CN116236516A - Traditional Chinese medicine composition for treating liver cancer, preparation and preparation method - Google Patents
Traditional Chinese medicine composition for treating liver cancer, preparation and preparation method Download PDFInfo
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- CN116236516A CN116236516A CN202310438835.4A CN202310438835A CN116236516A CN 116236516 A CN116236516 A CN 116236516A CN 202310438835 A CN202310438835 A CN 202310438835A CN 116236516 A CN116236516 A CN 116236516A
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Abstract
The embodiment of the invention discloses a traditional Chinese medicine composition for treating liver cancer, a preparation and a preparation method thereof; the preparation method comprises the following steps: (1) Taking a set amount of taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum fruits and rhodiola rosea; (2) Extracting the enzymatic hydrolysate of the yew with ethyl acetate, purifying and crystallizing the ethyl acetate extract to obtain the yew extract; (3) Crushing haematococcus pluvialis, and extracting with deionized water to obtain haematococcus pluvialis extract; (4) Extracting with ethanol to obtain Concha Ostreae dry extract, herba Silybi Mariani fruit extract and radix Rhodiolae extract respectively; (5) Mixing Concha Ostreae dry extract, herba Silybi Mariani extract, radix Rhodiolae extract and Haematococcus pluvialis extract to obtain extract mixtureA compound; subjecting the obtained extract mixture to CO 2 Supercritical extraction to obtain mixed extract; (6) The Chinese yew extract and the mixed extract are mixed according to the mass ratio of 1:3 to obtain the traditional Chinese medicine composition, which has high tumor inhibition rate on liver cancer.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and in particular relates to a traditional Chinese medicine composition for treating liver cancer, a preparation and a preparation method thereof.
Background
Liver cancer is a malignant tumor of liver and can be classified into primary and secondary. Primary liver malignancies originate in liver epithelium or mesenchymal tissue and are highly-developed, extremely-dangerous malignancies. Secondary liver cancer, also known as metastatic liver cancer, refers to malignant tumors of multiple organ origin throughout the body invading the liver.
The etiology and molecular mechanism of primary liver cancer are not clear, and the pathogenesis is generally considered to be a multi-factor and multi-step complex process, and is influenced by environmental and dietary factors. Epidemiology and experimental research data show that hepatitis B virus, hepatitis C virus infection, aflatoxin, drinking water pollution, alcohol, liver cirrhosis, sex hormone and nitrosamines are all related to liver cancer pathogenesis. Secondary liver cancer can develop disease through different pathways such as blood, lymph fluid transfer or direct infiltration of the liver.
Research on pathogenesis and pathogenic factors of liver cancer, and development of drugs or treatment methods capable of effectively treating liver cancer have been one of the research directions of those skilled in the art.
Disclosure of Invention
In view of this, some embodiments disclose a method for preparing a traditional Chinese medicine composition for treating liver cancer, comprising:
(1) Taking a set amount of taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum fruits and rhodiola rosea;
(2) Performing enzymolysis on the yew, extracting a yew enzymolysis product by using ethyl acetate, and purifying and crystallizing the ethyl acetate extraction product to obtain a yew extract;
(3) Crushing haematococcus pluvialis, and extracting the crushed haematococcus pluvialis with deionized water to obtain haematococcus pluvialis extract;
(4) Pulverizing Concha Ostreae, fructus Silybi Mariani and radix Rhodiolae respectively, and extracting with ethanol to obtain Concha Ostreae dry extract, herba Silybi Mariani extract and radix Rhodiolae extract respectively;
(5) Will beMixing Concha Ostreae dry extract, herba Silybi Mariani fruit extract, radix Rhodiolae extract and Haematococcus pluvialis extract to obtain extract mixture; subjecting the obtained extract mixture to CO 2 Supercritical extraction to obtain mixed extract of Concha Ostreae dry, herba Silybi Mariani, radix Rhodiolae and Haematococcus pluvialis extract;
(6) Mixing the Taxus chinensis extract and the mixed extract according to a mass ratio of 1:3 to obtain the traditional Chinese medicine composition.
Further, some embodiments disclose a preparation method of a traditional Chinese medicine composition for treating liver cancer, and the yew is subjected to enzymolysis under the action of cellulase.
The preparation method of the traditional Chinese medicine composition for treating liver cancer disclosed in some embodiments further purifies the taxus chinensis extract, and specifically comprises the following steps:
(2-1) mixing the Taxus chinensis extract with neutral alumina;
(2-2) subjecting the obtained mixture to chromatography by using a silica gel or alumina chromatographic column, eluting with petroleum ether and ethyl acetate to remove fat-soluble impurities, eluting with a mixed solution of alcohol and ethyl acetate, collecting a mixed solution fraction of alcohol and ethyl acetate, and removing alcohol and ethyl acetate under reduced pressure to obtain a primary purified product;
(2-3) using polymethyl acrylate resin column and polystyrene resin column layer as chromatographic column, using dichloromethane and acetone mixed solution as eluent, and sequentially performing chromatography to the primary purified product to obtain pure product of Taxus chinensis extract.
The preparation method of the traditional Chinese medicine composition for treating liver cancer disclosed in some embodiments comprises the step of further crystallizing and refining the pure taxus chinensis extract to obtain a refined taxus chinensis extract.
On the other hand, some embodiments disclose a traditional Chinese medicine composition for treating liver cancer, which is obtained by a preparation method of the traditional Chinese medicine composition for treating liver cancer.
Further, the traditional Chinese medicine composition for treating liver cancer disclosed in some embodiments comprises, by mass, 20-30 parts of taxus chinensis, 15-30 parts of haematococcus pluvialis, 15-25 parts of dried oyster, 20-35 parts of silybum marianum fruits and 5-15 parts of rhodiola rosea.
In yet another aspect, some embodiments disclose a traditional Chinese medicine preparation for treating liver cancer, including a traditional Chinese medicine composition for treating liver cancer.
Some embodiments disclose a traditional Chinese medicine preparation for treating liver cancer, and the traditional Chinese medicine preparation also comprises pharmaceutically acceptable auxiliary materials.
The traditional Chinese medicine preparation for treating liver cancer disclosed by some embodiments comprises the dosage forms of decoction, pills, ointment, powder, dew, pills, wine and lozenges.
The traditional Chinese medicine composition for treating liver cancer, the preparation and the preparation method disclosed by the embodiment of the invention have the advantages that the obtained traditional Chinese medicine composition and the preparation have no toxic or side effect and have high tumor inhibition rate on liver cancer.
Drawings
FIG. 1 example 2 tumor volume control schematic.
Detailed Description
The word "embodiment" as used herein does not necessarily mean that any embodiment described as "exemplary" is preferred or advantageous over other embodiments. Performance index testing in the examples of the present invention, unless otherwise specified, was performed using conventional testing methods in the art. It should be understood that the terminology used in the description of the embodiments of the invention presented is for the purpose of describing particular embodiments only, and is not intended to be limiting of the disclosure of the embodiments of the invention.
Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the invention belong; other test methods and techniques not specifically identified in the examples of the present invention are those generally employed by those skilled in the art.
The terms "substantially" and "about" are used herein to describe small fluctuations. For example, they may refer to less than or equal to ±5%, such as less than or equal to ±2%, such as less than or equal to ±1%, such as less than or equal to ±0.5%, such as less than or equal to ±0.2%, such as less than or equal to ±0.1%, such as less than or equal to ±0.05%. Numerical data presented or represented herein in a range format is used only for convenience and brevity and should therefore be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range. For example, a numerical range of "1 to 5%" should be interpreted to include not only the explicitly recited values of 1% to 5%, but also include individual values and sub-ranges within the indicated range. Thus, individual values, such as 2%, 3.5% and 4%, and subranges, such as 1% to 3%, 2% to 4% and 3% to 5%, etc., are included in this numerical range. The same principle applies to ranges reciting only one numerical value. Moreover, such an interpretation applies regardless of the breadth of the range or the characteristics being described.
In this document, including the claims, conjunctions such as "comprising," including, "" carrying, "" having, "" containing, "" involving, "" containing, "and the like are to be construed as open-ended, i.e., to mean" including, but not limited to. Only the conjunctions "consisting of … …" and "consisting of … …" are closed conjunctions.
Numerous specific details are set forth in the following examples in order to provide a better understanding of the present invention. It will be understood by those skilled in the art that the present invention may be practiced without some of these specific details. In the examples, some methods, means, instruments, devices, etc. well known to those skilled in the art are not described in detail in order to highlight the gist of the present invention.
On the premise of no conflict, the technical features disclosed by the embodiment of the invention can be combined at will, and the obtained technical scheme belongs to the disclosure of the embodiment of the invention.
Taxus chinensis, also called Taxus chinensis, is a well-known anticancer plant in the world. The natural secondary metabolite separated and extracted from the bark has good anti-tumor effect through clinical verification, and has special effects on ovarian cancer, uterine cancer, breast cancer and the like with high incidence rate of cancer.
As haematococcus pluvialis contains 1.5 to 10.0 percent of natural astaxanthin, the haematococcus pluvialis becomes the strongest element for delaying senescence so far. Since astaxanthin can significantly inhibit chemical-induced early canceration, it has antiproliferative and immunostimulating effects on epithelial cells exposed to carcinogens, such as inhibiting growth of bladder cancer, oral cancer, colon cancer, gastric cancer and breast cancer. Astaxanthin also reduces the number of aflatoxin-induced liver tumor cells.
Silybum marianum is a heat-clearing and toxicity-removing medicine, and the silibinin has obvious protection effect on liver injury caused by various liver toxicities. Silybin, silydianin and silychristin all have effects of stabilizing cell and intracellular biomembrane, and are a class of anti-liver virus active substances. In addition, silymarin alcohol also has bile secretion promoting effect.
Rhodiola rosea is a traditional Chinese medicine and has the effects of clearing heat and detoxicating, activating blood and stopping bleeding, clearing lung and relieving cough, tonifying kidney and nourishing heart. The dried oyster is a medicinal and edible Chinese medicine, has the effects of softening hardness and resolving hard mass when being used as a Chinese medicine, is compatible with other medicinal materials, and can also treat tumors caused by qi stagnation and blood stasis.
The inventor finds that the Chinese yew, haematococcus pluvialis, dried oyster, silybum marianum fruit and rhodiola rosea with certain mass ratio are compounded, the active ingredients in the raw medicines of the Chinese yew, haematococcus pluvialis, dried oyster, silybum marianum fruit and rhodiola rosea are extracted by different methods, and the active ingredients are compounded into a composition according to a certain proportion, so that the Chinese yew, haematococcus pluvialis, dried oyster, silybum marianum fruit and rhodiola rosea have good treatment effect on liver cancer.
In some embodiments, the method for preparing a traditional Chinese medicine composition for treating liver cancer comprises:
(1) Taking a set amount of taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum fruits and rhodiola rosea;
(2) Performing enzymolysis on the yew, extracting a yew enzymolysis product by using ethyl acetate, and purifying and crystallizing the ethyl acetate extraction product to obtain a yew extract;
(3) Crushing haematococcus pluvialis, and extracting the crushed haematococcus pluvialis with deionized water to obtain haematococcus pluvialis extract;
(4) Pulverizing Concha Ostreae, herba Silybi Mariani and radix Rhodiolae respectively, and extracting with ethanol to obtain Concha Ostreae dry extract, herba Silybi Mariani fruit extract and radix Rhodiolae extract respectively;
(5) Will be made of oysterMixing oyster dry extract, herba Silybi Mariani extract, radix Rhodiolae extract and Haematococcus pluvialis extract to obtain extract mixture; subjecting the obtained extract mixture to CO 2 Supercritical extraction to obtain mixed extract of Concha Ostreae dry, herba Silybi Mariani, radix Rhodiolae and Haematococcus pluvialis extract;
(6) Mixing the Taxus chinensis extract and the mixed extract according to a mass ratio of 1:3 to obtain the traditional Chinese medicine composition.
In some embodiments, the traditional Chinese medicine composition for treating liver cancer is prepared from the following raw materials, by mass, 20-30 parts of taxus chinensis, 15-30 parts of haematococcus pluvialis, 10-30 parts of dried oyster, 20-35 parts of silybum marianum fruits and 5-15 parts of rhodiola rosea.
In some preferred embodiments, the preparation method of the traditional Chinese medicine composition for treating liver cancer adopts raw medicines comprising, by mass, 22-28 parts of taxus chinensis, 18-25 parts of haematococcus pluvialis, 15-25 parts of dried oyster, 23-30 parts of silybum marianum fruits and 8-10 parts of rhodiola rosea.
In a more preferred embodiment, the preparation method of the traditional Chinese medicine composition for treating liver cancer adopts the raw medicine composition comprising, by mass, 25 parts of taxus chinensis, 20 parts of haematococcus pluvialis, 20 parts of dried oyster, 27 parts of silybum marianum and 80 parts of rhodiola rosea.
Some embodiments disclose a preparation method of a traditional Chinese medicine composition for treating liver cancer, and yew is subjected to enzymolysis under the action of cellulase.
The preparation method of the traditional Chinese medicine composition for treating liver cancer disclosed in some embodiments further purifies the taxus chinensis extract, and specifically comprises the following steps:
(2-1) mixing the Taxus chinensis extract with neutral alumina;
(2-2) subjecting the obtained mixture to chromatography by using a silica gel or alumina chromatographic column, eluting with petroleum ether and ethyl acetate to remove fat-soluble impurities, eluting with a mixed solution of alcohol and ethyl acetate, collecting a mixed solution fraction of alcohol and ethyl acetate, and removing alcohol and ethyl acetate under reduced pressure to obtain a primary purified product;
(2-3) using polymethyl acrylate resin column and polystyrene resin column layer as chromatographic column, using dichloromethane and acetone mixed solution as eluent, and sequentially performing chromatography to the primary purified product to obtain pure product of Taxus chinensis extract.
The preparation method of the traditional Chinese medicine composition for treating liver cancer disclosed in some embodiments comprises the step of further crystallizing and refining the pure taxus chinensis extract to obtain a refined taxus chinensis extract.
In some embodiments, the preparation method of the traditional Chinese medicine composition for treating liver cancer comprises the following steps:
(1) Taking a set amount of taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum fruits and rhodiola rosea;
(2) Grinding roots, stems and leaves of the red bean, sieving with a 100-200 mesh sieve, adding deionized water with the weight of 5-10 times and glacial acetic acid with the volume concentration of 0.1%, adding cellulase, and carrying out ultrasonic enzymolysis for 2-5 hours to obtain an enzymolysis product; adding an equal volume of ethyl acetate solvent into the enzymolysis product, and performing ultrasonic extraction for 0.5-2 hours; standing, filtering, layering, extracting water phase with organic solvent for two to three times, mixing organic phases, and removing organic solvent under reduced pressure to obtain extract of Taxus chinensis extract;
mixing the obtained extract with neutral alumina of equal mass, stirring, passing through 200-300 mesh silica gel or alumina chromatographic column, eluting with petroleum ether and ethyl acetate to remove liposoluble impurities, eluting with ethanol and ethyl acetate mixed solution with volume ratio of 1:1, monitoring, collecting ethanol and ethyl acetate fractions, and removing ethanol and ethyl acetate under reduced pressure to obtain primary purified product of Taxus chinensis;
sequentially eluting the obtained primary purification product of the Chinese yew by using polymethyl acrylate resin and polystyrene resin columns, and taking a mixed solution of dichloromethane and acetone with the mass ratio of 1:1-1:10 as an eluent to obtain a pure product of the Chinese yew extract;
dissolving pure Taxus chinensis extract product with 15 μg/ml methanol solution at 0-5deg.C for multiple times, crystallizing, and refining to obtain Taxus chinensis crystal.
(3) Cleaning Haematococcus pluvialis, oven drying, crushing into pieces, grinding into powder in a roller, adding 17.5mol/l glacial acetic acid into the powder, and adding 1000ml glacial acetic acid into 100g powder. Mixing to obtain a mixed solution, adding 1800ml of deionized water into the mixed solution, mixing to obtain uniform slurry, concentrating and drying the slurry, and preparing the haematococcus pluvialis extract.
(4) Pulverizing dried oyster, performing ultrasonic extraction, and performing ultrasonic extraction twice by using an ethanol aqueous solution with the mass content of 80-95%, wherein the extraction temperature is 30-40 ℃ and the extraction time is 30min; filtering the extract to obtain oyster dry extract;
performing ultrasonic extraction on the silybum marianum fruits and the rhodiola rosea according to the same method as that of oyster drying;
(5) Mixing oyster dry extract, silybum marianum fruit extract, rhodiola rosea extract and haematococcus pluvialis extract according to the mass ratio of mixing 1:1:2:3 to obtain an extraction mixture; subjecting the extract mixture to CO at 50deg.C under 10MPa 2 Supercritical extraction to extract substances from the mixture, and then reducing the density of supercritical fluid by reducing pressure to 2MPa or heating to 100deg.C to obtain extract and CO 2 The extract was isolated to give a powder.
(6) The obtained extract in powder state is mixed with yew crystal according to the mass ratio of 1:3 to prepare the traditional Chinese medicine composition. The obtained Chinese medicinal composition is in the form of powder solid.
Some embodiments disclose a traditional Chinese medicine composition for treating liver cancer, which is obtained by a preparation method of the traditional Chinese medicine composition for treating liver cancer. Generally, according to the preparation method of the traditional Chinese medicine composition for treating liver cancer, the traditional Chinese medicine composition is prepared by taking taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum and rhodiola rosea with set mass ratio as traditional Chinese medicine raw materials. The obtained Chinese medicinal composition is powdery solid.
The traditional Chinese medicine composition for treating liver cancer disclosed by some embodiments comprises the following traditional Chinese medicine raw materials in parts by mass: 20-30 parts of taxus chinensis, 15-30 parts of haematococcus pluvialis, 10-30 parts of dried oyster, 20-35 parts of silybum marianum fruit and 5-15 parts of rhodiola rosea.
Some preferred embodiments disclose a traditional Chinese medicine composition for treating liver cancer, which comprises the following traditional Chinese medicine raw materials in parts by mass: 22-28 parts of taxus chinensis, 18-25 parts of haematococcus pluvialis, 15-25 parts of dried oyster, 23-30 parts of silybum marianum fruits and 8-10 parts of rhodiola rosea.
Some more preferred embodiments disclose a traditional Chinese medicine composition for treating liver cancer, which comprises the following traditional Chinese medicine raw materials in parts by mass: 25 parts of taxus chinensis, 20 parts of haematococcus pluvialis, 20 parts of dried oyster, 27 parts of silybum marianum fruits and 80 parts of rhodiola rosea.
Some embodiments disclose a traditional Chinese medicine preparation for treating liver cancer, including a traditional Chinese medicine composition for treating liver cancer. Generally, a traditional Chinese medicine composition for treating liver cancer is a powdery solid, and the traditional Chinese medicine composition is taken as an effective ingredient and is usually a main component of a traditional Chinese medicine preparation. The traditional Chinese medicine preparation for treating liver cancer generally comprises a traditional Chinese medicine composition for treating liver cancer as an active ingredient.
Some embodiments disclose a traditional Chinese medicine preparation for treating liver cancer, and the traditional Chinese medicine preparation also comprises pharmaceutically acceptable auxiliary materials. The traditional Chinese medicine preparation for treating liver cancer generally comprises a traditional Chinese medicine composition for treating liver cancer as an active ingredient, and in order to prepare a proper dosage form, pharmaceutically acceptable auxiliary materials such as a forming agent, a stabilizer, a diluent, an absorbent, an adhesive, a colorant, a coating material and the like are also required to be added into the traditional Chinese medicine preparation.
The traditional Chinese medicine preparation for treating liver cancer disclosed by some embodiments comprises the dosage forms of decoction, pills, ointment, powder, dew, pills, wine and lozenges.
Further exemplary details are described below in connection with the embodiments.
The cellulases herein are purchased from Shandong Long Kete enzyme preparation Inc., commercial number unless otherwise noted: 3210090515. the fully automatic pressurized liquid extractor herein is produced by Shanghai Xingxing industry Co., ltd., product number: 310116002777077.
example 1
Preparation method of traditional Chinese medicine composition for treating liver cancer
(1) Taking 100g of roots, stems and leaves of the red bean, grinding, sieving with a 100-mesh sieve, adding deionized water with the weight being 5 times that of the roots, stems and leaves of the red bean and glacial acetic acid with the volume concentration of 0.1%, adding cellulase, and carrying out ultrasonic enzymolysis for 5 hours to obtain an enzymolysis product; adding an equal volume of ethyl acetate solvent into the enzymolysis product, and carrying out ultrasonic extraction for 2 hours; standing, filtering, layering, extracting water phase with organic solvent for two to three times, mixing organic phases, and removing organic solvent under reduced pressure to obtain extract of Taxus chinensis extract;
mixing the obtained extract with neutral alumina of equal mass, stirring, passing through 200 mesh silica gel or alumina chromatographic column, eluting with petroleum ether and ethyl acetate to remove liposoluble impurities, eluting with ethanol and ethyl acetate mixed solution with volume ratio of 1:1, monitoring, collecting ethanol and ethyl acetate fractions, and removing ethanol and ethyl acetate under reduced pressure to obtain primary purified product of Taxus chinensis;
sequentially eluting the primary purified product of the yew by using polymethyl acrylate resin and polystyrene resin columns, and taking a mixed solution of dichloromethane and acetone with the mass ratio of 1:1 as an eluent to obtain a pure product of the yew extract;
dissolving pure Taxus chinensis extract in 15 μg/ml methanol solution at 0-5deg.C for three times, crystallizing, and refining to obtain Taxus chinensis crystal.
(2) Cleaning and drying 100g haematococcus pluvialis, crushing into fragments, grinding into powder in a rolling machine, adding 1000mL of glacial acetic acid with the concentration of 17.5mol/l into the powder, mixing to obtain a mixed solution, adding 1800mL of deionized water into the mixed solution, mixing to obtain uniform slurry, concentrating and drying the slurry, and preparing the haematococcus pluvialis extract.
(3) Pulverizing dried oyster 100g, and performing ultrasonic extraction: ultrasonic extracting with 80% ethanol water solution for two times at 30deg.C for 30min; filtering the extract to obtain oyster dry extract;
performing ultrasonic extraction on the silybum marianum fruits and the rhodiola rosea according to the same method as that of oyster drying;
(4) Mixing Concha Ostreae dry extract, herba Silybi Mariani fruit extract, radix Rhodiolae extract and Haematococcus pluvialis extract at a mass ratio of 1:1:2:3, and performing CO 2 Supercritical extraction at 50deg.C under 10MPaSubsequently reducing the density of the supercritical fluid by reducing the pressure to 2MPa or increasing the temperature to 100 ℃, and reacting the extract with CO 2 Separating to obtain extract in powder state;
(5) The obtained extract in powder state is mixed with yew crystal according to the mass ratio of 1:3 to prepare the traditional Chinese medicine composition. The obtained Chinese medicinal composition is in the form of powder solid.
Example 2
1. Cell culture
HEP1-6 liver cancer cells are cultured in DMEM cell culture solution containing 10% fetal bovine serum, and supplemented with 100u/ml of each of cyan and streptomycin, and placed at 37deg.C and containing 5% CO 2 The medium was changed every 2 days. After confluence, digestion with pancreatin, centrifugation at 1000 r/min for 5 min, discarding supernatant, adding fresh culture medium, blowing up for passaging and collecting cells.
2. Tumor cell seeding and grouping
Seed protection mice: HEP1-6 hepatoma cells in logarithmic growth phase were digested with pancreatin, rinsed with Phosphate Buffered Saline (PBS), resuspended, and the cell suspension concentration was adjusted to 4X 107 cells/ml, inoculated under the right axilla of mice, 100. Mu.l/mouse, i.e., 4X 106 cells/mouse, tumor mice were established, and 8 mice per group were randomly grouped according to tumor volume of mice on the day of administration.
3. Experimental grouping and dosing regimen
Mice were divided into three groups, as listed in table 1, which were:
A. blank control group: the contrast agent is corn oil and is administrated by stomach irrigation according to the weight of 10 mu l/g, and the administration is carried out 2 times per day;
B. icaritin dispersion group: dispersing 350mg of traditional Chinese medicine icaritin in 50ml of corn oil to obtain a corn oil solution of icaritin; the administration is performed by gastric lavage according to the weight of 10 mu l/g, and the administration is performed 2 times a day;
C. the traditional Chinese medicine composition comprises the following components: 350mg of the Chinese medicinal composition obtained in example 1 was dispersed in a volume of 50ml of corn oil, and the obtained oil-soluble Chinese medicinal composition was administered to mice by gavage at a weight of 10. Mu.l/g, 2 times per day.
Table 1 example 2 experimental group
4. Tumor measurement and experimental index
The mice were weighed twice weekly, tumor volumes were measured, and the health status of the mice was observed. The change in body weight of mice was recorded as a function of time of administration. The survival and health conditions of the mice, such as general states of animal activity, feeding, etc. during administration, were also observed. 2 hours after the last dose, mice were euthanized, body weight after tumor removal, and tumor weight was recorded.
Experimental data are represented by x±s.
Tumor volume = 1/2ab 2 Tumor volume T/C (%) value = dosing group volume/control group volume 100%.
RTV: rtv=v relative to tumor volume t /V 0 。
V t I.e. the tumor volume at the end of an experimental period (typically around 30 days).
V 0 Tumor volume at the beginning of the experiment, intermediate time points were calculated, and one RTV was found for each of the experimental and control groups.
TGI: tumor growth inhibition rate. Tgi= [1-RTV (experimental group)/RTV (control group) ].
Tumor Growth Inhibition (TGI) = (control tumor volume-dosing tumor volume)/control tumor volume of 100%.
Tumor regrowth inhibition (TGI) = (control tumor weight-dosing tumor weight)/control tumor weight 100%.
The experimental results are shown in the comprehensive data statistics, tumor volumes, tumor weight distribution maps and related data analysis charts of each experimental group. Statistical analysis: graph.
5. On day 19 of dosing, the volume change of tumors is shown in table 2 below and fig. 1;
table 2 list of volume changes of tumors on day 19 in eight mice.
The icaritin dispersion is a medicament for treating liver cancer in clinical use at present, and the tumor volume list in Table 2 shows that the tumor volume of a mouse taking the icaritin dispersion is smaller than that of a mouse in a blank control group, so that the icaritin dispersion can effectively inhibit the growth of liver cancer, the Chinese medicinal composition disclosed in the embodiment 1 of the invention has a better inhibition effect on tumors, and the tumor volume is far smaller than that of the blank control group and also smaller than that of the icaritin dispersion control group.
6. On day 30 of administration, the effect of icaritin dispersion, control group, and Chinese medicinal composition on tumor weight of 8 mice was recorded. On day 30 of dosing, the weight change of the tumors is shown in table 3 below:
table 3 list of eight mice on day 30 tumor weight changes
The icaritin dispersion liquid is a medicine for treating liver cancer in clinical use at present, and the tumor weight list in table 3 shows that the weight of a mouse taking the icaritin dispersion liquid group is smaller than that of a mouse taking a blank control group, so that the icaritin dispersion liquid can effectively inhibit the growth of liver cancer, the tumor is smaller, the inhibition effect of the Chinese medicinal composition disclosed by the embodiment 1 of the invention on the tumor is better, and the weight of the tumor is far smaller than that of the blank control group and is also smaller than that of the icaritin dispersion liquid control group.
7. Calculating the tumor inhibition rate of icaritin dispersion liquid, control group and Chinese medicinal composition.
TABLE 4 tumor inhibition Rate of icaritin Dispersion, control group and Chinese medicinal composition
The traditional Chinese medicine composition has good liver cancer inhibition rate, and is obviously higher than icaritin.
The traditional Chinese medicine composition for treating liver cancer, the preparation and the preparation method disclosed by the embodiment of the invention have the advantages that the obtained traditional Chinese medicine composition and the preparation have no toxic or side effect and have high tumor inhibition rate on liver cancer.
The technical solutions disclosed in the embodiments of the present invention and the technical details disclosed in the embodiments of the present invention are only exemplary to illustrate the inventive concept of the present invention, and do not constitute a limitation on the technical solutions of the embodiments of the present invention, and all conventional changes, substitutions or combinations of the technical details disclosed in the embodiments of the present invention have the same inventive concept as the present invention, and are within the scope of the claims of the present invention.
Claims (9)
1. The preparation method of the traditional Chinese medicine composition for treating liver cancer is characterized by comprising the following steps:
(1) Taking a set amount of taxus chinensis, haematococcus pluvialis, dried oyster, silybum marianum fruits and rhodiola rosea;
(2) Performing enzymolysis on the yew, extracting a yew enzymolysis product by using ethyl acetate, and purifying and crystallizing the ethyl acetate extraction product to obtain a yew extract;
(3) Crushing haematococcus pluvialis, and extracting the crushed haematococcus pluvialis with deionized water to obtain haematococcus pluvialis extract;
(4) Pulverizing Concha Ostreae, fructus Silybi Mariani and radix Rhodiolae respectively, and extracting with ethanol to obtain Concha Ostreae dry extract, herba Silybi Mariani extract and radix Rhodiolae extract respectively;
(5) Mixing Concha Ostreae dry extract, herba Silybi Mariani fruit extract, radix Rhodiolae extract and Haematococcus pluvialis extract to obtain extract mixture, and using CO 2 Performing supercritical extraction on the obtained extract mixture to obtain a mixed extract of dried oyster, silybum marianum, rhodiola rosea and haematococcus pluvialis;
(6) Mixing the Taxus chinensis extract and the mixed extract according to a mass ratio of 1:3 to obtain the traditional Chinese medicine composition.
2. The method for preparing a Chinese medicinal composition for treating liver cancer according to claim 1, wherein the yew is enzymatically hydrolyzed by cellulase.
3. The method for preparing a Chinese medicinal composition for treating liver cancer according to claim 1, wherein the taxus chinensis extract is further purified, specifically comprising the steps of:
(2-1) mixing the Taxus chinensis extract product with neutral alumina;
(2-2) subjecting the obtained mixture to chromatography by using a silica gel or alumina chromatographic column, eluting with petroleum ether and ethyl acetate to remove fat-soluble impurities, eluting with a mixed solution of alcohol and ethyl acetate, collecting a mixed solution fraction of alcohol and ethyl acetate, and removing alcohol and ethyl acetate under reduced pressure to obtain a primary purified product;
(2-3) using polymethyl acrylate resin column and polystyrene resin column layer as chromatographic column, using dichloromethane and acetone mixed solution as eluent, and sequentially performing chromatography to the primary purified product to obtain pure product of Taxus chinensis extract.
4. The method for preparing a traditional Chinese medicine composition for treating liver cancer according to claim 3, comprising: further crystallizing and refining the pure Chinese yew extract to obtain refined Chinese yew extract.
5. A traditional Chinese medicine composition for treating liver cancer, which is characterized by being obtained by the preparation method of any one of claims 1-4.
6. The composition according to claim 5, wherein the amount of the Chinese yew is 20-30 parts, the amount of haematococcus pluvialis is 15-30 parts, the amount of dried oyster is 15-25 parts, the amount of the silybum marianum fruit is 20-35 parts, and the amount of the rhodiola rosea is 5-15 parts by mass.
7. A traditional Chinese medicine preparation for treating liver cancer, which is characterized by comprising the traditional Chinese medicine composition of claim 5 or 6.
8. The traditional Chinese medicine preparation for treating liver cancer according to claim 7, further comprising pharmaceutically acceptable auxiliary materials.
9. The traditional Chinese medicine preparation for treating liver cancer according to claim 7, wherein the dosage form of the traditional Chinese medicine preparation comprises decoction, pill, paste, powder, dew, pill, medicated wine and lozenge.
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| CN105017182A (en) * | 2015-07-06 | 2015-11-04 | 贵州玉福临科技开发有限公司 | Method for highly efficiently extracting paclitaxel from Chinese yew |
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| CN105017182A (en) * | 2015-07-06 | 2015-11-04 | 贵州玉福临科技开发有限公司 | Method for highly efficiently extracting paclitaxel from Chinese yew |
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