Pharmaceutical composition for treating lung cancer and application thereof

Abstract

The invention relates to the technical field of medicine application, and discloses a medicine composition for treating lung cancer and application thereof.The medicine composition is prepared from semen brassicae, radix ranunculi ternati, sculellaria barbata, selfheal, sophora tonkinensis, radix asparagi, sanguisorba officinalis, burdock, turmeric, dried balsam pear, earthworm, agilawood and the like, radix ranunculi ternati, sculellaria barbata and selfheal are used as ministerial drugs, sophora tonkinensis, radix asparagi, sanguisorba officinalis, burdock, turmeric, dried balsam pear, dandelion, radix angelicae pubescentis, sophora flower bud, radix angelicae, radix paeoniae rubra, radix puerariae, lumbricus and agilawood are used as adjuvant drugs, and liquorice is used as conductant drugs, so that the effect of treating lung cancer can be remarkably improved by combining all the drugs. Tests prove that the prepared pharmaceutical composition has the effects of awakening NK cells and inhibiting growth of lung cancer cells A549.

Description

Technical Field

[0001] The present invention relates to the technical field of drug application, and in particular to a drug composition for treating lung cancer and application thereof. Background Art

[0002] Lung cancer is a malignant tumor with a high mortality rate and is highly harmful to human health. Chemotherapy drugs are extremely harmful to the human body, and most patients will experience symptoms such as fatigue, phlebitis, hair loss, nausea, vomiting, and even heart dysfunction. Traditional Chinese medicine is low-cost, widely available, and has very little toxicity or side effects in clinical practice.

[0003] However, the existing Chinese medicine treatment course is long and the treatment effect needs to be improved. How to obtain better lung cancer treatment effects, especially how to find drugs with better treatment effects and lower toxic side effects for lung cancer cells, requires further research. Summary of the invention

[0004] In view of this, in order to solve the problem that the treatment methods for lung cancer in the prior art are relatively limited, the present invention provides a pharmaceutical composition for treating lung cancer and use thereof.

[0005] In a first aspect, the present invention provides a pharmaceutical composition for treating lung cancer.

[0006] In some specific embodiments, the raw materials of the pharmaceutical composition include, by weight:

[0007] 70-90 parts of white mustard seeds, 70-90 parts of cat's claw, 95-105 parts of Scutellaria barbata, 95-105 parts of Prunella vulgaris, 110-130 parts of Sophora japonica, 95-105 parts of Asparagus cochinchinensis, 125-135 parts of Sanguisorba officinalis, 70-90 parts of Arctium lappa, 80-100 parts of Curcuma longa, 70-90 parts of dried bitter melon, 70-90 parts of Taraxacum officinale, 55-65 parts of Angelica dahurica, 45-55 parts of Sophora japonica seeds, 35-45 parts of the Angelica dahurica, 30 parts of red peony root, 95-105 parts of Pueraria root, 55-65 parts of Pheretima, 45-55 parts of Aquilaria reticulata, and 45-55 parts of Liquorice.

[0008] In some specific embodiments, the raw material composition of the pharmaceutical composition is: 80 parts of white mustard seeds, 80 parts of cat's claw, 100 parts of bark scutellaria, 100 parts of selfheal, 120 parts of Vietnamese Sophora japonica, 100 parts of asparagus, 130 parts of sanguisorba officinalis, 80 parts of burdock seeds, 90 parts of turmeric, 80 parts of dried bitter melon, 80 parts of dandelion, 60 parts of angelica dahurica, 50 parts of sophora japonica seeds, 40 parts of angelica dahurica, 30 parts of red peony root, 100 parts of kudzu root, 60 parts of earthworms, 50 parts of agarwood, and 50 parts of licorice.

[0009] The raw material medicine of the present invention has the following effects:

[0010] White mustard contains sinapinic acid, sinapinic acid, myrosinase, fat, protein and mucus, and has the effects of dispersing nodules and relieving pain.

[0011] Cat's claw contains protoanemonin, ranunculin, polysaccharides, etc., and has the effects of resolving phlegm and dispersing nodules, detoxifying and reducing swelling.

[0012] Scutellaria barbata, containing flavonoids, has the effects of clearing away heat and detoxifying, dispersing nodules, and inhibiting tumor growth.

[0013] Prunella Vulgaris contains triterpenoid saponins, oleanolic acid, sugars, etc. It has the effects of clearing the liver and purging fire, dispersing nodules and reducing swelling, and anti-tumor.

[0014] Vietnamese Sophora japonica (Sophora japonica) contains oxymatrine, safflower oil, pterostilbene, sophora japonica, etc. It can fight tumors, inhibit leukemia cells, and increase the number of white blood cells.

[0015] Asparagus, containing asparagine, asparagine, citrulline, etc., has a strong inhibitory effect on diphtheria Corynebacterium, Staphylococcus aureus, Bacillus subtilis, Streptococcus and anthrax.

[0016] Sanguisorba officinalis, which contains sanguisorba officinalis saponins, ellagic acid, catechins, etc., has a killing effect on typhoid Salmonella, beta-hemolytic Streptococcus, and Escherichia coli, and has an inhibitory effect on the growth of uterine tumors.

[0017] Burdock seeds contain dietary fiber and arctiin, which have the effects of dispersing nodules and anti-tumor.

[0018] Turmeric, containing curcumin, has anti-tumor effects and has a strong inhibitory and killing effect on Gram-negative and Gram-positive bacteria (such as Escherichia coli, Staphylococcus aureus) and fungi (Candida albicans).

[0019] Dried bitter melon contains momordica glycoside, momordica extract, protein, potassium, etc. It can clear away heat and detoxify, and has strong anti-tumor and anti-HIV effects.

[0020] Dandelion mainly contains taraxasterol and inulin, which can clear away heat and detoxify, promote blood circulation and disperse nodules, and prevent the occurrence of tumors.

[0021] Angelica dahurica contains bergamotol, which has inhibitory and killing effects on various bacteria and has anti-tumor effects.

[0022] Sophora japonica seeds are rich in selenium and have the effects of cooling blood, anti-virus and anti-tumor growth.

[0023] Angelica dahurica contains coumarins and volatile oils, which have the effects of killing bacteria and preventing tumor growth.

[0024] Red peony root contains flavonoids, paeoniflorin, volatile oils, etc., and has bactericidal and anti-tumor effects.

[0025] Pueraria root, containing puerarin, puerarin, protein, amino acids, etc., has the effects of promoting blood circulation, removing blood stasis and anti-tumor growth.

[0026] Earthworms contain protein, succinic acid, etc., and have the effects of clearing away heat, detoxifying, and dispersing nodules.

[0027] Agarwood contains volatile oils (benzyl acetone, p-methoxybenzyl acetone, etc.), agarwood spirol, etc., which have the effects of promoting qi and relieving pain, promoting blood circulation and dispersing nodules.

[0028] Licorice contains alkaloids, coumarins, flavonoids, etc., which have the effects of clearing heat and detoxifying, anti-virus, and inhibiting tumor growth.

[0029] The white mustard seeds in the pharmaceutical composition of the present invention are the main drug and the raw material drug with the main effect. Cat's claw, Scutellaria barbata and Prunella vulgaris are the assistant drugs, which play a synergistic role and strengthen the therapeutic effect of the main drug; Vietnamese Sophora japonica, Asparagus cochinchinensis, Sanguisorba officinalis, Arctium lappa, Curcuma longa, Momordica charantia, Taraxacum officinale, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Pueraria root, Pheretima and Aquilaria sinensis are the adjuvant drugs, which play the role of assisting and supporting the main drug. Licorice is the guiding drug, which not only guides the drugs in the prescription to reach the lesion, but also has the function of coordinating the effects between the drugs.

[0030] In a second aspect, the present invention provides a pharmaceutical preparation.

[0031] The pharmaceutical preparation comprises the pharmaceutical composition and / or its active extract.

[0032] In some embodiments, the pharmaceutical preparation further comprises a pharmaceutically acceptable excipient;

[0033] Optionally, the auxiliary materials include one or more of solvents, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, stabilizers, flavoring agents, preservatives, suspending agents, coating materials, fragrances and thickeners.

[0034] Exemplarily, the raw materials are respectively passed through a 90-mesh sieve and then mixed into powders, and 54 g of refined honey is added to every 100 g of powders to prepare honeyed pills.

[0035] In some embodiments, the dosage form of the pre-drug preparation includes any pharmaceutically acceptable preparation;

[0036] Optionally, the dosage form of the pharmaceutical preparation is tablets, pills, decoctions, capsules, powders, drops, lyophilized products, granules, ointments, patches, suspensions, syrups, oral solutions, injections, and suppositories.

[0037] In a third aspect, the present invention provides a method for preparing the pharmaceutical preparation.

[0038] In some specific embodiments, the method for preparing the drug includes the steps of weighing a certain amount of bulk drug components for compounding or extracting the active ingredients of the bulk drug for compounding;

[0039] Optionally, the preparation method of the pharmaceutical preparation comprises:

[0040] (1) mixing white mustard seeds, asparagus cochinchinensis, dried bitter melon, kudzu root, earthworm, water and papain, performing enzymolysis, and performing solid-liquid separation to obtain liquid A;

[0041] Mixing cat's claw, selfheal, burdock, dandelion with water and cellulase, performing enzymolysis, and separating the enzymolysis product into solid and liquid to obtain liquid B;

[0042] Scutellaria barbata, Sophora vietnamese, Sanguisorba officinalis, Curcuma longa, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Aquilaria sinensis and Licorice root are extracted with water, and the solid-liquid separation is performed to obtain a medicinal solution C;

[0043] The drug solution A, the drug solution B and the drug solution C are mixed and then distilled to remove impurities to obtain a puree;

[0044] (2) mixing soybean flour and part of the puree obtained in step (1) and fermenting them to obtain baking powder;

[0045] (3) fermenting the remaining puree obtained in step (1) and the fermentation powder obtained in step (2), filtering, and obtaining a fermentation liquid;

[0046] (4) adding lysozyme to the fermentation broth obtained in step (3) and mixing to form the pharmaceutical preparation.

[0047] The inventors found that the water extract of the raw material medicine contains more antibacterial components, which makes the subsequent fermentation process slow. In order to increase the fermentation speed and promote the growth of fermentation bacteria, the present invention uses papain to hydrolyze white mustard, asparagus, dried bitter melon, kudzu root, and earthworm to produce more protein and amino acids, and uses cellulase to hydrolyze cat's claw, selfheal, burdock, and dandelion to produce more polysaccharides; the products after enzymatic hydrolysis can promote the growth of fermentation bacteria, and more components that inhibit the growth of lung cancer cells are produced after adding lysozyme.

[0048] In some specific embodiments, in step (1), the raw materials are first soaked and boiled in water before enzymolysis, and the amount of water used for soaking is 6-10 times the total weight of the raw materials to be soaked; the soaking time is 1-3 hours, the boiling temperature is 90-100° C., and the boiling time is 50-60 minutes;

[0049] And / or, the amount of water used for the water extraction is 6-10 times the total weight of the raw material to be extracted; the temperature of the water extraction is 90-100° C., and the time of the water extraction is 50-60 minutes;

[0050] And / or, the enzymatic activity of the papain is 80000-120000 U / g, the added amount of the papain is 0.003-0.01% of the total weight of white mustard, asparagus, dried bitter melon, kudzu root and earthworm; the enzymatic hydrolysis time of the papain is 1-10 hours, and the enzymatic hydrolysis temperature is 55-65° C.;

[0051] And / or, the enzymatic activity of the cellulase is 8000-15000 U / g, the added amount of the cellulase is 0.1-0.3% of the total weight of cat's claw, selfheal, burdock and dandelion; the enzymatic hydrolysis time of the cellulase is 5-24 hours, and the enzymatic hydrolysis temperature is 28-32° C.;

[0052] And / or, the distillation temperature is 60-80°C.

[0053] In some specific embodiments, the mass ratio of the soybean flour to the puree in step (2) is 1:(1-3);

[0054] And / or, in step (3), lysozyme is added to the fermentation broth and mixed until saturated; 2-4 g lysozyme is added per 100 ml of fermentation broth.

[0055] The fermentation temperature is 40-50°C.

[0056] The enzymatic activity of lysozyme is 2-4 million / g; in the fermentation broth, the amount of lysozyme added is 2-5g / 100ml of the fermentation broth;

[0057] The enzymatic activity of papain is 80000-120000U / g, and the added amount is 0.003-0.01% of the total weight of white mustard, asparagus, dried bitter melon, kudzu root and earthworm;

[0058] The enzymatic activity of the cellulase is 8000-15000U / g, and the added amount is 0.1-0.3% of the total weight of cat's claw, selfheal, burdock and dandelion;

[0059] The effects of lysozyme in the present invention are as follows: 1. Antibacterial and antiviral, it can hydrolyze the insoluble mucopolysaccharides on the cell walls of Gram-positive bacteria in the pharmaceutical preparation, dissolve the bacteria, and release more effective ingredients, thereby achieving better anti-inflammatory and antiviral effects; 2. Promote tissue repair in the body; 3. If the pharmaceutical preparation of the present invention is used in combination with antibiotics, lysozyme can also enhance the efficacy of antibiotics.

[0060] In some specific embodiments, the distillation temperature is 70-80°C;

[0061] And / or, the fermentation temperature in step (2) is 40-50°C;

[0062] And / or, the fermentation temperature in step (3) is 50-60°C.

[0063] In a fourth aspect, the present invention provides a use of the pharmaceutical composition and / or its active extract or the pharmaceutical preparation for preparing a drug for preventing or treating lung cancer.

[0064] In some specific embodiments, the pharmaceutical composition and / or its active extract or the pharmaceutical preparation is used to prepare a drug that can awaken NK cells and / or inhibit the growth of lung cancer cells A549.

[0065] In some embodiments, the drug also has the effect of dispersing nodules.

[0066] In some embodiments, the subjects of application include humans and animals other than humans.

[0067] In some specific embodiments, the drug preparation is used orally; the adult dosage is 5 g once, twice a day, for three months or as directed by a doctor.

[0068] Compared with the prior art, the present invention has the following beneficial effects:

[0069] The pharmaceutical composition of the present invention is composed of white mustard, cat's claw, Scutellaria barbata, Prunella vulgaris, Vietnamese Sophora japonica, asparagus, burdock, turmeric, dried bitter melon, dandelion, angelica dahurica, sophora japonica seed, angelica dahurica, red peony root, liquorice, pueraria root, earthworm and agarwood, wherein white mustard is the main drug, which is the main raw material drug; cat's claw, Scutellaria barbata, Prunella vulgaris are ministerial drugs, which play a synergistic role and strengthen the therapeutic effect of the main drug. Vietnamese Sophora japonica, asparagus, burdock, turmeric, dried bitter melon, dandelion, angelica dahurica, red peony root, pueraria root, earthworm and agarwood are adjuvant drugs, which play the role of assisting and supporting the main drug. Licorice is a guiding drug, which not only guides the drugs in the prescription to reach the lesion, but also has the function of coordinating the effects between the drugs. The combination of the drugs can significantly improve the effect of treating lung cancer. It is proved by experiments that the prepared pharmaceutical composition has the effect of awakening NK cells and inhibiting the growth of lung cancer cells A549.

[0070] The pharmaceutical composition of the present invention has an inhibition rate of 97.1% on human lung cancer cells at a concentration of 0.1 g / ml, and a half-inhibitory growth concentration IC50 of 0.0018 g / ml on human lung cancer cells A549.

[0071] The preparation method of the pharmaceutical preparation of the present invention adopts papain enzymolysis to produce more proteins and amino acids, and cellulase enzymolysis to produce more polysaccharides; the product after enzymolysis can promote the growth of fermentation bacteria, and more components that inhibit the growth of lung cancer cells are produced after adding lysozyme. The prepared pharmaceutical preparation can reach the nanometer level, has very good permeability, can decompose lung cancer cell protein, and convert it into beneficial protein for absorption and utilization. The effective ingredients therein can penetrate the cell membrane of lung cancer cells, destroy its cell wall and cell membrane system, make it lose its barrier function, inhibit its growth and reproduction, and at the same time destroy its genetic material or cell microstructure, hinder the replication of genetic information, and inhibit the decomposition of lung cancer cells to produce harmful substances, and convert lung cancer cells into nutrients for damaged cells to absorb and then perform self-repair. At the same time, the whole process does not produce any harmful substances, and has high safety. Moreover, the pharmaceutical composition of the present invention has been proved to be non-toxic through experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0073] Figure 1 Comparison of OD values ​​measured in each test group in Experimental Example 1 of the present invention;

[0074] Figure 2 The results of the effects of the pharmaceutical composition at concentrations of 0, 0.63 μl / ml, 0.125 μl / ml, 0.5 μl / ml, and 1 μl / ml on the epigenetic state of lung cancer cells in Experimental Example 1 of the present invention;

[0075] Figure 3 The results of the effects of the pharmaceutical composition at concentrations of 2 μl / ml, 4 μl / ml, 8 μl / ml, and 16 μl / ml on the epigenetic state of lung cancer cells in Experimental Example 1 of the present invention;

[0076] Figure 4 The results of the logarithmic linear regression curve of probability unit-sample concentration in Experimental Example 2 of the present invention. DETAILED DESCRIPTION

[0077] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0078] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0079] In the present invention, the selection scope involving "and / or", "or / and", and "and / or" includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, and the any and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items.

[0080] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0081] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0082] The present invention is further described in detail below in conjunction with specific embodiments. These embodiments should not be construed as limiting the scope of protection claimed by the present invention.

[0083] The papain used in the following experiments has an enzyme activity of 100,000 U / g, model FDG-2203, the cellulase has an enzyme activity of 10,000 U / g, model FFY-0673, and the lysozyme has an enzyme activity of 3 million U / g, model SDG-2447, all of which were purchased from Xiasheng Biotechnology Development Co., Ltd.

[0084] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0085] Example 1

[0086] This embodiment provides a pharmaceutical composition for treating lung cancer, comprising the following raw materials:

[0087] 80g white mustard seeds, 80g cat's claw, 100g bark scutellaria, 100g selfheal, 120g Vietnamese sophora, 100g asparagus, 130g burdock, 80g burdock seeds, 90g turmeric, 80g dried bitter melon, 80g dandelion, 60g angelica, 50g sophora japonica seeds, 40g angelica dahurica, 30g red peony root, 100g kudzu root, 60g earthworm, 50g agarwood, and 50g licorice.

[0088] The preparation method is as follows:

[0089] (1) The above raw materials are crushed into fine powder, white mustard seeds, asparagus cochinchinensis, dried bitter melon, kudzu root and earthworm are weighed according to the proportion, and mixed to form a mixture A; 0.007% papain is weighed based on the weight of the mixture A;

[0090] Add 8 times the weight of water to the mixture A and soak for 2 hours, then boil at 95°C for 55 minutes; cool the boiled liquid to 60°C, add papain for enzymolysis for 5 hours at a temperature of 60°C, separate the solid and liquid, and obtain liquid A;

[0091] Weigh cat's claw, selfheal, burdock and dandelion according to the proportion, and mix to form a mixture B; weigh 0.2% of the weight of the mixture B in cellulase; add 8 times the weight of water to the mixture B and soak for 2 hours, then boil at 90-100° C. for 50-60 minutes; cool the boiled liquid to 30° C., add cellulase for enzymolysis for 5-24 hours, the enzymolysis temperature is 28-32° C., and separate the enzymolysis product into solid and liquid to form liquid B;

[0092] Weigh Scutellaria barbata, Sophora vietnamese, Sanguisorba officinalis, Curcuma longa, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Aquilaria sinensis and Licorice according to the proportion, mix, add 8 times the weight of water, soak for 2 hours, then boil at 95°C for 55 minutes, separate the solid and liquid, and form liquid C;

[0093] The drug solution A, the drug solution B and the drug solution C are mixed and then distilled at 70° C. to remove impurities to obtain a puree;

[0094] (2) adding 2 times the weight of the puree obtained in step (1) to 10 g of soybean flour, mixing and fermenting at 45° C. for 4 days to obtain baking powder;

[0095] (3) fermenting the remaining puree obtained in step (1) and the baking powder obtained in step (2) at 45° C. for 9 hours, filtering through a 200-mesh sieve to obtain a fermentation liquid;

[0096] (4) Add lysozyme to the fermentation broth obtained in step (3) and mix until saturated, adding 3 g of lysozyme per 100 ml of fermentation broth to form a pharmaceutical preparation.

[0097] Example 2

[0098] This embodiment provides a pharmaceutical composition for treating lung cancer, comprising the following raw materials:

[0099] 80g white mustard seeds, 80g cat's claw, 100g bark scutellaria, 100g selfheal, 120g Vietnamese sophora, 100g asparagus, 130g burdock, 80g burdock seeds, 90g turmeric, 80g dried bitter melon, 80g dandelion, 60g angelica, 50g sophora japonica seeds, 40g angelica dahurica, 30g red peony root, 100g kudzu root, 60g earthworm, 50g agarwood, and 50g licorice.

[0100] The preparation method is as follows:

[0101] (1) The above raw materials are crushed into fine powder, white mustard seeds, asparagus cochinchinensis, dried bitter melon, kudzu root and earthworm are weighed according to the proportion, and mixed to form a mixture A; 0.003% papain is weighed based on the weight of the mixture A;

[0102] Add 10 times the weight of water to the mixture A and soak for 1 hour, then boil at 100°C for 50 minutes; cool the boiled liquid to 55°C, add papain for enzymolysis for 10 hours at a temperature of 55°C, separate the solid and liquid, and obtain liquid A;

[0103] Weigh cat's claw, selfheal, burdock and dandelion according to the proportion, and mix to form a mixture B; weigh 0.3% of the weight of the mixture B in cellulase; add 6 times the weight of water to the mixture B and soak for 3 hours, then boil at 90° C. for 60 minutes; cool the boiled liquid to 28° C., add cellulase for enzymolysis for 24 hours, the enzymolysis temperature is 28° C., and the enzymolysis product is separated into solid and liquid to form liquid B;

[0104] Weigh Scutellaria barbata, Sophora vietnamese, Sanguisorba officinalis, Curcuma longa, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Aquilaria sinensis and Licorice according to the proportion, mix, add 10 times the weight of water, soak for 1 hour, then boil at 90°C for 60 minutes, separate the solid and liquid, and form liquid C;

[0105] The drug solution A, the drug solution B and the drug solution C are mixed and then distilled at 60° C. to remove impurities to obtain a puree;

[0106] (2) adding 30 g of the puree obtained in step (1) to 10 g of soybean flour, mixing and fermenting at 50° C. for 2 days to obtain baking powder;

[0107] (3) fermenting the remaining puree obtained in step (1) and the baking powder obtained in step (2) at 50° C. for 6 hours, filtering through a 200-mesh sieve to obtain a fermentation liquid;

[0108] (4) adding lysozyme to the fermentation broth obtained in step (3) and mixing until saturated to form a pharmaceutical preparation, and freeze-drying to form a pharmaceutical composition.

[0109] Example 3

[0110] This embodiment provides a pharmaceutical preparation for treating lung cancer, comprising the following raw materials:

[0111] 80g white mustard seeds, 80g cat's claw, 100g bark scutellaria, 100g selfheal, 120g Vietnamese sophora, 100g asparagus, 130g burdock, 80g burdock seeds, 90g turmeric, 80g dried bitter melon, 80g dandelion, 60g angelica, 50g sophora japonica seeds, 40g angelica dahurica, 30g red peony root, 100g kudzu root, 60g earthworm, 50g agarwood, and 50g licorice.

[0112] The preparation method is as follows:

[0113] (1) The above raw materials are crushed into fine powder, white mustard seeds, asparagus cochinchinensis, dried bitter melon, kudzu root and earthworm are weighed according to the proportion, and mixed to form a mixture A; 0.01% papain is weighed based on the weight of the mixture A;

[0114] Add 6 times the weight of water to the mixture A and soak for 3 hours, then boil at 90°C for 60 minutes; cool the boiled liquid to 65°C, add papain for enzymolysis for 1 hour at a temperature of 65°C, separate the solid and liquid, and obtain liquid A;

[0115] Weigh cat's claw, selfheal, burdock and dandelion according to the proportion, and mix to form a mixture B; weigh 0.1% of the weight of the mixture B in cellulase; add 10 times the weight of water to the mixture B and soak for 1 hour, then boil at 9100℃ for 50 minutes; cool the boiled liquid to 32℃, add cellulase for enzymolysis for 5 hours, the temperature of enzymolysis is 32℃, and the enzymolysis product is separated into solid and liquid to form liquid B;

[0116] Weigh Scutellaria barbata, Sophora vietnamese, Sanguisorba officinalis, Curcuma longa, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Aquilaria sinensis and Licorice according to the proportion, mix, add 6 times the weight of water, soak for 3 hours, then boil at 100° C. for 50 minutes, separate the solid and liquid, and form a medicinal solution C;

[0117] The drug solution A, the drug solution B and the drug solution C are mixed and then distilled at 80° C. to remove impurities to obtain a raw paste;

[0118] (2) adding 10 g of the puree obtained in step (1) to 10 g of soybean flour, mixing and fermenting at 40° C. for 6 days to obtain baking powder;

[0119] (3) fermenting the remaining puree obtained in step (1) and the baking powder obtained in step (2) at 40° C. for 12 hours, filtering through a 200-mesh sieve to obtain a fermentation liquid;

[0120] (4) adding lysozyme to the fermentation broth obtained in step (3) and mixing until saturated to form a pharmaceutical preparation.

[0121] Example 4

[0122] This embodiment provides a pharmaceutical preparation for treating lung cancer, comprising the following raw materials:

[0123] 80g white mustard seeds, 80g cat's claw, 100g bark scutellaria, 100g selfheal, 120g Vietnamese sophora, 100g asparagus, 130g burdock, 80g burdock seeds, 90g turmeric, 80g dried bitter melon, 80g dandelion, 60g angelica, 50g sophora japonica seeds, 40g angelica dahurica, 30g red peony root, 100g kudzu root, 60g earthworm, 50g agarwood, and 50g licorice.

[0124] The preparation method is as follows:

[0125] (1) Weigh the above raw materials according to the ratio and crush them into fine powders, mix them, add 6 times the weight of water, soak them for 3 hours, and then boil them at 100° C. for 50 minutes to separate the solid and liquid to form a drug solution;

[0126] The drug solution is distilled at 80° C. to remove impurities and obtain a puree;

[0127] (2) adding 10 g of the puree obtained in step (1) to 10 g of soybean flour, mixing and fermenting at 40° C. for 6 days to obtain baking powder;

[0128] (3) fermenting the remaining puree obtained in step (1) and the baking powder obtained in step (2) at 40° C. for 12 hours, filtering through a 200-mesh sieve to obtain a fermentation liquid;

[0129] (4) adding lysozyme to the fermentation broth obtained in step (3) and mixing until saturated to form a pharmaceutical preparation.

[0130] Example 5

[0131] This embodiment provides a pharmaceutical preparation for treating lung cancer, comprising the following raw materials:

[0132] 80g white mustard seeds, 80g cat's claw, 100g bark scutellaria, 100g selfheal, 120g Vietnamese sophora, 100g asparagus, 130g burdock, 80g burdock seeds, 90g turmeric, 80g dried bitter melon, 80g dandelion, 60g angelica, 50g sophora japonica seeds, 40g angelica dahurica, 30g red peony root, 100g kudzu root, 60g earthworm, 50g agarwood, and 50g licorice.

[0133] The preparation method is as follows:

[0134] (1) Weigh the above raw materials according to the ratio and crush them into fine powders, mix them, add 6 times the weight of water, soak them for 3 hours, and then boil them at 100° C. for 50 minutes to separate the solid and liquid to form a drug solution;

[0135] The drug solution is distilled at 80° C. to remove impurities and obtain a puree;

[0136] (2) adding 10 g of the puree obtained in step (1) to 10 g of soybean flour, mixing and fermenting at 40° C. for 6 days to obtain baking powder;

[0137] (3) fermenting the remaining puree obtained in step (1) and the baking powder obtained in step (2) at 40° C. for 12 hours, filtering through a 200-mesh sieve, and forming a pharmaceutical preparation.

[0138] Example 6

[0139] This embodiment provides a pharmaceutical composition for treating lung cancer, comprising the following raw materials:

[0140] 90g white mustard seeds, 70g cat's claw, 105g barbata, 95g selfheal, 130g Vietnamese Sophora japonica, 105g asparagus, 135g sanguisorba officinalis, 70g burdock seeds, 100g turmeric, 70g dried bitter melon, 90g dandelion, 65g angelica dahurica, 45g sophora japonica seed, 45g angelica dahurica, 30g red peony root, 105g kudzu root, 55g earthworm, 55g agarwood, and 45g licorice.

[0141] The preparation method is as follows:

[0142] The above raw materials are crushed into fine powder, passed through a 90-mesh sieve, mixed evenly, and pressed into tablets to form tablets.

[0143] Example 7

[0144] This embodiment provides a pharmaceutical composition for treating lung cancer, comprising the following raw materials:

[0145] 70g white mustard seeds, 90g cat's claw, 95g bark scutellaria, 105g selfheal, 110g Vietnamese sophora, 95g asparagus, 125g burdock, 90g burdock seeds, 80g turmeric, 90g dried bitter melon, 70g dandelion, 55g angelica dahurica, 55g sophora japonica seed, 35g angelica dahurica, 30g red peony root, 95-105g kudzu root, 55-65g earthworm, 45-55g agarwood, and 45-55g licorice.

[0146] The preparation method is as follows:

[0147] The above raw materials are crushed into fine powder, passed through a 200-mesh sieve, mixed evenly, made into a soft material with an appropriate amount of 80% ethanol, granulated, dried, and made into capsules.

[0148] Experimental Example 1 CCK-8 kit to determine the inhibitory effect of the drug composition on A549 cell growth

[0149] 1. A549 cell culture

[0150] 1.1 Cells

[0151] Human lung cancer cell line A549 (Shanghai Meixuan Biotechnology Co., Ltd., MXC026)

[0152] 1.2 Instruments and reagents

[0153] The details are as follows Table 1:

[0154] Table 1 Instruments and reagents

[0155]

[0156] 1.3 Cell culture

[0157] (1) Take out the cells from the liquid nitrogen tube, quickly put the cryotube into a water bath preheated to 37°C to thaw quickly, and shake it continuously to melt the liquid in the tube quickly. After about 1mL of the liquid in the cryotube is completely dissolved, take out the cells under sterile conditions and inoculate them into 10mL of complete culture medium (containing 90% DMEM high glucose or RPMI1640 + 10% fetal bovine serum), place it in a 37°C, CO2 incubator, replace the culture medium the next day, continue culturing, and observe the growth.

[0158] (2) After the cells have grown to full size, subculture them. Discard the culture medium and wash twice with PBS (without calcium and magnesium ions); add 1 mL of digestion solution (0.25% Trypsin-0.53 mM EDTA) to the culture bottle and place it upside down in a 37°C incubator for 3 minutes to preheat. After that, flip the culture bottle for 30 seconds and observe the cell digestion under an inverted microscope. If most of the cells become round, quickly take it back to the operating table, tap the culture bottle a few times, and add a small amount of complete culture medium to terminate the digestion.

[0159] (3) Add complete culture medium at a dosage of 10 mL / bottle, gently stir to mix, then aspirate half of it and transfer it to a new culture bottle.

[0160] 2.CCk8 detection

[0161] 2.1 Trial Grouping

[0162] Drug group: Take 0.63 μl of the drug preparation of Example 4, add sterile water to 1 ml, and obtain a concentration of 0.63 μl / ml (drug group 1); according to the above method, the drug preparation of Example 4 was diluted to concentrations of 0.125 μl / ml, 0.5 μl / ml, 1 μl / ml, 2 μl / ml, 4 μl / ml, 8 μl / ml, and 16 μl / ml (drug groups 2-8); no drug preparation was added to the blank control group; an equal volume of sterile water was used in the negative control group; three replicates were used in each group.

[0163] 2.2 Experimental procedures

[0164] (1) Digest cells, count cells, and adjust the cell concentration to 2×104 Pieces / ml.

[0165] (2) Prepare 200 μL of cell suspension in a 96-well plate and culture the plate in an incubator at 37° C. and 5% CO 2 for 24 h.

[0166] (3) According to different groups, the corresponding concentrations of drug preparation solutions (drug groups 1-8) were added, and the negative control group was added with an equal volume of sterile water and cultured for 48 h; this step was omitted for the blank control group;

[0167] (4) After the culture, the cells were washed twice with PBS and 10 μL of CCK-8 solution was added to each well;

[0168] (5) Incubate the culture plate for 1 hour; measure the absorbance at 490 nm using an ELISA reader.

[0169] 3. Test results

[0170] The OD values ​​of each test group were as follows: Figure 1 As shown;

[0171] The results of the effects of the drug composition at concentrations of 0 (negative control group), 0.63 μl / ml, 0.125 μl / ml, 0.5 μl / ml, and 1 μl / ml on the epigenetic state of lung cancer cells are shown in Figure 2 The results of the effects of the drug composition at concentrations of 2 μl / ml, 4 μl / ml, 8 μl / ml, and 16 μl / ml on the epigenetic state of lung cancer cells are shown in Figure 3 .

[0172] This test is conducted by Fuda Testing Center. Figure 1-3 The test results show that the drug composition of different concentrations has a good dose-effect relationship on the proliferation inhibition of A549 cells. Under the microscope, it can be seen that the adherent cells fall off to varying degrees and are suspended in the culture medium. Under the light microscope, the cells are swollen, and more particles and vacuoles can be seen in the cytoplasm. The nucleus is condensed. The number of cells decreases significantly with the increase of drug concentration. The cells in the control group are polygonal, adherent to the wall, and almost do not fall off. Under the light microscope, the cells are intact, the cytoplasm is full, and the adjacent cells grow and fuse into pieces. It is suggested that the drug composition of the present invention has a good inhibitory effect on human lung cancer cells A549.

[0173] Experimental Example 2: Half-maximal inhibitory growth concentration IC50 of the drug composition on human lung cancer A549 cells

[0174] 2.1 Cells

[0175] Human lung adenocarcinoma A549 cells (Manufacturer: BNCC, No.: BNCC337696)

[0176] 2.2 Reagents

[0177] DMEM culture medium (manufacturer: Biological Industries);

[0178] FBS (manufacturer: AUSGENEX);

[0179] Double antibody (manufacturer: GIBCO);

[0180] Phosphate buffer (manufacturer: GIBCO);

[0181] Pancreatin (manufacturer: GIBCO);

[0182] MTT (Manufacturer: GIBCO).

[0183] 2.3 Test methods

[0184] (1) Preparation of serum-containing culture medium: 88% DMEM, 10% FBS, 2% double antibody;

[0185] (2) Preparation of cell suspension:

[0186] Take a dish of monolayer cells in good growth condition with a near-confluence of 90%, wash it twice with PBS, remove cells in poor condition, digest them with trypsin until the cells fall off the plate, resuspend the cells in the prepared culture medium containing 10% fetal bovine serum to obtain a cell suspension, count the live cells with a cell counter, and adjust the live cell concentration of the cell suspension to 50,000 / mL with serum-containing culture medium.

[0187] (3) Preparation of sample solution

[0188] 0.5 g of the pharmaceutical preparation sample of Example 1 of the present invention was mixed with 5 mL of serum-containing culture medium to obtain a 0.1 g / mL sample solution.

[0189] (4) Half inhibition test:

[0190] Seed plate: prepare a 96-well plate, add 100 μl of the cell suspension obtained in step (2) to each well, and then place it in a 37°C ± 1°C, 5% CO2 incubator for 24 hours.

[0191] Sampling: Take the drug preparation sample obtained in step (3) and dilute it with sterile water to obtain sample dilutions with concentrations of 0.1 g / mL, 0.05 g / mL, 0.035 g / mL, 0.0125 g / mL, 0.00625 g / mL, 0.00313 g / mL, 0.00157 g / mL, 0.00078 g / mL, and 0.00039 g / mL.

[0192] After culturing the 96-well plate with A549 cells for 24 hours, take it out of the incubator, remove the supernatant, add 100 μL of the corresponding sample dilution, set up 5 replicates for each concentration, and use an equal volume of culture medium containing 10% fetal bovine serum as a negative control. Then place it in a 37℃±1℃, 5% CO2 incubator for 22 hours.

[0193] (5) MTT incubation: After 22 hours of incubation, the 96-well plate was removed, the supernatant in the wells was removed, and then 50 μL of 1 mg / mL MTT solution (solvent MEM culture medium) was added to each well. The plate was then placed in an incubator at 37°C ± 1°C and 5% CO2 for 2 hours.

[0194] (6) Reading absorbance: After the incubation, remove the 96-well plate, remove the MTT reagent, add 100 μL of isopropanol to each well to dissolve the reaction product, shake for 15 min, and then read the absorbance of each well at 570 nm and 650 nm, respectively.

[0195] (7) Calculation:

[0196] The cell growth inhibition rate was calculated according to the following formula:

[0197]

[0198] Where:

[0199] <OD 570 -OD 650 ) T : indicates the absorbance value of the test group

[0200] <OD 570 -OD 650 ) K : indicates the absorbance value of the negative control group

[0201] The IC50 (half inhibitory concentration) was calculated as follows:

[0202] The average growth inhibition rate of the five replicate wells was calculated and used as the growth inhibition rate of the sample at that concentration. The average was then converted into a probability unit. The probability unit was linearly correlated with the logarithm of the sample concentration. A quadratic equation of the probability unit-logarithm of the sample concentration was obtained by linear regression. The probability unit was taken as 5 to obtain the logarithm of IC50, which was then converted into IC50.

[0203] (9) Results:

[0204] The results of the linear regression curve of probability unit-sample concentration logarithm are as follows Figure 4The concentrations applicable to the probability unit method are 0.0125, 0.00625, 0.00313, 0.00157, 0.00078, and 0.00039 g / ml. Figure 4 The results showed that under the experimental conditions, the half-maximal inhibitory growth concentration IC50 of the pharmaceutical composition of the present invention on human lung cancer cells A549 was 0.0018 g / ml, indicating that the pharmaceutical composition of the present invention has a good inhibitory effect on human lung cancer cells A549.

[0205] Test Example 3 Acute inhalation toxicity test

[0206] 1. Experimental Animals

[0207] 10 clean grade Kunming mice, weighing 18-22g, half male and half female. Provided by Shanghai Slake Experimental Animal Co., Ltd., animal qualification certificate number SCXK (Shanghai) 2013-0003. The temperature of the breeding room is 20-25℃, and the relative humidity is 40-70%. Mouse feed is provided by Suzhou Shuangshi Experimental Animal Feed Technology Service Co., Ltd., registration certificate number: Su (2009) 05032.

[0208] 2. Methods:

[0209] 1. Inspection basis: Ministry of Health "Disinfection Technical Specifications" 2002 edition.

[0210] 2. Dosage grouping: 10 animals, half male and half female, both male and female were given a single dose of 10000 mg / m 3 Poisoning.

[0211] 3. Inspection method: at 1m 3 Ten male and female mice were placed in the poisoning cabinet, and the drug preparation of Example 1 was placed on an electric heater in the cabinet to be heated and volatilized. The animals were statically inhaled for 2 hours, and the sample consumption was 10000 mg. After 2 hours, the animals were taken out and moved to the observation room, and the symptoms and death number of the animals during and within two weeks after the poisoning were observed and recorded. Acute toxicity classification was performed according to the acute toxicity classification standard.

[0212] 3. The test results are shown in Table 4:

[0213] Table 4

[0214]

[0215] As can be seen from the above table, during the test period, no obvious changes were observed in the gross pathology of the animals, and the pharmaceutical composition of the present invention is non-toxic.

[0216] In summary, this experiment used human lung cancer cells A549 to conduct pharmacodynamic evaluation of the pharmaceutical composition of the present invention. First, the CCK-8 detection method was used to determine that the pharmaceutical composition of the present invention can inhibit the growth of human lung cancer cells A549; then the MTT method was used to determine that the half-maximal inhibitory growth concentration IC50 of the pharmaceutical composition of the present invention on human lung cancer cells A549 was 0.0018 g / ml, indicating that the pharmaceutical composition of the present invention has a good inhibitory effect on human lung cancer cells A549.

[0217] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A pharmaceutical composition for treating lung cancer, characterized in that: The raw materials of the pharmaceutical composition include, by weight: 70-90 parts of white mustard seeds, 70-90 parts of cat's claw, 95-105 parts of Scutellaria barbata, 95-105 parts of Prunella vulgaris, 110-130 parts of Sophora japonica, 95-105 parts of Asparagus cochinchinensis, 125-135 parts of Sanguisorba officinalis, 70-90 parts of Arctium lappa, 80-100 parts of Curcuma longa, 70-90 parts of dried bitter melon, 70-90 parts of Taraxacum officinale, 55-65 parts of Angelica dahurica, 45-55 parts of Sophora japonica seeds, 35-45 parts of the Angelica dahurica, 30 parts of red peony root, 95-105 parts of Pueraria root, 55-65 parts of Pheretima, 45-55 parts of Aquilaria reticulata, and 45-55 parts of Liquorice.

2. The pharmaceutical composition according to claim 1, characterized in that The raw material composition of the pharmaceutical composition is: 80 parts of Semen Sinapis, 80 parts of Cat's Claw, 100 parts of Scutellaria barbata, 100 parts of Spica Prunellae, 120 parts of Sophora japonica, 100 parts of Asparagus cochinchinensis, 130 parts of Sanguisorba officinalis, 80 parts of Fructus Arcti, 90 parts of Curcuma longa, 80 parts of Dried Momordica charantia, 80 parts of Herba Taraxaci, 60 parts of Angelica dahuricae, 50 parts of Flos Sophorae, 40 parts of the Angelicae Dahuricae, 30 parts of Red Peony Roots, 100 parts of Radix Puerariae, 60 parts of Pheretima, 50 parts of Aquilariae Resinatum, 50 parts of Radix Glycyrrhizae.

3. A pharmaceutical preparation, characterized in that The pharmaceutical preparation comprises the pharmaceutical composition according to claim 1 or 2 and / or its active extract.

4. The pharmaceutical preparation according to claim 3, characterized in that The pharmaceutical preparation further includes pharmaceutically acceptable excipients; wherein the excipients include one or more of solvents, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, stabilizers, flavoring agents, preservatives, suspending agents, coating materials, fragrances and thickeners; And / or, the dosage form of the pharmaceutical preparation includes tablets, pills, decoctions, capsules, powders, drops, lyophilized products, granules, ointments, patches, suspensions, syrups, oral solutions, injections, and suppositories.

5. A method for preparing the pharmaceutical preparation according to claim 3 or 4, characterized in that: The preparation method of the pharmaceutical preparation comprises the steps of weighing a certain amount of raw material drug components for compounding or extracting the active ingredients of the raw material drug for compounding; Optionally, the preparation method of the pharmaceutical preparation comprises: (1) mixing white mustard seeds, asparagus cochinchinensis, dried bitter melon, kudzu root, earthworm, water and papain, performing enzymolysis, and performing solid-liquid separation to obtain liquid A; Mixing cat's claw, selfheal, burdock, dandelion with water and cellulase, performing enzymolysis, and separating the enzymolysis product into solid and liquid to obtain liquid B; Scutellaria barbata, Sophora vietnamese, Sanguisorba officinalis, Curcuma longa, Angelica dahurica, Sophora japonica seed, Angelica dahurica, Paeonia lactiflora, Aquilaria sinensis and Licorice root are extracted with water, and the solid-liquid separation is performed to obtain a medicinal solution C; The drug solution A, the drug solution B and the drug solution C are mixed and then distilled to remove impurities to obtain a puree; (2) mixing soybean flour and part of the puree obtained in step (1) and fermenting them to obtain baking powder; (3) fermenting the remaining puree obtained in step (1) and the fermentation powder obtained in step (2), filtering, and obtaining a fermentation liquid; (4) adding lysozyme to the fermentation broth obtained in step (3) to form the pharmaceutical preparation.

6. The preparation method according to claim 5, characterized in that: In step (1), the raw materials are first soaked and boiled in water before enzymolysis, the amount of water used for soaking is 6-10 times the total weight of the raw materials to be soaked; the soaking time is 1-3 hours, the boiling temperature is 90-100° C., and the boiling time is 50-60 minutes; And / or, the amount of water used for the water extraction is 6-10 times the total weight of the raw material to be extracted; the temperature of the water extraction is 90-100° C., and the time of the water extraction is 50-60 minutes; And / or, the enzymatic activity of the papain is 80000-120000 U / g, the added amount of the papain is 0.003-0.01% of the total weight of white mustard, asparagus, dried bitter melon, kudzu root and earthworm; the enzymatic hydrolysis time of the papain is 1-10 hours, and the enzymatic hydrolysis temperature is 55-65° C.; And / or, the enzymatic activity of the cellulase is 8000-15000 U / g, the added amount of the cellulase is 0.1-0.3% of the total weight of cat's claw, selfheal, burdock and dandelion; the enzymatic hydrolysis time of the cellulase is 5-24 hours, and the enzymatic hydrolysis temperature is 28-32° C.; And / or, the distillation temperature is 60-80°C.

7. The preparation method according to claim 5, characterized in that: The mass ratio of the soybean flour to the puree in step (2) is 1:(1-3); And / or, lysozyme is added to the fermentation broth in step (3) and mixed until saturated; The fermentation temperature is 40-50°C.

8. Use of the pharmaceutical composition and / or active extract thereof according to claim 1 or 2 or the pharmaceutical preparation according to any one of claims 3 to 5 in preparing a drug for preventing or treating lung cancer.

9. The pharmaceutical composition and / or active extract thereof according to claim 1 or 2 or the pharmaceutical preparation according to any one of claims 3 to 5 is used for preparing a drug having the effect of awakening NK cells and / or inhibiting the growth of lung cancer cells A549.

10. The use according to claim 9, characterized in that The medicine also has the effect of dispersing nodules.

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