CN106074586A - Dangshen astragalus composition and preparation method and application - Google Patents
Dangshen astragalus composition and preparation method and application Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7032—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a polyol, i.e. compounds having two or more free or esterified hydroxy groups, including the hydroxy group involved in the glycosidic linkage, e.g. monoglucosyldiacylglycerides, lactobionic acid, gangliosides
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
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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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/34—Campanulaceae (Bellflower family)
- A61K36/344—Codonopsis
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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Abstract
The invention provides a kind of dangshen astragalus composition, this dangshen astragalus composition includes: lobetyolin 0.54mg 1.28mg, calycosin 7 O β D pyranglucoside 0.65mg 0.87mg, Radix Astragali saponin III 1.02mg 2.41mg, Radix Astragali saponin II 0.71mg 1.67mg, Radix Astragali saponin I 0.47mg 1.11mg, astragaloside 1.02mg 2.4mg, pratensein 7 O β D pyranglucoside 0.72mg 1.7mg, brachyoside B 0.055mg 0.13mg, atractylenoide Ⅲ 0.36mg 0.86mg；The dangshen astragalus composition active constituent content that the present invention provides is high, steady quality.
Description
Technical field
The present invention is about a kind of dangshen astragalus composition and preparation method and application, particularly a kind of from Chinese crude drug Radix Codonopsis,
Answering in the pharmaceutical preparatioies such as compositions that astragalus extraction obtains and preparation method thereof improves immunity with it in preparation, antitumor
With.
Background technology
The Chinese medicine preparation that effective component extracting is made from Chinese crude drug Radix Codonopsis, the Radix Astragali, its effective ingredient is to anti-in organism
The generation of body has obvious facilitation, can strengthen reticuloendothelial system phagocytic function, can make leukocyte and apocyte in blood
Dramatically increase；Macrophage phagocytic percentage rate and phagocytic index significantly rise；There is promotion to make to humoral immunization and cellular immunization
With, Radix Codonopsis also has simultaneously increases erythrocyte and the effect of hematochrome.
Twentieth century related personnel's seventies, Radix Codonopsis, the proportioning of the Radix Astragali, dosage form are studied, it is believed that according to Radix Codonopsis with
The ratio of the Radix Astragali is to carry out extraction obtain extract and be configured to vein injection (20ml) clinical efficacy best at 1: 1, is used for
The auxiliary treatment of the postoperative Radiotherapy chemotherapy of digestive tract tumor, can make the patient of 100% favorably accomplish Radiotherapy chemotherapy overall process.Ginseng stilbene
Compositions mainly has following effect: improve symptom, extends the life cycle of late tumor patient；Safeguard that hemopoietic function, minimizing are put, changed
Leukopenia during treatment；Improve cytophagous phagocytic function；Tumor is had certain inhibitory action；There is blood circulation promoting and blood stasis dispelling
Effect.
But, the most traditional Radix codonopsis and Radix Astragali composition (abbreviation dangshen astragalus composition) lacks mostly to wherein effective ingredient
Quantitative analysis, ginseng stilbene preparation on the market, such as ginseng astragalus pill agent, powder, injection (containing lyophilized powder) its working substance the most uncontrollable
Content, it is impossible to its main component total saponins, polysaccharide, the actual content of flavone are described, is simply simply defined as Radix Codonopsis water extract
It is quantity for quantity, alcohol extraction thing, thus in actual production, causes the difference between last consignment of product and next group product
Different very big, portioned product does not reaches content and the state of an illness does not reaches therapeutic effect, and the product of too high levels then compromises patient's
Healthy.
In order to solve traditional Radix codonopsis and Radix Astragali composition (abbreviation dangshen astragalus composition), effective ingredient is lacked quantitative analysis, main
The problem wanting composition actual content not name, for the content of effective ingredient in specification ginseng stilbene pharmaceutical preparation, industry related personnel is to ginseng
The chemical composition of stilbene extract has carried out substantial amounts of analysis and research.
Part research is each chemical composition of qualitative isolation identification dangshen astragalus composition, such as " SHENQI FUZHENG ZHUSHEYE
Chemical constitution study " (Gu Hong is gorgeous, " Chinese patent medicine ", 7 phases in 2013) uses silica gel, ODS, Sephadex LH-20 and
The chemical composition of RP-HPLC isochromatic spectrum method separation Shenqi extraction product SHENQI FUZHENG ZHUSHEYE, passes through physicochemical property
And the structure of Spectrum Analysis authenticating compound.Result shows 15 lists of isolated from SHENQI FUZHENG ZHUSHEYE crude drug extractum
Body compound, is identified respectively as Radix Astragali saponin I, II, III, different Radix Astragali saponin II, astragaloside, 6 "-O-acetyl group Mao Ruiyi Huangs
Ketone etc.." in SHENQI FUZHENG ZHUSHEYE, Radix Codonopsis composition UHPLC-MS analyzes " (thank to Potiria pectinifera (Mukller et Tro Sehel), etc. " Chinese crude drug ". the 8th phase of volume 37) public
6 kinds of monomer chemical combination in 12 batches of Shenqi extraction product SHENQI FUZHENG ZHUSHEYE that Livzon Group Limin Pharmaceutical Factory produces are opened
The content of thing.
Part 3 kinds of research dangshen astragalus composition or 4 kinds of characteristic compositions and content such as astragaloside, calycosin and
Total solid matterss etc. also disclose that a kind of party as different batches quality control standard, such as Chinese patent ZL 200510064551.5
Ginseng Radix Astragali extract, is wherein no less than 0.325 gram containing total solid matters in every 1 gram of extract, containing total sugar in every 1 gram of this total solid matters
(in terms of anhydrous glucose) 0.08-0.15 gram, total saponins (in terms of astragaloside) 7-10 milligram, flavonoid glycoside 5-8 milligram.This patent
Disclosed technology determines Contents of Main Components in Radix codonopsis and Radix Astragali composition, to some extent solves tradition ginseng stilbene and extract
The problem of content difference between thing product batches, so that therapeutic effect is more reliable and more stable.
Chemical composition for dangshen astragalus composition is qualitative and quantitative study advances the system joining stilbene pharmaceutical preparation quality standard
Fixed, between the different ginseng stilbene pharmaceutical preparation batches of specification product quality, but the dangshen astragalus composition of existing ginseng stilbene pharmaceutical preparation
Extractive technique makes each chemical composition extraction efficiency on the low side, causes the some chemical properties in Radix Codonopsis and astragalus extraction garbage
Waste；Radix Codonopsis and Radix Astragali raw material medical material provenance and production year is further limit plus the new strict quality specifications formulated
Limit, causes raw material medical supply nervous.It is thus desirable to Optimized Extraction Process, improve Radix Codonopsis and Radix Astragali utilization rate；Find more
Add abundant raw material medical material Liaoyuan, work along both lines and solve the problem that raw material supply is nervous.
But it is former that the simple raising Radix Codonopsis utilizing modern preparation technology and astragalus extraction efficiency can not directly solve Chinese medicine
The some chemical properties waste of material medical material.Modern preparation technology specific aim compared with traditional handicraft, to related chemical constituents
By force, such as organic solvent extraction, steam distillation, supercritical fluid extraction, ultrafiltration, acid-alkali treatment method etc., extraction
Material composition composition extracts with traditional decocting in water or has occurred that bigger change, such as animal drugs compared with water extract-alcohol precipitation extraction method
Material Solenognathus, Hippocampus etc., take tradition extraction process by water, and most of composition of extraction is protein, aminoacid etc., and uses organic molten
Agent is solvent when extracting, then can by fatty acid constituents extraction out, and chemical composition spectrum there occurs bigger compared with the former
Change.In general, most of chemical composition extraction efficiencies of modern Chinese medicine extraction and purification process extract work far above tradition
Skill, and the chemical composition that cannot extract in the past also can be extracted, improve the utilization rate of Chinese herb medical material, have
Being beneficial to save material, (height enters medical material, Lu Peng, Dai Huiqing, etc. Chinese medicine modern times preparation technology on the impact of untoward reaction and relevant
Countermeasure [J]. Chinese experimental pharmacology of Chinese medical formulae magazine, 2014,20 (14): 52-55).
When modern Chinese medicine extraction and purification process improves extraction efficiency, have also been changed the chemical composition composition in Chinese medicine medicine,
Being probably and change chemical composition ratio in original Chinese medicine medicine, may add new chemical composition, these changes may
Improve the drug effect of Chinese medicine medicine, it is also possible to bring the untoward reaction improving Chinese medicine medicine.As used according to reports such as Liu Shenlin
The Fructus Cnidii compound preparation of alcohol extraction technique is compared with using tradition water extraction process, and toxicity untoward reaction substantially increases (Liu
Shen Lin, Xiong Ningning, Liu Fang, etc. the Ethic review [J] of Compound Fructus Cnidium preparation clinical trial untoward reaction. Chinese Clinical pharmacology
Learn and therapeutics, 2004,8 (2): 238).
It is thus desirable to modern times preparation technology and pharmacological evaluation are combined, filter out efficient and safe preparation technology, with
Obtain active constituent content dangshen astragalus composition high, stay-in-grade.
Summary of the invention
The present invention is the further research in CN1686404A correlation technique.Inventor is through long-term research, and leads to
Cross the checking repeatedly of zoopery and clinical experiment, work out a kind of active constituent content dangshen astragalus composition high, stay-in-grade.
On the one hand, the invention provides a kind of dangshen astragalus composition, this dangshen astragalus composition includes:
It is furthermore preferred that the invention provides a kind of dangshen astragalus composition, this dangshen astragalus composition includes:
According to specific embodiments of the present invention, the dangshen astragalus composition of the present invention also includes:
According to specific embodiments of the present invention, the dangshen astragalus composition of the present invention also includes syringic acid, (6aR, 11aR)-
3-hydroxy-9,10-dimethoxy pterocarpane, (3R)-7,2 '-dihydroxy-3 ', in 4 '-dimethoxy isoflavan, Flos Caryophylli fat glycosides
One or more.
The Chinese herb medical material that dangshen astragalus composition of the present invention relates to is Radix Codonopsis and the Radix Astragali, described Radix Codonopsis
(Codonopsis Pilosula (Franch.) Nannf) is the dry root of Campanulaceae and relevant medicinal effects, described Huang
Stilbene is the pod membrane Radix Astragali (Astraglus membranaceus (Fisch.) Bge.) or Radix Astagali (Astraglus
Membranaceus (Fisch.) Bge.Var.mongholicus (Bge.) Hsiao) dry root.
Chinese herb medical material Radix Codonopsis due to the emetic saying of Rhizoma Ginseng, the drug effect of Radix Codonopsis and Radix Ginseng and medicinal part phase
When, common usage is to remove use after Radix Codonopsis reed head.But there are some researches show now that Radix Codonopsis reed head becomes with Radix Codonopsis dry root chemistry
Split-phase seemingly, although medicinal ingredient such as total polysaccharides and total saponin content are less than root, but the content of total amino acids, liposoluble constituent
Higher than root (Qin Lei, Zhu Huifeng, Wang Tao, etc. Radix Codonopsis reed head and root chemical composition comparative analysis [J]. Food Science .2015,
135 (14): 135-139).In order to enrich the source of Radix Codonopsis raw material medical material, the reed head of Radix Codonopsis is joined Radix Codonopsis and does by the present invention
In dry, investigate medicinal part and change the impact on dangshen astragalus composition drug effect.Present invention discover that prepared by the Radix Codonopsis not removing reed head
The drug effect of dangshen astragalus composition better than the dangshen astragalus composition prepared of Radix Codonopsis removing reed head, it may be possible to the composition one in Radix Codonopsis reed head
Aspect have impact on the extraction efficiency of astragalus root components, the on the other hand accident of the composition in Radix Codonopsis reed head and other dangshen astragalus composition compositions
There occurs the synergism of forward, jointly improve the drug effect of dangshen astragalus composition.
Radix Codonopsis reed head is about 10%-20% with the weight ratio of Radix Codonopsis dry root, and the present invention is not except removing LU, by Radix Codonopsis reed
Head and Radix Codonopsis dry root use together as raw material medical material, and experiment proves that the dangshen astragalus composition drug effect of gained is enhanced, may
It is composition in Radix Codonopsis reed head and the composition of former dangshen astragalus composition there occurs forward synergism.Radix Codonopsis reed head may be used for joining stilbene
The result of compositions, expands the source of raw material medical material, and reduces the cost of raw material medical material, has great meaning
Justice.
Dangshen astragalus composition of the present invention can use water to carry, and-ol is heavy, water carry-chitosan is heavy, water proposes-resin separation etc.
Shenqi extraction is prepared in method extraction, and then Shenqi extraction is prepared as dangshen astragalus composition further.
Dangshen astragalus composition of the present invention can use water to put forward the heavy method of-ol and extract preparation, and water carries 2-3 time, precipitate with ethanol
2-3 time.
Dangshen astragalus composition of the present invention can use water to put forward the heavy method of-ol and extract preparation, concretely comprises the following steps: take party
Joining and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.Extracting solution is concentrated into every 1ml containing raw
Medicine about 1.2g, adds ethanol and makes alcohol content reach 75%, stand, filter, and filtrate recycling ethanol is also concentrated into every 1ml about 2g Han crude drug,
Adding ethanol again makes alcohol content reach 85%, stands, filters, and filtrate recycling ethanol is also concentrated into every 1ml about 2.5g Han crude drug, then add second
Alcohol makes alcohol amount reach 88%, stands, alcohol deposit fluid disk centrifuge centrifugation, then adds filtering with microporous membrane with filter paper, and filtrate is returned
Receive ethanol and be concentrated into every 1ml about 1.0g Han crude drug, obtaining Shenqi extraction.
Dangshen astragalus composition of the present invention can use water carry-method that chitosan is heavy extracts preparation, concretely comprise the following steps:
Taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.Extracting solution is concentrated into every 1ml
Containing crude drug about 1.2g, adding chitosan 17% (w/v), temperature maintains 80 DEG C, stirs 10 minutes, uses disk centrifuge after standing
Centrifugation, then add filtering with microporous membrane with filter paper, filtrate is also concentrated into every 1ml about 1.0g Han crude drug, obtains Shenqi extraction.
Dangshen astragalus composition of the present invention can use water carry-resin separate method extract preparation, concretely comprise the following steps:
Taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.After three times extracting solution filters
Merge, be concentrated into liquor strength and make a living concentration 0.5g/mL.D101 macroporous resin on medicinal liquid, uses 70% ethanol, 5 times of volumes to wash
De-, collect mother solution and eluent；Glucosan resin on mother solution, uses 70% ethanol, 5 times of volume eluting, collects mother solution and eluent,
Merge D101 macroporous resin and the eluent of glucosan resin, be concentrated into every 1ml about 1.0g Han crude drug, obtain Shenqi extraction.
Dangshen astragalus composition of the present invention can use water carry-resin separate method extract preparation, concretely comprise the following steps:
Taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.After three times extracting solution filters
Merge, be concentrated into liquor strength and make a living concentration 0.5g/mL.D101 macroporous resin on medicinal liquid, uses 70% ethanol, 5 times of volumes to wash
De-, collect mother solution and eluent；Mother liquor concentrations, to every 1ml about 1.2g Han crude drug, adds ethanol and makes alcohol content reach 75%, stand, filter
Crossing, filtrate recycling ethanol is also concentrated into every 1ml about 2g Han crude drug, then adds ethanol and make alcohol content reach 85%, stands, filters, filtrate
Reclaiming ethanol and be also concentrated into every 1ml about 2.5g Han crude drug, then add ethanol and make alcohol amount reach 88%, standing, alcohol deposit fluid dish-style separates
Machine centrifugation, then add filtering with microporous membrane with filter paper, filtrate recycling ethanol is also concentrated into every 1ml about 1.0g Han crude drug, obtains ginseng
Stilbene extract.
Detailed description of the invention
It is further elucidated with the present invention below in conjunction with specific embodiment.Should be understood that these embodiments are merely to illustrate the present invention
Rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally according to conventional strip
Part, or according to the condition proposed by manufacturer.
Embodiment 1
(1) taking Radix Codonopsis 400g and Radix Astragali 400g, rinse well with deionized water, add 6400ml deionized water, addition carries
After taking 1 hour, release extracting solution；Adding 4800ml deionized water, heating extraction, after 1 hour, releases extracting solution；Add
4800ml deionized water, after heating 0.5 hour, releases extracting solution.Merge after three times extracting solution filters, be concentrated into 1600ml, obtain
Liquor strength make a living concentration 0.5g/mL.
(2) take D101 macroporous resin on step (1) medicinal liquid, use 70% ethanol, 5 times of volume eluting, collect mother solution and eluting
Liquid；
(3) mother solution extracting solution is concentrated into every 1ml about 1.5g Han crude drug, adds ethanol and makes alcohol content reach 75%, stands, filter,
Filtrate recycling ethanol is also concentrated into every 1ml about 3g Han crude drug, then adds ethanol and make alcohol content reach 85%, stands, filters, and filtrate is reclaimed
Ethanol is also concentrated into every 1ml about 3.8g Han crude drug, then adds ethanol and make alcohol amount reach 88%, stands, centrifugal point of seperator of alcohol deposit fluid
From, the eluent of filtrate with step (2) gained being merged, then adds filtering with microporous membrane with filter paper, filtrate recycling ethanol also concentrates
To every 1ml about 5g Han crude drug.
(4) medicinal liquid of step (3) gained injects and makes every 1ml about 2g Han crude drug with water, heating, adds total crude drug amount
The pin charcoal of 0.35%-0.45% (g/g), boils, and filters, and filtrate cold preservation, after separated machine centrifugation, through 0.2 μm micropore
Filter filters, then filters through ultrafiltration system, is concentrated into every 1ml (40mg Radix Codonopsis and 40mg Radix Astragali) Han crude drug about 80mg,
Ginseng stilbene extracts compound.
(5) Shenqi extraction injects with water appropriate, boils 20 minutes, adds pin charcoal 0.15g, boils 10 minutes, cold
But, filtering, filtrate is standby.Take sodium chloride 6.5g, add appropriate water for injection and make dissolving, add pin charcoal 0.25g, boil, cooling,
Filtering, filtrate merges with above-mentioned standby filtrate, adds disodium edetate 0.2g, sodium pyrosulfite 0.5g and dissolves, injects and adjust with water
Whole total amount to 6.0-6.4, adds pin charcoal 0.15g-0.30g to 1000ml, regulation pH value, filters, fill, every bottle of 250ml, go out
Bacterium, obtaining crude drug content is the 20g (dangshen astragalus composition of (10g Radix Codonopsis and the 40g Radix Astragali).
Take respectively and do not remove reed head and remove the Radix Codonopsis of reed head as raw material medical material in step (1), prepare ginseng stilbene
Compositions 1 and dangshen astragalus composition 2.
Take 10ml dangshen astragalus composition 1 and 2 respectively, respectively the lobetyolin in detection by quantitative dangshen astragalus composition 1 and 2, Mao Rui
Isoflavone 7-O-β-D pyranglucoside, Radix Astragali saponin III, Radix Astragali saponin II, Radix Astragali saponin I, astragaloside, Herba Trifolii Pratentis
Element-7-O-β-D-pyranglucoside, brachyoside B and the content of atractylenoide Ⅲ, the results are shown in Table lattice 1.
Form 1: dangshen astragalus composition component
Conclusion: not removing the dangshen astragalus composition 1 of reed head compared with the composition of the dangshen astragalus composition 2 removing reed head, content is deposited
In certain difference.Owing to the content of the lobetyolin in Radix Codonopsis reed head is lower than the lobetyolin's content in Radix Codonopsis dry root, because of
Low than in dangshen astragalus composition 2 of this component in this dangshen astragalus composition 1, with existing document disclosed in data be consistent.Ginseng stilbene group
In compound 1, pratensein-7-O-β-D-pyranglucoside, brachyoside B, atractylenoide Ⅲ content are significantly higher than
Content in dangshen astragalus composition 1.Radix Astragali saponin III, Radix Astragali saponin II, Radix Astragali saponin I and astragaloside are at dangshen astragalus composition 1 and 2
Middle changes of contents does not has obvious rule.Illustrate that not removing reed head can affect carrying of some compositions of the Radix Astragali with the Radix Codonopsis removing reed head
Taking, result causes the constituent content difference of dangshen astragalus composition 1 and 2.
Embodiment 2 pharmacodynamic experiment:
Experiment: the Radix codonopsis and Radix Astragali composition of the present invention and China's application 03123045.8, China's application
200510064551.5 Radix codonopsis and Radix Astragali in extracts the contrast of compositions antitumor action
Laboratory animal: BALB/C mice 56, SP F level, male, body weight 18～20g, the Chinese Academy of Medical Sciences test
Institute of Botany provides.
Tumor strain: S180 cell strain, Institute Of Chinese Materia Medica Of China Academy of Chinese Medical Sciences provides.
Experimental agents: the Radix codonopsis and Radix Astragali of medicine A: the present invention extracts compositions, according to embodiments of the present invention the ginseng stilbene of 1 preparation
Compositions 1；
Medicine B: the dangshen astragalus composition 2 of 1 preparation according to embodiments of the present invention；
Medicine C: the Radix codonopsis and Radix Astragali in China's application 03123045.8 embodiment 1 extracts compositions；
Medicine D: the Radix codonopsis and Radix Astragali in China's application 200510064551.5 embodiments 1 extracts compositions；
Medicine E: ginseng stilbene syrup (Jiangxi Jimin Kexin Drug Industry Co., Ltd's production)；
Experimental technique:
1) preparation of tumor cell suspension: take well-grown S180 and pass on mice, extracts mouse ascites under aseptic condition,
With normal saline dilution, 1500r min-1Centrifugal 5min, is repeated 2 times, is then dispelled by cell to individual cells, at microscope
Cell is also adjusted 1 × 10 by lower counting6mL-1。
2) it is grouped and is administered: mice is randomly divided into 6 groups, often group 8, respectively matched group, group medicine A experimental group, medicine
Thing B experimental group, medicine C experimental group, medicine D experimental group, medicine E experimental group.
Lotus tumor matched group gives physiological saline solution, according to 0.02mL/g intraperitoneal injection, and continuous 14 days, every day one
Secondary；
Get it filled thing A, medicine B, medicine C, medicine D and medicine E respectively, adds normal saline dilution, filters, and prepares every
1ml is containing the medicine A of crude drug about 40mg, medicine B, medicine C, medicine D and the medicine of medicine E experimental group, according to 0.02mL/g abdominal cavity
Drug administration by injection, continuous 14 days, once a day；
3) 12h, right armpit skin routine disinfection, subcutaneous vaccination S180 oncocyte liquid 0.2mL after the 3rd injection medicine
Only.Last be administered after 24h, by mouse weights, de-neck is put to death, and dissects immediately, takes out tumor, sucks blood, remove fat,
Fascia, weighs its weight in wet base with electronic balance, by following equation calculating tamor index and tumour inhibiting rate:
Tamor index (%)=tumor weight/body weight × 100%；
Tumour inhibiting rate (%)=(matched group tumor weight-experimental group tumor weight)/matched group tumor weight × 100%.
Experimental result: be shown in Table lattice 2
Form 2: medicine A, medicine B, medicine C, medicine D and medicine E experimental mice lotus tumor data
Conclusion: the data of form 2 show, the tumor of medicine A, medicine B, medicine C and medicine D experimental group is heavily substantially less than lotus tumor
Matched group (p ＜ 0.05), the tumor of medicine E experimental group does not heavily have significance (p ＞ 0.05) less than lotus tumor matched group.And medicine
The tumor of thing A experimental group is heavily substantially less than other drug experimental group (p ＜ 0.05), illustrates that the suppression tumor effect of medicine A is better than it
The medicine of his group；The tumor of medicine B experimental group be heavily substantially less than medicine C, medicine D experimental group and medicine E experimental group ((p ＜ 0.05),
The suppression tumor effect of medicine B is better than medicine C, medicine D and medicine E.
Therefore the dangshen astragalus composition 1 not removing reed head of the embodiment of the present invention 1 preparation suppresses the effect of tumor to be better than removing
Dangshen astragalus composition 2 prior art of reed head is disclosed using Radix Codonopsis and the Radix Astragali etc. as the product of raw material medical material.
Embodiment 3 dangshen astragalus composition rat long term toxicity test
Experiment: the Radix codonopsis and Radix Astragali of the present invention extracts compositions rat chronic toxicity test
Laboratory animal: SPF level SD kind rat 144, male and female half and half, body weight 80～120g, Mus age 5～7 weeks, by middle traditional Chinese medical science
Institute of lab animals of subject institute provides.
Experimental agents: the Radix codonopsis and Radix Astragali of the present invention extracts compositions, the according to embodiments of the present invention dangshen astragalus composition of 1 preparation
1；
Experimental technique:
1) it is grouped and is administered: being randomly divided into normal saline group, high dose group respectively by sex and body weight: 33.6g crude drug/kg
Dangshen astragalus composition group, middle dosage group: 16.8g crude drug/kg dangshen astragalus composition group, low dose group: 8.4g crude drug/kg dangshen astragalus composition
Group, often group 36 rats, test before male and female sub-cage rearing, 3, every cage, adaptability observation begin with after 7 days lumbar injection to
Medicine, drug withdrawal 1 time in every 7 days, successive administration 90 days, drug withdrawal 14 days.
Take the dangshen astragalus composition 1 of the embodiment of the present invention 1 preparation, add normal saline dilution, filter, prepare every 1ml and contain
The medicine of crude drug about 336mg, 168mg and 84mg.
33.6g crude drug/kg dangshen astragalus composition group, 16.8g crude drug/kg dangshen astragalus composition group, 8.4g crude drug/kg ginseng stilbene combination
Thing group is equivalent to draft clinical the 90 of people's dosage, 60,40 times.
During whole experiment, observe rat outward appearance ingest the general symptoms such as body weight growth behavior activity respectively in administration 90
My god (often group 20) and drug withdrawal 14 days (often group 16) detect that rat blood serum is biochemical, organ coefficient index, and carry out system corpse
Inspection, carries out organ coefficient detection and organs and tissues pathological examination.
2) Testing index:
Serological biochemical markers: albumin (ALB), total protein (TP), aspartate amino transferase (AST).
Organ coefficient and histopathology index: kidney, adrenal gland, testis, epididymis, uterus, ovary.
Experimental result:
1) impact on general symptom: be administered phase and convalescent period, each group rat has no because giving what campanumaea pilosula composition caused
Toxic reaction, rat all survives, and outward appearance sign, hair behavior, movable body of gland, secretion breathing, excretion etc. show no obvious abnormalities.
2) compare the change of rat food ration with matched group and have no substantially fluctuating, be shown in Table lattice 3；Male female rats each dosage group exists
In 4th week to the 8th week, there is very fast growth (being shown in Table lattice 4) in body weight, compares tool with matched group and is significantly increased (p ＜ 0.05).Push away
It is relevant that survey has benefiting action to Radix codonopsis and Radix Astragali composition.
Form 3: dangshen astragalus composition on the impact of rat food ration (unit: g/100gBW/ days,)
Form 4: dangshen astragalus composition on the impact of rat body weight (unit: g,)
3) liver function test: be administered 90 days, it is seen that male and male and female whole rat low dose group AST pole significantly reduces (p ＜
0.001), but without clinical meaning.Male rat high dose group TP, A LB significantly reduces (p ＜ 0.001), and shows dosage and depend on
Lai Xing.Middle dosage group and following dosage thereof have no obvious abnormal liver function.Drug withdrawal checks for 14 days: no abnormality seen.The results are shown in Table lattice
5。
Form 5: dangshen astragalus composition 90 days be administered after impact on rat blood serum biochemical indexes
| Group | TP(g/L) | ALB | AST(U/L) |
| Normal saline group | 55.3±1.5 | 34.1±0.6 | 77±15 |
| Low dose group | 52.3±1.3 | 30.8±1.3 | 71±16 |
| Middle dosage group | 51.1±2.1 | 29.1±1.1 | 72±15 |
| High dose group | 49.2±2.1 | 25.6±1.2 | 71±12 |
4) system becomes celestial and organ coefficient inspection: after being administered 90 days: no abnormality seen；Organ coefficient checks drug withdrawal inspection in 14 days
Look into: no abnormality seen.The results are shown in Table lattice 6,7.
Form 6: dangshen astragalus composition 90 days be administered after on the impact of Rats Organs and Tissues coefficient (g/100gBW/ days,)
Form 7: dangshen astragalus composition on the impact of convalescent period Rats Organs and Tissues coefficient (g/100gBW/ days,)
5) histopathologic examination: after being administered 90 days: check that matched group and high dose group respectively find 1 example lung local inflammation,
High dose group compares not statistically significant (P ＞ 0.05) with matched group；There is 1 example renal pelvis local inflammation in high dose group, and compares
Group compares not statistically significant (p ＞ 0.05):
Matched group and high dose group respectively find 2 examples and 1 example trachea local inflammation, but high dose group and matched group compare without system
Meaning (P ＞ 0.05) learned by meter；Remaining tissue changes without pathologic.
Drug withdrawal 14 days checks: matched group finds 1 example lung local inflammation: matched group finds 1 example trachea local inflammation: remaining group
Knit and change without pathologic.Illustrate that rat avirulence is damaged by dangshen astragalus composition drug withdrawal for 14 days.
Long term toxicity test conclusion: dangshen astragalus composition rat chronic toxicity test mainly shows as routine blood test regulating liver-QI function part
Point index occurs that physiological fluctuates, and body weight increases very fast, recovers normal, have no retardance toxic reaction after drug withdrawal.
The constituent content of existing Radix codonopsis and Radix Astragali composition product in embodiment 4 prior art
Experimental agents: medicine 1: according to Zhang Laihua, etc. the chitosan impact on ginseng stilbene decocting liquid component purification, Shi Zhenguo
Doctor's traditional Chinese medicines, the Radix codonopsis and Radix Astragali that 2010,21 (6): 1316-1318 prepare extracts compositions；
Medicine 2: according to the Radix codonopsis and Radix Astragali composition of ZL200710065396.5 embodiment 8 preparation；
Medicine 3: according to Yang Hao, etc. SHENQI FUZHENG injectable powder polishing purification technical study. Chinese Journal of New Drugs, 2009,18
(18): the method for 1771-1776 prepares Radix codonopsis and Radix Astragali composition；
The results are shown in Table lattice 8.
The contrast table (all in terms of 20g crude drug) of table 8. prior art Radix codonopsis and Radix Astragali composition based article
In upper table, the prescription (being converted into every gram) of all products is Radix Codonopsis: the Radix Astragali=1: 1.
Conclusion: the ginseng astragalus pill, medicine 1, medicine 2 and medicine 3 reference example 2 method that relate in the present embodiment carry out drug effect
Learning experiment, test result indicate that, the tumor of medicine A experimental group is heavily substantially less than ginseng astragalus pill, medicine 1, medicine 2, medicine 3 and lotus tumor pair
Matched group experimental group (p ＜ 0.05), illustrates that the suppression tumor effect of medicine A is better than ginseng astragalus pill, medicine 1, medicine 2 and medicine 3.
Embodiment 5
(1) take Radix Codonopsis (not removing reed head) 400g and Radix Astragali 400g, rinse well with deionized water, add 6400ml go from
Sub-water, adds after extracting 1 hour, releases extracting solution；Adding 4800ml deionized water, heating extraction, after 1 hour, is released and is extracted
Liquid；Add 4800ml deionized water, after heating 0.5 hour, release extracting solution.Merge after three times extracting solution filters, be concentrated into
1600ml, it is thus achieved that liquor strength make a living concentration 0.5g/mL.
(2) take D101 macroporous resin on step (1) medicinal liquid, use 70% ethanol, 5 times of volume eluting, collect mother solution and eluting
Liquid；
(3) mother solution extracting solution is concentrated into every 1ml about 1.5g Han crude drug, adds ethanol and makes alcohol content reach 75%, stands, filter,
Filtrate recycling ethanol is also concentrated into every 1ml about 3g Han crude drug, then adds ethanol and make alcohol content reach 85%, stands, filters, and filtrate is reclaimed
Ethanol is also concentrated into every 1ml about 3.8g Han crude drug, then adds ethanol and make alcohol amount reach 88%, stands, centrifugal point of seperator of alcohol deposit fluid
From, the eluent of filtrate with step (2) gained being merged, then adds filtering with microporous membrane with filter paper, filtrate recycling ethanol also concentrates
To every 1ml about 5g Han crude drug.
(4) medicinal liquid of step (3) gained injects and makes every 1ml about 2g Han crude drug with water, heating, adds total crude drug amount
The pin charcoal of 0.35%-0.45% (g/g), boils, and filters, and filtrate cold preservation, after separated machine centrifugation, through 0.2 μm micropore
Filter filter, then through ultrafiltration system filter, inject with water obtain every 1ml containing crude drug 1g Shenqi extraction.
(5) Shenqi extraction injects with water appropriate, boils 20 minutes, adds pin charcoal 0.15g, boils 10 minutes, cold
But, filtering, filtrate is standby.Take sodium chloride 6.5g, add appropriate water for injection and make dissolving, add pin charcoal 0.25g, boil, cooling,
Filtering, filtrate merges with above-mentioned standby filtrate, adds disodium edetate 0.2g, sodium pyrosulfite 0.5g and dissolves, injects and adjust with water
Whole total amount to 6.0-6.4, adds pin charcoal 0.15g-0.30g to 1000ml, regulation pH value, filters, fill, every bottle of 120ml, go out
Bacterium, obtaining crude drug content is the 20g (dangshen astragalus composition of (10g Radix Codonopsis and the 40g Radix Astragali).
Taking a dangshen astragalus composition, the related substances of detection by quantitative dangshen astragalus composition, content is as follows: lobetyolin
0.54mg, calycosin 7-O-β-D pyranglucoside 0.65mg, Radix Astragali saponin III1.62mg, Radix Astragali saponin II
0.71mg, Radix Astragali saponin I 0.47mg, astragaloside 1.02mg, pratensein-7-O-β-D-pyranglucoside 0.72mg,
Brachyoside B 0.055mg and atractylenoide Ⅲ 0.36mg.
Note: the pretreatment of resin weighs dry Di01 macroporous resin with dress post, with filling post after 95% soak with ethanol 24h,
Then with 95% ethanol eluting repeatedly, it is washed till eluent and mixes with water (1: 2) and do not produce muddiness, then with water eluting repeatedly, to washing
De-liquid is without ethanol taste, and resin no longer declines at once.
Embodiment 6
Take Shenqi extraction in 10ml embodiment 5, inject with water appropriate, add sodium chloride 2.25g, adjust with water for injection
Entirety is amassed to 125ml, adds appropriate injection charcoal, boils 20 minutes, lets cool, and filters, and sterilization is filled in low boron by 1ml volume integral
In glass ampoule, cool drying, obtain lyophilized injectable powder, obtaining crude drug content is the 20g (present invention of (10g Radix Codonopsis and the 40g Radix Astragali)
Described dangshen astragalus composition.
Taking a dangshen astragalus composition freeze-dried powder, the related substances of detection by quantitative dangshen astragalus composition, content is as follows: Radix Codonopsis alkynes
Glycosides 0.54mg, calycosin 7-O-β-D pyranglucoside 0.87mg, Radix Astragali saponin III2.41mg, Radix Astragali saponin
II1.67mg, Radix Astragali saponin I 0.47mg, astragaloside 1.53mg, pratensein-7-O-β-D-pyranglucoside
0.72mg, brachyoside B 0.13mg and atractylenoide Ⅲ 0.86mg.Laricin 0.023mg, different Radix Astragali saponin II
0.019mg, pricklyfruit licorice glycosides D 0.017mg.
Embodiment 7
In Example 5, Shenqi extraction concentrating under reduced pressure becomes extractum, relative proportion to be 1.28-1.30 (60 DEG C of surveys), is placed in
In vacuum drying oven, 50-80 DEG C of drying under reduced pressure 48-96 hour, takes out, pulverizes, cross 60-80 mesh sieve；Medicated powder is mixed with adjuvant
Uniformly, crossing 60-80 mesh sieve, make soft material with 70-90% ethanol, 20-30 mesh sieve is pelletized, and 50-80 DEG C of drying, 20-30 mesh sieve is whole
Grain, adds the magnesium stearate mix homogeneously of 0-1.0%, fills capsule, and the crude drug content controlling every seed lac capsule is 20g, has both obtained this
Invent described dangshen astragalus composition.
Taking a dangshen astragalus composition capsule, the related substances of detection by quantitative dangshen astragalus composition, content is as follows: lobetyolin
1.28mg, calycosin 7-O-β-D pyranglucoside 0.87mg, Radix Astragali saponin III 2.41mg, Radix Astragali saponin II
1.67mg, Radix Astragali saponin I 1.11mg, astragaloside 2.4mg, pratensein-7-O-β-D-pyranglucoside 1.7mg,
Brachyoside B 0.13mg, atractylenoide Ⅲ 0.86mg, laricin 0.057mg, different Radix Astragali saponin II 0.036mg, thorn
Really liquirtin D 0.035mg.
Embodiment 8
In Example 5, Shenqi extraction concentrating under reduced pressure becomes extractum, relative proportion to be 1.28-1.30 (60 DEG C of surveys), is placed in
In vacuum drying oven, 50-80 DEG C of drying under reduced pressure 48-96 hour, takes out, pulverizes, cross 60-80 mesh sieve；Medicated powder is mixed with adjuvant
Uniformly, being placed in granulator, add 6% binding agent, pelletize, rush rotary tablet machine tabletting, subpackage, the content of every is about
20g crude drug, both dangshen astragalus composition of the present invention.
Taking a piece of dangshen astragalus composition tablet, the related substances of detection by quantitative dangshen astragalus composition, content is as follows: lobetyolin
1.00mg, calycosin 7-O-β-D pyranglucoside 0.70mg, Radix Astragali saponin III 2.00mg, Radix Astragali saponin II
1.40mg, Radix Astragali saponin I 0.89mg, astragaloside 2.00mg, pratensein-7-O-β-D-pyranglucoside 1.40mg,
Brachyoside B 0.099mg and atractylenoide Ⅲ 0.65mg.
Embodiment 9
(1) taking Radix Codonopsis 400g and Radix Astragali 400g, rinse well with deionized water, add 6400ml deionized water, addition carries
After taking 1 hour, release extracting solution；Adding 4800ml deionized water, heating extraction, after 1 hour, releases extracting solution；Add
4800ml deionized water, after heating 0.5 hour, releases extracting solution.Merge after three times extracting solution filters, be concentrated into 1600ml, obtain
Liquor strength make a living concentration 0.5g/mL.
(2) take D101 macroporous resin on step (1) medicinal liquid, use 70% ethanol, 5 times of volume eluting, collect mother solution and eluting
Liquid；
(3) mother solution extracting solution is concentrated into every 1ml about 1.5g Han crude drug, adds ethanol and makes alcohol content reach 75%, stands, filter,
Filtrate recycling ethanol is also concentrated into every 1ml about 3g Han crude drug, then adds ethanol and make alcohol content reach 85%, stands, filters, and filtrate is reclaimed
Ethanol is also concentrated into every 1ml about 3.8g Han crude drug, then adds ethanol and make alcohol amount reach 88%, stands, centrifugal point of seperator of alcohol deposit fluid
From, the eluent of filtrate with step (2) gained being merged, then adds filtering with microporous membrane with filter paper, filtrate recycling ethanol also concentrates
To every 1ml about 5g Han crude drug.
(4) medicinal liquid of step (3) gained injects and makes every 1ml about 2g Han crude drug with water, heating, adds total crude drug amount
The pin charcoal of 0.35%-0.45% (g/g), boils, and filters, and filtrate cold preservation, after separated machine centrifugation, through 0.2 μm micropore
Filter filters, then filters through ultrafiltration system, is concentrated into every 1ml (40mg Radix Codonopsis and 40mg Radix Astragali) Han crude drug about 80mg,
Ginseng stilbene extracts compound.
(5) Shenqi extraction injects with water appropriate, boils 20 minutes, adds pin charcoal 0.15g, boils 10 minutes, cold
But, filtering, filtrate is standby.Take sodium chloride 6.5g, add appropriate water for injection and make dissolving, add pin charcoal 0.25g, boil, cooling,
Filtering, filtrate merges with above-mentioned standby filtrate, adds disodium edetate 0.2g, sodium pyrosulfite 0.5g and dissolves, injects and adjust with water
Whole total amount to 6.0-6.4, adds pin charcoal 0.15g-0.30g to 1000ml, regulation pH value, filters, fill, every bottle of 250ml, go out
Bacterium, obtaining crude drug content is the 20g (dangshen astragalus composition of (10g Radix Codonopsis and the 40g Radix Astragali).
Taking 1 bottle of dangshen astragalus composition tablet, the related substances of detection by quantitative dangshen astragalus composition, content is as follows: lobetyolin
1.20mg, calycosin 7-O-β-D pyranglucoside 0.80mg, Radix Astragali saponin III 2.40mg, Radix Astragali saponin II
1.60mg, Radix Astragali saponin I 1.01mg, astragaloside 2.20mg, pratensein-7-O-β-D-pyranglucoside 1.60mg,
Brachyoside B 0.11mg and atractylenoide Ⅲ 0.77mg.
Claims (8)
1. a dangshen astragalus composition, this dangshen astragalus composition includes:
2. dangshen astragalus composition as claimed in claim 1, it is characterised in that described compositions includes:
3. dangshen astragalus composition as claimed in claim 1, it is characterised in that described compositions also includes:
Laricin 0.023mg-0.057mg
Different Radix Astragali saponin II 0.019mg-0.036mg.
4. the dangshen astragalus composition as described in claim 1 or 3, it is characterised in that described compositions also includes:
Pricklyfruit licorice glycosides D 0.017mg-0.035mg.
5. dangshen astragalus composition as claimed in claim 1, it is characterised in that described compositions also includes: syringic acid, (6aR,
11aR)-3-hydroxy-9,10-dimethoxy pterocarpane, (3R)-7,2 '-dihydroxy-3 ', 4 '-dimethoxy isoflavan, Flos Caryophylli fat
One or more in glycosides.
6. the dangshen astragalus composition as described in any one of Claims 1 to 5, it is characterised in that the preparation method of described dangshen astragalus composition
Including: taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.Three extracting solution mistakes
Merge after filter, be concentrated into liquor strength and make a living concentration 0.5g/mL.D101 macroporous resin on medicinal liquid, uses 70% ethanol, 5 times of bodies
Long-pending eluting, collects mother solution and eluent；Mother liquor concentrations, to every 1ml about 1.2g Han crude drug, adds ethanol and makes alcohol content reach 75%, quiet
Putting, filter, filtrate recycling ethanol is also concentrated into every 1ml about 2g Han crude drug, then adds ethanol and make alcohol content reach 85%, stands, filtration,
Filtrate recycling ethanol is also concentrated into every 1ml about 2.5g Han crude drug, then adds ethanol and make alcohol amount reach 88%, stands, alcohol deposit fluid dish-style
Seperator centrifugation, then add filtering with microporous membrane with filter paper, filtrate recycling ethanol is also concentrated into every 1ml about 1.0g Han crude drug,
Obtain Shenqi extraction.
7. the dangshen astragalus composition as described in any one of Claims 1 to 5, it is characterised in that the preparation method of described dangshen astragalus composition
Including: taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.Three extracting solution mistakes
Merge after filter, be concentrated into liquor strength and make a living concentration 0.5g/mL.D101 macroporous resin on medicinal liquid, uses 70% ethanol, 5 times of bodies
Long-pending eluting, collects mother solution and eluent；Glucosan resin on mother solution, uses 70% ethanol, 5 times of volume eluting, collects mother solution and wash
De-liquid, merges D101 macroporous resin and the eluent of glucosan resin, is concentrated into every 1ml about 1.0g Han crude drug, must join stilbene and extract
Thing.
8. the dangshen astragalus composition as described in any one of Claims 1 to 5, it is characterised in that the preparation method of described dangshen astragalus composition
Including: taking Radix Codonopsis and the Radix Astragali, extracting in water three times, extraction time is respectively 1 hour, 1 hour, 0.5 hour.Extracting solution is concentrated into
Every 1ml about 1.2g Han crude drug, adds chitosan 17% (w/v), and temperature maintains 80 DEG C, stirs 10 minutes, uses dish-style after standing
Seperator centrifugation, then add filtering with microporous membrane with filter paper, filtrate is also concentrated into every 1ml about 1.0g Han crude drug, must join stilbene and carry
Take thing.
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| CN1546152A (en) * | 2003-12-02 | 2004-11-17 | 张正生 | Freeze-dried 'Shenqifuzheng' powder for injection and its preparing process |
| CN1686404A (en) * | 2005-04-13 | 2005-10-26 | 丽珠集团利民制药厂 | Dangshen astragalus composition for regulating immunity and its preparation method |
| CN101181619A (en) * | 2007-11-19 | 2008-05-21 | 上海复星临西药业有限公司 | Method for preparing bolus for reinforcing middle-jiao and replenishing qi |
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| CN1546152A (en) * | 2003-12-02 | 2004-11-17 | 张正生 | Freeze-dried 'Shenqifuzheng' powder for injection and its preparing process |
| CN1686404A (en) * | 2005-04-13 | 2005-10-26 | 丽珠集团利民制药厂 | Dangshen astragalus composition for regulating immunity and its preparation method |
| CN101181619A (en) * | 2007-11-19 | 2008-05-21 | 上海复星临西药业有限公司 | Method for preparing bolus for reinforcing middle-jiao and replenishing qi |
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