achieve the above objectives, the present invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Composition 1, Composition 2, and Composition 3; Composition 1 comprises, by weight parts: Astragalus membranaceus 20-45 parts, Polygonatum odoratum 15-30 parts, Ophiopogon japonicus 8-18 parts, Coix lacryma-jobi 20-45 parts, Phragmites communis 20-45 parts, and Glycyrrhiza uralensis 3-6 parts; Composition 2 comprises, by weight parts: Clematis armandii 9-15 parts, Actinidia chinensis root 15-30 parts, Scutellaria barbata 10-22 parts, Pinellia ternata 6-13 parts, Fritillaria thunbergii 6-13 parts, and Citrus reticulata peel 6-13 parts; Composition 3 comprises, by weight parts: Buthus martensii 3-8 parts.

[0006] Preferably, the mass ratio of composition one, composition two, and composition three is (1~2):(1~2):(1~2).

[0007] This invention also provides a method for preparing a traditional Chinese medicine composition for treating lung cancer, comprising the following steps: S1. Preparation of Composition 1: Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis are extracted with alcohol to obtain residue and extract. The extract is enriched and purified by macroporous adsorption resin and vacuum dried to obtain alcohol extract. The residue is extracted with water and precipitated with alcohol and vacuum dried to obtain water-extracted alcohol precipitate. The alcohol extract and water-extracted alcohol precipitate are mixed to obtain Composition 1. S2. Preparation of Composition 2: Take three leaves of *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. After alcohol extraction and enrichment and purification with macroporous adsorption resin, the mixture is dried under vacuum to obtain Composition 2. S3. Preparation of Composition 3: Take whole scorpions and perform enzymatic hydrolysis. After enzyme inactivation and solid-liquid separation, collect the supernatant and freeze-dry it under vacuum to obtain Composition 3. S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three evenly to obtain the traditional Chinese medicine composition.

[0008] Preferably, in S1, the alcohol extraction is performed by using 70%-90% ethanol as a solvent, extracting by heating and reflux 1-3 times at a material-to-liquid ratio of 1:(6~12) g / mL, each time for 0.5-2 hours, combining the extracts and concentrating.

[0009] Preferably, in S1, the enrichment and purification process is as follows: after the extract is loaded onto a macroporous adsorption resin, it is first eluted with water, then eluted with ethanol of 50%-95% by volume, the alcohol eluent is collected, and concentrated under reduced pressure.

[0010] Preferably, in S1, the water extraction and alcohol precipitation are performed as follows: after evaporating the ethanol from the residue, water is used as the solvent, and the mixture is boiled and extracted 1-3 times at a material-to-liquid ratio of 1:(6~12) g / mL, each time for 0.5-2 hours. The extracts are combined, concentrated to 0.2-1.0 g crude drug / mL, 2-6 times the amount of anhydrous ethanol is added, the mixture is refrigerated for 8-24 hours, centrifuged, and the precipitate is collected.

[0011] Preferably, in S2, the alcohol extraction is performed by using 60%-85% ethanol as solvent, with a material-to-liquid ratio of 1:(6~12) g / mL, heating and refluxing for 1-3 times, each time for 0.5-2 hours, combining the extracts and concentrating.

[0012] Preferably, in S2, the enrichment and purification process is as follows: after adsorbing onto a macroporous adsorption resin, the resin is first eluted with water, then eluted with ethanol at a volume fraction of 50%-95%, the alcohol eluent is collected, and concentrated under reduced pressure.

[0013] Preferably, in S3, the enzymatic hydrolysis is performed by adding 2000-5000U of trypsin, adjusting the pH to 6.5-8.0, and hydrolyzing at 40-60℃ for 4-10 hours.

[0014] Preferably, in S3, enzyme inactivation is performed by heating the enzyme hydrolysate to 85-100℃ and maintaining it for 15-30 minutes.

[0015] Therefore, the present invention employs the above-mentioned traditional Chinese medicine composition for treating lung cancer and its preparation method, which has the following beneficial effects: (1) The present invention focuses on the core pathogenesis of lung cancer, which is based on deficiency of both Qi and Yin and phlegm and blood stasis. It strictly follows the principle of supporting the body's resistance and eliminating pathogens, combining the principles of monarch, minister, assistant and guide, and attacking and tonifying. It organically combines Qi-tonifying and Yin-nourishing, heat-clearing and detoxifying, phlegm-resolving and nodule-dispersing, blood-activating and stasis-removing, and meridian-clearing and toxin-attacking, so as to achieve the goal of supporting the body's resistance without leaving pathogens and eliminating pathogens without harming the body's resistance. It overcomes the defects of traditional Chinese medicine compound prescriptions such as single efficacy and unreasonable combination.

[0016] (2) The first composition of the present invention uses a combination of alcohol extraction and water extraction with alcohol precipitation to simultaneously enrich fat-soluble total saponins and water-soluble total polysaccharides, taking into account both direct anticancer and immune enhancement effects; the second composition uses alcohol extraction combined with macroporous adsorption resin purification to highly enrich anticancer active ingredients such as total flavonoids and total alkaloids; the third composition uses a mild trypsin enzymatic hydrolysis process to convert the large molecular protein of whole scorpion into easily absorbed and highly active small molecular polypeptides, retaining biological activity to the maximum extent, with stable and controllable process, good repeatability and high retention rate of effective ingredients.

[0017] (3) The composition of the present invention can strengthen the body’s immune system, improve the tumor immune microenvironment, and block tumor immune escape through the synergistic effect of multiple components, multiple targets and multiple pathways. It can also directly inhibit the proliferation of lung cancer cells, induce tumor cell apoptosis, reduce cell adhesion ability, and significantly inhibit the invasion and migration of lung cancer cells and block distant metastasis. The overall anti-lung cancer effect is significantly better than that of a single component, with a broader anti-cancer spectrum and more stable effect.

[0018] (4) This invention selects authentic medicinal materials with mild medicinal properties and few toxic side effects. While ensuring strong anti-lung cancer efficacy, it can reduce adverse reactions such as gastrointestinal reactions and bone marrow suppression caused by radiotherapy and chemotherapy, improve the quality of life of patients, and is suitable for adjuvant treatment and long-term conditioning of non-small cell lung cancer. It has good clinical translational value and market application prospects.

[0019] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0020] Figure 1 This is a comparison chart showing the effects of Example 1 and Comparative Examples 1-3 of the present invention on the survival rate of A549 lung cancer cells; Figure 2 This is a comparison diagram of the effects of Example 1 and Comparative Examples 1-3 of the present invention on the adhesion ability of A549 lung cancer cells; Figure 3 This is a comparison diagram of the effects of Example 1 and Comparative Examples 1-3 of the present invention on the invasive ability of A549 lung cancer cells; Figure 4This is a comparison chart showing the effects of Example 1 and Comparative Examples 1-3 of the present invention on the migration ability of A549 lung cancer cells. Detailed Implementation

[0021] The present invention will be further described below with reference to the accompanying drawings and embodiments. Unless otherwise defined, the technical or scientific terms used in this invention should be understood in their ordinary sense by those skilled in the art. The features mentioned above or in the specific examples mentioned in this invention can be combined arbitrarily, and these specific embodiments are only used to illustrate the invention and are not intended to limit the scope of the invention.

[0022] This invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Composition 1, Composition 2, and Composition 3. Composition 1 comprises, by weight, 20-45 parts of Astragalus membranaceus, 15-30 parts of Polygonatum odoratum, 8-18 parts of Ophiopogon japonicus, 20-45 parts of Coix lacryma-jobi, 20-45 parts of Phragmites communis, and 3-6 parts of Glycyrrhiza uralensis. Composition 2 comprises, by weight, 9-15 parts of Clematis armandii, 15-30 parts of Actinidia chinensis root, 10-22 parts of Scutellaria barbata, 6-13 parts of Pinellia ternata, 6-13 parts of Fritillaria thunbergii, and 6-13 parts of Citrus reticulata peel. Composition 3 comprises, by weight, 3-8 parts of Buthus martensii.

[0023] Preferably, the mass ratio of composition one, composition two, and composition three is (1~2):(1~2):(1~2).

[0024] The compatibility principle of this invention: This invention takes the deficiency of both Qi and Yin in lung cancer as its root cause and the accumulation of phlegm and blood stasis as its symptoms as its core pathogenesis. It strictly follows the principles of supporting the body's resistance and eliminating pathogens, the principle of combining the effects of the principal, assistant, and adjuvant herbs, and the principle of attacking and tonifying simultaneously, so as to achieve the goal of supporting the body's resistance without leaving pathogens and eliminating pathogens without harming the body's resistance. With *Trifolium repens*, *Actinidia chinensis* root, and *Scutellaria barbata* as the principal herbs, these three herbs work together as the core of the formula to dispel pathogens, achieving the effects of clearing heat and detoxifying, resolving blood stasis and detoxifying, and eliminating masses and nodules. It is specifically targeted at blood stasis, stagnant heat, and lingering turbid pathogens, and is often used for nodules, lumps, and chronic inflammatory lesions formed by phlegm and blood stasis. *Astragalus membranaceus* is sweet and warm, tonifying qi, strengthening the spleen and lungs, and protecting the body's vital energy, enhancing the body's disease resistance, counteracting the spleen-damaging effects of other cold and cooling herbs, supporting qi and blood, and treating the root cause of deficiency. *Polygonatum odoratum*, *Ophiopogon japonicus*, and *Phragmites communis* are sweet and cold, nourishing yin and generating fluids, clearing deficiency heat, moistening the lungs and stomach, clearing residual heat, nourishing the yin of the lungs and stomach, relieving dryness, dry mouth, and yin fluid depletion, and balancing the dryness and coldness of the entire formula. *Coix lacryma-jobi* strengthens the spleen and eliminates dampness, clears heat and turbidity, resolves phlegm and dissipates nodules, and also benefits the body. The formula protects the spleen and eliminates the source of phlegm and dampness. Pinellia ternata and Citrus reticulata peel, with their pungent and warm properties, dry dampness, regulate qi, strengthen the spleen, harmonize the stomach, and resolve phlegm. They regulate the qi mechanism of the middle jiao, resolve internal obstruction of phlegm and dampness, abdominal distension, and phlegm stagnation, and prevent cold and cooling medicines from hindering the stomach and causing qi stagnation. Fritillaria thunbergii clears heat, resolves phlegm, and relieves stagnation, effectively treating phlegm-heat stagnation and phlegm nodules. Combined with heat-clearing medicines, it enhances the ability to soften and disperse nodules. Scorpion clears the collaterals, breaks up blood stasis, disperses nodules, and dispels wind, penetrating deep into the collaterals to expel deep-seated stagnation and stubborn phlegm, targeting chronic diseases that have entered the collaterals and stubborn, difficult-to-resolve nodules. Licorice harmonizes the various medicines, tonifies qi, harmonizes the middle jiao, relieves urgency, and protects the stomach. It moderates the harshness of insect-based and bitter-cold medicines, coordinating the cold and heat, tonification and purgation properties of the entire formula, and protecting the spleen and stomach.

[0025] The formula combines tonification and purgation: Astragalus, Polygonatum, and Ophiopogon japonicus tonify Qi, nourish Yin, and strengthen the body; Trifolium repens, Actinidia chinensis root, and Scutellaria barbata clear heat and eliminate pathogens, combining tonification and purgation, suitable for chronic illnesses with mixed deficiency and excess. It balances cold and heat: Actinidia chinensis root, Trifolium repens, Scutellaria barbata, Phragmites communis root, and Fritillaria thunbergii are slightly cooling; Pinellia ternata, Citrus reticulata peel, and Astragalus membranaceus are slightly warming, achieving a balance of cold and heat, and long-term use will not harm the spleen and stomach. It treats phlegm and blood stasis simultaneously: Pinellia ternata, Citrus reticulata peel, and Coix lacryma-jobi dry dampness and resolve phlegm; Scorpion unblocks the meridians and breaks up blood stasis; Fritillaria thunbergii and Actinidia chinensis root soften hardness and disperse nodules, directly targeting the core pathogenesis of phlegm, dampness, blood stasis, and toxins. It addresses both upper and lower body imbalances: clearing heat stagnation in the upper burner and nourishing lung and stomach Yin; strengthening the spleen and stomach in the middle burner and resolving internally generated phlegm and dampness; clearing meridian blockages and regulating the body's Qi.

[0026] The traditional Chinese medicine composition of this invention combines the effects of strengthening the body's resistance and eliminating pathogens, invigorating qi and dispersing nodules, and resolving phlegm and unblocking the meridians, thus achieving the combined effects of invigorating qi and nourishing yin, detoxifying and dispersing nodules, and resolving phlegm and removing blood stasis.

[0027] Pharmacological effects of each component of this invention: Composition 1: Strengthens the body's resistance, enhances immunity, improves the microenvironment, and inhibits tumor immune escape.

[0028] Astragalus contains astragaloside A, astragalus polysaccharides, and flavonoids. It invigorates qi and strengthens the exterior, strengthens the spleen and replenishes the middle jiao, significantly enhances the body's immune function, improves the tumor immune microenvironment, and inhibits the proliferation and migration of lung cancer cells.

[0029] Polygonatum odoratum contains polysaccharides and steroidal saponins, which nourish yin and moisten dryness without being greasy or irritating to the stomach. It promotes macrophage phagocytosis and lymphocyte proliferation, enhances humoral and cellular immunity, and improves immunosuppression. It also inhibits the proliferation of cells such as liver cancer, lung cancer, and breast cancer.

[0030] Ophiopogon japonicus: Contains Ophiopogon japonicus polysaccharides and steroidal saponins, which nourish yin and promote body fluid production, moisten the lungs and clear the heart, improve qi and yin deficiency in lung cancer, enhance immunity and inhibit the inflammatory microenvironment.

[0031] Job's tears: Contains Job's tears oil, Job's tears ester, and polysaccharides, which inhibit immune escape and malignant progression of lung adenocarcinoma, directly suppress cancer, reduce inflammation, and improve the tumor microenvironment.

[0032] Reed rhizome: Contains polysaccharides and amino acids, inhibits the TGF-β signaling pathway, reduces lung inflammation and damage, and promotes airway repair.

[0033] Licorice contains triterpenoid saponins, flavonoids, and polysaccharides, which promote antigen presentation and CD8+ T cell infiltration, inhibit lung cancer progression, and harmonize various drugs.

[0034] Composition 2: Clears heat and detoxifies, resolves phlegm and dissipates nodules, inhibits proliferation, promotes apoptosis, and inhibits invasion.

[0035] Three-leaf green: Contains flavonoids, polysaccharides, and phenolic acids. It clears heat and detoxifies, removes phlegm and promotes blood circulation. It inhibits NF-κB, TNF-α, and IL-6. It has an improving effect on respiratory and digestive tract inflammation, inhibits tumor cell proliferation, induces apoptosis, and inhibits invasion and metastasis.

[0036] Actinidia chinensis root contains triterpenoids, flavonoids, anthraquinones, etc., which downregulate the AKT / JNK pathway, activate Caspase-3 / 9, upregulate Bax, downregulate Bcl-2 / survivin, and promote programmed cell death in tumor cells. It can also relieve cancer pain.

[0037] Scutellaria barbata: Contains flavonoids and polysaccharides, which inhibit the proliferation, migration, and invasion of lung cancer cells, promote apoptosis, clear heat and detoxify, and promote diuresis.

[0038] Pinellia ternata: Contains alkaloids and polysaccharides, regulates the PI3K / AKT pathway, resolves phlegm and nodules, and inhibits the progression of lung cancer.

[0039] Fritillaria thunbergii: Contains fritillary A and fritillary B, which clear heat and resolve phlegm, disperse nodules and reduce swelling, inhibit lung cancer cell proliferation and induce apoptosis.

[0040] Dried tangerine peel: Contains hesperidin and volatile oils, regulates qi and strengthens the spleen, dries dampness and resolves phlegm, and helps to disperse nodules and improve the inflammatory microenvironment of tumors.

[0041] Composition 3: Unblocks the meridians, eliminates pathogens, inhibits metastasis, breaks up masses and disperses nodules.

[0042] Whole scorpion: After enzymatic hydrolysis, active polypeptides and amino acids are obtained, which can clear the meridians, attack toxins, break up masses and dissipate nodules, effectively inhibit tumor invasion and metastasis, and enhance the overall effect of the formula in inhibiting metastasis and dissipating stubborn nodules.

[0043] The principle of monarch-minister-assistant-guide compatibility in this invention: The principal herbs are: Trifoliate orange, Actinidia chinensis root, and Scutellaria barbata, which have the functions of clearing heat and detoxifying, eliminating masses and dissipating nodules, and promoting diuresis and reducing swelling.

[0044] The accompanying herbs are: Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Phragmites communis, and Coix lacryma-jobi. They are used to invigorate qi and nourish yin, strengthen the spleen and kidneys, and support the body's vital energy.

[0045] The adjuvant herbs are Pinellia ternata, Citrus reticulata peel, Fritillaria thunbergii, and Scorpion. They regulate qi, resolve phlegm, clear heat and detoxify, and when combined with Scorpion, they can clear the meridians and break up blood stasis.

[0046] The guiding herb is licorice, which invigorates qi, strengthens the middle jiao, and harmonizes the other herbs, ensuring that the entire formula is nourishing without causing stagnation and purging without causing harm.

[0047] In this invention, composition one is rich in total polysaccharides and total saponins, which can comprehensively regulate the body's immune network. Composition two is rich in total flavonoids, which can directly act on lung cancer cells. Composition three, with its active peptides and amino acids obtained through enzymatic hydrolysis, is the core component for attacking the virus and inhibiting metastasis, responsible for inhibiting invasion and distant metastasis. This invention's herbal compositions follow the modern principles of traditional Chinese medicine: multi-component, multi-target, multi-pathway, and synergistic effects. Using total polysaccharides, total saponins, total flavonoids, and active peptides as the core material basis, through the complementarity and synergy of different types of effective components, a holistic anti-lung cancer effect is formed, encompassing immune regulation, direct cancer inhibition, anti-inflammatory repair, and metastasis inhibition.

[0048] This invention also provides a method for preparing a traditional Chinese medicine composition for treating lung cancer, comprising the following steps: S1. Preparation of Composition 1: Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis are extracted with alcohol to obtain residue and extract. The extract is enriched and purified by macroporous adsorption resin and vacuum dried to obtain alcohol extract. The residue is extracted with water and precipitated with alcohol and vacuum dried to obtain water-extracted alcohol precipitate. The alcohol extract and water-extracted alcohol precipitate are mixed to obtain Composition 1. S2. Take three leaves of *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. After alcohol extraction and enrichment and purification with macroporous adsorption resin, vacuum drying is used to obtain composition two. S3. Preparation of Composition 3: Take whole scorpions and perform enzymatic hydrolysis. After enzyme inactivation and solid-liquid separation, collect the supernatant and freeze-dry it under vacuum to obtain Composition 3. S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three evenly to obtain the traditional Chinese medicine composition.

[0049] Preferably, in S1, the ethanol extraction is performed as follows: using 70%-90% ethanol as the solvent, at a material-to-liquid ratio of 1:(6~12) g / mL, the mixture is heated under reflux for 1-3 times, each time for 0.5-2 hours. The extracts are then combined and concentrated. Using ethanol as the extraction solvent allows for the selective extraction of lipid-soluble components with high anti-cancer activity, such as total saponins and astragaloside A. These components can rapidly enter cells, regulate signaling pathways, and inhibit the proliferation and migration of lung cancer cells.

[0050] Preferably, in S1, the enrichment and purification process involves: after the extract is loaded onto a macroporous adsorption resin, it is first eluted with water, then eluted with 50%-95% ethanol (v / v), and the ethanol eluent is collected and concentrated under reduced pressure. Macroporous adsorption resin purification selectively removes impurities such as polysaccharides, pigments, and tannins, and highly enriches total saponins, astragaloside A, and other immune-enhancing and anticancer components, improving the efficacy per unit dose and reducing interference from ineffective impurities.

[0051] Preferably, in S1, the water extraction and alcohol precipitation process is as follows: After evaporating the ethanol from the residue, water is used as the solvent, and the mixture is decocted and extracted 1-3 times at a material-to-liquid ratio of 1:(6~12) g / mL, each time for 0.5-2 hours. The extracts are combined and concentrated to 0.2-1.0 g crude drug / mL. 2-6 times the amount of anhydrous ethanol is added, and the mixture is refrigerated for 8-24 hours. After centrifugation, the precipitate is collected. Water-soluble extraction can fully obtain macromolecular total polysaccharides, while alcohol precipitation can remove impurities such as proteins and starches. High-purity polysaccharides are core substances for immune regulation and improving the tumor microenvironment. They can activate macrophages, promote CD8+ T cell infiltration, and block tumor immune escape.

[0052] In this invention, alcohol extract and water-extracted alcohol precipitate are combined to achieve a perfect blend of total saponins (lipid-soluble) and total polysaccharides (water-soluble), so that the composition can simultaneously have dual pharmacological effects of direct anticancer and immune enhancement, thereby maximizing the efficacy of strengthening the body's resistance and addressing both the symptoms and the root cause.

[0053] Preferably, in S2, the ethanol extraction is performed using 60%-85% ethanol as the solvent, at a material-to-liquid ratio of 1:(6~12) g / mL, by heating and reflux extraction 1-3 times, each time for 0.5-2 hours, combining the extracts and concentrating. This method can efficiently dissolve the total flavonoids from *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, and tangerine peel, as well as the alkaloids from *Fritillaria thunbergii*, which are key components for inhibiting the PI3K / AKT pathway, inducing tumor apoptosis, anti-inflammation, and inhibiting invasion.

[0054] Preferably, in S2, the enrichment and purification process involves: after adsorption onto a macroporous adsorption resin, elution with water followed by elution with 50%-95% ethanol (v / v), collection of the ethanol eluent, and concentration under reduced pressure. This precisely removes water-soluble impurities, enriches high-purity total flavonoids and alkaloids, and significantly enhances the pharmacological activities of the second composition in clearing heat and detoxifying, resolving phlegm and dissipating nodules, and directly inhibiting cancer. This ensures that the second composition can effectively inhibit the proliferation, migration, and invasion of lung cancer cells and promote apoptosis.

[0055] Preferably, in S3, the enzymatic hydrolysis is performed by adding 2000-5000U of trypsin, adjusting the pH to 6.5-8.0, and hydrolyzing at 40-60℃ for 4-10 hours. Using trypsin at near-physiological pH and temperature gently breaks down the whole scorpion protein, releasing small-molecule active peptides and amino acids, avoiding the destruction of efficacy by high temperature and pressure. Large scorpion protein molecules are highly toxic and poorly absorbed; the small-molecule peptides generated after enzymatic hydrolysis are rapidly absorbed and have strong penetrability, reaching deep into the lung channels, inhibiting tumor metastasis, and breaking down masses, making them potent anti-metastasis components.

[0056] Preferably, in S3, enzyme inactivation is performed by heating the enzyme hydrolysate to 85-100℃ and maintaining it for 15-30 minutes. Maintaining a low temperature throughout the process maximizes the preservation of polypeptide bioactivity, avoids inactivation, and ensures the stable and potent pharmacological effects of composition 3 in promoting blood circulation, attacking toxins, and inhibiting metastasis.

[0057] Example 1 This invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Composition 1, Composition 2, and Composition 3 in a mass ratio of 1:1:1; Composition 1 comprises, by weight, 30 parts of Astragalus membranaceus, 20 parts of Polygonatum odoratum, 12 parts of Ophiopogon japonicus, 30 parts of Coix lacryma-jobi, 30 parts of Phragmites communis, and 4 parts of Glycyrrhiza uralensis; Composition 2 comprises, by weight, 12 parts of Clematis armandii, 20 parts of Actinidia chinensis root, 15 parts of Scutellaria barbata, 9 parts of Pinellia ternata, 9 parts of Fritillaria thunbergii, and 9 parts of Citrus reticulata peel; Composition 3 comprises, by weight, 5 parts of Buthus martensii.

[0058] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps: S1. Preparation of Composition One: S1.1 Take Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis. Use 80% ethanol as solvent and extract twice by heating and reflux at a material-to-liquid ratio of 1:8 g / mL for 1 hour each time. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load onto AB-8 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then elute with 4 column bed volumes (BV) of 80% ethanol. Collect the eluent, concentrate under reduced pressure, and dry under vacuum to obtain the ethanol extract.

[0059] S1.2 After removing the ethanol from the residue obtained after extraction in S1.1, use water as a solvent and decoct twice at a material-to-liquid ratio of 1:8 g / mL, each time for 1 hour. Combine the extracts, concentrate to 0.5 g crude drug / mL, add 4 times the amount of anhydrous ethanol, refrigerate for 12 hours, centrifuge, collect the precipitate, and vacuum dry to obtain the water-ethanol precipitate.

[0060] S1.3. Mix the alcohol extract of S1.1 with the water extract and alcohol precipitate of S1.2 to obtain composition one.

[0061] S2. Preparation of Composition 2: Take *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. Use 70% ethanol as solvent and extract twice by heating and reflux at a material-to-liquid ratio of 1:8 g / mL, each time for 1 hour. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load the extract onto HPD417 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then wash with 4 column bed volumes (BV) of 80% ethanol and vacuum dry to obtain Composition 2.

[0062] S3. Preparation of Composition 3: Take whole scorpion, add 3000U of trypsin, adjust the pH to 7, and enzymatically hydrolyze at 50℃ for 6 hours. Heat the hydrolysate to 90℃ and maintain for 20 minutes. Separate the solid and liquid, collect the supernatant, and freeze-dry under vacuum to obtain Composition 3.

[0063] S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three in a mass ratio of 1:1:1 to obtain the traditional Chinese medicine composition.

[0064] Example 2 This invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Composition 1, Composition 2, and Composition 3 in a mass ratio of 2:1:1; Composition 1 comprises, by weight, 20 parts of Astragalus membranaceus, 15 parts of Polygonatum odoratum, 8 parts of Ophiopogon japonicus, 20 parts of Coix lacryma-jobi, 20 parts of Phragmites communis, and 3 parts of Glycyrrhiza uralensis; Composition 2 comprises, by weight, 9 parts of Clematis armandii, 15 parts of Actinidia chinensis root, 10 parts of Scutellaria barbata, 6 parts of Pinellia ternata, 6 parts of Fritillaria thunbergii, and 6 parts of Citrus reticulata peel; Composition 3 comprises, by weight, 3 parts of Buthus martensii.

[0065] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps: S1. Preparation of Composition One: S1.1 Take Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis. Use 70% ethanol as solvent and extract by heating and reflux three times at a material-to-liquid ratio of 1:6 g / mL for 0.5 h each time. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load onto AB-8 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then elute with 4 column bed volumes (BV) of 50% ethanol. Collect the eluent, concentrate under reduced pressure, and vacuum dry to obtain the ethanol extract.

[0066] S1.2 After removing the ethanol from the residue obtained after extraction in S1.1, use water as a solvent and decoct and extract three times at a material-to-liquid ratio of 1:6 g / mL, each time for 0.5 h. Combine the extracts, concentrate to 0.5 g crude drug / mL, add 4 times the amount of anhydrous ethanol, refrigerate for 8 h, centrifuge, collect the precipitate, and vacuum dry to obtain the water-ethanol precipitate.

[0067] S1.3. Mix the alcohol extract of S1.1 with the water extract and alcohol precipitate of S1.2 to obtain composition one.

[0068] S2. Preparation of Composition 2: Take *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. Use 60% ethanol as solvent and extract by heating and reflux three times at a material-to-liquid ratio of 1:6 g / mL, each time for 0.5 h. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load the extract onto HPD417 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then wash with 4 column bed volumes (BV) of 50% ethanol and vacuum dry to obtain Composition 2.

[0069] S3. Preparation of Composition 3: Take whole scorpion, add 2000U of trypsin, adjust the pH to 6.5, and enzymatically hydrolyze at 40℃ for 4h. Heat the hydrolysate to 85℃ and maintain for 30min. Separate the solid and liquid, collect the supernatant, and freeze-dry under vacuum to obtain Composition 3.

[0070] S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three in a mass ratio of 2:1:1 to obtain the traditional Chinese medicine composition.

[0071] Example 3 This invention provides a traditional Chinese medicine composition for treating lung cancer, comprising Composition 1, Composition 2, and Composition 3 in a mass ratio of 1:2:1; Composition 1 comprises, by weight, 45 parts of Astragalus membranaceus, 30 parts of Polygonatum odoratum, 18 parts of Ophiopogon japonicus, 45 parts of Coix lacryma-jobi, 45 parts of Phragmites communis, and 6 parts of Glycyrrhiza uralensis; Composition 2 comprises, by weight, 15 parts of Clematis armandii, 30 parts of Actinidia chinensis root, 22 parts of Scutellaria barbata, 13 parts of Pinellia ternata, 13 parts of Fritillaria thunbergii, and 13 parts of Citrus reticulata peel; Composition 3 comprises, by weight, 8 parts of Buthus martensii.

[0072] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps: S1. Preparation of Composition One: S1.1 Take Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis. Use 90% ethanol as solvent and extract once by heating and reflux at a material-to-liquid ratio of 1:12 g / mL for 2 hours each time. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load onto AB-8 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then elute with 4 column bed volumes (BV) of 95% ethanol. Collect the eluent, concentrate under reduced pressure, and dry under vacuum to obtain the ethanol extract.

[0073] S1.2 After removing the ethanol from the residue obtained after extraction in S1.1, use water as a solvent and decoct once for 2 hours each time at a material-to-liquid ratio of 1:12 g / mL. Combine the extracts, concentrate to 0.5 g crude drug / mL, add 6 times the amount of anhydrous ethanol, refrigerate for 24 hours, centrifuge, collect the precipitate, and vacuum dry to obtain the water-ethanol precipitate.

[0074] S1.3. Mix the alcohol extract of S1.1 with the water extract and alcohol precipitate of S1.2 to obtain composition one.

[0075] S2. Preparation of Composition 2: Take *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. Use 85% ethanol as solvent and extract by heating and reflux three times at a material-to-liquid ratio of 1:12 g / mL, each time for 2 hours. Combine the extracts and concentrate to 0.5 g crude drug / mL. Load the extract onto HPD417 macroporous adsorption resin and wash with 4 column bed volumes (BV) of water to remove impurities. Then wash with 4 column bed volumes (BV) of 95% ethanol and vacuum dry to obtain Composition 2.

[0076] S3. Preparation of Composition 3: Take whole scorpion, add 2000U of trypsin, adjust the pH to 8.0, and enzymatically hydrolyze at 60℃ for 10h. Heat the hydrolysate to 100℃ and maintain for 30min. Separate the solid and liquid, collect the supernatant, and freeze-dry under vacuum to obtain Composition 3.

[0077] S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three in a mass ratio of 1:2:1 to obtain the traditional Chinese medicine composition.

[0078] Comparative Example 1 This is composition one from Example 1.

[0079] Comparative Example 2 This is composition two from Example 1.

[0080] Comparative Example 3 This is composition three from Example 1.

[0081] Test and experiment: Experimental groups: blank control group (containing 0.1% DMSO (dimethyl sulfoxide)), composition group 1 (50 μg / mL), composition group 2 (50 μg / mL), composition group 3 (50 μg / mL), and the traditional Chinese medicine composition group from Example 1 (hereinafter referred to as the traditional Chinese medicine composition group), for a total of 5 groups. The powder of each composition was dissolved in DMSO and then diluted with cell culture medium to the required concentration, with DMSO content ≤ 0.1%.

[0082] CCK-8 assay for cell viability: A549 cells (human alveolar basal epithelial cells for lung cancer) in logarithmic growth phase were digested with trypsin to prepare a single-cell suspension, and the cell concentration was adjusted to 5 × 10⁻⁶ cells / cells. 4Cells were seeded at a density of 100 μl / ml into 96-well plates, with 6 replicates per group. After incubation at 37°C and 5% CO2 for 24 h, the plates were treated with the drug and cultured for another 48 h. The drug-containing medium was then discarded, and freshly prepared medium containing CCK8 reagent was added to each well. The plates were then incubated for another 2 h. The OD value (optical density) at 450 nm was measured using a microplate reader, and the cell viability (%) was calculated. Cell viability = (OD value of drug-treated group / OD value of normal group) × 100%.

[0083] The results are as follows Figure 1 As shown in the figure The difference was statistically significant compared to the blank control group. Compared with the blank control group, the 48-hour cell survival rate of each group was significantly reduced (P<0.01), and the cell survival rate of the traditional Chinese medicine composition group was significantly lower than that of composition group 1, composition group 2 and composition group 3. This indicates that the traditional Chinese medicine composition of the present invention has a significant synergistic anti-cancer effect and its anti-lung cancer effect is better than that of the single component.

[0084] Cell adhesion assay to detect cell adhesion ability: A549 cells in the logarithmic growth phase were harvested and their concentration adjusted to 1×10⁻⁶. 5 pcs·mL -1 2.5 mL was seeded into each well of a 6-well plate, and the drug was added for 48 hours after 24 hours. Diluted fibronectin (10 μg / mL) was then seeded into pre-cooled 96-well plates. -1 After allowing the gel to solidify naturally overnight, place the coated 96-well plate in an incubator for 3-4 hours to activate it before the experiment. Add 2 × 10⁶ cells after treating with the above-mentioned drug for 48 hours. 5 pcs·mL -1 Add 100 μL to each well, incubate for 45 min, wash away unadhered cells, add CCK8 solution, incubate for 3-4 h, and detect OD value at 450 nm using a microplate reader.

[0085] The results are as follows Figure 2 As shown in the figure The difference was statistically significant compared to the blank control group. Compared with the blank control group, the cell adhesion ability of composition group 1, composition group 2, composition group 3 and traditional Chinese medicine composition group was significantly reduced (P<0.01). The traditional Chinese medicine composition group showed the most significant reduction effect, indicating that the traditional Chinese medicine composition of the present invention can significantly inhibit the adhesion of lung cancer cells to the extracellular matrix and reduce tumor colonization and local infiltration.

[0086] Transwell assay for cell invasion: Take a Transwell chamber and add 1 mg / mL to the upper chamber. -1The matrix gel was prepared in batches of 100 μL per well and cured at 37°C before use. A549 cells in the logarithmic growth phase were digested, centrifuged, resuspended in serum-free solution, and diluted to 1 × 10⁻⁶. 5 pcs·mL -1 Cell suspensions were added to the lower chamber of a Transwell plate with 500 μL of complete culture medium containing 20% ​​FBS (fetal bovine serum), and 200 μL of cell suspension was added to the upper chamber. Drug intervention was also added, and the plate was incubated for 24 h. The chambers were then removed, the culture medium was washed off with PBS (phosphate-buffered saline), fixed with 4% paraformaldehyde at room temperature for 20 min, stained with 0.25% crystal violet for 20 min, rinsed with PBS, and photographed in five randomly selected fields under an optical microscope. Invasive cell counts were performed using ImageJ image analysis software, and the invasive ability of each group of cells was compared.

[0087] The results are as follows Figure 3 As shown in the figure The difference was statistically significant compared to the blank control group. Compared with the blank control group, the number of transmembrane invasive cells in each treatment group was significantly reduced (P<0.01). The number of transmembrane invasive cells in the traditional Chinese medicine composition group was significantly less than that in the single component group, indicating that the traditional Chinese medicine composition of the present invention can effectively inhibit the invasive ability of A549 lung cancer cells and block the tumor from penetrating the stromal barrier.

[0088] Transwell assay for cell migration ability: A549 cells in logarithmic growth phase were digested, centrifuged, resuspended in serum-free solution, and diluted to 1×10⁻⁶. 5 pcs·mL -1 Cell suspensions were collected and transwell chambers were prepared. 500 μL of complete culture medium containing 20% ​​FBS was added to the lower chamber, and 200 μL of cell suspension was added to the upper chamber. Simultaneously, drug intervention was introduced, and the cells were cultured for 24 h. The chambers were then removed, the culture medium was washed off with PBS, and the cells were fixed with 4% paraformaldehyde at room temperature for 20 min, stained with 0.25% crystal violet for 20 min, rinsed with PBS, and photographed in five randomly selected fields under an optical microscope. Migrating cell counts were performed using ImageJ image analysis software, and the migration ability of each group of cells was compared.

[0089] The results are as follows Figure 4 As shown in the figure The difference was statistically significant compared to the blank control group. Compared with the blank control group, the cell migration ability of each treatment group was significantly reduced (P<0.01). The traditional Chinese medicine composition group showed the most prominent inhibitory effect on migration, indicating that the traditional Chinese medicine composition of the present invention can significantly inhibit the migration of lung cancer cells and reduce the risk of distant metastasis.

[0090] Therefore, the present invention employs the above-mentioned traditional Chinese medicine composition for anti-lung cancer and its preparation method. The formulation is scientific, the process is stable, and the anti-lung cancer synergistic effect is significant. It can effectively inhibit the proliferation, invasion and metastasis of lung cancer cells, while strengthening the body's resistance and reducing toxic side effects, and has good clinical application prospects.

[0091] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A traditional Chinese medicine composition for treating lung cancer, characterized in that: Includes Composition 1, Composition 2, and Composition 3; Composition 1, by weight, includes: Astragalus membranaceus 20-45 parts, Polygonatum odoratum 15-30 parts, Ophiopogon japonicus 8-18 parts, Coix lacryma-jobi 20-45 parts, Phragmites communis 20-45 parts, and Glycyrrhiza uralensis 3-6 parts; Composition 2, by weight, includes: Clematis armandii 9-15 parts, Actinidia chinensis root 15-30 parts, Scutellaria barbata 10-22 parts, Pinellia ternata 6-13 parts, Fritillaria thunbergii 6-13 parts, and Citrus reticulata peel 6-13 parts; Composition 3, by weight, includes: Buthus martensii 3-8 parts.

2. The traditional Chinese medicine composition for treating lung cancer according to claim 1, characterized in that: The mass ratio of Composition 1, Composition 2, and Composition 3 is (1~2):(1~2):(1~2).

3. A method for preparing a traditional Chinese medicine composition for treating lung cancer, characterized in that: The preparation of a traditional Chinese medicine composition for treating lung cancer as described in any one of claims 1-2 comprises the following steps: S1. Preparation of Composition 1: Astragalus membranaceus, Polygonatum odoratum, Ophiopogon japonicus, Coix lacryma-jobi, Phragmites communis, and Glycyrrhiza uralensis are extracted with alcohol to obtain residue and extract. The extract is enriched and purified by macroporous adsorption resin and vacuum dried to obtain alcohol extract. The residue is extracted with water and precipitated with alcohol and vacuum dried to obtain water-extracted alcohol precipitate. The alcohol extract and water-extracted alcohol precipitate are mixed to obtain Composition 1. S2. Preparation of Composition 2: Take three leaves of *Trifolium repens*, *Actinidia chinensis* root, *Scutellaria barbata*, *Pinellia ternata*, *Fritillaria thunbergii*, and tangerine peel. After alcohol extraction and enrichment and purification with macroporous adsorption resin, the mixture is dried under vacuum to obtain Composition 2. S3. Preparation of Composition 3: Take whole scorpions and perform enzymatic hydrolysis. After enzyme inactivation and solid-liquid separation, collect the supernatant and freeze-dry it under vacuum to obtain Composition 3. S4. Preparation of traditional Chinese medicine composition: Mix composition one, composition two, and composition three evenly to obtain the traditional Chinese medicine composition.

4. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S1, the alcohol extraction is performed by using 70%-90% ethanol as the solvent, with a material-to-liquid ratio of 1:(6~12) g / mL, and heating and refluxing for 1-3 times, each time for 0.5-2 hours. The extracts are then combined and concentrated.

5. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S1, the enrichment and purification process is as follows: after the extract is loaded onto a macroporous adsorption resin, it is first eluted with water, then eluted with ethanol at a volume fraction of 50%-95%, the alcohol eluent is collected, and concentrated under reduced pressure.

6. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S1, the water extraction and alcohol precipitation are performed as follows: after evaporating the ethanol from the residue, water is used as the solvent, and the mixture is boiled and extracted 1-3 times at a material-to-liquid ratio of 1:(6~12) g / mL, each time for 0.5-2 hours. The extracts are combined and concentrated to 0.2-1.0 g crude drug / mL. 2-6 times the amount of anhydrous ethanol is added, and the mixture is refrigerated for 8-24 hours. After centrifugation, the precipitate is collected.

7. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S2, the alcohol extraction is performed by using 60%-85% ethanol as the solvent, with a solid-liquid ratio of 1:(6~12) g / mL, and heating and refluxing for 1-3 times, each time for 0.5-2 hours. The extracts are then combined and concentrated.

8. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S2, the enrichment and purification process is as follows: after adsorption onto macroporous resin, the resin is first eluted with water, then eluted with ethanol at a volume fraction of 50%-95%, and the ethanol eluent is collected and concentrated under reduced pressure.

9. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S3, the enzymatic hydrolysis is performed by adding 2000-5000U of trypsin, adjusting the pH to 6.5-8.0, and hydrolyzing at 40-60℃ for 4-10 hours.

10. The method for preparing a traditional Chinese medicine composition for treating lung cancer according to claim 3, characterized in that: In S3, enzyme inactivation is performed by heating the enzyme hydrolysate to 85-100℃ and maintaining it for 15-30 minutes.

Images