Traditional Chinese medicine composition for treating lung tumors, method of manufacturing the same, and use

Abstract

This invention provides a Kampo (traditional Japanese herbal medicine) composition for treating lung tumors, a method for producing the same, and a method for using the same. [Solution] This is a Kampo medicine composition for treating lung tumors, characterized by being manufactured using the following raw materials: 5% to 15% by weight of Chrysanthemum, 2% to 12% by weight of Honeysuckle, 5% to 15% by weight of Peppermint, 5% to 15% by weight of Poria, 10% to 20% by weight of Atractylodes Rhizome, 0.5% to 3% by weight of Cinnamon, 0.5% to 3% by weight of Lumpsucker's Tears, 3% to 12% by weight of Astragalus, 5% to 15% by weight of Reishi mushroom, 10% to 20% by weight of Ligusticum chuanxiong, 5% to 12% by weight of Saussurea rhizome, and 2% to 10% of Licorice. Adopting the above technical solution has the advantage of significantly preventing tumor growth.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine composition for treating lung tumors, its manufacturing method and use.

Background Art

[0002] The occurrence of cancer in the human body is mostly caused by the decline of the human body's immunity. As a result, the balance of the immune function is disrupted, and the growth of cancer cells becomes uncontrollable.

[0003] Immunotherapy is a new cancer treatment method in recent years. Western medicine immunotherapy is usually carried out in combination with chemotherapy or radiotherapy, which has a certain therapeutic effect, but has side effects and drug resistance.

[0004] Therefore, in the Chinese invention patent with the publication number CN108567820B, a traditional Chinese medicine composition for treating lung tumors is researched and developed, and this technical problem is solved by blending 13 kinds of crude drugs. However, this traditional Chinese medicine composition has room for improvement in the inhibitory effect on the growth of lung tumors.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The object of the present invention is to provide a traditional Chinese medicine composition for treating lung tumors, its manufacturing method and use, which have the advantage of significantly preventing the growth of tumors against the defects and drawbacks of the prior art.

Means for Solving the Problems

[0006] To achieve the above objectives, the technical solutions used in the present invention are as follows: A Kampo medicine composition for treating lung tumors, manufactured using as raw materials: 5% to 15% by weight of Chrysanthemum japonica, 2% to 12% by weight of Lonicera japonica, 5% to 15% by weight of Mentha arvensis, 5% to 15% by weight of Poria cocos, 10% to 20% by weight of Atractylodes macrocephala, 0.5% to 3% by weight of Cinnamon, 0.5% to 3% by weight of Lactuca indica, 3% to 12% by weight of Astragalus membranaceus, 5% to 15% by weight of Ganoderma lucidum, 10% to 20% by weight of Ligusticum chuanxiong, 5% to 12% by weight of Saussurea rhizome, and 2% to 10% by weight of Licorice.

[0007] In a further embodiment of the present invention, the herbal medicine composition for treating the lung tumor is manufactured using the following raw materials: 5% to 10% by weight of Chrysanthemum japonica, 3% to 8% by weight of Lonicera japonica, 5% to 10% by weight of Mentha arvensis, 5% to 10% by weight of Poria cocos, 12% to 20% by weight of Atractylodes macrocephala, 0.5% to 3% by weight of Cinnamon, 0.5% to 3% by weight of Lactuca indica, 5% to 10% by weight of Astragalus membranaceus, 5% to 10% by weight of Reishi mushroom, 11% to 15% by weight of Ligusticum chuanxiong, 8% to 12% by weight of Saussurea rhizome, and 3% to 8% by weight of Licorice.

[0008] In a further embodiment of the present invention, the herbal medicine composition for treating the lung tumor is manufactured using the following raw materials: 6% to 9% by weight of Chrysanthemum, 3% to 6% by weight of Honeysuckle, 6% to 9% by weight of Peppermint, 6% to 9% by weight of Poria, 15% to 18% by weight of Atractylodes, 1% to 3% by weight of Cinnamon, 0.5% to 1.5% by weight of Lactuca, 6% to 10% by weight of Astragalus, 6% to 10% by weight of Reishi mushroom, 12% to 15% by weight of Ligusticum chuanxiong, 8% to 11% by weight of Saussurea rhizome, and 3% to 6% by weight of Licorice.

[0009] In a further embodiment of the present invention, the dosage form of the herbal medicine composition for treating the lung tumor is an injection, a honey-based pill, a water-based pill, a capsule, a tablet, a drop pill, a powder, an oral liquid, a gel, an extract, or a film.

[0010] To achieve the above objectives, another technical solution employed by the present invention is as follows: A method for producing a Kampo medicine composition for treating lung tumors, Step (1) involves weighing each herbal medicine component in the aforementioned weight ratio, crushing each until the particle size is between 10 mesh and 100 mesh, sieving, and mixing them uniformly. Step (2) involves preparing an ethanol solution with a concentration of 40% to 95%, soaking a uniformly mixed herbal medicine in the prepared ethanol solution with a concentration of 40% to 95% for 10 to 48 hours, adding the soaked herbal medicine mixture to a percolation container in multiple batches, and lightly pressing and leveling it. Step (3) involves adding the ethanol solution with a concentration of 40% to 95% prepared above to the percolation container in step (2), immersing the herbal medicine to a depth of 1 to 2 cm above the surface for a predetermined time, then percolating with the remaining ethanol, retaining the residue, concentrating the resulting alcohol solution under reduced pressure, and drying to obtain the herbal medicine complex powder A. The method includes step (4) adding 2 to 10 times the amount of water to the drug residue, decocting it in water for 0.5 to 2 hours to extract, centrifuging and filtering, concentrating the drug solution under reduced pressure and drying to obtain herbal medicine complex powder B, and uniformly mixing herbal medicine complex powder A and herbal medicine complex powder B to obtain a herbal medicine composition.

[0011] In a further embodiment of the present invention, in step (1), the various crude drugs are pulverized to a particle size of 20 to 40 mesh, and the concentration of the ethanol prepared in steps (2) and (3) is 60% to 80%.

[0012] In a further embodiment of the present invention, in step (2), the uniformly mixed herbal medicine is infused for 18 to 30 hours, and in step (3), the immersion time is 2 to 48 hours.

[0013] To achieve the above objectives, another technical solution employed by the present invention is as follows: A method for producing a Kampo medicine composition for treating lung tumors, Step (1) involves weighing out Cnidium officinale, Atractylodes lancea, Saussurea chinensis, Carnation, Cinnamon, and Peppermint in the aforementioned weight ratio, adding an appropriate amount of water to extract the volatile oil, then adding cyclodextrin to encapsulate it, filtering the decoction and collecting it in another container to prepare it for use as residual medicinal solution, and Step (2) involves adding water to the drug residue from which the volatile oil has been extracted and to Poria cocos, Astragalus membranaceus, Chrysanthemum laciniata, Reishi mushroom, Honeysuckle, and Licorice, weighed in the aforementioned weight ratio, decocting the mixture, filtering the liquid, and combining the filtered liquid with the residual liquid from which the volatile oil has been extracted to obtain a mixed herbal medicine solution. Step (3) involves concentrating the obtained herbal medicine mixture in a water bath until it becomes a clear ointment, precipitating with alcohol, and adding ethanol until the alcohol content reaches 50% to 80% to obtain an herbal medicine mixture alcohol solution. The process includes step (4) of concentrating the aforementioned mixed alcohol solution of herbal medicines under reduced pressure and drying it, and then adding a volatile oil cyclodextrin inclusion complex to obtain the herbal medicine composition.

[0014] In a further embodiment of the present invention, in step (1), the volatile oil extraction solvent is water in an amount 4 to 15 times the amount of the crude drug, and the extraction time is 2 to 12 hours. The resulting volatile oil undergoes cyclodextrin inclusion at temperatures below 40°C, with an inclusion time of 1 to 10 hours.

[0015] To achieve the above objectives, another technical solution employed by the present invention is as follows: the use of a Kampo medicine composition, the Kampo medicine composition being used in combination with cyclophosphamide for the treatment of lung tumors. [Effects of the Invention]

[0016] By adopting the above technical solutions, the beneficial effects of the present invention are as follows.

[0017] 1. In this invention, Reishi mushroom nourishes the five internal organs, strengthens the spleen and qi, and improves the body's immunity. Astragalus strengthens the spleen and lungs, replenishes qi and yang, and improves the body's immunity. Reishi mushroom and Astragalus can inhibit the proliferation of tumor telomerase. Atractylodes lancea, Poria cocos, and Licorice strengthen the spleen and remove dampness. Cinnamon, Saussurea rhizome, and Cinnamon bark are pungent and warming, improve the flow of qi, warm the middle burner, dissipate cold, and relieve pain. Honeysuckle removes heat and detoxifies. Chrysanthemum verum purifies the liver, improves eyesight, and detoxifies. Peppermint detoxifies with its pungent and cold taste, improves the circulation of liver qi, alleviates symptoms such as sighing, dryness, depression, and irritability caused by liver qi stagnation, and regulates the function of the autonomic nervous system. Ligusticum chuanxiong improves the flow of qi and blood and relieves pain. This formula uses Reishi and Astragalus to strengthen the spleen and lungs, nourish the five internal organs, support the function of vital energy, strengthen the foundation of the constitution, prevent tumors, and function as the main ingredient. Atractylodes lancea, Poria cocos, and Licorice improve the function of the spleen and expel dampness. Cinnamon, Saussurea rhizome, and Cinnamon bark improve the flow of qi, warm the middle burner, dissipate cold, and relieve pain, functioning as auxiliary ingredients. Chrysanthemum chinense and Lonicera japonica remove heat and detoxify, functioning as auxiliary ingredients. Ligusticum chuanxiong and Mentha arvensis purify the liver, improve blood flow, relieve pain, and function as a driving force ingredient.

[0018] 2. The herbal medicine composition of the present invention has a clear preventive effect against tumor growth in A-549 tumor-bearing nude mice.

[0019] 3. The herbal medicine composition of the present invention can increase the number of white blood cells, neutrophils, lymphocytes, and monocytes, whether used alone or in combination with cyclophosphamide.

[0020] To more clearly describe the embodiments of the present invention or the technical solutions in the prior art, the drawings necessary for describing the embodiments or the prior art will be briefly described below. However, the drawings in the following description are only a few of the embodiments of the present invention, and it will be obvious to those skilled in the art that other drawings can be obtained based on these drawings without any creative effort. [Brief explanation of the drawing]

[0021] [Figure 1] It is the tumor growth curve. [Figure 2] It is the tumor growth curve. [Figure 3] It is the weight change curve. [Figure 4] It is the weight gain rate curve. [Figure 5] These are photographs of the tumor tissues of each group.

MODE FOR CARRYING OUT THE INVENTION

[0022] Hereinafter, the present invention will be described in more detail with reference to the drawings.

[0023] This specific embodiment is only for explaining the present invention and does not limit the present invention. Those skilled in the art can modify this embodiment as necessary without making creative contributions after reading this description, but they are protected by the patent law as long as they are within the scope of the claims of the present invention.

EXAMPLE

[0024] The herbal medicine composition for treating lung tumors is produced with 9% by weight of Chrysanthemum morifolium Ramat., 5% by weight of Carthamus tinctorius L., 7% by weight of Mentha haplocalyx Briq., 8% by weight of Atractylodes lancea (Thunb.) DC., 16% by weight of Bupleurum falcatum L., 2% by weight of Paeonia lactiflora Pall., 1% by weight of Glycyrrhiza glabra L., 10% by weight of Alpinia officinarum Hance, 12% by weight of Ziziphus jujuba Mill., 13% by weight of Rehmannia glutinosa Libosch., 9% by weight of Aucklandia lappa Decne., and 8% by weight of Licorice as raw materials.

[0025] The method for producing this herbal medicine composition for treating lung tumors is as follows: weigh each herbal medicine raw material in the above weight ratio, grind each until the particle size is 20 mesh, sift and mix uniformly; prepare a 70% ethanol solution; soak the uniformly mixed herbal medicine in the prepared 70% ethanol solution for at least 24 hours; add the soaked herbal medicine mixture to a percolation container in multiple batches and lightly press and spread it evenly; and pour the prepared 70% ethanol solution so that it is 1-2 cm above the surface of the crude drugs. The process includes the steps of: soaking for 8 to 30 hours (most preferably 20 hours), adding the ethanol solution to a percolation container, percolation, retaining the drug residue, concentrating the resulting alcohol solution under reduced pressure, drying it, and obtaining a herbal medicine complex powder A; adding the drug residue to 2 to 10 times (most preferably 8 times) the amount of water relative to the amount of drug residue, decocting with water for 0.5 to 2 hours (most preferably 1 hour), centrifuging and filtering, concentrating the drug solution under reduced pressure and drying it, and obtaining a herbal medicine complex powder B; and uniformly mixing the herbal medicine complex powder A and the herbal medicine complex powder B to obtain a herbal medicine composition. [Examples]

[0026] A Kampo medicine composition for treating lung tumors is manufactured using the following raw materials: Chrysanthemum (8% by weight), Honeysuckle (7% by weight), Peppermint (7% by weight), Poria (9% by weight), Atractylodes (15% by weight), Cinnamon (2% by weight), Lactuca (2% by weight), Astragalus (11% by weight), Reishi (10% by weight), Ligusticum chuanxiong (14% by weight), Saussurea (10% by weight), and Licorice (5% by weight).

[0027] The method for producing this herbal medicine composition involves weighing out Cnidium officinale, Atractylodes lancea, Saussurea rhizome, Cinnamon jasminoides, and Mentha arvensis in the aforementioned mass ratio, adding an appropriate amount of volatile oil extraction solvent, which is 6 times the amount of water relative to the amount of crude drugs, and extracting the volatile oil for 2 to 12 hours (most preferably 10 hours), and after extracting the volatile oil, performing cyclodextrin inclusion at 40°C or below for 6 hours, filtering the decoction and collecting the filtrate in another container, and mixing the residue from which the volatile oil has been extracted with Poria cocos, Astragalus membranaceus, Chrysanthemum sieboldii, Ganoderma lucidum, Lonicera japonica, and Glycyrrhiza uralensis, which are weighed out in the aforementioned weight ratio, and adding 10 to 15 times (most preferably 12 times) the amount of crude drugs. The process includes the steps of: adding water and decocting for 1 to 2 hours (most preferably 1.5 hours) 2 to 5 times (most preferably 3 times); filtering the medicinal liquid obtained by decocting, and combining the filtered medicinal liquid with the filtrate from which volatile oil has been extracted to obtain a mixed herbal medicine solution; concentrating the obtained mixed herbal medicine solution in a water bath to make a clear paste with a relative density of 1.06 to 1.08, adding ethanol until the alcohol content reaches 60%, and letting it stand for 12 hours or more (most preferably 48 hours) to obtain a mixed herbal medicine alcohol solution; and concentrating the mixed herbal medicine alcohol solution under reduced pressure, drying it, and adding a volatile oil cyclodextrin inclusion complex to obtain a herbal medicine composition. [Examples]

[0028] A Kampo medicine composition for treating lung tumors is manufactured using the following raw materials: 7% by weight of Chrysanthemum, 6% by weight of Honeysuckle, 8% by weight of Peppermint, 11% by weight of Poria, 17% by weight of Atractylodes, 1.5% by weight of Cinnamon, 0.5% by weight of Lactuca, 9% by weight of Astragalus, 9% by weight of Reishi mushroom, 16% by weight of Ligusticum chuanxiong, 9% by weight of Saussurea rhizome, and 6% by weight of Licorice.

[0029] The method for producing this herbal medicine composition includes the steps of: weighing each herbal medicine raw material in the aforementioned weight ratio; preparing a 60% concentration ethanol solution; adding the uniformly mixed herbal medicines to six times the amount of the ethanol solution prepared above; extracting by heating under reflux twice for 1 to 2 hours each (most preferably 1.5 hours) (heating until slightly boiling, then refluxing and extracting); combining the two extracts, recovering the ethanol under reduced pressure; concentrating and drying to obtain the herbal medicine composition. [Examples]

[0030] A Kampo medicine composition for treating lung tumors is manufactured using the following raw materials: 9% by weight of Chrysanthemum, 7% by weight of Honeysuckle, 7% by weight of Peppermint, 8% by weight of Poria, 13% by weight of Atractylodes, 2% by weight of Cinnamon, 1% by weight of Lactuca, 10% by weight of Astragalus, 15% by weight of Reishi mushroom, 13% by weight of Ligusticum chuanxiong, 7% by weight of Saussurea rhizome, and 8% by weight of Licorice.

[0031] The method for producing this herbal medicine composition involves weighing each herbal medicine raw material in the above weight ratio, grinding each until the particle size is 30 mesh, sieving and mixing them uniformly, preparing a 65% ethanol solution, soaking the uniformly mixed herbal medicine in the prepared 65% ethanol solution for 36 hours, adding the soaked herbal medicine mixture to a percolation container in multiple batches and lightly pressing and leveling it, and pouring the 65% ethanol solution prepared above to a depth of 1-2 cm above the surface of the crude drugs. The process includes the steps of: adding the ethanol solution to a percolation container, immersing for 18 hours, then percolating with the remaining ethanol, retaining the drug residue, concentrating the resulting alcohol solution under reduced pressure and drying it to obtain a herbal medicine complex powder A; and adding five times the amount of water relative to the drug residue to the drug residue, decocting with water for 1 to 2 hours (most preferably 1.5 hours) to extract, centrifuging and filtering, concentrating the drug solution under reduced pressure and drying it to obtain a herbal medicine complex powder B; and uniformly mixing the herbal medicine complex powder A and the herbal medicine complex powder B to obtain a herbal medicine composition. [Examples]

[0032] The herbal medicine composition for maintaining health is manufactured using the following raw materials: 8% by weight of Chrysanthemum japonica, 3% by weight of Lonicera japonica, 9% by weight of Mentha arvensis, 9% by weight of Poria cocos, 16.5% by weight of Atractylodes macrocephala, 1% by weight of Cinnamon, 1% by weight of Lactuca indica, 9% by weight of Astragalus membranaceus, 9% by weight of Reishi mushroom, 13.5% by weight of Ligusticum chuanxiong, 10% by weight of Saussurea rhizome, and 11% by weight of Licorice.

[0033] The method for producing this herbal medicine composition involves weighing out Cnidium officinale, Atractylodes lancea, Saussurea rhizome, Cinnamon jasminoides, and Mentha arvensis in the aforementioned weight ratio, adding an appropriate amount of volatile oil extraction solvent, which is four times the amount of water relative to the amount of crude drugs, and extracting the volatile oil for 2 to 12 hours (most preferably 10 hours), then, after extracting the volatile oil, performing cyclodextrin inclusion at 40°C or below for 5 hours, filtering the decoction and collecting the filtrate in another container, and finally, adding 10 to 15 times the amount of water (most preferably 12 times) relative to the amount of crude drugs to the drug residue from which the volatile oil has been extracted, along with Poria cocos, Astragalus membranaceus, Chrysanthemum sacchariflorum, Ganoderma lucidum, Lonicera japonica, and Glycyrrhiza uralensis, which are weighed out in the aforementioned weight ratio. The process includes the steps of: 1) simmering the mixture for 1 to 2 hours (most preferably 1.5 hours) 2 to 5 times (most preferably 3 times); 2) filtering the medicinal liquid obtained by simmering, and combining the filtered medicinal liquid with the filtrate from which the volatile oil has been extracted to obtain a mixed herbal medicine solution; 3) concentrating the obtained mixed herbal medicine solution in a water bath to obtain a clear ointment with a relative density of 1.06 to 1.08, adding ethanol until the alcohol content reaches 60%, and letting it stand for 12 hours or more (most preferably 24 hours) to obtain a mixed herbal medicine alcohol solution; and 4) concentrating the mixed herbal medicine alcohol solution under reduced pressure and drying it, and then adding a volatile oil cyclodextrin inclusion complex to obtain a herbal medicine composition.

[0034] Pharmacodynamics Test To verify the effects of the present invention in tumor treatment, the following efficacy tests were conducted using the herbal medicine composition prepared by the method of Example 1.

[0035] 1. Test materials 1.1 Test drug The herbal medicine composition containing Sophora flavescens (hereinafter simply referred to as the original formula), the herbal medicine composition prepared by the method of Example 1 (hereinafter simply referred to as the present formula), properties: dark brown viscous liquid, dosage and administration: orally, the recommended clinical dose of the original formula is 7.6g crude drug / 60kg / d, and the recommended clinical dose of the present formula is 7.0g crude drug / 60kg / d. Sample drug content: 1g crude drug / ml, storage conditions: refrigerated, caution: before use, heat the drug solution to 35-40°C and shake well.

[0036] 1.2 Positive control drug Cyclophosphamide for injection, Manufacturing Unit: Jiangsu Shengdi Pharmaceutical Co., Ltd., Indications: This product is a widely used anticancer agent that is highly effective against malignant lymphoma, acute or chronic lymphocytic leukemia, and multiple myeloma, and also has some effect on lung cancer and breast cancer. Batch number: 17020625, Manufacturing date: 2017.02.06, Expiration date: 2019.02.05. Ingredients: Cyclophosphamide, Appearance: White crystals or crystalline powder, Specifications: 0.2g / bottle, 10 bottles / box, Dosage and administration: For adults, 500-1000mg / m² per dose depending on body surface area. 2 Administer intravenously once a week for two consecutive injections, followed by a 1-2 week break before repeating. Storage conditions: Store in a light-shielded, sealed container at 30°C or below.

[0037] 1.3 Test animals [Table A]

[0038] 1.4 Tumor cell lines [Table B]

[0039] 1.5 Test Reagents [Table C]

[0040] 1.6 Test equipment [Table D]

[0041] 1.7 Main consumables for testing [Table E]

[0042] 2. Test Method 2.1 Dose design 2.1.1 Original formula: The recommended clinical dose for adults is 7.6 g of crude drug / 60 kg / d. In this study, when converted to the body surface area of ​​humans and animals, the unit dose for mice is 11 times the unit dose for humans, so the dose for mice is (7.6 / 60) × 11 = 1.4 g of crude drug / kg / d. The concentration of the drug solution submitted to the study is 1 g of crude drug / ml, so when converted to volume, the dose for mice is 1.4 / 1 = 1.4 ml / kg / d. Therefore, the high dose and low dose of the original formula are 2.8 ml / kg / d and 1.4 ml / kg / d (corresponding to twice and equal to the clinical dose for humans, respectively). For this prescription, the recommended clinical dose for adults is 7.0 g of herbal medicine per 60 kg / day. In this study, when converted to the body surface area of ​​humans and animals, the unit dose for mice is 11 times the unit dose for humans, so the dose for mice is (7.0 / 60) × 11 = 1.3 g of herbal medicine / kg / day. The concentration of the drug solution submitted to the study is 1 g of herbal medicine / ml, so when converted to volume, the dose for mice is 1.3 / 1 = 1.3 ml / kg / day. Therefore, the high and low doses of this prescription are 2.6 ml / kg / day and 1.3 ml / kg / day (corresponding to twice and the same as the clinical dose for humans, respectively). 2.1.2 Cyclophosphamide for injection: The recommended clinical dose for adults is 1000 mg / m². 2 The average body surface area of ​​an adult is 1.6246 m². 2 The dosage is per 60kg, and the converted recommended clinical dose for adults is 1624.6 mg / 60kg. In the study, when converted to the body surface area of ​​humans and animals, the unit dose for mice was calculated as 11 times the unit dose for humans, and the dose for mice was (1624.6 / 60) × 11 = 297.84 mg / kg / dose = 0.3 g / kg / dose.

[0043] 2.2 Culture of human lung adenocarcinoma cell line (A549) The cell line was cultured in complete medium (DMEM medium + 10% fetal bovine serum, pH 7.2), placed in a 37°C, 5% CO2 constant temperature incubator, the medium was changed daily, and the cells were cultured to a confluence rate of 80% to 90%. After digestion with 0.25% trypsin, the cells were centrifuged and subcultured, and once a certain number of cells had been cultured, they were diluted with PBS to form a tumor cell suspension, which was then inoculated into the bodies of test animals.

[0044] 2.3 Construction of tumor lines in A549 tumor-bearing nude mice 1) Construction of first-generation A549 tumor-bearing nude mice: Conventionally cultured A549 cell lines were digested with trypsin-EDTA, centrifuged at 1000 rpm, the supernatant was discarded, and the cells were resuspended in DMEM high-glucose medium to a cell concentration of 10 6 ~10 7 The cell concentration was adjusted to 1 / ml. Three BALB / c nude mice were subcutaneously inoculated with 0.4 ml of A549 cell suspension into the right axilla. After inoculation, the mice were reared under normal conditions, and their survival status and tumor growth were observed. 2) Construction of Passage A549 Tumor-Bearing Nude Mice: After the tumors of the first-generation tumor-bearing nude mice grew to a diameter of approximately 1 cm, one nude mouse with good survival status and well-developed tumors was selected. Under sterile conditions, the tumor was removed and placed in a glass homogenizer. 1.5 ml of sterile physiological saline was added to prepare a homogenate, which was then filtered through a 70 μm cell strainer and the filtered homogenate solution was stored. 0.4 ml of the homogenate solution was subcutaneously inoculated into the right axilla of each of three BALB / c nude mice. After inoculation, the mice were reared under normal conditions, and their survival status and tumor growth were observed.

[0045] 2.4 Creation of a human lung cancer transplant tumor model Eighty-eight nude mice (half male, half female) weighing 18-20g were adaptively reared at an animal laboratory for five days. After the tumors of the serially tumor-bearing nude mice grew to a diameter of approximately 1.5cm, tumors were removed under sterile conditions from three nude mice that were in good condition and had well-developed tumors. 3.0g of each tumor was weighed out, placed in a glass homogenizer, and 20ml of sterile physiological saline was added to prepare a homogenate. The homogenate was filtered through a 70μm cell strainer, and the filtered homogenate solution was stored. Eight nude mice were randomly selected as a blank control group, and 0.2 ml of homogenate solution was subcutaneously inoculated into the right axilla of the remaining 80 nude mice. After inoculation, the mice were divided into the model control group, cyclophosphamide group (0.3 g / kg), high-dose group of the original formula (2.8 g / kg), low-dose group of the original formula (1.4 g / kg), high-dose group of the current formula (2.6 g / kg), and low-dose group of the current formula (1 The animals were randomly divided into 10 groups of 8 each: a high-dose group of cyclophosphamide (0.3g / kg) + original formula (2.8g / kg), a low-dose group of cyclophosphamide (0.3g / kg) + original formula (1.4g / kg), a high-dose group of cyclophosphamide (0.3g / kg) + this formula (2.6g / kg), and a low-dose group of cyclophosphamide (0.3g / kg) + this formula (1.3g / kg).

[0046] 2.5 Administration Administration began on the second day after vaccination, with 0.2 ml / 10 g administered intragastricly. The blank control group and model control group were given distilled water under the same conditions. Tumors were approximately 50 mm in size. 3 When the animal reached maturity, cyclophosphamide was started to be administered intraperitoneally at a dose of 0.1 ml / 10 g.

[0047] 2.6 Material collection On the evening of the final administration, the mice were fasted, their weight was measured the following morning, and the longest and shortest diameters of the tumor were measured. Then, the eyeballs were removed from the mice, and whole blood (anticoagulant) and serum were collected. The weights of the tumor mass, liver, heart, spleen, lungs, and kidneys were then measured and weighed in the following order: liver, heart, spleen, lungs, kidneys, and tumor mass.

[0048] 2.7 Observational Indicators 1) Body weight: The body weight was measured once a week, and the dosage was adjusted according to the body weight. 2) Tumor volume: When tumor growth was observed, the longest diameter (a) and the shortest diameter (b) of the tumor were measured using a digital caliper, and the tumor volume (TV) was measured twice a week using the formula: TV = (1 / 2)ab2. 3) Tumor inhibition rate and organ index: The tumor mass, liver, heart, spleen, lung, and kidney were excised, weighed in the order of liver, heart, spleen, lung, kidney, and tumor mass, and the tumor inhibition rate and each organ index were calculated.

Number

[0049] 2.8 Evaluation of the effect of drug combination Using King's formula q = E(a + b) / (Ea + Eb - Ea×Eb), evaluate whether there is a synergistic effect or an antagonistic effect in the combination of cyclophosphamide with this prescription or the original prescription. Here, E(a + b) is the effective rate of the drug combination, and Ea and Eb are the effective rates of the drugs alone. When q < 0.85, the two drugs are antagonistic; when q > 1.15, the two drugs are synergistic; when 0.85 < q < 1.15, the effects of the two drugs are simply additive.

[0050] 2.9 Statistical analysis All data are presented as mean ± standard deviation

Number

[0051] 3 Test results 3.1 Effect on the increase in tumor volume in a nude mouse model of human lung adenocarcinoma cell line (A549) As shown in Table 1 and Figures 1 and 2, after inoculating nude mice with human lung adenocarcinoma (A549), prophylactic administration was started on day 2. From day 27 (first measurement) to day 50, both the low-dose and high-dose groups of this formulation showed a significant inhibitory effect on tumor growth in A-549-bearing nude mice, with a statistically significant difference compared to the model control group (p<0.05, p<0.01). The tumor inhibition rate in the high-dose group was 90.62%. When the original prescription and the present prescription were used in combination with cyclophosphamide, and the value of q was calculated using King's formula, the therapeutic effect on A549 tumors when cyclophosphamide is used in combination with a low dose of the present prescription was simply additive (q=0.97). When cyclophosphamide is used in combination with a low dose of the original prescription (q=0.27), cyclophosphamide with a high dose of the original prescription (q=0.34), or cyclophosphamide with a high dose of the present prescription (q=-3.47), the therapeutic effects on A549 tumor weight were all antagonistic. The original low-dose and high-dose formulations, when administered alone or in combination with cyclophosphamide, did not demonstrate a preventive effect against the development of human lung adenocarcinoma. Figures 3, 4, and 5 show that during the administration period, the body weight of mice in the low-dose and high-dose groups of the original formulation, as well as in the low-dose and high-dose groups of the current formulation, increased significantly compared to the model control group. When administered in combination with phosphoramide, the body weight of each group decreased significantly after 4 weeks of administration.

[0052] [Table 1] In Table 1 and the drawings in the specification above (TIFF2026076908000010.tif238170), the formulation without kudzu is this formulation.

[0053] 3.2 Effects of human lung adenocarcinoma cell line (A549) nude mice on liver function As shown in Table 2, no significant changes were observed in liver index, serum ALT, serum AST, ALT / AST, serum TBi1, or serum TP content in the model control group, cyclophosphamide group, or each treatment group.

[0054] [Table 2] TIFF2026076908000012.tif51170

[0055] 3.3 Effects of human lung adenocarcinoma cell line (A549) nude mice on renal function As shown in Table 3, there were no significant changes in renal index, serum BUN, serum UA, or serum CRE content in the model control group, the cyclophosphamide group, or each treatment group.

[0056] [Table 3]

[0057] 3.4 Effects of human lung adenocarcinoma cell lines on the heart, spleen, and lung indices in nude mice (A549) Table 4 shows the results: 1. Cardiac index: Compared to the blank control group, the cardiac index of the model control group tended to increase. In all groups, including the high-dose group of the original prescription, the low-dose and high-dose groups of this prescription, and the low-dose and high-dose groups of cyclophosphamide + original prescription, the cardiac index decreased significantly, and there was a significant difference compared to the model control group and the cyclophosphamide group (p<0.05, p<0.01). 2. No statistically significant differences were observed in the spleen index and lung index in the model control group, the cyclophosphamide group, and each administration group.

[0058] [Table 4]

[0059] 3.5 Effects of human lung adenocarcinoma cell line (A549) nude mice on blood routines The results in Table 5 show that the total number of white blood cells, neutrophils, lymphocytes, and monocytes in the model control group were significantly reduced, and there was a significant difference compared to the blank control group (p<0.05, p<0.01). In all cases—the high-dose group of the original prescription, the low-dose and high-dose groups of the present prescription, and the combination of cyclophosphamide with the low-dose and high-dose groups of the original prescription, and the low-dose and high-dose groups of the present prescription—the number of white blood cells, neutrophils, lymphocytes, and monocytes was increased, with significant differences compared to the model control group (p<0.05, p<0.01). When cyclophosphamide was used in combination with the high-dose group of the original prescription and the low-dose group of the current prescription, the effect of increasing the number of white blood cells, neutrophils, lymphocytes, and monocytes was superior to that of each drug alone, and there was a statistically significant difference compared to the cyclophosphamide control group (p<0.05, p<0.01). In the model control group, basophils increased significantly, and this was observed in all cases: when cyclophosphamide, the current formulation, and the original formulation were used alone, and when administered in combination with cyclophosphamide. The red blood cell, platelet, hemoglobin, and eosinophil content in the normal control group, model control group, and each treatment group varied within the normal range.

[0060] [Table 5] TIFF2026076908000016.tif255168TIFF2026076908000017.tif68170

[0061] 4 Conclusion 1) Both low and high doses of this formulation alone showed a clear preventive effect against tumor growth in A-549 tumor-bearing nude mice, and at high doses, the tumor inhibition rate was 90.62%. 2) The combination of a low dose of this prescription with cyclophosphamide resulted in a purely additive effect on the treatment of A549 tumors (q=0.97). 3) Both the low and high doses of the original formulation, as well as the low and high doses of this formulation alone, have the effect of significantly increasing the body weight of tumor model mice. 4) High doses of the original prescription, low and high doses of this prescription, and low and high doses of the original prescription in combination with cyclophosphamide can significantly reduce the cardiac index. 5) In all cases, whether using the high dose of the original prescription, the low and high doses of this prescription, or the low and high doses of the original prescription, or the low and high doses of this prescription in combination with cyclophosphamide, the number of white blood cells, neutrophils, lymphocytes, and monocytes can be increased. 6) The combination of the high dose of the original prescription and the low dose of this prescription with cyclophosphamide was more effective than each drug alone in increasing the number of white blood cells, neutrophils, lymphocytes, and monocytes. 7) In any case, whether the original formula, this formula alone, or in combination with cyclophosphamide, a significant increase in basophils can be achieved. 8) When each treatment group was administered alone or in combination, there were no significant changes in the levels of liver index, kidney index, spleen index, lung index, serum BUN, serum UA, or serum CRE. There was also no effect on the levels of serum ALT, serum AST, ALT / AST, serum TBil, or serum TBil or serum TP. 9) When each treatment group was administered alone or in combination, the content of red blood cells, platelets, hemoglobin, and eosinophils varied within the normal range.

[0062] The formulation of the herbal medicine composition of the present invention is as follows. component: Reishi (Giant pine cone fungus and Pine pine cone fungus) Ougi Atractylodes lancea, Licorice, Cinnamon, Saussurea, Poria cocos, Scutellaria Gold and silver goose Mint and Cnidium You: Reishi (Large-leaved pine mushroom and pine nut fungus) Ougi Minister: Sojutsu, Kanzo, Nikkei, Mokkou, Bukuryo, Chokou Sa: Gold and silver gourd Use: Cnidium officinale Indications: Cancer patients with spleen and lung qi deficiency, qi and blood stagnation, phlegm and dampness, and weakened immunity. Function: Strengthens the spleen and lungs, disperses stagnation and accumulation, increases immune cells such as white blood cells, neutrophils, and lymphocytes, and improves immunity. Analysis: Reishi mushroom nourishes the five internal organs, strengthens the spleen and lungs, and improves the body's immunity. Astragalus strengthens the spleen and lungs, replenishes qi and yang, and improves the body's immunity. Reishi and Astragalus can inhibit the proliferation of tumor telomerase. Atractylodes lancea, Poria cocos, and Licorice strengthen the spleen and remove dampness. Cinnamon, Saussurea rhizome, and Cinnamon bark are pungent and warming, improve the flow of qi, warm the middle burner, dissipate cold, and relieve pain. Honeysuckle removes heat and detoxifies. Chrysanthemum purifies the liver, improves eyesight, and detoxifies. Peppermint detoxifies with its pungent and cool taste, improves the circulation of liver qi, alleviates symptoms such as sighing, dryness, depression, and irritability caused by liver qi stagnation, and regulates the function of the autonomic nervous system. Ligusticum chuanxiong improves the flow of qi and blood and relieves pain.

[0063] This formula uses Reishi and Astragalus to strengthen the spleen and lungs, nourish the five internal organs, support the function of vital energy, strengthen the foundation of the constitution, prevent tumors, and function as the main ingredient. Atractylodes lancea, Poria cocos, and Licorice improve the function of the spleen and expel dampness. Cinnamon, Saussurea rhizome, and Cinnamon bark improve the flow of qi, warm the middle burner, dissipate cold, and relieve pain, functioning as auxiliary ingredients. Chrysanthemum chinense and Lonicera japonica remove heat and detoxify, functioning as auxiliary ingredients. Ligusticum chuanxiong and Mentha arvensis purify the liver, improve blood flow, relieve pain, and function as a driving force ingredient.

[0064] The foregoing is used solely to illustrate, and not to limit, the technical solutions of the present invention, and any other modifications or equivalent substitutions made by those skilled in the art to the technical solutions of the present invention should all be included within the scope of the claims of the present invention, as long as they do not depart from the spirit and scope of the technical solutions of the present invention.

Claims

1. A Kampo medicine composition for treating lung tumors, characterized by being manufactured using 5% to 15% by weight of Chrysanthemum japonica, 2% to 12% by weight of Lonicera japonica, 5% to 15% by weight of Mentha arvensis, 5% to 15% by weight of Poria cocos, 10% to 20% by weight of Atractylodes macrocephala, 0.5% to 3% by weight of Cinnamon, 0.5% to 3% by weight of Lactuca indica, 3% to 12% by weight of Astragalus membranaceus, 5% to 15% by weight of Reishi mushroom, 10% to 20% by weight of Ligusticum chuanxiong, 5% to 12% by weight of Saussurea rhizome, and 2% to 10% by weight of Licorice.

2. The herbal medicine composition for treating lung tumors according to claim 1, characterized in that it is manufactured using 5% to 10% by weight of chrysanthemum, 3% to 8% by weight of honeysuckle, 5% to 10% by weight of mint, 5% to 10% by weight of porcini, 12% to 20% by weight of rosary, 0.5% to 3% by weight of cinnamon, 0.5% to 3% by weight of saffron, 5% to 10% by weight of astragalus, 5% to 10% by weight of reishi mushroom, 11% to 15% by weight of lilac, 8% to 12% by weight of Saussurea root, and 3% to 8% by weight of licorice.

3. The herbal medicine composition for treating lung tumors according to claim 1, characterized in that it is manufactured using as raw materials 6% to 9% by weight of Chrysanthemum japonica, 3% to 6% by weight of Lonicera japonica, 6% to 9% by weight of Mentha arvensis, 6% to 9% by weight of Poria cocos, 15% to 18% by weight of Atractylodes macrocephala, 1% to 3% by weight of Cinnamomum cassia, 0.5% to 1.5% by weight of Leptospermum asiaticum, 6% to 10% by weight of Astragalus membranaceus, 6% to 10% by weight of Reishi mushroom, 12% to 15% by weight of Ligusticum chuanxiong, 8% to 11% by weight of Saussurea rhizome, and 3% to 6% by weight of Licorice.

4. A Kampo medicine composition for treating lung tumors according to any one of claims 1 to 3, characterized in that it is in the form of an injection, honey paste pill, aqueous pill, capsule, tablet, drop pill, powder, oral liquid medicine, gel, extract, or film.

5. Step (1) involves weighing each herbal medicine component in the aforementioned weight ratio, crushing each until the particle size is between 10 mesh and 100 mesh, sieving, and mixing them uniformly. Step (2) involves preparing an ethanol solution with a concentration of 40% to 95%, soaking a uniformly mixed herbal medicine in the prepared ethanol solution with a concentration of 40% to 95% for 10 to 48 hours, adding the soaked herbal medicine mixture to a percolation container in multiple batches, and lightly pressing and leveling it. Step (3) involves adding the ethanol solution with a concentration of 40% to 95% prepared above to the percolation container in step (2) up to 1 to 2 cm above the surface of the crude drug, immersing it for a predetermined time, then percolating with the remaining ethanol, retaining the drug residue, concentrating the resulting alcohol solution under reduced pressure, and drying it to obtain the herbal medicine complex powder A. A method for producing a Kampo medicine composition for treating lung tumors according to any one of claims 1 to 3, comprising the step (4) of adding 2 to 10 times the amount of water relative to the amount of the drug residue, decocting with water and extracting for 0.5 to 2 hours, centrifuging and filtering, concentrating the drug solution under reduced pressure and drying to obtain Kampo medicine complex powder B, and uniformly mixing Kampo medicine complex powder A and Kampo medicine complex powder B to obtain a Kampo medicine composition.

6. The method for producing a Kampo medicine composition for treating lung tumors according to claim 5, characterized in that in step (1), various crude drugs are pulverized to a particle size of 20 to 40 mesh, and the concentration of the ethanol prepared in steps (2) and (3) is 60% to 80%.

7. The method for producing a Kampo medicine composition for treating lung tumors according to claim 6, characterized in that in step (2), a uniformly mixed Kampo medicine is impregnated for 18 to 30 hours, and in step (3), the immersion time is 2 to 48 hours.

8. Step (1) involves weighing out Cnidium officinale, Atractylodes lancea, Saussurea chinensis, Carnation, Cinnamon, and Peppermint in the aforementioned weight ratio, adding an appropriate amount of water to extract the volatile oil, then adding cyclodextrin to encapsulate it, filtering the decoction and collecting it in another container to prepare it for use as residual medicinal solution, and Step (2) involves adding water to the drug residue from which the volatile oil has been extracted and to Poria cocos, Astragalus membranaceus, Chrysanthemum laciniata, Reishi mushroom, Honeysuckle, and Licorice, weighed in the aforementioned weight ratio, decocting the mixture, filtering the liquid, and combining the filtered liquid with the residual liquid from which the volatile oil has been extracted to obtain a mixed herbal medicine solution. Step (3) involves concentrating the obtained herbal medicine mixture in a water bath until it becomes a clear ointment, precipitating with alcohol, and adding ethanol until the alcohol content reaches 50% to 80% to obtain an alcoholic herbal medicine mixture, A method for producing a Kampo medicine composition for treating lung tumors according to any one of claims 1 to 3, comprising the step (4) of concentrating the Kampo medicine mixed alcohol solution under reduced pressure and drying it, and then adding a volatile oil cyclodextrin inclusion complex to obtain a Kampo medicine composition.

9. In step (1) above, the volatile oil extraction solvent is water in an amount 4 to 15 times the amount of the crude drug, and the extraction time is 2 to 12 hours. The method for producing a Kampo medicine composition for treating lung tumors according to claim 8, characterized in that the obtained volatile oil undergoes cyclodextrin inclusion at 40°C or below, with an inclusion time of 1 to 10 hours.

10. The use of a Kampo medicine composition, characterized in that the Kampo medicine composition is used in combination with cyclophosphamide for the treatment of lung tumors, and the Kampo medicine composition is a Kampo medicine composition for the treatment of lung tumors as described in any one of claims 1 to 3.

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