CN113456785A - Traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy, and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy, and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy, which is prepared from astragalus root, pilose asiabell root, mitsubishi, zedoary, leech, centipede, medicinal cyathula root, white peony root, red peony root, oldenlandia diffusa, barbed skullcap herb, scutellaria root and moxibustion licorice. The traditional Chinese medicine composition can also prevent and treat tumor-related thrombosis, reduce the incidence rate of liver cancer of patients with alpha-fetoprotein positive chronic hepatitis, and has obvious synergistic attenuation effects in combination with chemotherapeutic drugs.
Description
Technical Field
The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy.
Background
Cancer is a main disease threatening human health, and the 'killing type' treatment methods such as radiotherapy and chemotherapy of a heavy focus and a light whole body are beneficial to focus control in a short time, but the contribution to improving the overall life cycle of a patient is limited, and the cost such as huge medical cost and reduction of life quality is needed. The tumor is a metabolic disease, metabolic reprogramming is a core characteristic of the tumor, but the metabolic reprogramming of the tumor has complex system characteristics including flexible metabolic plasticity, metabolic coupling, heterozygous metabolic phenotype, temporal-spatial heterogeneity and the like, and the ideal effect cannot be achieved in a modern medical treatment mode only taking 'point and plane' as metabolic intervention targets. The Chinese medicine element takes 'holistic view' and 'dialectical treatment' as essences, and the balanced formula principle of 'monarch, minister, assistant and guide' and 'channel guide' endows the Chinese medicine with the 'natural' advantages of multiple ways, multiple targets and space-time cooperative regulation and control of the complex metabolic system of the human body, and is the main force of the future anti-tumor metabolic treatment.
The clinical application finds that: although the medicine or compound prescription for promoting blood circulation and removing blood stasis has better inhibiting effect on tumor, the risk of inducing tumor metastasis is as high as 30-40%, so that only a few anti-tumor Chinese patent medicines mainly promoting blood circulation and removing blood stasis, such as rhubarb and hibernating pill, are clinically used at present. Although the side effect of promoting cancer metastasis in the process of treating tumors by promoting blood circulation and removing blood stasis is always well regarded in the medical field of China, the mechanism of promoting cancer metastasis while inhibiting tumors by the blood circulation and removing blood stasis medicines is not clarified, the indications of the blood circulation and removing blood stasis treatment of tumors and the early warning markers for promoting cancer metastasis are not clarified, and the 'treatment by syndrome differentiation', 'cautious use' and 'combined promotion of blood circulation and pathogen expelling' and the like become 'escape-type' choices for treating the cancer metastasis of the medicines, so that the clinical application of the blood circulation and removing blood stasis medicines of tumors is greatly limited.
The blood circulation promoting and blood stasis removing is one of three major methods for treating tumors in traditional Chinese medicine, has definite treatment effect on the tumors, is easy to induce tumor metastasis, and is a thousand-year problem in the treatment of the tumors by promoting blood circulation and removing blood stasis. 20% of tumor patients die from tumor-associated thrombi. At present, tumor-related thrombosis is difficult to prevent, anticoagulant methods such as heparin and warfarin are mainly adopted in the aspect of treatment, and patients who are used for a long time have bleeding and are not suitable for long-time use.
Disclosure of Invention
Technical problem to be solved
The invention aims to solve the technical problems of side effects of promoting cancer metastasis of the traditional Chinese medicines for promoting blood circulation, removing blood stasis and resisting tumors, solve the problem of low curative effect of tumor anti-metabolism treatment, prevent and treat tumor-related thrombus, reduce the incidence rate of liver cancer of patients with alpha-fetoprotein positive chronic hepatitis, and have obvious synergistic and toxicity-reducing effects by combining chemotherapy.
(II) technical scheme
In order to solve the problems in the prior art, the invention provides a blood circulation promoting and blood stasis removing traditional Chinese medicine composition for anti-tumor metabolic therapy, which mainly intervenes 8 targets of tumor lipid metabolic reprogramming, inhibits the signal transmission pathway of the tumor metabolic reprogramming PI3K-AKT-mTOR, has obvious inhibition effect on the growth of various tumors (such as gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer, breast cancer and the like), does not promote tumor metastasis after long-term use, can prevent and treat tumor-related thrombus, particularly can be combined with entecavir for application, can ensure that the negative conversion rate of patients with alpha-fetoprotein-positive chronic hepatitis is as high as 95 percent, and plays a role in preventing liver cancer.
The invention aims at providing a traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy, which is prepared from 20-40 parts of astragalus root, 12-20 parts of codonopsis pilosula, 9-12 parts of pedicellus et pericarpium Trapae, 5-12 parts of curcuma zedoary, 3-8 parts of leech, 2-6 parts of centipede, 10-20 parts of medicinal cyathula root, 6-16 parts of white peony root, 10-25 parts of red peony root, 20-30 parts of oldenlandia diffusa, 10-15 parts of barbed skullcap herb, 10-20 parts of scutellaria baicalensis and 6-18 parts of moxibustion licorice.
The invention also aims to provide a preparation method of the traditional Chinese medicine composition, which comprises the following steps:
(1) extracting radix astragali with 4-12 times of 40-80% ethanol under reflux to obtain radix astragali ethanol extractive solution;
(2) volatilizing ethanol from the residue, adding into the rest twelve medicinal materials, adding 4-12 times of water, mixing, heating in hot water in constant temperature water bath, extracting, mixing the extractive solutions, filtering, concentrating the filtered extractive solution under reduced pressure to obtain fluid extract with density of 1.1-1.3, and freeze drying to obtain granule powder.
The invention also aims to provide a traditional Chinese medicine preparation which comprises the traditional Chinese medicine composition and pharmaceutic adjuvants.
The fourth purpose of the invention is to provide the application of the traditional Chinese medicine composition in preparing antitumor drugs, preferably in treating gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
The fifth purpose of the invention is to provide a pharmaceutical composition, which comprises the traditional Chinese medicine composition and a chemotherapeutic drug, wherein the chemotherapeutic drug is preferably 5-fluorouracil.
The invention also provides application of the pharmaceutical composition containing the traditional Chinese medicine composition and a chemotherapeutic drug in preparing an antitumor drug, and the pharmaceutical composition is preferably used for treating gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
The seventh purpose of the invention is to provide a pharmaceutical composition, which comprises the traditional Chinese medicine composition and entecavir.
The invention also provides the application of the medicinal composition containing the traditional Chinese medicine composition and entecavir in preparing anti-liver cancer medicaments.
(III) advantageous effects
The technical scheme of the invention has the following advantages:
(1) the invention provides a traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy based on the rule of tumor metabolic reprogramming, and experimental results show that the traditional Chinese medicine composition has a definite anti-tumor curative effect and can not cause tumor metastasis.
(2) The traditional Chinese medicine composition for anti-tumor metabolic therapy and promoting blood circulation to remove blood stasis has a regulating effect on multiple targets and multiple signal conduction paths of tumor metabolic reprogramming, is high in curative effect and wide in anti-tumor spectrum, has an inhibiting effect on multiple tumors such as gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer and breast cancer, and is not easy to generate drug resistance.
(3) The traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy can improve the peripheral blood hypercoagulable state of a tumor patient and prevent the occurrence of tumor-related thrombus of the tumor patient due to the reduction of the expression of tumor surface tissue factor receptors while playing an anti-tumor role, has a therapeutic effect on the tumor-related thrombus, has small side effect, and can promote blood circulation without causing bleeding.
(4) The traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy, combined with entecavir, can enable the negative conversion rate of a patient with chronic hepatitis with positive alpha fetoprotein to be as high as 95% (the negative conversion rate is only about 30% when the entecavir is used for treating alone), obviously reduces the risk of liver cancer of the patient, and plays a role in preventing liver cancer.
(5) The traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy has obvious synergistic attenuation effects by combining chemotherapy, can effectively inhibit tumor growth and proliferation, prevent tumor invasion and metastasis, improve clinical symptoms of tumor patients, improve the survival quality of cancer patients and prolong the survival period of the cancer patients.
Drawings
FIG. 1 shows the 293T cell inhibitory effect of the Chinese medicinal composition of example 1;
FIG. 2 shows the inhibitory effect of the Chinese medicinal composition of example 1 on HCT-116 cells of colorectal cancer;
FIG. 3 shows the inhibitory effects of the Chinese medicinal composition of example 1 on colorectal cancer SW-480, gastric cancer cell SGC-7901, pancreatic cancer cell BXPC-3 and PANC-1;
FIG. 4 shows the inhibitory effect of the Chinese medicinal composition of example 1 on hepatoma carcinoma cells SMMC-7721-3 and the IC50 concentration thereof;
fig. 5 is a graph of the experimental effect of the Chinese medicinal composition of embodiment 1 on anti-tumor animals, wherein fig. 5A: tumor volume changes in control (PBS fed) and treatment groups; FIG. 5B: graphs of tumor weight change for control (PBS feed) and treatment groups; FIG. 5C: tumor size comparison of control (PBS fed) and treatment groups; FIG. 5D: control group tumor-bearing mice; FIG. 5E: treatment group tumor-bearing mice; in the figure, JC734 is the traditional Chinese medicine composition of example 1;
FIG. 6 is a graph showing the experimental effects of the combination of the single-use and the chemotherapeutic agents of the Chinese medicinal composition of example 1 on the anti-tumor animals; wherein FIG. 6A: a control group, the Chinese medicinal composition treatment group of example 1, the 5-Fu treatment group, and the Chinese medicinal composition combination 5-Fu treatment group of example 1; FIG. 6B: body weight change plots for four groups of mice; FIG. 6C: four sets of tumor size comparison plots; in the figure, JC734 is the traditional Chinese medicine composition of example 1;
fig. 7 shows the effect of the Chinese medicinal composition of example 1 on the major organs of the body in experimental study on anti-tumor cells, wherein JC734 is the Chinese medicinal composition of example 1.
FIG. 8 shows a qPCR experiment to study the effect of the Chinese medicinal composition of example 1 on TFEB, PPAR α, SREBF-1 and mTOR of HCT-116 cells, wherein JC734 is the Chinese medicinal composition of example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
The quality of the traditional Chinese medicine material of the invention is in accordance with each relevant regulation under the corresponding medicinal material item of the first part of the Chinese pharmacopoeia 2007 edition.
Radix astragali is dried root of Astragalus membranaceus Bunge of Astragalus of Leguminosae.
Radix Codonopsis is dried root of Codonopsis pilosula of Campanulaceae.
Radix Paeoniae Rubra is dried root of Ranunculaceae plant Radices paeoniae rubra.
Scutellaria barbata is the whole herb of Scutellaria barbata of Scutellaria of Labiatae.
Mitsubishi is a dried tuber of Sparganium stolonii Erum, Buch. -ham.
The Curcumae rhizoma is dried rhizome of Curcuma zedoaria Rosb of Zingiberaceae.
The Hirudo is dried whole body of Whitm. ania Pigra Whitman of Hirudinidae.
The Scolopendra is dried product of Scolopendra subspinipes mutilans L.Koch of Scolopendra family.
Radix Cyathulae is the dried root of Cyathula officinalis Kuan of Amaranthaceae.
Oldenlandia diffusa is the whole plant of Hedyotis diffusa Willd (Oldenlandia diffusa (Willd.)) of the genus Oldenlandia of the family rubiaceae.
Radix Paeoniae alba is dried root of Paeonia lactiflora pall.
The Scutellariae radix is dried root tuber of perennial herb of Scutellaria Baicalensis Georgi of Scutellaria of Labiatae.
The Glycyrrhrizae radix is dried root and rhizome of Glycyrrhiza uralensis Fisch, Glycyrrhiza inflata Bat, or Glycyrrhiza glabra L.
5-Fu is 5-fluorouracil.
The traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy is prepared from 20-40 parts of astragalus membranaceus, 12-20 parts of codonopsis pilosula, 9-12 parts of pedicellus et pericarpium Trapae, 5-12 parts of curcuma zedoary, 3-8 parts of leech, 2-6 parts of centipede, 10-20 parts of radix cyathulae, 6-16 parts of radix paeoniae alba, 10-25 parts of radix paeoniae rubra, 20-30 parts of oldenlandia diffusa, 10-15 parts of herba scutellariae barbatae, 10-20 parts of scutellaria baicalensis and 6-18 parts of moxibustion liquorice.
The traditional Chinese medicine composition is preferably prepared from 30 parts of astragalus membranaceus, 16 parts of codonopsis pilosula, 9 parts of mitsubishi, 9 parts of curcuma zedoary, 5 parts of leech, 4 parts of centipede, 20 parts of medicinal cyathula root, 12 parts of white paeony root, 20g of red paeony root, 20 parts of spreading hedyotis herb, 10 parts of barbed skullcap herb, 16 parts of scutellaria baicalensis and 12 parts of moxibustion liquorice.
The preparation method of the traditional Chinese medicine composition comprises the following steps:
(1) extracting radix astragali with 4-12 times of 40-80% ethanol under reflux to obtain radix astragali ethanol extractive solution;
(2) volatilizing ethanol from the residue, adding into the rest twelve medicinal materials, adding 4-12 times of water, mixing, heating in hot water in constant temperature water bath, extracting, mixing the extractive solutions, filtering, concentrating the filtered extractive solution under reduced pressure to obtain fluid extract with density of 1.1-1.3, and freeze drying to obtain granule powder.
The preparation method of the traditional Chinese medicine composition preferably comprises the following steps:
(1) extracting radix astragali with 4-12 times of 40-80% ethanol under reflux for 1-4 times (each time for 1 hr) to obtain radix astragali ethanol extractive solution;
(2) volatilizing ethanol from the residue, adding into the rest twelve medicinal materials, adding 4-12 times of water, mixing, heating in 80 deg.C hot water with thermostatic water bath for 3 times, each time for 1 hr, mixing the 3 times of Chinese medicinal extractive solutions, filtering, concentrating under reduced pressure until the fluid density is 1.2, and freeze drying to obtain granule powder.
The invention also provides a traditional Chinese medicine preparation which comprises the traditional Chinese medicine composition and pharmaceutic adjuvants.
The preparation method of the traditional Chinese medicine preparation comprises the following steps: adding medicinal adjuvants into the extract powder, and making into granule, tablet, capsule, effervescent tablet or liquid preparation.
The pharmaceutic adjuvants used in the invention comprise dextrin, lactose, sodium bicarbonate, citric acid, sodium hydroxymethyl starch, aerosil, starch, soluble starch and magnesium stearate, which are all pharmaceutic specification adjuvants.
The preparation method of the traditional Chinese medicine preparation comprises the following steps: adding 4-16Kg of dextrin into the obtained extract powder, uniformly mixing, performing dry granulation, finishing granules, and putting the granules into a composite aluminum-plastic bag after passing inspection to obtain a finished product.
The preparation method of the traditional Chinese medicine tablet comprises the following steps: adding sodium hydroxymethyl starch, silica gel micropowder, soluble starch, lactose, and magnesium stearate, mixing, and making into granule or tablet with 95% ethanol.
The preparation method of the traditional Chinese medicine capsule comprises the following steps: adding appropriate amount of silica gel micropowder, mixing, and making into capsule.
The preparation method of the traditional Chinese medicine tablet comprises the following steps: adding citric acid and starch into part of the fine powder, granulating, adding sodium bicarbonate and starch into the rest fine powder, granulating, adding magnesium stearate into the two granules, mixing, and tabletting.
The liquid preparation of the present invention can be prepared according to a conventional method.
The invention also provides a pharmaceutical composition, which comprises the traditional Chinese medicine composition and a chemotherapeutic drug. The chemotherapeutic agent is preferably 5-fluorouracil.
The invention also provides application of the traditional Chinese medicine composition in preparing an anti-tumor medicine, and the traditional Chinese medicine composition is preferably used for treating gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
The invention also provides application of a pharmaceutical composition containing the traditional Chinese medicine composition and a chemotherapeutic drug in preparing an antitumor drug, and the pharmaceutical composition is preferably used for treating gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
The invention also provides a pharmaceutical composition, which comprises the traditional Chinese medicine composition and entecavir.
The invention also provides application of a pharmaceutical composition containing the traditional Chinese medicine composition and entecavir in preparation of anti-liver cancer drugs.
In order to better understand the present invention, the following further explains or illustrates the present invention by specific examples, but these examples should not be construed as limiting the scope of the present invention.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
In order to better understand the present invention, the following further explains or illustrates the present invention by specific examples, but these examples should not be construed as limiting the scope of the present invention.
Example 1: preparation of Chinese medicinal composition
The bulk drugs comprise: 30g of astragalus, 16g of codonopsis pilosula, 9 g of pedicellus et pericarpium Trapae, 9 g of curcuma zedoary, 5g of leech, 4 g of centipede, 20g of medicinal cyathula root, 12 g of white paeony root, 20g of red paeony root, 20g of spreading hedyotis herb, 10g of barbed skullcap herb, 16g of baical skullcap root and 12 g of moxibustion licorice.
Extracting radix astragali with 4 times of 40% ethanol under reflux for 1 time, each time for 1 hr to obtain radix astragali ethanol extractive solution; evaporating ethanol from the residue, adding into the rest twelve medicinal materials, adding 4 times of water, heating in 80 deg.C hot water with constant temperature water bath for 3 times, each time for 1 hr, mixing the 3 times of Chinese medicinal extractive solutions, filtering, concentrating under reduced pressure until the fluid density is 1.2, and freeze drying for 48 hr to obtain granule.
Example 2: preparation of Chinese medicinal composition
The bulk drugs comprise: 20g of astragalus, 12 g of codonopsis pilosula, 10g of pedicellus et pericarpium Trapae, 5g of curcuma zedoary, 3 g of leech, 2 g of centipede, 10g of medicinal cyathula root, 6g of white paeony root, 10g of red paeony root, 20g of spreading hedyotis herb, 10g of barbed skullcap herb, 10g of baical skullcap root and 6g of moxibustion licorice.
The preparation method is the same as example 1.
Example 3: preparation of Chinese medicinal composition
The bulk drugs comprise: 40 g of astragalus root, 20g of codonopsis pilosula, 12 g of pedicellus et pericarpium Trapae, 12 g of curcuma zedoary, 6g of leech, 6g of centipede, 20g of medicinal cyathula root, 16g of white peony root, 25 g of red peony root, 30g of spreading hedyotis herb, 15g of barbed skullcap herb, 20g of scutellaria root and 12 g of moxibustion licorice.
The preparation method is the same as example 1.
Example 4: preparation of Chinese medicinal composition
The bulk drugs comprise: 30g of astragalus, 20g of codonopsis pilosula, 12 g of pedicellus et pericarpium Trapae, 10g of curcuma zedoary, 8 g of leech, 6g of centipede, 20g of medicinal cyathula root, 10g of white paeony root, 16g of red paeony root, 20g of spreading hedyotis herb, 10g of barbed skullcap herb, 20g of baical skullcap root and 18 g of moxibustion licorice.
The preparation method is the same as example 1.
Example 5: EXAMPLE 1 Experimental study of antitumor cells of the Chinese medicinal composition
(1) Example 1 study of in vitro inhibitory Effect of the Chinese medicinal composition on human 293T cells and human colorectal cancer cells
(2) Example 1 study of in vitro inhibitory effect of the Chinese medicinal composition on hepatoma cell line SMMC-7721 and determination of IC50
Culturing the liver cancer cell strain SMMC-7721 in vitro, adding the traditional Chinese medicine composition of the embodiment 1 with different concentrations for co-culture when the growth is vigorous, and observing the inhibition rate of the traditional Chinese medicine composition of the embodiment 1 with different concentrations on liver cancer cells under the co-culture for 24h, 48h and 72 h. As a result, the inhibition rate of 1mg/ml of the traditional Chinese medicine composition of the embodiment 1 on liver cancer cells is 98% (48h), the inhibition rate of 0.8mg/ml of the traditional Chinese medicine composition of the embodiment 1 on liver cancer cells also reaches 78% (24h) and 82% (48h), and the IC of the traditional Chinese medicine composition of the embodiment 1 on liver cancer cells is500.6232, it is suggested that the Chinese medicinal composition of example 1 has a very significant inhibitory effect on liver cancer.
(3) EXAMPLE 1 in vitro cell assay study of the Chinese medicinal composition for inhibiting different tumors
Colorectal cancer SW480, gastric cancer SGC-7901, pancreatic cancer BXPC-3 and PANC-1 cells, breast cancer cell strains MDA-MB-231, human lung adenocarcinoma cell strains A549, human cervical cancer cell strains AKG-IIIA, ovarian cancer cell strains SKOV-3 and liver cancer cell strains SMMC-7721 are cultured in vitro, the traditional Chinese medicine composition in the embodiment 1 with different concentrations is added for co-culture when the growth is vigorous, and the inhibition rate of the traditional Chinese medicine composition in the embodiment 1 with different concentrations on the cells under the condition of co-culture for 48h is observed. As a result, the 2mg/ml inhibition ratio of the traditional Chinese medicine composition of example 1 to each tumor cell is respectively 64.5% (colorectal cancer), 73.7% (gastric cancer), 67.4% (pancreatic cancer BXPC-3), 45.2% (pancreatic cancer PANC-1), 70.7% (breast cancer), 78.9% (lung cancer), 65.6% (cervical cancer), 54.6% (ovarian cancer) and 98% (liver cancer), and the effect on liver cancer is most obvious.
Example 6: example 2-4 Experimental study of antitumor cells of the Chinese medicinal composition
And (3) carrying out in-vitro culture on the colorectal cancer SW480, adding 200 microliters of different prescription extracts with the concentration of 2mg/ml for co-culture when the colorectal cancer SW grows vigorously, and observing the inhibition rate of the drugs with different concentrations on cells under the co-culture for 48 h. As a result, the inhibition rate of the standard prescription on each tumor cell is respectively as follows: 45.6% for example 2, 68.9% for example 3 and 56.7% for example 4.
Example 7: EXAMPLE 1 antitumor animal Experimental study of the Chinese medicinal composition
(1) Experimental study of the Chinese medicinal composition of example 1 on colorectal cancer treatment in animals: 15 colorectal cancer-bearing mice were randomized into 2 groups after stratification by tumor volume: the control group, 7 mice (fed PBS), and the treatment group (fed 5g/kg of the Chinese medicinal composition of example 1), were sacrificed 18 days after continuous feeding. Animal experiment results show that the traditional Chinese medicine composition in the example 1 can obviously relieve the body weight loss caused by experiments, and the inhibition rate of the traditional Chinese medicine composition in the example 1 to colorectal cancer is 43% after the traditional Chinese medicine composition is fed for 1 time every day at the speed of 5 g/kg.
(2) Experimental study of animal having colorectal cancer treated with the Chinese medicinal composition of example 1 alone or in combination with 5-Fu: 16 colorectal cancer-bearing mice were randomized into 4 groups after stratification by tumor volume: a control group, a 5-Fu treated group, the Chinese medicinal composition treated group of example 1, and a 5-Fu combined Chinese medicinal composition treated group of example 1, each group containing 4 animals. At the beginning of the experiment, the difference between the groups in terms of body weight, mean tumor volume, etc. was not significant. Adding 7.5mg/kg of physiological saline into 5-Fu of the treatment group, diluting, and performing abdominal injection chemotherapy 1 time per week; the chemotherapy method of the treatment group of 5-Fu combined with the traditional Chinese medicine composition of the embodiment 1 is the same as that of the chemotherapy group of the traditional Chinese medicine composition of the embodiment 1, and meanwhile, 200mg of the traditional Chinese medicine composition of the embodiment 1 is diluted by normal saline and is perfused into the stomach, 1 time a day and 4ml of the traditional Chinese medicine composition each time; the treatment group of the traditional Chinese medicine composition of the embodiment 1 is prepared by diluting the traditional Chinese medicine composition of the embodiment 1 with normal saline and irrigating the stomach 1 time a day, 4ml each time; the control group was intragastrically administered with 4ml of physiological saline daily. Tumor growth up to 100m3The experiment started treatment by size, and 16 days later, the mice were sacrificed. Animal experiment results show that the traditional Chinese medicine compound preparation can obviously relieve the body weight loss caused by chemotherapy; the inhibition rate of the traditional Chinese medicine composition for treating the colorectal cancer in the embodiment 1 is 57%, the inhibition rate of 5-Fu for treating the colorectal cancer is 73%, and the inhibition rate of 5-Fu combined with the traditional Chinese medicine composition in the embodiment 1 for treating the colorectal cancer is 94%.
Example 8: clinical test of the Chinese medicinal composition of example 1
1. Case selection
120 gastrointestinal tumor patients in Beijing century bed hospital, a university of capital medicine between 1990 and 2004 between 9 and 11 months, have pathology proved to be primary gastrointestinal malignant tumor stage I-IV.
Inclusion criteria were: (1) patients with gastrointestinal tumors confirmed clinically and pathologically have an expected life cycle of greater than 3 months; (2) neoadjuvant chemotherapy, transformation therapy or palliative therapy is planned using the mflfox 6 regimen; (3) the age is more than 16 years old and less than 80 years old; (4) the traditional Chinese medicine tumor pathogenesis identifies tumor patients with deficiency, stagnation, stasis and toxicity; (5) patients with no bleeding tendency; (6) can evaluate the focus or tumor marker; (7) patients who completed at least 4-8 cycles of treatment were included in the efficacy assessment.
Exclusion criteria: (1) drug allergy to the traditional Chinese medicine composition of example 1; (2) can not be orally taken with the traditional Chinese medicine decoction with the obstruction of digestive tract; (3) patients taking part in other drug trials or taking other traditional Chinese medicines orally; (4) with organ active bleeding; (5) patients receiving chemotherapy with other regimens; (6) combined with lipid metabolism diseases such as hyperlipoproteinemia, lipid storage diseases, ketoacidosis, etc.
The traditional Chinese medicine syndrome identification method comprises the following steps: the traditional Chinese medicine tumor pathogenesis identification is divided into 6 pathogenesis types of yang deficiency, yin deficiency, phlegm dampness, heat toxin, blood stasis and qi stagnation, all questions in the 'traditional Chinese medicine tumor pathogenesis identification scale' are answered according to experience and feeling of nearly 1 year, each question is graded according to 5 grades (1 grade: none; 2 grade: one point; 3 grade: sometimes or a few; 4 grade: often; 5 grade: very obvious), and each pathogenesis type identification score is not less than 10 to diagnose the pathogenesis type to be positive (Table 1).
TABLE 1 Chinese medicine tumor pathogenesis differentiation Scale (6 points of the law)
Note that: 1 minute: none; and 2, dividing: there is a point; and 3, dividing: sometimes or some; and 4, dividing: frequently; and 5, dividing: is very obvious.
2. Grouping
60 gastrointestinal malignant tumor chemotherapy patients (mFoLFoX6 scheme) treated by the traditional Chinese medicine composition of the embodiment 1 are taken as a treatment group, and 60 gastrointestinal tumor chemotherapy patients are taken as a control group. The age, pathological type, clinical stage and syndrome differentiation type of the two groups of patients are tested by composition ratio, and the difference has no statistical significance and is comparable (as shown in Table 2).
TABLE 2120 basic condition table for gastrointestinal tumor patients
The treatment scheme comprises the following steps: the Chinese medicinal composition of example 1 is diluted with warm water to 50ml, and administered with empty stomach for 1 day and 2 times, 10 days is 1 treatment cycle, and the second treatment cycle is started after 1 week of rest. Control group: oxaliplatin 100mg/m2Intravenous drip for 2h, day 1; calcium folinate 400mg/m2Intravenous drip for 2h, day 1; fluorouracil 400mg/m2Bolus intravenous injection, day 1; fluorouracil 2400mg/m2Intravenous drip for 46 h; repeat 1 time every 2 weeks; treatment groups: the Chinese medicinal composition of example 1 is orally administered 1 day before chemotherapy, and is continuously administered 6 days during and after chemotherapy, wherein 1 treatment cycle is defined as 10 days.
Observation indicator and detection method
(1) And (3) evaluating the curative effect of the tumor: 120 patients with gastrointestinal tumors completed an average of 5.7 cycles of treatment. 12 patients reached CR, mainly with colorectal cancer with neoadjuvant chemotherapy, of which 3 in the control group (1 in each of complete clinical remission and complete pathological remission), 3 in the treatment group with complete pathological remission and 6 in the clinical remission; 35 patients with PR, 14 control groups and 21 treatment groups; 40 patients with SD, 17 controls, 23 treatments; 33 PD patients, of which 26 were in the control group and 7 were in the treatment group (table 3). Clinical benefit rate control group: the treatment group equals 56.7:88.3(p < 0.01).
TABLE 3 comparison of the efficacy of the control and treatment groups
Note that: CBR (Clinical benefit rate) ═ CR + PR + SD/total cases.
(2) Effect on tumor markers: tumor marker detection is carried out before chemotherapy of a patient in each cycle, and the items comprise: alpha-fetoprotein, carbohydrate antigen-CA 724, neuron-specific enolase, cytokeratin 19 fragment, carbohydrate antigen-CA 199, carbohydrate antigen-CA 125, carbohydrate antigen-CA 153, carbohydrate antigen-CA 50, total prostate specific antigen, free prostate specific antigen, carcinoembryonic antigen, squamous cell carcinoma-associated antigen, and the like. The study was conducted mainly on digestive tract tumors, and representative digestive tract tumor markers, cancer blank antigen, carbohydrate antigen-CA 724 and carbohydrate antigen-CA 199, were selected for comparison (Table 4). The cancer germ antigen, CA724 and CA199 before treatment of the control group are respectively 86.6 +/-17.8, 209.0 +/-32.1 and 705.4 +/-89.7, the antigen content is respectively reduced to 50.3 +/-6.1, 134.2 +/-26.0 and 354.6 +/-76.2 after treatment, and the comparison before and after treatment has significant difference (p is less than 0.001); the cancer blank antigen, CA724 and CA199 before treatment in the treatment group are 84.4 +/-15.2, 231.4 +/-32.8 and 749.5 +/-94.4 respectively, the reduction after treatment is 29.8 +/-7.5, 66.6 +/-16.9 and 167.5 +/-35.3, the comparison before and after treatment is obviously different (p is less than 0.01), the reduction ratio of the control group to the treatment group is also obviously different (p is less than 0.01), and the reduction of tumor markers in the treatment group is more obvious.
TABLE 4 Effect of control and treatment groups on markers
(3) Effects on the coagulation system: four indexes of D-Dime, FDP, PT-INR and APTT are selected for comparison, and the patient is detected before chemotherapy in each period. D-Dime, FDP, PT-INR and APTT of a control group are respectively 401.3 +/-96.5, 11.5 +/-4.4, 31.3 +/-4.0 and 14.0 +/-0.8 before treatment, and are respectively 387.4 +/-36.9, 13.4 +/-6.3, 33.3 +/-8.2 and 13.7 +/-1.2 after treatment, and the comparison of the four indexes before and after treatment has no significant difference; in the treatment groups, the D-Dime, FDP, PT-INR and APTT before treatment are 387.4 +/-33.2, 11.5 +/-4.4, 28.0 +/-8.8 and 15.2 +/-2.0 respectively, and after treatment are 256.6 +/-44.8, 6.2 +/-1.7, 26.5 +/-2.6 and 16.0 +/-4.5 respectively, the D-Dime and FDP (p is less than 0.01) of abnormal coagulation indexes can be obviously reduced, but the D-Dime and FDP have no obvious influence on bleeding indexes (APTT and PT-INR) (p is more than 0.05), namely, the traditional Chinese medicine composition in the example 1 has the effect of activating blood on the high coagulation state of tumors, but does not cause bleeding risk (Table 5).
TABLE 5 mFOLFOX6 regimen chemotherapeutic group mFOLFOX6 in combination with JC724 treatment group has an effect on clotting function
Note: FDP ═ fibrin degradation products; D-Dimer (plasma D-Dimer); APTT ═ activated partial prothrombin time; PT-INR is an international normalized ratio. *: p < 0.05; **: p <0.01.
(4) Regulation of lipid metabolism: the Chinese medicinal composition of example 1 has a regulating effect on lipid metabolism. There were no significant changes in Total Cholesterol (TC), Triglycerides (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein AI (apoA-I), and apolipoprotein B (apoB) in the peripheral blood lipid metabolism of the control group (Table 6). TC, HDL-C, LDL-C, and apoB were significantly elevated after treatment in the treatment group (p <0.001), but TG and apoA-I had no significant effect (p > 0.05).
TABLE 6 Effect of the Chinese medicinal composition of example 1 on the lipid metabolism of a patient
*：P<0.05；**：P<0.01.
(5) Therapeutic effects of thrombi, cancer emboli and new metastases: the 7 cases of the thrombus in the simple chemotherapy group are all lower limb intramuscular vein thrombus and 2 cases of mesenteric vein cancer thrombus. The intramuscular venous thrombosis has no therapeutic pointer of antithrombotic drugs, only dynamic reexamination and observation are carried out, and the thrombosis persists but progresses in the experimental process. About 3X 2cm for 1 case of mesenteric vein cancer embolus, 2X 1.8cm for cancer embolus after 6 cycles of chemotherapy with mFOLFOX6 regimen, and 1 case of mesenteric vein cancer embolus patients died due to ileus. In the simple chemotherapy group, 60 patients have new lesions in 7 cases during treatment period (4 cases of liver metastasis, 1 case of lung metastasis, and 1 case of liver and lung metastasis). 9 cases of the combined medicine group lower limb intramuscular venous thrombosis and 1 case of the portal vein combined superior mesenteric vein cancer embolism. All of the 9 cases of intramuscular venous thrombosis were cured during the combination treatment, with an average treatment period of 3.6 cycles, without the use of antithrombotic agents. The patient with the portal vein and the superior mesenteric vein cancer embolus is treated for 5 cycles and then is checked again by the color Doppler to find that the cancer embolus disappears. No new metastatic lesions were found during the course of the 60 combination therapy patient trials.
The statistical method comprises the following steps: data are expressed as x + -s, and statistical analysis was performed using the One-way ANOVA procedure of SPSS 20.0, with P <0.05 being statistically significant.
The results show that: the traditional Chinese medicine compound preparation is an effective treatment method for treating gastrointestinal tumors, does not promote tumor diffusion, does not cause bleeding, and can play an anti-tumor role by regulating lipid metabolism.
Example 9: clinical test of the Chinese medicinal composition of example 1
1. Case selection
The prospective and random multicenter clinical trial study was included in 68 patients with AFP-positive CHB from Beijing century bed Hospital, the second medical center of the general Hospital of people Release force in China, and the eighth Hospital in Guangzhou, between 2016 (7) and 2020 (7), and all patients received examinations such as blood routine, HBsAg, HBeAg, HBV-DNA, AFP, biochemical complete set, blood coagulation function, abdominal ultrasound, computed tomography and/or magnetic resonance imaging before they were enrolled. Exclusion criteria: infection with human immunodeficiency virus, hepatitis C virus or hepatitis D virus; there has been a history of antiviral drug treatment over the last 3 years; or a proven HCC. 62 patients were randomly divided into a control group and a test group, and the control group was treated with entecavir (0.5 mg, oral administration, once a day, manufactured by Shandong Lu anti-drug Co., Ltd.) for 12 months; test group the Chinese medicinal composition of example 1 was treated with entecavir, and prepared into powder, 1g each time, taken with water 2 times a day, and after 2 weeks of continuous administration, the powder was left for 1 week for 12 months, and the entecavir treatment method was the same as that of the control group. The tested patients are subjected to symptomatic support treatment such as liver protection and diuresis by conventional administration for complicating the decompensation of the cirrhosis.
Clinical baseline data of patients including age, sex, cirrhosis Child stage and liver color Doppler are recorded at the beginning of treatment, laboratory data include blood routine, HBsAg, HBeAg, HBV-DNA, AFP, blood coagulation function and biochemical complete set (including liver and kidney function, electrolyte, blood sugar, blood fat and the like), all patients review the items every 2 months and follow-up visit are carried out during treatment, and finally the follow-up visit time is 7, 7 and 31 days in 2020. The AFP level of serum is determined by chemiluminescence immunoassay (Roche diagnosis), and AFP positivity is defined as AFP positivity of more than or equal to 7ng/mL in initial diagnosis, because the AFP level of serum is quantitatively determined by a Roche AFP kit, and the 95% confidence interval is 7 ng/mL. During the trial, if patients were found: firstly, new nodules with the diameter of more than 2cm appear in the liver or original nodules grow rapidly; ② newly appearing 3 or more cirrhosis nodules with the diameter less than 2 cm; ③ infiltrative HCC cannot be excluded; fourthly, if the AFP level is gradually increased after more than 2 times of dynamic detection, abdominal computed tomography or magnetic resonance imaging examination is added.
All statistical analyses were performed using SPSS 20.0 software, with continuous variables expressed as mean ± standard deviation, and data analysis using t-test and chi-square test, with P values less than 0.05 considered significant differences and P values less than 0.01 considered very significant differences.
2. Results
2.1 patient Baseline characteristics
65 patients met the queue entry standard and entered the test, 2 patients received other traditional Chinese medicine treatment automatic termination test during the test period, 1 patient lost visit, and 62 patients (30 patients in the control group and 32 patients in the test group) completed 12 months of treatment and visit. Table 1 summarizes the baseline characteristics of clinical and laboratory examinations of 62 patients, and when they were enrolled, the baseline information, AFP, liver function, virology, blood coagulation function, blood lipid, etc. of both groups of patients were not significantly different (p values were all greater than 0.05), and the study was comparable (see table 7). The average age of these patients was 42.4 ± 14 years, the male was 56.5%, and 23 cases of cirrhosis (37.1%).
Table 7 baseline characteristics of patients enrolled
2.2 AFP, virology and liver function reactions
After 12 months of treatment, the AFP response rate of 32 patients in a test group reaches 100%, 30 patients (93.8%) recover AFP normally, the average recovery time is 153.2 +/-123 days, in addition, the AFP of 2 patients before treatment is 163.4 ng/ml and 815.2ng/ml, after 12 months of treatment, the AFP is respectively reduced to 42.3 ng/ml and 117.5ng/ml, and then the treatment of the traditional Chinese medicine composition of the embodiment 1 and the entecavir is continuously adopted, so that the AFP response rate is respectively recovered to normal in 14 months and 17 months of treatment; in 30 patients in the control group 24 patients had a drop in AFP after 12 months of treatment and a response rate of 80.0% (24/30), wherein 16 patients (53.3%) had a return to normal AFP with an average recovery time of 206.6. + -. 117 days, 8 patients had a drop in AFP but not to normal levels after 12 months of treatment, 6 patients had an overall tendency to increase in AFP during treatment, and the control group had very significant differences in AFP response rate, AFP recovery rate and average recovery time (p values were all less than 0.01) compared to the test group. The virological response rates of the test group and the control group at 12 months are respectively 100% (32/32) and 90.3% (28/30), and the two groups have no significant difference (p is more than 0.05); HBV-DNA before and after treatment of the control group is respectively 5.28 +/-1.5 ㏒ IU/mL and 4.1 +/-1.2 ㏒ IU/mL, HBV-DNA before and after treatment of the test group is respectively 5.07 +/-2.1 ㏒ IU/mL and 2.44 +/-1.0 ㏒ IU/mL, and the HBV-DNA inhibition effect of the test group is obviously better than that of the control group (P < 0.01). The ALT response rate of the liver function of the control group and the test group is 100 percent (32/32) and 93.3 percent (28/30), respectively, and compared with the two groups, the ALT response rate of the control group and the test group has no significant difference (P >0.05), but the ALT improvement degree of the test group is better than that of the control group (P < 0.05).
TABLE 8 results of liver function, HBV-DNA, AFP changes
2.3 HCC onset
During the 12-month treatment period, 32 patients in the experimental group did not develop HCC; of the 6 control patients with CHB with a sustained increase in AFP, 3 (10%) had converted to HCC over a 12-month treatment period: 1 example of CHB patients treated by AFP 106.4ng/ml at the beginning, AFP slowly rises during treatment, AFP rises to 415.8ng/ml after 8 months of treatment, CT examination finds that two nodules with the size of 3cm appear on the right lobe of the liver, puncture examination proves that the liver is HCC, after liver protection treatment, surgical resection is performed, postoperative recovery is smooth, and after 12 months of treatment, the AFP slowly falls to 128.8ng/ml after entecavir is continuously administered; when 1 CHB patient is grouped, 427.6ng/ml of AFP is obtained, the AFP is minimally reduced to 310.4ng/ml in the first 8 months of treatment and recheck, but the AFP is detected to be 502.0ng/ml after 10 months of treatment, the enhanced CT indicates that the left lobe of the liver is a 2 cm-sized nodule, the condition is stable after the radio frequency ablation treatment of HCC (liver cancer) lines through the puncture biopsy, the AFP is reduced after the entecavir treatment is continued, and the AFP is normal in the second 12 months of recheck. (ii) a When another CHB patient is grouped, AFP is 408.4ng/ml, AFP is reduced to 47.6ng/ml in a recheck in the 4 th month after treatment, but after 10 months of treatment, AFP is increased to 328.2ng/ml, 1 node with the size of 2cm is newly added in the right lobe of the liver in a CT check, the condition is stable after HCC line radiofrequency ablation treatment as proved by CT enhancement and puncture check, AFP is reduced after entecavir treatment is continued, and AFP is reduced by 24.8 ng/ml in a recheck in the 12 th month.
2.4 Effect on blood lipids and coagulation function
When patients are treated, PT of 18 patients in a treatment group and a control group is prolonged by more than 4s compared with a normal value, D-Dime and PT of the control group are not significantly different before and after treatment, D-Dime and PT of a test group are respectively 1.0 +/-0.84 mg/L and 17.6 +/-11.2 s before and after treatment, and D-Dime and PT of a test group are respectively 0.67 +/-0.39 mg/L and 16.5 +/-9.0 s after treatment, the traditional Chinese medicine composition in the embodiment 1 can obviously reduce blood coagulation index D-Dime (p is less than 0.01) which is abnormally increased by CHB patients, but has no obvious influence on bleeding index PT (p is more than 0.05), namely, the traditional Chinese medicine composition in the embodiment 1 of the blood activating and stasis removing medicine has the effect of activating blood circulation on hypercoagulable states of CHB patients, but does not cause bleeding risks (Table 3). The traditional Chinese medicine composition in example 1 has certain influence on blood lipid metabolism, no significant difference exists before and after treatment of the control group of total peripheral blood cholesterol (TC), Triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) (p is greater than 0.05), and TC, TG and LDL-C are obviously increased after treatment of the test group (p is less than 0.001), but HDL-C has no obvious influence (p is greater than 0.05).
TABLE 9 Effect on blood lipids and coagulation function (n ═ 31)
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A traditional Chinese medicine composition for promoting blood circulation and removing blood stasis for anti-tumor metabolic therapy is characterized by being prepared from 20-40 parts of astragalus membranaceus, 12-20 parts of codonopsis pilosula, 9-12 parts of pedicellus et pericarpium Trapae, 5-12 parts of curcuma zedoary, 3-8 parts of leech, 2-6 parts of centipede, 10-20 parts of radix cyathulae, 6-16 parts of radix paeoniae alba, 10-25 parts of radix paeoniae rubra, 20-30 parts of oldenlandia diffusa, 10-15 parts of sculellaria barbata, 10-20 parts of scutellaria baicalensis and 6-18 parts of radix glycyrrhizae preparata.
2. The traditional Chinese medicine composition according to claim 1, which is prepared from 30 parts of astragalus membranaceus, 16 parts of codonopsis pilosula, 9 parts of pedicellus et pericarpium Trapae, 9 parts of curcuma zedoary, 5 parts of leech, 4 parts of centipede, 20 parts of medicinal cyathula root, 12 parts of radix paeoniae alba, 20g of red paeony root, 20 parts of herba Hedyotidis Diffusae, 10 parts of herba Scutellariae Barbatae, 16 parts of scutellaria baicalensis and 12 parts of radix glycyrrhizae preparata.
3. A method for preparing the Chinese medicinal composition of claim 1, comprising the steps of:
(1) extracting radix astragali with 4-12 times of 40-80% ethanol under reflux to obtain radix astragali ethanol extractive solution;
(2) volatilizing ethanol from the residue, adding into the rest twelve medicinal materials, adding 4-12 times of water, mixing, heating in hot water in constant temperature water bath, extracting, mixing the extractive solutions, filtering, concentrating the filtered extractive solution under reduced pressure to obtain fluid extract with density of 1.1-1.3, and freeze drying to obtain granule powder.
4. A Chinese medicinal preparation comprising the Chinese medicinal composition of claim 1 or 2 and a pharmaceutic adjuvant.
5. Use of a Chinese medicinal composition according to claim 1 for the preparation of an antitumor medicament, preferably for the treatment of gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
6. A pharmaceutical composition comprising a Chinese medicinal composition according to claim 1 or 2 and a chemotherapeutic agent, preferably 5-fluorouracil.
7. Use of a pharmaceutical composition comprising a Chinese medicinal composition according to claim 1 or 2 and a chemotherapeutic agent for the preparation of an anti-tumor medicament, preferably for the treatment of gastric cancer, intestinal cancer, liver cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer or breast cancer.
8. The use according to claim 7, wherein the chemotherapeutic agent is 5-fluorouracil.
9. A pharmaceutical composition comprising the Chinese medicinal composition of claim 1 or 2 and entecavir.
10. Use of a pharmaceutical composition comprising a Chinese medicinal composition according to claim 1 or 2 and entecavir in the preparation of a medicament for treating liver cancer.
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| CN1101559A (en) * | 1993-10-14 | 1995-04-19 | 刘彦彬 | Traditional Chinese medicine oral administration liquid for anti-cancer and its preparing method |
| CN106177792A (en) * | 2016-08-29 | 2016-12-07 | 王建斌 | A kind of Chinese medicine composition treating tumor and preparation method thereof |
| CN107970299A (en) * | 2017-11-16 | 2018-05-01 | 饶本强 | A kind of clearing heat and detoxicating anti-tumor compound preparation |
| CN109331146A (en) * | 2018-11-23 | 2019-02-15 | 厦门市中医院 | A kind of herbal mixture for treating liver cancer |
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| CN106177792A (en) * | 2016-08-29 | 2016-12-07 | 王建斌 | A kind of Chinese medicine composition treating tumor and preparation method thereof |
| CN107970299A (en) * | 2017-11-16 | 2018-05-01 | 饶本强 | A kind of clearing heat and detoxicating anti-tumor compound preparation |
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