CN106728908A - It is a kind of to treat compound Chinese medicinal preparation of colorectal cancer and preparation method thereof - Google Patents
It is a kind of to treat compound Chinese medicinal preparation of colorectal cancer and preparation method thereof Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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Abstract
The present invention relates to field of medicaments, disclose it is a kind of treat compound Chinese medicinal preparation of colorectal cancer and preparation method thereof, the compound Chinese medicinal preparation is made up of the bulk drug that following weight parts are matched:Glutinous rehmannia:50~400 parts, the fruit of glossy privet:45~300 parts, saline cistanche:30~240 parts, mock-strawberry:50~400 parts, edible tulip:75~420 parts, fiveleaf akebia fruit:15~180 parts.In embodiments of the present invention, above-mentioned each bulk drug produces the effect of Synergistic, can effectively treat colorectal cancer, particularly obvious to Advanced Colorectal Cancer or Postoperative carcinoma speen-kidney dificiency card patient outcomes.
Description
Technical field
It is more particularly to a kind of to treat compound Chinese medicinal preparation of colorectal cancer and preparation method thereof the present invention relates to field of medicaments.
Background technology
Colorectal cancer is one of clinically common gastrointestinal cancer, and the incidence of disease occupy the 3rd in world wide.China
The people of the death rate of colorectal cancer about 25/,100,000, accounts for the 4th of urban area malignant tumour.With the improvement of compatriots' diet structure, greatly
The illness rate of intestinal cancer shows the trend for rising year by year.Found according to research, the incidence of disease of Shanghai City colorectal cancer apparently higher than its
He is regional, the morbidity statistics of malignant tumour in 1997, and incidence of colorectal male is 37.2/10 ten thousand, and women is 35.5/10
Ten thousand, the incidence of disease occupy the 3rd.From 2003, colorectal cancer leapt to second common cancer in Areas in Shanghai City, Nan Xing great
Then apparently higher than women, its ratio is about 1.6 to the incidence of disease of intestinal cancer:1.The treatment of current colorectal cancer is the synthesis based on operation
Treatment, the resection rate of operation can reach 60%~70%, and postoperative 5 years survival rates of large Case report are 50%~60% left side
The right side, belongs to reduced levels.It is seen that, continuing the scope of expansion operation cannot also improve the benefit factor of patient.Over nearly 20 years, greatly
The postoperative study on prevention of intestinal cancer although existing significant progress, treatment means improve constantly (operation, radiotherapy, chemotherapy, biological respinse tune
Section treatment etc.), but 5 years survival rates are not improved significantly, and in addition, research finds, cancer patients recurrence and metastatic rate is high
It is its major causes of death.
In China, Chinese traditional treatment has turned into the part in colorectal cancer complex treatment.Substantial amounts of clinic shows, middle treatment
Treatment can improve patient to the tolerance performed the operation, and increase the completion rate of chemicotherapy, mitigate the adverse reaction of chemicotherapy, in colorectal cancer
Root value criterion prevents recurrence and transfer and is played in patients with terminal life quality, the treatment of extension life cycle is improved to lift foot
The effect of weight.But definite effect of the Chinese medicine in colorectal cancer relapse and metastasis are prevented and treated and its contacted with modern molecular biology,
Not yet it is fully apparent from present.Additionally, document report cancer patients card type is divided owes specification, experience composition is more, and therapeutic scheme is received
The influence of the factor such as doctor's level and academic school and it is diversified, therapeutic evaluation is uneven.
In sum, there is provided it is a kind of can effectively treat colorectal cancer and preventing and treating colorectal-cancer postoperative recurrence transfer medicine be mesh
The problem of preceding urgent need to resolve.
The content of the invention
The purpose of embodiment of the present invention is that offer is a kind of treats compound Chinese medicinal preparation of colorectal cancer and preparation method thereof,
Colorectal cancer can be effectively treated, it is particularly obvious to Advanced Colorectal Cancer or Postoperative carcinoma speen-kidney dificiency card patient outcomes.
In order to solve the above technical problems, embodiments of the present invention provide a kind of Chinese medicine compound prescription system for treating colorectal cancer
Agent, the compound Chinese medicinal preparation is made up of the bulk drug that following weight parts are matched:
Glutinous rehmannia:50~400 parts
The fruit of glossy privet:45~300 parts
Saline cistanche:30~240 parts
Mock-strawberry:50~400 parts
Edible tulip:75~420 parts
Fiveleaf akebia fruit:15~180 parts.
In terms of existing technologies, embodiments of the present invention are under the guidance of traditional Chinese medical science traditional theory, according in clinic
The syndrome performance of advanced CRC or Postoperative carcinoma patient, using Chinese medicine glutinous rehmannia (Rehmannia glutinosa (Gaetn.)
Libosch.ex Fisch.et), the fruit of glossy privet (Fructus Ligustri Lucidi), saline cistanche (Cistanche
Deserticola Ma), mock-strawberry (Duchesnea indica (Andr.) Focke), edible tulip (Asarum
Sagittarioides C.F.Liang), fiveleaf akebia fruit (Fructus Akebiae) etc. be made compound Chinese medicinal preparation, to middle and advanced stage
Colorectal cancer or Postoperative carcinoma patient are treated, and can significantly improve life in patients, mitigate the toxicity of chemicotherapy,
Extension patient survival, takes safe and reliable, and without any side effects.
Specifically, colorectal cancer belongs to " ovarian cyst ", " dysentery ", " anorectal cancer ", " lower blood ", " diarrhea ", " product in traditional Chinese medicine
It is poly- ", the category such as " under stagnant "." healthy tendency deposits interior, and heresy can not be done " is total summary pathogenetic to disease, and colorectal cancer is no exception.It is existing
Forefathers' understanding and modern clinical experience are referred and synthesized for traditional Chinese medical science doctor, it is believed that ferritic healthy tendency virtual loss, in addition eating and drinking without temperance (clean) usually,
Deficiency of spleen and stomach, interior raw phlegm wet, is stranded in enteron aisle, and mechanism of qi is obstructed, and operation is not smooth, hinders and forms stasis of blood product in train of thought, ultimately forms large intestine
Cancer.Colorectal cancer is the card of asthenia in origin and asthenia in superficiality, and insufficiency of the spleen, deficiency of the kidney, positive unsaturated vapor are disease sheet, and it is the mark of disease, two that damp and hot, cold-dampness, the stasis of blood are stagnant
Person's reciprocal causation, product is caused by empty, and because of product, benefit is empty, then gradually big long and body is more empty.With the further development of the state of an illness, wet poison
The stasis of blood is stagnant more to contain, the toxic and side effect of the method such as operation, chemotherapy, radiotherapy in addition, and in the medical method such as clearing heat and detoxicating, promoting blood circulation and removing blood stasis attack
Cut down too, make patient's healthy tendency virtual loss day very.Therefore, internal organs weakness, healthy tendency, without heresy is resisted strenuously, are the basic of metastases.It is big at present
The Syndrome Differentiation of Traditional Chinese Medicine of intestinal cancer still disunity, but can be divided into that stagnation of blood stasis, liver-stomach disharmony, the deficiency of liver-yin and kidney-yin, enteron aisle be damp and hot and spleen kidney
Two empty 5 types, wherein speen-kidney dificiency card are the common TCM Syndrome Types of clinically colorectal cancer.Studies have reported that to domestic at present clinical
Intestinal cancer TCM Syndrome Type statistics thinks:Weakened body resistance is that the domination of pathogen is with heat toxin based on insufficiency of the spleen (89.66%), kidney deficiency (63.79%)
(82.76%), hemostasis (75.86%), based on damp and hot (72.4l%).Glutinous rehmannia enriching yin in side in compound Chinese medicinal preparation of the invention
Cool blood, kidney tonifying, essence replenishing is monarch, and the fruit of glossy privet, saline cistanche invigorate the kidney and stop nocturnal emission, and warm yang transforming qi is minister, two medicines and glutinous rehmannia compatibility, treating YANG within YIN,
Treating YIN within YANG, it is biochemical more than.The clearing heat and detoxicating dissipating bind of edible tulip, mock-strawberry is sweet cold, and clearing heat and detoxicating, cool blood detumescence goes large intestine to flee liver
The evil poison of lung;Fiveleaf akebia fruit qi-regulating disappears stagnant to make, and full side's kidney tonifying, essence replenishing, clearing heat and detoxicating, the effect of mass dissipating and swelling eliminating, medicine letter power turns, and mends
Without stagnant, get rid of evils without wound just.All formulas are matched, gathers clearing heat and promoting diuresis altogether, removing toxicity for detumescence, the effect of right supporting anticancer.
Preferably, the compound Chinese medicinal preparation is made up of the bulk drug that following weight parts are matched:
Glutinous rehmannia:100~200 parts
The fruit of glossy privet:90~150 parts
Saline cistanche:60~120 parts
Mock-strawberry:100~200 parts
Edible tulip:150~210 parts
Fiveleaf akebia fruit:30~90 parts.
It is further preferred that the compound Chinese medicinal preparation is made up of the bulk drug that following weight parts are matched:
Glutinous rehmannia:150 parts
The fruit of glossy privet:120 parts
Saline cistanche:90 parts
Mock-strawberry:150 parts
Edible tulip:180 parts
Fiveleaf akebia fruit:60 parts.
In addition, the compound Chinese medicinal preparation can also add the one kind or many in the bulk drug that following weight parts are matched when preparing
Kind:
100~1000 parts of the Radix Astragali
50~200 parts of the bighead atractylodes rhizome
1~100 part of maggot
50~250 parts of ginseng
50~250 parts of Poria cocos
50~250 parts of Psoralea corylifolia
50~200 parts of Fructus Corni
100~600 parts of hairy grape stem
100~600 parts of Chinese actinidia root.
In addition, the compound Chinese medicinal preparation is oral medicinal herb preparation.Specifically, the compound Chinese medicinal preparation can be electuary,
Pulvis, capsule, tablet, syrup, supensoid agent, water decoction or oral liquid.
Embodiments of the present invention additionally provide this kind of preparation method of compound Chinese medicinal preparation, comprise the following steps:According to
Above-mentioned weight weighs bulk drug mixing, adds water to cook, and water decoction is concentrated into stiff shape;Absolute ethyl alcohol is added to make dense
Liquid precipitation after contracting, takes supernatant, and Distillation recovery ethanol stands after concentration, filtering precipitation;By precipitation concentration to thick paste, do
Powdery compound Chinese medicinal preparation is obtained final product after dry.
Carried out 3 times in above-mentioned preparation method, the step of add water to cook:
8 times of water of amount of bulk drug weight are added for the first time, after boiling, after being cooked by slow fire certain hour, leach liquid;Second
It is secondary to add 7 times of water of amount of bulk drug weight, after boiling, after being cooked by slow fire certain hour, leach liquid;Third time adds bulk drug
6 times of water of amount of weight, after boiling, after being cooked by slow fire certain hour, leach liquid;
The liquid that above-mentioned 3 times are leached merges, and obtains water decoction.
In addition, adding the weight of absolute ethyl alcohol and the liquid after concentrating to be equal in weight.
In addition, adding, absolute ethyl alcohol precipitates liquid, the method that takes supernatant is:Fully mix, stand overnight, next day takes
Supernatant.
Brief description of the drawings
Fig. 1 is that the compound Chinese medicinal preparation in experimental example of the present invention 3 is shown human colon carcinoma HCT-116 cell inhibitory effects
It is intended to;
Fig. 2 is shadow of the compound Chinese medicinal preparation in experimental example of the present invention 5 to HCT-116 cell HIPK2 protein expressions
Ring schematic diagram;
Fig. 3 is the influence of compound Chinese medicinal preparation in experimental example of the present invention 5 to HCT-116 cell p53 protein expressions
Schematic diagram.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing to each reality of the invention
The mode of applying is explained in detail.However, it will be understood by those skilled in the art that in various embodiments of the present invention, being
The reader is set to more fully understand the application and propose many ins and outs.But, even if without these ins and outs and being based on
The many variations of following implementation method and modification, it is also possible to realize the application technical scheme required for protection.
Involved raw medicinal material records Chinese medicine for one by China's coastal port in the embodiment of the present invention, and meets
Pharmacopoeial quality requirement.In addition, bulk drug involved in the embodiment of the present invention can be bought by market obtaining.
Embodiment 1
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 2
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 120, the fruit of glossy privet 90, saline cistanche 60, mock-strawberry 120, edible tulip 150, fiveleaf akebia fruit 45
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 60 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 55 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 50 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 3
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 180, the fruit of glossy privet 150, saline cistanche 120, mock-strawberry 180, edible tulip 210, fiveleaf akebia fruit 90
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 55 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 50 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 45 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 4
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 75, the fruit of glossy privet 60, saline cistanche 45, mock-strawberry 75, edible tulip 90, fiveleaf akebia fruit 30
Above-mentioned raw materials medicine half amount is taken, is added water 3 times and is measured, after immersion half an hour, decoct 45min, add water 3 times of amounts, decocted
30min, filtering, merging decoction liquor, is concentrated under reduced pressure, and drying under reduced pressure obtains dry extract, is ground into fine powder (crossing 80 mesh).Second half crude drug
80 mesh were crushed, was sterilized (irradiation dose 8k) with CO60, mixed with extract powder, added following appropriate pharmaceutic adjuvant:Mannitol, breast
Sugar, cane sugar powder, Aspartame and menthol, particle, every bag 9 are made after being well mixed with 60~70% ethanol as adhesive
Gram.
Embodiment 5
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 400, the fruit of glossy privet 300, saline cistanche 240, mock-strawberry 400, edible tulip 300, fiveleaf akebia fruit 180
Above-mentioned raw materials medicine is added into 50% ethanol, 1h is decocted, with beginning timing of seething with excitement, extract solution is poured out, the dregs of a decoction add 50% second
Alcohol, decocts 1h, pours out extract solution, merges extract solution twice, and filtering is concentrated under reduced pressure to give medicinal extract;Plus starch, superfine silica gel powder,
Sodium carboxymethyl starch, is well mixed, and adds water, softwood processed, crosses 18 eye mesh screens, and granulation is dried, 14 mesh sieve whole grains, is added following suitable
Amount pharmaceutic adjuvant:Sodium carboxymethyl starch, magnesium stearate is tabletted, every weight 500mg, or encapsulated, every capsule 0.5
Gram.
Embodiment 6
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60
By the medicinal water boiling and extraction of above-mentioned raw materials 2 times, the 1500ml that adds water is decocted for the first time, decocting time is 60 minutes, is taken
Remaining juice 150ml, decocts the 1000ml that adds water for the second time, and decocting time is 45 minutes, and third time decocts the 1000ml that adds water, decoction
Time is 45 minutes, takes remaining juice 150ml, daily oral secondary, each 150ml.
Embodiment 7
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, ginseng 50, Poria cocos 50,
Psoralea corylifolia 50, Fructus Corni 50, hairy grape stem 100, Chinese actinidia root 100
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 8
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, ginseng 250, Poria cocos
250th, Psoralea corylifolia 250, Fructus Corni 200, hairy grape stem 600, Chinese actinidia root 600
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 9
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, ginseng 150, Poria cocos 150
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 10
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, Psoralea corylifolia 150, mountain Zhu
Cornel 150
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 11
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, hairy grape stem 300, rattan
Pears root 300
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 12
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, ginseng 150, Psoralea corylifolia
150th, hairy grape stem 300
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 13
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60, Poria cocos 250, Fructus Corni
200th, Chinese actinidia root 600
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 14
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60,100 parts of the Radix Astragali, the bighead atractylodes rhizome
50 parts, 1 part of maggot.
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Embodiment 15
Medicine material (gram) is weighed by following portions by weight proportioning
Glutinous rehmannia 150, the fruit of glossy privet 120, saline cistanche 90, mock-strawberry 150, edible tulip 180, fiveleaf akebia fruit 60,1000 parts of the Radix Astragali, the bighead atractylodes rhizome
200 parts, 100 parts of maggot.
By above-mentioned Chinese medicine, consumption is mixed up and be placed in pot for boiling chinese traditional medicine together in proportion, decocts 3 times, and medicine is added for the first time
8 times of water of weight sum, after boiling, are cooked by slow fire 50 minutes, leach liquid;7 times of water of drug weight sum are added for the first time, are boiled
Afterwards, it is cooked by slow fire 45 minutes, leaches liquid;Third time adds 6 times of water of drug weight sum, after boiling, is cooked by slow fire 40 minutes, leaches
Liquid;Three gained liquids are merged, liquid evaporation is concentrated into by stiff shape using slow fire.The absolute ethyl alcohol of equivalent is added,
Fully mix, stand overnight, precipitate it, next day takes its supernatant, and Distillation recovery ethanol stands 24h, makes precipitation complete after concentration
The 80 DEG C of water-baths of precipitation rotary evaporator are concentrated into thick paste (being 1.4/25 DEG C with respect to proportion) by full filtering, and 50 DEG C are vacuum dried,
Obtain powdery Chinese medicinal compound extract, pack, 9 grams every bag.
Drug dose in above-described embodiment, is experimentally verified that, its pharmacological action is identical.
In addition, the consumption and the course for the treatment of of compound Chinese medicinal preparation in the embodiment of the present invention can be according to formulation, the age of patient, diseases
The light and heavy degree of disease is appropriately adjusted, but generally daily oral 2~3 times, every time 2~4 (grain), 0.5 gram of every (grain), or
1 bag every time (9g/ bags), with 3 months for a course for the treatment of, can continuously use 2 courses for the treatment of or longer time.
The beneficial effect of the embodiment of the present invention is:The compound Chinese medicinal preparation that embodiments of the present invention are provided, to suppression
Colorectal-cancer postoperative recurrence transfer processed obtains preferable clinical efficacy, and in animal and cell experiment, the compound Chinese medicinal preparation is to big
Colon-cancer cell growth, propagation, invasion and attack and transfer also show that obvious inhibitory action.Show through clinical and experimental study result, this
Invention compound Chinese medicinal preparation can effectively solve the problems, such as the treatment of clinically colorectal cancer, especially to Advanced Colorectal Cancer or colorectal cancer
Postoperative speen-kidney dificiency card patient outcomes are obvious.
Further, beneficial effects of the present invention are further elaborated below by way of experimental example:
The clinical test of experimental example 1
1. case source experiment case comes from Shuguang Hospital and Shanghai Chinese Medicine Hospital.Institute
There is case to be mCRC (Metastatic Colorectal Cancer) patient of colorectal cancer root value criterion, totally 380.According to admission time successively to trouble
Person numbers, and Applied Digital table method presses 1:1 is randomly divided into two groups.Two groups of patient baseline's data compare, and difference is not statistically significant,
With comparativity.
2. include and exclusion standard
2.1 Western medicine diagnostic criteria clinical diagnosises were with reference to 2009《Practical clinical practice》The diagnostic criteria of middle colorectal cancer;Tissue
Pathology, imageological examination turn out to be Metastatic Colorectal Cancer, and it is by stages anti-with reference to american cancer joint committee (AJCC)/world
Cancer alliance (UICC) colorectal cancer TNM stage system (7 editions).
2.2 TCM syndrome diagnostic criteria references《New Chinese medicine guideline of clinical investigations》And《Shanghai City Chinese medical disease
Conventional treatment》, with reference to clinical experience.
1. 2.3 inclusive criterias previously received the colorectal cancer of radical correction, had clear and definite postoperative pathological to diagnose;2. through pathology
Learn confirmation and there occurs DISTANT METASTASES IN, and at least 1 measurable focus;3. first 6 months non-row antineoplastons are tested or is taken
The medicine of other influences observed result;4. without chemotherapy contraindication, be not in the mood for, liver, kidney and hemopoietic system serious disease, KPS scorings >=
60 points, it is contemplated that life cycle >=3 month, age >=18 year old;5. traditional Chinese medical science syndrome of dampness-heat due to spleen deficiency CM syndrome differentiation criterion is met;6. informed consent is signed
Book person.
1. 2.4 exclusion standards are associated with the systemic diseases such as severe cardiac, lung, liver, kidney, blood and marrow function exception person;②
Age < 18 years old;3. pregnant woman or women breast-feeding their children.
1. 2.5 rejecting standards there is intolerable serious adverse reaction during chemotherapy;2. sb.'s illness took a turn for the worse person during observing,
Can not continue to receive chemotherapy person;3. the curers such as targeting, radiotherapy are received during treating again.
3. treatment method according to colorectal cancer NCCN updated Guidelines using conventional FOLFIRI, XELOX, FOLFOX4,
FOLFOX6 schemes carry out chemotherapy, the same control group for the treatment of group's chemotherapy regimen, while plus with the Chinese medicine compound prescription in the embodiment of the present invention 1
Preparation, daily 1 dose, decocting.
4. observational technique
4.1 evaluation of clinical curative effect judge curative effect using RECIST1.1 solid tumors the standard of curative effect evaluation, are divided into:Complete incidence graph
(partial response, PR), stabilization (stable disease, SD) are alleviated in (complete response, CR), part
With progress (progressive disease, PD), total effective rate (CR+PR, ORR), disease control rate (CR+PR+SD, DCR).
Progression free survival phase (progress free survival, PF S) be since treatment start to tumor focus occur be in progress when
Between.Overall survival (overall survival, OS) is time of the treatment beginning to dead or last follow-up.
4.2 qualities of life treat fore-and-aft observing life quality scores (KPS), are estimated by Karnofsky standards.Carry
It is high:Score value improves 10 points or more after treatment;Reduce:Score value reduces 10 points or more after treatment;Stabilization:Score value increases after treatment
Or reduce less than 10 points.
4.3 adverse reactions are according to WHO《The performance of acute and subacute toxicity and indexing standard》Evaluation adverse reaction.
5. statistical method carries out statistical analysis using SPSS 19.0.Measurement data is checked using t;Enumeration data uses χ
2 inspections, ranked data use Wilcoxon rank tests.With P < 0.05 for difference has statistical significance.
As a result
1. influence treatment group patient of the Chinese medicine combined chemotherapy to mcRC short term effects is without CR, PR 61 (30.5%), SD
94 (47%);PD 45 (22.5%), ORR are that 30.5%, DCR is 77.5%.Control group is without CR, PR 50
(27.7%), SD 68 (37.7%), PD 62 (34.4%), ORR is that 27.7%, DCR is 65.6%.Two groups objective effective
Rate, disease control rate no statistical difference difference, are shown in Table 1.
Influence (example) of the Chinese medicine combined chemotherapy of table 1. to Metastatic Colorectal Cancer short term effect
2. influence of the Chinese medicine to mcRC life cycles survives follow-up in addition to the case that comes off, 200 treatment groups and 180 control groups
Patient's middle position PFS is respectively 11.2,7.96 months (P=0.012), and 200 treatment groups and 180 control group middle position OS are respectively
26.3rd, 19.8 months (P=0.027).Treatment group middle position PFS and middle position OS are considerably longer than control group (P < 0.05).
3.KPS situations of change treatment group, control group KPS improve situation and compare, and KPS improves 50 after treatment group's treatment, surely
It is fixed 30, reduce by 15;KPS improves 24 after control group treatment, stablizes 29, reduces by 34, the statistically significant (P of difference
< 0.05), it is shown in Table 2.
KPS scorings are compared (example) before and after 2. two groups of patient's treatments of table
4. in adverse reaction hematology adverse reaction control, treatment group, control group are in blood leukocytes reduction, neutrophil leucocyte
Generation aspect is reduced, difference is statistically significant (P < 0.05), is shown in Table 3.
3 two groups of Metastatic Colorectal Cancer blood samples of patients toxicity of table compare
The zoopery of experimental example 2
Human Colonic Tumor in Nude Mice Transplanted tumor model is prepared according to document, the human colon carcinoma HCT-116 cells in growth period of taking the logarithm,
0.25% Trypsin Induced, 1 × 10 is diluted to 0.01mol/LPBS7Cell/mL single cell suspensions.Take 4-6 week old,
18-20gBalb/c nude mices 3, the nearly right armpit side skin of back of routine disinfection nude mice, people's intestinal cancer HCT-116 is drawn with 1mL syringes
Single cell suspension, by 0.2mL/ (2 × 106Cell), be inoculated into nude mice right fore armpit it is subcutaneous after, with ethanol cotton balls oppress pin
Hole in case cell flows out, conventinal breeding under the conditions of SPF, when nude mice by subcutaneous tumour it is long to 1cm or so when, choose the good knurl of growth conditions
Nude mice of the body without diabrosis takes off neck and puts to death as knurl mouse is supplied, and knurl body is peeled off under aseptic condition.Blood stains, removal are cleaned with physiological saline
Slough and fibr tissue, take borderline tumor flesh tissue and are cut into 2mm3Size, nude mice right fore armpit is inoculated in by tumor mass
It is subcutaneous, pass on 3 times in this way, take 4-6 week old, 18-20g Balb/c nude mices 30 after 3 generations, choose growth conditions it is good, without diabrosis
Knurl body, be inoculated in the nude mice subcutaneous conventinal breeding of right fore armpit.
After nude inoculation subcutaneous tumors, when tumour growth to 200-300mm3When, it is randomly divided into 5 groups:Model group (0.9% chlorine
Change sodium solution group), the basic, normal, high dosage group of Chinese medicine (200,400,800mg/ (kgd)), 5-FU groups (1mg/ (kgd)), often
Group 6, Chinese medicine gastric infusion, 5-FU intraperitoneal injections, 0.9% isometric sodium chloride solution of model group gavage.Administration the
14 days nude mices take out knurl body respectively, measure tumor size.
The computing formula of gross tumor volume (TV) is:TV=ab2/ 2, wherein a, b represents the length and width of knurl body respectively, calculates swollen
Knurl volume inhibiting rate:Gross tumor volume inhibiting rate %=(1- experimental groups mean tumour volume/model group mean tumour volume) ×
100%.
Experimental result shows, each group tumor volume no difference of science of statistics (P before administration>0.05), give Chinese medicine and 5-FU is controlled
After treating 2W, tumor volume inhibiting rate is calculated, as a result as shown in table 1:The basic, normal, high dosage group of Chinese medicine and 5-FU groups show not
Same tumor killing effect, Chinese medicine low dose group tumor volume (P statistically significant relative to model group difference<0.05), middle and high dose
Amount group and 5-FU groups tumor volume are statistically significant (P relative to model group difference<0.01).After treatment Chinese medicine it is low,
The tumor volume inhibiting rate of middle and high dosage group and 5-FU groups be respectively 28.83%, 39.98%, 47.67% and 54.26% (see
Table 4), and the dependence of dosage is presented.
The Chinese medicine of table 4. to the growth inhibition effect of Human Colonic Tumor in Nude Mice transplantable tumor (N=6)
* vs model groups p<0.05, * * vs model groups p<0.01
The cell experiment 1 of experimental example 3
The human colon carcinoma HCT-116 cells that cellar culture is in exponential phase are taken, with containing 10% hyclone after digestion
RPMI-1640 nutrient solutions diluted concentration to 5 × 104/ mL, per the μ L of hole 100 in 96 porocyte culture plates of addition, cellar culture.
The adherent situation of observation of cell after 6h, the difference for adding 100 μ L to be prepared by the RPMI-1640 nutrient solutions containing 10% hyclone per hole
The Chinese medicine alcohol extract of concentration, activity is 10,20,40,60,80,100,120 μ g/mL totally 7 dosage, and each dosage sets 6
Multiple holes, and it is Normal group to set not dosing cell, continues to cultivate.After culture 48h, the old nutrient solution of 96 orifice plates is abandoned, added per hole
The CCK solution (lucifuge) of 180 μ l serum-free RPMI-1640 nutrient solutions and 20 μ l, continues to cultivate.After 1-4h, in enzyme detector
(dual wavelength 490nm/630nm) detects light absorption value (OD).Life of the Chinese medicine to people's intestinal cancer HCT-116 cells is calculated according to testing result
Inhibiting rate (Growth Inhibitor Rate, GIR) long, and calculation of half inhibitory concentration (IC50).
CCK-8 testing results display kidney tonifying and detoxicating dissipating bind side can suppress the propagation of human colon carcinoma HCT-116 cells, and be in
Dosage-time-dependent relation.Kidney tonifying and detoxicating dissipating bind side is respectively to the IC50 of human colon carcinoma HCT-116 cells 24h, 48h, 72h
88.12 μ g/ml, 63.23 μ g/ml, 47.12 μ g/ml, as shown in Figure 1.Wherein, in Fig. 1, curve from top to bottom is followed successively by
The testing result made under 24h, 48h, 72h.
The cell experiment 2 of experimental example 4
Take the logarithm growth period HCT-116 cell 5 × 103Paving 60mmm culture dishes, after cell patch 24h after add it is low in it is highly concentrated
(20,40,60 μ g/mL) Chinese medicine culture of degree 14 days, changes band Chinese-medicinal culture medium for every two days, after culture terminates, discards culture medium,
With methyl alcohol is added after PBS, after fixed 15min, fixer is discarded, add haematoxylin eosin stains, observe Clone formation feelings
Condition.
According to CCK-8 method experimental results, selection 20,40,60 μ g/mL, 3 concentration are follow-up experiment condition.Result shows
Show, with the raising of middle concentration, the inhibitory action to HCT-116 cell clonal formations constantly strengthens, in dose-dependence.
The cell experiment 3 of experimental example 5
After the Chinese medicine treatment 48h of various concentrations is added in human colon carcinoma HCT-116 cells, protein extraction is carried out, and use
BCA kits are quantified;Configure SDS-PAGE glue and carry out electricity using BioRad electrophoretic apparatus after adding sample and Marker
Swimming is separated;Transferring protein closes 2h after transferring film on pvdf membrane with 5% BSA, adds primary antibody, 4 DEG C of overnight incubations;TBST
Wash 3 times, after each 5-10min, add secondary antibody to be incubated 2h, then washed 3 times with TBST;Developer solution is finally added dropwise, chemiluminescence is put into
Imaging system is developed, and analyzes its gray value.
Various concentrations Chinese medicine (20,40,60 μ g/mL), after effect HCT-116 cells 48h, Western Blot results show
Show, compared with control group, Chinese medicine each group can raise human colon carcinoma HCT-116 cells in HIPK2 expressing quantities (such as Fig. 2 institutes
Show), dose-dependant effect is presented, difference has statistical significance (p<0.05, p<0.01).Wherein, in fig 2, A represents mould
Type group;B represents Chinese medicine low dose group;C represents Chinese medicine middle dose group;D represents Chinese medicine high dose group.Compare * p with A groups<
0.05, * * p<0.01.
Various concentrations Chinese medicine (20,40,60 μ g/mL), after effect HCT-116 cells 48h, Western Blot results show
Show, compared with control group, Chinese medicine each group can raise human colon carcinoma HCT-116 cells in p53 expressing quantities (such as Fig. 3 institutes
Show), dose-dependant effect is presented, difference has statistical significance (p<0.05, p<0.01).Wherein, in fig. 3, A represents mould
Type group;B represents Chinese medicine low dose group;C represents Chinese medicine middle dose group;D represents Chinese medicine high dose group, and * p are compared with A groups<
0.05, * * p<0.01.
It will be understood by those skilled in the art that the various embodiments described above are to realize specific embodiment of the invention, and
In actual applications, can to it, various changes can be made in the form and details, without departing from the spirit and scope of the present invention.
Claims (10)
1. a kind of compound Chinese medicinal preparation for treating colorectal cancer, it is characterised in that the bulk drug matched by following weight parts is made:
Glutinous rehmannia:50~400 parts
The fruit of glossy privet:45~300 parts
Saline cistanche:30~240 parts
Mock-strawberry:50~400 parts
Edible tulip:75~420 parts
Fiveleaf akebia fruit:15~180 parts.
2. compound Chinese medicinal preparation according to claim 1, it is characterised in that the bulk drug system matched by following weight parts
Into:
Glutinous rehmannia:100~200 parts
The fruit of glossy privet:90~150 parts
Saline cistanche:60~120 parts
Mock-strawberry:100~200 parts
Edible tulip:150~210 parts
Fiveleaf akebia fruit:30~90 parts.
3. compound Chinese medicinal preparation according to claim 2, it is characterised in that the bulk drug system matched by following weight parts
Into:
Glutinous rehmannia:150 parts
The fruit of glossy privet:120 parts
Saline cistanche:90 parts
Mock-strawberry:150 parts
Edible tulip:180 parts
Fiveleaf akebia fruit:60 parts.
4. compound Chinese medicinal preparation according to claim 1, it is characterised in that following weight parts proportioning is additionally added during preparation
One or more in bulk drug:
100~1000 parts of the Radix Astragali
50~200 parts of the bighead atractylodes rhizome
1~100 part of maggot
50~250 parts of ginseng
50~250 parts of Poria cocos
50~250 parts of Psoralea corylifolia
50~200 parts of Fructus Corni
100~600 parts of hairy grape stem
100~600 parts of Chinese actinidia root.
5. the compound Chinese medicinal preparation according to any one of Claims 1-4, it is characterised in that the compound Chinese medicinal preparation is
Oral medicinal herb preparation.
6. compound Chinese medicinal preparation according to claim 5, it is characterised in that the compound Chinese medicinal preparation be electuary, pulvis,
Capsule, tablet, syrup, supensoid agent, water decoction or oral liquid.
7. the preparation method of the compound Chinese medicinal preparation described in any one of Claims 1-4, it is characterised in that comprise the following steps:
The bulk drug mixing being weighed by weight ratio, being added water to cook, water decoction is concentrated into stiff shape;Add anhydrous second
Alcohol precipitates the liquid after concentration, takes supernatant, and Distillation recovery ethanol stands after concentration, filtering precipitation;By precipitation concentration to thick
Cream, powdery compound Chinese medicinal preparation is obtained final product after drying.
8. preparation method according to claim 7, it is characterised in that described to carry out 3 times the step of add water to cook:
8 times of water of amount of bulk drug weight are added for the first time, after boiling, after being cooked by slow fire certain hour, leach liquid;Add for second
Enter 7 times of water of amount of bulk drug weight, after boiling, after being cooked by slow fire certain hour, leach liquid;Third time adds bulk drug weight
6 times of water of amount, after boiling, after being cooked by slow fire certain hour, leach liquid;
The liquid that above-mentioned 3 times are leached merges, and obtains water decoction.
9. preparation method according to claim 7, it is characterised in that the weight of the addition absolute ethyl alcohol and the concentration
Rear liquid is equal in weight.
10. preparation method according to claim 7, it is characterised in that the addition absolute ethyl alcohol makes liquid precipitate, take
The method of clear liquid is:Fully mix, stand overnight, next day takes supernatant.
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CN116392553A (en) * | 2023-03-07 | 2023-07-07 | 上海中医药大学附属曙光医院 | Traditional Chinese medicine compound preparation for treating colorectal cancer metastasis |
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CN109674925A (en) * | 2019-01-31 | 2019-04-26 | 蒋先仲 | A kind of advanced colorectal cancer Chinese medicine composition and preparation method |
CN116392553A (en) * | 2023-03-07 | 2023-07-07 | 上海中医药大学附属曙光医院 | Traditional Chinese medicine compound preparation for treating colorectal cancer metastasis |
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