A traditional Chinese medicine composition, its preparation method, and its application in the preparation of drugs with anti-gastric cancer effects.

Abstract

This invention relates to the field of traditional Chinese medicine (TCM) technology, specifically disclosing a TCM composition, its preparation method, and its application in the preparation of drugs with anti-gastric cancer effects. The TCM composition comprises the following TCM components in parts by weight: 20-30 parts of Salvia miltiorrhiza; 10-20 parts of Astragalus membranaceus; 10-20 parts of Curcuma zedoaria; 8-15 parts of Smilax glabra; and 4-8 parts of Glycyrrhiza uralensis. Studies have shown that the TCM composition prepared from these five herbs has anti-gastric cancer effects; therefore, using the TCM composition of this invention as an active ingredient in the preparation of drugs with anti-gastric cancer effects has significant application value.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine composition, its preparation method, and its application in the preparation of drugs with anti-gastric cancer effects. Background Technology

[0002] Stomach cancer is a malignant tumor originating from the gastric mucosal epithelium. In the early stages, most patients with stomach cancer experience no obvious symptoms, making it difficult to attract sufficient attention. As the tumor grows, patients in the late stages of stomach cancer typically experience anemia, weight loss, and malnutrition. Stomach cancer exhibits significant regional differences in my country; for example, the incidence rate is significantly higher in the northwest and eastern coastal regions than in the south. Furthermore, stomach cancer is one of the most common cancers in my country.

[0003] Currently, the main treatments for gastric cancer include surgery, chemotherapy, targeted therapy, radiotherapy, and traditional Chinese medicine. However, gastric cancer patients are prone to developing drug resistance when using a single chemotherapy drug or traditional Chinese medicine for a long period of time. Therefore, developing more anti-gastric cancer drugs for patients to choose from is of great significance. Summary of the Invention

[0004] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a traditional Chinese medicine composition and its preparation method and its application in the preparation of a drug with anti-gastric cancer effect.

[0005] The technical problem to be solved by the present invention is achieved through the following technical solution:

[0006] This invention first provides a traditional Chinese medicine composition, the raw materials of which comprise the following traditional Chinese medicine components in parts by weight:

[0007] Salvia miltiorrhiza 20-30 parts; Astragalus membranaceus 10-20 parts; Curcuma zedoaria 10-20 parts; Smilax china 8-15 parts; Glycyrrhiza uralensis 4-8 parts.

[0008] This invention provides a novel traditional Chinese medicine composition. Studies have shown that the traditional Chinese medicine composition prepared from five herbs, namely Danshen, Huangqi, Ezhu, Smilax, and Gancao, has an anti-gastric cancer effect.

[0009] Most preferably, the raw materials for preparing the traditional Chinese medicine composition contain the following traditional Chinese medicine components in parts by weight:

[0010] Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts.

[0011] Preferably, the raw materials for preparing the traditional Chinese medicine composition also include Brucea javanica and Podophyllum hexandrum.

[0012] More preferably, the weight parts of Brucea javanica and Podophyllum hexandrum in the raw materials for preparing the traditional Chinese medicine composition are as follows: 10-20 parts of Brucea javanica and 8-15 parts of Podophyllum hexandrum.

[0013] Most preferably, the weight parts of Brucea javanica and Podophyllum hexandrum in the raw materials for preparing the traditional Chinese medicine composition are as follows: 15 parts of Brucea javanica and 10 parts of Podophyllum hexandrum.

[0014] Further research by the inventors revealed that a traditional Chinese medicine composition prepared by adding two more herbs, namely Brucea javanica and Podophyllum hexandrum, to five herbs—Salvia miltiorrhiza, Astragalus membranaceus, Curcuma zedoaria, Smilax china, and Glycyrrhiza uralensis—significantly improved its anti-gastric cancer effect compared to a composition prepared solely from these five herbs.

[0015] The present invention also provides a method for preparing the above-mentioned traditional Chinese medicine composition, which includes the following steps:

[0016] After mixing and pulverizing the Chinese herbal raw materials, they are extracted with an ethanol aqueous solution. After extraction, the extract is separated, concentrated and dried to obtain an ethanol aqueous solution extract of the Chinese herbal medicine. The Chinese herbal composition is obtained by taking the ethanol aqueous solution extract of the Chinese herbal medicine.

[0017] Preferably, the ratio of the total weight of the Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 5-20 L.

[0018] Most preferably, the ratio of the total weight of the Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L.

[0019] Preferably, the ethanol aqueous solution refers to an ethanol aqueous solution with an ethanol volume fraction of 70% to 95%.

[0020] Preferably, the extraction refers to reflux extraction, and the extraction time is 1 to 2 hours.

[0021] More preferably, the preparation method further includes the following steps:

[0022] The ethanol-water solution extract of the traditional Chinese medicine is suspended in water, and then extracted with a mixed extractant. The extract obtained by the mixed extractant is concentrated and dried to obtain the traditional Chinese medicine extract. The traditional Chinese medicine composition is obtained by taking the traditional Chinese medicine extract.

[0023] Further research by the inventors revealed that the preparation method of the traditional Chinese medicine composition of the present invention is crucial; in the preparation process of the traditional Chinese medicine composition, the traditional Chinese medicine extract obtained by further extracting the traditional Chinese medicine ethanol aqueous solution extract through the mixed extractant described in the present invention has a significantly higher anti-gastric cancer effect than the traditional Chinese medicine ethanol aqueous solution extract.

[0024] Preferably, the mixed extractant is composed of n-hexane, diethyl ether, and ethyl acetate.

[0025] Further research by the inventors revealed that the selection of the extractant is crucial. Studies have shown that when using a single extractant such as n-hexane, diethyl ether, or ethyl acetate, the anti-gastric cancer effect of the extracted traditional Chinese medicine is not significantly improved compared to that of the ethanol-water solution extract. Only when the extractant is composed of n-hexane, diethyl ether, and ethyl acetate can the anti-gastric cancer effect be significantly improved compared to that of the ethanol-water solution extract.

[0026] More preferably, the volume ratio of hexane, diethyl ether, and ethyl acetate in the mixed extractant is 8–12:3–6:1–3.

[0027] Most preferably, the volume ratio of n-hexane, diethyl ether, and ethyl acetate in the mixed extractant is 10:5:2.

[0028] Preferably, the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water, and mixed extractant is 1g:5-10mL:5-10mL.

[0029] Preferably, the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water, and mixed extractant is 1g:8mL:8mL.

[0030] More preferably, the preparation method further includes the following steps: eluting the herbal extract onto a silica gel column, first using a mixed organic solvent of chloroform and methanol in a volume ratio of 1000:30:50 (3-6 column volumes) to remove impurities; then eluting again using a mixed organic solvent of chloroform and methanol in a volume ratio of 1000:150:180 (3-6 column volumes), collecting the eluent obtained by elution with the mixed organic solvent of chloroform and methanol in a volume ratio of 1000:150:180, concentrating and drying to obtain the herbal composition.

[0031] Most preferably, the preparation method further includes the following steps: taking the herbal extract and loading it onto a silica gel column, first eluting it with a mixed organic solvent consisting of chloroform and methanol in a volume ratio of 1000:45 (3-6 column volumes) to remove impurities; then eluting it with a mixed organic solvent consisting of chloroform and methanol in a volume ratio of 1000:170 (3-6 column volumes), collecting the eluent eluted with the mixed organic solvent consisting of chloroform and methanol in a volume ratio of 1000:170, concentrating and drying it to obtain the herbal composition.

[0032] In further research, the inventors discovered that, in the preparation process of the traditional Chinese medicine composition of the present invention, the traditional Chinese medicine composition prepared by further eluting the traditional Chinese medicine extract under the silica gel column elution conditions described above has a significantly enhanced anti-gastric cancer effect compared to the traditional Chinese medicine extract.

[0033] The inventors also discovered in their research that the silica gel column elution conditions of the present invention are crucial; only when the Chinese herbal extract is further prepared under the silica gel column elution conditions of the present invention can the anti-gastric cancer effect of the Chinese herbal extract be significantly improved compared to that of the Chinese herbal extract; however, the anti-gastric cancer effect of the Chinese herbal extract prepared under other silica gel column elution conditions cannot be significantly improved compared to that of the Chinese herbal extract.

[0034] The present invention also provides the application of the above-mentioned traditional Chinese medicine composition in the preparation of a drug with anti-gastric cancer effect.

[0035] Beneficial effects: This invention provides a novel traditional Chinese medicine composition. Studies have shown that the traditional Chinese medicine composition prepared from five herbs, namely Danshen, Huangqi, Ezhu, Smilax, and Gancao, has anti-gastric cancer effects. Therefore, using the traditional Chinese medicine composition described in this invention as an active ingredient to prepare drugs with anti-gastric cancer effects has important application value. Detailed Implementation

[0036] The present invention will be further explained below with reference to specific embodiments, but the embodiments do not limit the present invention in any way.

[0037] Example 1: Preparation of the Traditional Chinese Medicine Composition

[0038] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts.

[0039] Preparation method: The above-mentioned parts by weight of Chinese herbal raw materials are mixed and pulverized through a 100-mesh sieve. Then, the mixture is heated to boiling with an ethanol aqueous solution and refluxed for 1 hour. After extraction, the extract is filtered and separated. The extract is concentrated and dried to obtain the Chinese herbal ethanol aqueous solution extract. The Chinese herbal composition is obtained by taking the Chinese herbal ethanol aqueous solution extract. The ratio of the total weight of the Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L. The ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%.

[0040] Example 2: Preparation of the Traditional Chinese Medicine Composition

[0041] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0042] Preparation method: Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them and pass them through a 100-mesh sieve. Then, heat the ethanol aqueous solution to boiling and reflux for 1 hour. After extraction, filter and separate the extract. Concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract. Take the Chinese herbal ethanol aqueous solution extract to obtain the Chinese herbal composition.

[0043] The ratio of the total weight of the Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%.

[0044] Example 3: Preparation of the Traditional Chinese Medicine Composition

[0045] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0046] Preparation method:

[0047] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0048] (2) Take the ethanol aqueous solution extract of traditional Chinese medicine, add water to suspend it, and then extract it with a mixed extractant. After the extract obtained by the mixed extractant is concentrated and dried, the traditional Chinese medicine extract is obtained. Take the traditional Chinese medicine extract to obtain the traditional Chinese medicine composition.

[0049] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and the mixed extractant is 1g:8mL:8mL; the mixed extractant is composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0050] Example 4: Preparation of Traditional Chinese Medicine Composition

[0051] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0052] Preparation method:

[0053] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0054] (2) Take the Chinese herbal ethanol aqueous solution extract, add water to suspend it, and then extract it with a mixed extractant. The extract obtained by the mixed extractant is concentrated and dried to obtain the Chinese herbal extract. The ratio of the Chinese herbal ethanol aqueous solution extract, water and mixed extractant is 1g:8mL:8mL. The mixed extractant is composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0055] (3) Take the Chinese herbal extract and place it on a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of silica gel is 40 times the weight of the Chinese herbal extract). First, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:45 (5 column volumes) to remove impurities. Then, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:170 (5 column volumes) to elute. Collect the eluent eluted by the mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:170, concentrate and dry it to obtain the Chinese herbal composition.

[0056] Preparation of Traditional Chinese Medicine Composition in Comparative Example 1

[0057] The weight composition of the Chinese herbal raw materials is the same as in Example 3.

[0058] Preparation method:

[0059] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0060] (2) Take the ethanol aqueous solution extract of the Chinese medicine, add water to suspend it, and then extract it with an extractant. After the extract obtained by the extractant is concentrated and dried, the Chinese medicine extract is obtained. The Chinese medicine composition is obtained by taking the Chinese medicine extract.

[0061] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and extractant is 1g:8mL:8mL; the extractant is n-hexane.

[0062] The difference between Comparative Example 1 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 1 uses n-hexane as a single extractant; while Example 3 uses a mixed extractant composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0063] Preparation of Traditional Chinese Medicine Composition in Comparative Example 2

[0064] The weight composition of the Chinese herbal raw materials is the same as in Example 3.

[0065] Preparation method:

[0066] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0067] (2) Take the ethanol aqueous solution extract of the Chinese medicine, add water to suspend it, and then extract it with an extractant. After the extract obtained by the extractant is concentrated and dried, the Chinese medicine extract is obtained. The Chinese medicine composition is obtained by taking the Chinese medicine extract.

[0068] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and extractant is 1g:8mL:8mL; the extractant is diethyl ether.

[0069] The difference between Comparative Example 2 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 2 uses a single diethyl ether as the extractant; while Example 3 uses a mixed extractant consisting of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0070] Preparation of the traditional Chinese medicine composition in Comparative Example 3

[0071] The weight composition of the Chinese herbal raw materials is the same as in Example 3.

[0072] Preparation method:

[0073] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0074] (2) Take the ethanol aqueous solution extract of the Chinese medicine, add water to suspend it, and then extract it with an extractant. After the extract obtained by the extractant is concentrated and dried, the Chinese medicine extract is obtained. The Chinese medicine composition is obtained by taking the Chinese medicine extract.

[0075] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and extractant is 1g:8mL:8mL; the extractant is ethyl acetate.

[0076] The difference between Comparative Example 3 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 3 uses ethyl acetate as the single extractant; while Example 3 uses a mixed extractant composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0077] Preparation of the traditional Chinese medicine composition in Comparative Example 4

[0078] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0079] Preparation method:

[0080] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0081] (2) Take the ethanol aqueous solution extract of traditional Chinese medicine, add water to suspend it, and then extract it with a mixed extractant. After the extract obtained by the mixed extractant is concentrated and dried, the traditional Chinese medicine extract is obtained. Take the traditional Chinese medicine extract to obtain the traditional Chinese medicine composition.

[0082] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and the mixed extractant is 1g:8mL:8mL; the mixed extractant is composed of n-hexane and diethyl ether in a volume ratio of 10:5.

[0083] The difference between Comparative Example 4 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 4 uses a mixed extractant consisting of n-hexane and diethyl ether in a volume ratio of 10:5; while Example 3 uses a mixed extractant consisting of n-hexane, diethyl ether, and ethyl acetate in a volume ratio of 10:5:2.

[0084] Preparation of the traditional Chinese medicine composition in Comparative Example 5

[0085] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0086] Preparation method:

[0087] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0088] (2) Take the ethanol aqueous solution extract of traditional Chinese medicine, add water to suspend it, and then extract it with a mixed extractant. After the extract obtained by the mixed extractant is concentrated and dried, the traditional Chinese medicine extract is obtained. Take the traditional Chinese medicine extract to obtain the traditional Chinese medicine composition.

[0089] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and the mixed extractant is 1g:8mL:8mL; the mixed extractant is composed of n-hexane and ethyl acetate in a volume ratio of 10:2.

[0090] The difference between Comparative Example 5 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 5 uses a mixed extractant consisting of n-hexane and ethyl acetate in a volume ratio of 10:2; while Example 3 uses a mixed extractant consisting of n-hexane, diethyl ether, and ethyl acetate in a volume ratio of 10:5:2.

[0091] Preparation of the Traditional Chinese Medicine Composition in Comparative Example 6

[0092] Composition of Chinese medicinal materials by weight: Salvia miltiorrhiza 25 parts; Astragalus membranaceus 15 parts; Curcuma zedoaria 15 parts; Smilax china 10 parts; Glycyrrhiza uralensis 5 parts; Brucea javanica 15 parts; Podophyllum hexandrum 10 parts.

[0093] Preparation method:

[0094] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0095] (2) Take the ethanol aqueous solution extract of traditional Chinese medicine, add water to suspend it, and then extract it with a mixed extractant. After the extract obtained by the mixed extractant is concentrated and dried, the traditional Chinese medicine extract is obtained. Take the traditional Chinese medicine extract to obtain the traditional Chinese medicine composition.

[0096] In step (2), the ratio of the amount of the Chinese herbal ethanol aqueous solution extract, water and the mixed extractant is 1g:8mL:8mL; the mixed extractant is composed of diethyl ether and ethyl acetate in a volume ratio of 5:2.

[0097] The difference between Comparative Example 6 and Example 3 lies in the choice of extractant during the preparation of the traditional Chinese medicine composition; Comparative Example 6 uses a mixed extractant consisting of diethyl ether and ethyl acetate in a volume ratio of 5:2; while Example 3 uses a mixed extractant consisting of n-hexane, diethyl ether, and ethyl acetate in a volume ratio of 10:5:2.

[0098] Preparation of the traditional Chinese medicine composition in Comparative Example 7

[0099] The weight composition of the Chinese herbal raw materials is the same as in Example 4.

[0100] Preparation method:

[0101] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0102] (2) Take the Chinese herbal ethanol aqueous solution extract, add water to suspend it, and then extract it with a mixed extractant. The extract obtained by the mixed extractant is concentrated and dried to obtain the Chinese herbal extract. The ratio of the Chinese herbal ethanol aqueous solution extract, water and mixed extractant is 1g:8mL:8mL. The mixed extractant is composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0103] (3) Take the Chinese herbal extract and place it on a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of silica gel is 40 times the weight of the Chinese herbal extract). First, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:150 and 5 times the column volume to remove impurities. Then, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:300 and 5 times the column volume to elute. Collect the eluent eluted by the mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:300, concentrate and dry it to obtain the Chinese herbal composition.

[0104] The difference between Comparative Example 7 and Example 4 lies in the different elution conditions of the silica gel column.

[0105] Preparation of the traditional Chinese medicine composition in Comparative Example 8

[0106] The weight composition of the Chinese herbal raw materials is the same as in Example 4.

[0107] Preparation method:

[0108] (1) Mix the above-mentioned parts by weight of Chinese herbal raw materials, pulverize them through a 100-mesh sieve, then heat them to boiling with an ethanol aqueous solution and reflux for 1 hour. After extraction, filter and separate the extract, concentrate and dry the extract to obtain the Chinese herbal ethanol aqueous solution extract; wherein, the ratio of the total weight of Chinese herbal raw materials to the amount of ethanol aqueous solution is 1 kg: 10 L; the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 95%;

[0109] (2) Take the Chinese herbal ethanol aqueous solution extract, add water to suspend it, and then extract it with a mixed extractant. The extract obtained by the mixed extractant is concentrated and dried to obtain the Chinese herbal extract. The ratio of the Chinese herbal ethanol aqueous solution extract, water and mixed extractant is 1g:8mL:8mL. The mixed extractant is composed of n-hexane, diethyl ether and ethyl acetate in a volume ratio of 10:5:2.

[0110] (3) Take the Chinese herbal extract and place it on a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of silica gel is 40 times the weight of the Chinese herbal extract). First, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:300 and 5 times the column volume to remove impurities. Then, use a mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:500 and 5 times the column volume to elute. Collect the eluent eluted by the mixed organic solvent consisting of chloroform and methanol with a volume ratio of 1000:500, concentrate and dry it to obtain the Chinese herbal composition.

[0111] The difference between Comparative Example 8 and Example 4 lies in the different elution conditions of the silica gel column.

[0112] Experimental Example 1: Anti-gastric cancer trial

[0113] The experimental method is as follows: Gastric cancer cells were added to 96-well plates containing RPMI 1640 medium (2 × 10⁶ cells / well per well). 4Cells were cultured in RPMI 1640 medium (0.01–100 μg / mL) at 37°C for 24 hours, and the medium was then removed. Next, the experimental group was added to RPMI 1640 medium containing the test sample (concentration of the test sample was 0.01–100 μg / mL), while the blank control group was added to fresh RPMI 1640 medium. The cells were then cultured in RPMI 1640 medium at 37°C for 48 hours. Then, 20 μL of MTT (5 mg / mL) was added to each well, and the cells were cultured for another 4 hours, after which the supernatant was removed. Next, 100 μL of DMSO was added to each well, and the cells were incubated in the dark for 10 minutes. The absorbance at 570 nm was then measured using a microplate reader. Cell viability was calculated based on the absorbance, and the half-inhibitory concentration (IC50) was calculated based on the dose-response curve. 50 Each treatment had 3 replicate wells. Cell viability (%) = ΔOD 药物处理组 / ΔOD 空白对照组 ×100; Experimental results are shown in Table 1.

[0114] The gastric cancer cells used in this experiment were human gastric cancer cells BGC-823.

[0115] The samples used in this experiment were the traditional Chinese medicine compositions prepared in Examples 1-4 and Comparative Examples 1-8.

[0116] Table 1. Results of the anti-gastric cancer test of the traditional Chinese medicine composition of the present invention.

[0117] Sample to be tested <![CDATA[IC 50 Value The traditional Chinese medicine composition prepared in Example 1 32.3 μg / mL The traditional Chinese medicine composition prepared in Example 2 25.9 μg / mL The traditional Chinese medicine composition prepared in Example 3 10.5 μg / mL The traditional Chinese medicine composition prepared in Example 4 1.1 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 1 18.4 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 2 20.6 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 3 22.3 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 4 19.7 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 5 20.2 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 6 21.7 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 7 8.8 μg / mL The traditional Chinese medicine composition prepared in Comparative Example 8 16.1 μg / mL

[0118] As can be seen from the experimental results in Table 1, the traditional Chinese medicine composition prepared in Example 1 showed an IC50 response against gastric cancer cells. 50 The value was 32.3 μg / mL, showing a certain anti-gastric cancer effect; this indicates that the traditional Chinese medicine composition prepared by the present invention using five Chinese medicinal herbs, namely Danshen, Huangqi, Ezhu, Smilax, and Gancao, has an anti-gastric cancer effect.

[0119] As can be seen from the experimental results in Table 1, the traditional Chinese medicine composition prepared in Example 2 showed an IC50 response against gastric cancer cells. 50 The value was significantly lower than that of the traditional Chinese medicine composition prepared in Example 1; this indicates that the traditional Chinese medicine composition prepared by further adding two traditional Chinese medicine raw materials, namely Brucea javanica and Podophyllum hexandrum, to the five traditional Chinese medicine raw materials of Salvia miltiorrhiza, Astragalus membranaceus, Curcuma zedoaria, Smilax china and Glycyrrhiza uralensis, has a significantly improved anti-gastric cancer effect compared with the traditional Chinese medicine composition prepared by using the five traditional Chinese medicine raw materials of Salvia miltiorrhiza, Astragalus membranaceus, Curcuma zedoaria, Smilax china and Glycyrrhiza uralensis.

[0120] As can be seen from the experimental results in Table 1, the traditional Chinese medicine composition prepared in Example 3 showed an IC50 response against gastric cancer cells. 50The value is significantly lower than that of the traditional Chinese medicine composition prepared in Example 2; this indicates that the preparation method of the traditional Chinese medicine composition of the present invention is crucial; in the preparation process of the traditional Chinese medicine composition, the traditional Chinese medicine extract obtained by further extracting the traditional Chinese medicine ethanol aqueous solution extract through the mixed extractant described in the present invention has a significantly higher anti-gastric cancer effect than the traditional Chinese medicine ethanol aqueous solution extract.

[0121] As can be seen from the experimental results in Table 1, the traditional Chinese medicine compositions prepared in Comparative Examples 1-3 showed high IC50 against gastric cancer cells. 50 Compared with the traditional Chinese medicine composition prepared in Example 2, the value was not significantly reduced, and the reduction was much smaller than that of the traditional Chinese medicine composition prepared in Example 3. This indicates that the anti-gastric cancer effect of the traditional Chinese medicine extract obtained by using a single hexane, ether, or ethyl acetate as an extractant during the preparation process is not significantly improved compared with the traditional Chinese medicine ethanol aqueous solution extract. Only the traditional Chinese medicine extract obtained by using an extractant composed of hexane, ether, and ethyl acetate can achieve a significant improvement in anti-gastric cancer effect compared with the traditional Chinese medicine ethanol aqueous solution extract.

[0122] As can be seen from the experimental results in Table 1, the traditional Chinese medicine compositions prepared in Comparative Examples 4-6 showed high IC50 against gastric cancer cells. 50 Compared with the traditional Chinese medicine composition prepared in Example 2, the value was not significantly reduced, and the reduction was also significantly smaller than that of the traditional Chinese medicine composition prepared in Example 3. This indicates that, in the preparation process, the traditional Chinese medicine extract obtained by using any combination of two of hexane, diethyl ether, and ethyl acetate as the extractant does not have a significantly improved anti-gastric cancer effect compared with the traditional Chinese medicine ethanol aqueous solution extract. Only when the traditional Chinese medicine extract is extracted by using an extractant composed of hexane, diethyl ether, and ethyl acetate simultaneously can its anti-gastric cancer effect be significantly improved compared with the traditional Chinese medicine ethanol aqueous solution extract.

[0123] As can be seen from the experimental results in Table 1, the traditional Chinese medicine composition prepared in Example 4 showed an IC50 response against gastric cancer cells. 50 The value is significantly lower than that of the traditional Chinese medicine composition prepared in Example 3; it has excellent anti-gastric cancer effect. This shows that in the preparation process of the traditional Chinese medicine composition of the present invention, the traditional Chinese medicine composition prepared by further eluting the traditional Chinese medicine extract under the above-mentioned silica gel column elution conditions of the present invention has a significantly improved anti-gastric cancer effect compared with the traditional Chinese medicine extract.

[0124] As can be seen from the experimental results in Table 1, the traditional Chinese medicine composition prepared in Comparative Example 7 showed an IC50 response against gastric cancer cells. 50Compared with the traditional Chinese medicine composition prepared in Example 3, the IC50 value was not significantly reduced, and the reduction was much smaller than that of the traditional Chinese medicine composition prepared in Example 4; the traditional Chinese medicine composition prepared in Comparative Example 8 showed a lower IC50 value against gastric cancer cells. 50 Compared with the traditional Chinese medicine composition prepared in Example 3, the value not only did not decrease, but actually increased. This indicates that the silica gel column elution conditions of the present invention are crucial; only when the traditional Chinese medicine extract is further prepared under the above-mentioned silica gel column elution conditions of the present invention can the anti-gastric cancer activity of the traditional Chinese medicine composition be significantly improved compared with that of the traditional Chinese medicine extract; however, the anti-gastric cancer activity of the traditional Chinese medicine composition prepared under other silica gel column elution conditions cannot be significantly improved compared with that of the traditional Chinese medicine extract.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of a drug with anti-gastric cancer activity, characterized in that, Its preparation raw materials consist of the following traditional Chinese medicine components in parts by weight: Salvia miltiorrhiza 20-30 parts; Astragalus membranaceus 10-20 parts; Curcuma zedoaria 10-20 parts; Smilax china 8-15 parts; Glycyrrhiza uralensis 4-8 parts; Brucea javanica 10-20 parts; Podophyllum hexandrum 8-15 parts; The method for preparing the traditional Chinese medicine composition includes the following steps: After mixing and pulverizing the Chinese herbal raw materials, they are extracted with an ethanol aqueous solution. After extraction, the extract is separated, concentrated and dried to obtain the Chinese herbal ethanol aqueous solution extract. The ethanol-water extract of the traditional Chinese medicine was suspended in water, and then extracted with a mixed extractant. The extract obtained by the mixed extractant was concentrated and dried to obtain the traditional Chinese medicine extract. The traditional Chinese medicine composition was obtained by taking the traditional Chinese medicine extract. The mixed extractant is composed of n-hexane, diethyl ether, and ethyl acetate; The volume ratio of hexane, diethyl ether, and ethyl acetate in the mixed extractant is 8–12:3–6:1–3.

2. The application according to claim 1, characterized in that, The ratio of the total weight of Chinese herbal raw materials to the amount of ethanol-water solution used is 1 kg: 5-20 L.

3. The application according to claim 1, characterized in that, The ratio of the total weight of the Chinese herbal raw materials to the amount of ethanol aqueous solution used is 1 kg: 10 L.

4. The application according to claim 1, characterized in that, The ethanol-water solution mentioned refers to an ethanol-water solution with an ethanol volume fraction of 70% to 95%.