CN101816702B - Lung cancer preventing and treating composition extracted from selfheal and/or garden orache and application thereof in preparation of lung cancer preventing and treating medicine - Google Patents
Lung cancer preventing and treating composition extracted from selfheal and/or garden orache and application thereof in preparation of lung cancer preventing and treating medicine Download PDFInfo
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Abstract
The invention relates to lung cancer preventing and treating composition extracted from selfheal and/or garden orache and application thereof in preparation of lung cancer preventing and treating medicine. The composition comprises phenolic acid component, flavone component and triterpene component and is characterized in that the weight ratio of the phenolic acid component, flavone component and triterpene component is 5-70: 2-60: 0.5-40. In optimum proposal, the phenolic acid component takes caffeic acid and rosmarinic acid as typical ingredients; the flavone component takes rutin and quercetin as typical ingredients; the triterpene component takes ursolic acid and oleanolic acid as typical ingredients; the weight ratio of the caffeic acid, rosmarinic acid, rutin, quercetin, ursolic acid and oleanolic acid is 0-20: 5-50: 2-40: 0-20: 0.25-20: 0.25-20. The invention overcomes the defect of control is difficult in the prior art, active component is separated, purified and enriched, process is simple; preventing and treating activity on lung cancer and related diseases is strong; and mass production can be carried out.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition, be specifically related to a kind of take from Spica Prunellae and/or french spinach prevent and treat the lung cancer drugs compositions, and said composition is prevented and treated the application in the lung cancer drugs in preparation.
Background technology
According to the data that IARC announces, pulmonary carcinoma is modal disease in the interior cancer of global range.Research shows that the pulmonary carcinoma mean survival time is 6.2 months.5 years survival rates of non-operative treatment patient are 7.8%.Surgical groups patient's mean survival time is 33 months, and patients undergoing chemotherapy is 5.7 months.The sickness rate of pulmonary carcinoma and mortality rate are in propradation always, and annual speed with 0.5% increases, and go out this average level in that most of developing countries are taller.In China, pulmonary carcinoma accounts for the first place of common cancer in the city, accounts for 17.6% of all cancer mortalities.Over nearly 20 years, along with urban economy develops and the going from strength to strength of scale fast, environment runs down, and lung cancer mortality has risen more than one times.In addition, along with the increase of smoking in adolescents number, lung cancer morbidity rate of China and mortality rate continue to rise.The famous oncologist Peto prediction of Britain, if the untimely control of China's smoking and air pollution, by 2025, the annual lung cancer morbidity number of China will become the first in the world pulmonary carcinoma big country above 1,000,000.Thus, in the quite a long time from now on, the task of preventing and treating of China's pulmonary carcinoma is quite arduous.
It is labiate that Spica Prunellae belongs to medical material, and having relieves inflammation or internal heat makes eye bright, and the effect of hard masses softening and resolving mainly is used as medicine with its drying and ripening fruit ear clinically.Modern pharmacology research proof Spica Prunellae belongs to medical material and has pharmacological action widely, like antitumor, immunocompetence, antiviral, antioxidation and removing free radical etc.
Spica Prunellae belongs to medical material and contains multiple pharmacological component, mainly comprises compositions such as terpenoid and saponins thereof, phenolic acids, flavonoid, Coumarins, organic acid, volatile oil, saccharide, and wherein terpenoid, phenolic acids and flavones ingredient are the abundantest.The bibliographical information Spica Prunellae contains 28 kinds of terpenoids and saponin thereof altogether at present, is main with oleanolic acid and ursolic acid; Be main with rosmarinic acid and caffeic acid in the phenolic acid compound; Be main with rutin and Quercetin in the flavone compound.
Spica Prunellae belongs to medical material and is widely used determined curative effect as a kind of antitumor drug for a long time clinical.Spica Prunellae has multiple pharmacologically active, but its effective substance of bringing into play different pharmacological actions is also different.In addition, the material base of the effect of the anti-curing oncoma of Spica Prunellae performance is not that certain element of the first species constitutes separately, but through multi-class composition/component synergism, brings into play curative effect jointly through the many target spots of multicomponent.Therefore; The present invention is from the angle of Chinese medicine globality; Under the guidance of Chinese medical theory,, Prunella plant is prevented and treated the material base of pulmonary carcinoma and confirmed in conjunction with Chinese herb prevention pulmonary carcinoma screening system; And the compatibility proportion relation of each component in the material base carried out quality controling research, reach the best pulmonary carcinoma effect of preventing and treating.
Aspect patent documentation, publication number is that CN1559519A discloses " a kind of Spica Prunellae extract and preparation method thereof and the method for preparing that also relates to above-mentioned composition with this invention with its in treatment cyclomastopathy, thyroid, pulmonary tuberculosis, coronary heart disease, angina pectoris, hypertension, hyperglycemia, cervical erosion, application in antibiotic ".
Publication number is that CN1965896A discloses " a kind of Prunella plant extract and method for preparing and medical usage and application "; The disclosed Spica Prunellae of this patent belongs in the extract; Its active component is the prunellin compounds that contains the tuberculosis effect, and this patent also discloses this extract as the application in the tuberculosis disease medicines such as treatment pulmonary tuberculosis, bone tuberculosis, lymphoid tuberculosis.
At present, be used as pulmonary carcinoma control medicine for phenolic acids, flavonoid and triterpenes compositions in the Spica Prunellae and method of quality control is not also seen bibliographical information.
Summary of the invention
The purpose of this invention is to provide a kind of take from that Spica Prunellae belongs to Spica Prunellae and/or french spinach prevent and treat the lung cancer drugs compositions, and said composition is prevented and treated the application in the lung cancer drugs in preparation.
The scheme of accomplishing above-mentioned first invention task is: a kind of extract prevented and treated the lung cancer drugs compositions from Spica Prunellae and/or french spinach; Include phenolic acid component, flavonoid component and triterpene component in the said composition; It is characterized in that the weight ratio of said phenolic acid component, flavonoid component and triterpene component is 5～70: 2～60: 0.5～40.
Wherein, described phenolic acid component is representative composition with caffeic acid and rosmarinic acid; Described flavonoid component is representative composition with rutin and Quercetin; Described triterpene component is representative composition with ursolic acid and oleanolic acid;
Described caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 0～20: 5～50: 2～40: 0～20: 0.25～20: 0.25～20.
Further optimize, phenolic acid among the present invention: flavone: the weight ratio of triterpene is 7～30: 2～40: 2～30;
Caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 0～10: 7～20: 2～25: 0～15: 2～15: 0.25～15;
More optimize with scheme more specifically and be: phenolic acid: flavone: the weight ratio of triterpene is 8～19: 3～15: 2.5～12;
Caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 0～3: 8～16: 3～10: 0～5: 2.5～8: 0.25～4;
The optimal proportion that the application recommends is phenolic acid: flavone: the weight ratio of triterpene is 15.8: 4.8: 7.4 or 15.7: 4.9: 5.8 or 9.5: 6.4: 5.0 or 14.4: 9.5: 3.6;
Caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: oleanolic acid=1.0: 14.8: 4.0: 0.8: 5.6: 1.8 or 1.0: 14.7: 3.9: 1.0: 4.4: 1.4 or 1.0: 8.5: 5.8: 0.6: 4.1: 0.9 or 1.0: 13.4: 8.0: 1.5: 2.8: 0.8.
Crude drug described in the present invention " Spica Prunellae and/or french spinach " is meant: Spica Prunellae belongs to Spica Prunellae and/or french spinach medical material.The application recommends, and the crude drug described in the present invention " Spica Prunellae and/or french spinach " is meant that the Spica Prunellae that originates from Anhui, Guangxi, Hubei belongs to Spica Prunellae and/or originates from the french spinach medical material in Jiangsu, Henan.
The scheme of accomplishing the 2nd invention of the application task is that said extracted is prevented and treated the application in the lung-cancer medicament from the lung cancer drugs compositions of preventing and treating of Spica Prunellae and/or french spinach in preparation.
The dosage that the application recommends is: the Spica Prunellae or the french spinach extract that whenever are equivalent to 50g crude drug amount are processed the pharmaceutics common formulations and are taken.
The applicant utilizes cell and animal model that each place of production Spica Prunellae of different component structure ratio is belonged to medicinal material composition and separates the component compatibility compositions that obtains through macroporous resin and carried out medicine efficacy screening; The result all explains: Spica Prunellae and/or french spinach compositions with component structure ratio of the present invention all have pulmonary carcinoma prevention effect preferably; Mechanism of action has comprised antioxidation, cell cycle arrest and apoptosis-induced.Spica Prunellae and/or french spinach compositions with this component structure ratio will be a kind of effectively preventing lung-cancer medicaments.
Following experimental data has proved purposes and the mechanism of action of compositions of the present invention as the control lung-cancer medicament, compositions of the present invention can be applicable to pulmonary carcinoma and with the control of pulmonary carcinoma relevant disease.
1 different component structure belongs to medical material than Spica Prunellae and prevents and treats the pulmonary carcinoma effect
Choose component structure and belong to medical material than different Spica Prunellaes and prevent and treat lung cancer activity screening (seeing table 1), nine kinds of Spica Prunellaes selecting for use derive from the different genera and the different places of production respectively, prepare by same method for distilling.
Method for preparing: each each 4kg of place of production Spica Prunellae, respectively measure 95% ethanol, 60% ethanol, 30% ethanol, water reflux, extract, (2 times, 2h/ time) with 10 times successively, obtain 95% alcohol extraction, 60% alcohol extraction, 30% alcohol extraction, water and carry position solution.Each position is respectively at 60 ℃ of following concentrating under reduced pressure and reclaim solvent, and concentrated solution is in 60 ℃ of drying under reduced pressure of vacuum drying oven.Get each extract powder by the calculating of crude drug amount and mix, be prepared into need testing solution, get the different Spica Prunellae of each component structure ratio and belong to medicinal substances extract.
The table 1 nine kinds of different proportion Spica Prunellaes kind and the place of production
A cell model medicine efficacy screening
A549 anthropogenic pulmonary carcinoma cell strain, SPC-A-1 anthropogenic pulmonary carcinoma cell: all available from Chinese Academy of Sciences's Shanghai cell biological institute cell bank.
The different proportion Spica Prunellae belongs to medical material MTT experiment: take the logarithm trophophase A549 cell and SPC-A-1 cell belong to medical material with the Spica Prunellae that variable concentrations gives the different structure ratio respectively; With the positive contrast of cisplatin; Calculate growth inhibition ratio (by following formula), calculate the IC50 value.More than the experiment repetition is 3 times.
Inhibitory rate of cell growth=(matched group OD average-experimental group OD average)/matched group OD average * 100%.
As can beappreciated from fig. 1, behind the Spica Prunellae administration 48h, the shrinkage of cell cell space, refractivity weakens, and the cell endoparticle increases, and along with the increase of administration concentration, normal cell is fewer and feweri in the visual field.The negative control group cell is tight growth, and cell cell space refractivity is good.The positive controls cell presents broken state basically.
Table 2 A549 cell and SPC-A-1 cell drug effect evaluation result
Can find out that from table 2 structure is very bigger than the IC50 difference to A549 cell and SPC-A-1 cell that each different place of production Spica Prunellaes belongs to medical material.The plant extract cell screening experiment IC50≤50 μ g extract/ml that formulate according to South America antitumor drug research and development office think effective standard; Phenolic acid, flavone, triterpene component structure ratio are respectively the Spica Prunellae extract that derived from Jiangsu, Anhui, Henan, four places of production, Guangxi in 15.8: 4.8: 7.4,15.7: 4.9: 5.8,9.5: 6.4: 5.0,14.4: 9.5: 3.6 and have better inhibited pulmonary carcinoma cultivation effect, and other component structures are more not obvious than the Spica Prunellae antitumous effect that derives from Guizhou, Zhejiang, Jiangxi, four places of production, Sichuan.Can find out that from the result phenolic acid, flavone, triterpene component structure exist than big-difference than the Spica Prunellae medical material anti-lung cancer activity in each different places of production, having phenolic acid, flavone, the similar component structure of triterpene is that to derive from the french spinach and the Spica Prunellae medical material in Jiangsu, Anhui, Henan, four places of production, Guangxi in 15.8: 4.8: 7.4,15.7: 4.9: 5.8,9.5: 6.4: 5.0,14.4: 9.5: 3.6 the strongest to the proliferation inhibition activity of A549 and SPC-A-1 lung carcinoma cell than component.
Can find out from two cell model results of screening; Phenolic acid, flavone, that the triterpene component structure belongs to activity of drug ingredients than different Spica Prunellaes is inconsistent; Difference is bigger; Wherein preventing and treating lung cancer activity is to derive from the best french spinach and the Spica Prunellae medical material in Jiangsu, Anhui, Henan, four places of production, Guangxi in 15.8: 4.8: 7.4,15.7: 4.9: 5.8,9.5: 6.4: 5.0,14.4: 9.5: 3.6 for having phenolic acid, flavone, the similar component structure of triterpene than component preferably; Wherein had 15.8: 4.8: 7.4 and the Jiangsu french spinach and the Anhui Spica Prunellae of 15.7: 4.9: 5.8 component structure ratios, both ratios are the most close, and the control lung cancer activity is the strongest.Spica Prunellae and/or french spinach compositions that prompting has this phenolic acid, flavone, triterpene similar proportion have the effect of better inhibited proliferation of lung cancer cells.
B animal model medicine efficacy screening
Choose component structure than different Anhui Spica Prunellaes, Jiangsu french spinach, Sichuan bristle Spica Prunellae and Hubei Spica Prunellae, further investigate the relation of preventing and treating between pulmonary carcinoma drug effect and the component structure ratio with animal model.
(1) lewis bearing mouse model
Select the 6-8 C57 BL/6J mice (Shanghai Si Laike animal center) in age in week for use, oxter inoculation lewis lung carcinoma cell becomes tumor-bearing mice, and routine is raised the Experimental Animal Center in the Jiangsu Prov. Research Inst. Traditional Chinese Medical.Mice with tumor is divided into 11 groups at random, 8 every group, respectively behind the successive administration 14d, pull out that eyeball is got blood and dislocation is put to death, peel off the oxter tumor and weigh, calculate and respectively organize tumour inhibiting rate.
The mensuration of SOD, MDA and TNF-α in the blood plasma: measure according to the test kit recommend method.
Table 3 different component structure than Spica Prunellae compositions to lewis tumor-bearing mice drug effect result
Can find out from table 3; Compare with clinical antitumor drug cyclophosphamide group; The tumour inhibiting rate of Anhui high dose administration group, Jiangsu high dose administration group is there was no significant difference with it, and it is suitable to the tumor killing effect of lewis tumor-bearing mice with cyclophosphamide administration group to show that Anhui and the place of production, Jiangsu Spica Prunellae belong to high dose administration group; Other each administration groups therewith three groups utmost point significant difference (p≤0.01) is more all arranged.
Can find out that four places of production are followed successively by the strong drug action weak ordering of lewis tumor-bearing mice from lewis tumor-bearing mice tumour inhibiting rate experimental result relatively: (Anhui Spica Prunellae and Jiangsu french spinach)＞Sichuan bristle Spica Prunellae＞Hubei Spica Prunellae shows that also Jiangsu french spinach (1.0: 14.8: 4.0: 0.8: 5.6: 1.8) and Anhui Spica Prunellae (1.0: 14.7: 3.9: 1.0: 4.4: 1.4) with similar component structure ratio are that each place of production Spica Prunellae of different proportion belongs in the medical material best in the drug effect of lewis mice model of lung cancer.The mechanism of action of the anti-pulmonary carcinoma of this compositions also comprises enhance SOD vigor, reduces the antioxidant activity of MDA content and induces TNF-α generation etc.
(2) A/J mice chemoprophylaxis model
Select the 6-8 A/J mice in age in week, with 10mg/kg body weight dosage lumbar injection 3,4-benzopyrene-Semen Maydis oil solution is induced generation pulmonary carcinoma.Be divided into normal saline group and administration group, the administration group is irritated stomach 0.2ml with 5g Anhui Spica Prunellae compositions (1.0: 14.7: 3.9: 1.0: 4.4: 1.4)/kg body weight every day, continues medication 5 months.Take off neck and put to death mice, open the thoracic cavity, after 10% neutral formalin perfusion pulmonary, take out and be soaked in 10% neutral formalin fixedly 24h, the surperficial tuberosity number of calculating lung.
Table 4 Spica Prunellae is prevented and treated lung cancer group compound A/J mice chemoprophylaxis result
Can find out from A/J mice chemoprophylaxis model result (table 4); The pulmonary carcinoma that the Spica Prunellae compositions that had phenolic acid, flavone, triterpene component structure ratio and be 15.7: 4.9: 5.8 can effectively prevent chemical inducer to bring out takes place, and explains that it has the effect of good chemical prevention pulmonary carcinoma.
C cell cycle and apoptosis detect
The cell cycle detection kit, Annexin V-FITC apoptosis test regent box (Nanjing Kai Ji Bioisystech Co., Ltd), flow cytometer (U.S. Beeton-Diekinson company).
Get pulmonary carcinoma SPC-A-1 cell, press established procedure and cultivate, add Spica Prunellae solution (phenolic acid, flavone, triterpene component structure ratio are 15.7: 4.9: 5.8) with variable concentrations respectively, effect 72h.Collecting cell is measured cell cycle respectively by the test kit requirement and is changed and apoptosis.
Obvious variation has taken place in the cell cycle of the SPC-A-1 cell behind the Spica Prunellae effect 72h.The SPC-A-1 cell strain of administration group increased than the matched group G0/G1 phase, and the S phase reduces, and can see tangible hypodiploid peak, and along with the increase of administration concentration, apoptotic peak is more and more obvious, and matched group does not see that the hypodiploid apoptotic peak forms.Explain that Spica Prunellae can effectively influence the G1/S check point in the cell cycle, make tumor cell stagnate the phase, do not carry out dna replication dna, thereby suppress tumor proliferation and do not get into the S phase in G0/G1.The administration group obvious apoptosis peak occurred before the G1 phase, and presented dose-effect relationship, and administration group apoptosis rate rises to 28.57% from 2.07%.
The result show have phenolic acid, flavone, triterpene component structure ratio be that 15.7: 4.9: 5.8 Spica Prunellae can be through influencing cell cycle and apoptosis-induced performance antitumor action.
2 component compatibility proportionings are prevented and treated the pulmonary carcinoma effect
(1) proliferation of lung cancer cells experiment: by same procedure, separate flavone phenolic acids component and the triterpenes component that obtains after getting macroporous resin, measure each component to the SPC-A-1 cell inhibitory rate.
After acting on SPC-A-1 cell 48h, the suppression ratio result shows that the IC50 of terpenoid and phenolic acid flavonoid component is respectively 918.13 μ g crude drug/mL in the component separately, and 833.94 μ g crude drug/mL have good inhibitory effect to the SPC-A-1 cell.Both with phenolic acid, flavone, 15.7: 4.9: 5.8 ratio compatibility of triterpene component after; IC50 is 613.87 μ g crude drug/mL; Promptly strengthen in the suppression ratio to lung carcinoma cell behind this ratio compatibility; Component structure in Spica Prunellae among the present invention and/or the french spinach compositions is described than not being only applicable to extract, additive method separates these three types of components that obtain and regulates so far that ratio also has very strong control lung cancer activity.
(2) inhibition of Lewis lung cancer growth in the C57BL/6J mice body
Press preceding method and observe Spica Prunellae phenolic acids, flavonoid, triterpenes component tumour inhibiting rate the lewis tumor-bearing mice; The result can find out; When independent component; Triterpenes component 1, phenolic acid flavonoid component 2 high dose group all demonstrate good control tumor effect to the pulmonary carcinoma tumor-bearing mice, and tumour inhibiting rate is respectively 46.83 ± 8.38%, 44.91 ± 5.73%.When in after with phenolic acid, flavone, 15.7: 4.9: 5.8 ratio compatibility of triterpene component; Pulmonary carcinoma tumor-bearing mice control lung cancer activity is better than independent component; Tumour inhibiting rate is respectively 55.27 ± 5.34%, discloses and is better than one-component by behind this compatibility the pulmonary carcinoma tumor-bearing mice being prevented and treated activity.
Can find out that from the result of cell and animal model triterpenes component, the independent effect of phenolic acid flavonoid component also have certain control function of tumor.IC50 to the SPC-A-1 cell after with phenolic acid, flavone, 15.7: 4.9: 5.8 ratio of triterpene component three types of components being carried out compatibility is 613.87 μ g crude drug/mL; Tumour inhibiting rate to the lewis tumor-bearing mice is 55.27 ± 5.34%, and the Spica Prunellae/french spinach compositions that discloses with this ratio compatibility has good control activity to pulmonary carcinoma.The component structure that further specifies phenolic acid in Spica Prunellae among the present invention and/or the french spinach compositions, flavone, triterpene is not than being only applicable to extract, is applicable to that additive method separates the compositions that these three types of components that obtain ratio so far of regulating obtains yet.
The present invention to the sign chromatogram of the chemical fingerprint of Spica Prunellae and/or french spinach compositions and main active all available from the C-18 reverse-phase HPLC system.Spica Prunellae is belonged in the medical material six kinds of main active carried out the qualitative, quantitative sign.Chemical compound 1: caffeic acid; 2. rosmarinic acid; 3. rutin; 4. Quercetin; 5. oleanolic acid; 6. ursolic acid is seen Fig. 2.
The assay of caffeic acid, rosmarinic acid, rutin, Quercetin:
Instrument: Agilent 1200 high performance liquid chromatographs; Chromatographic column: Alltima C
18(4.6 * 250mm, 5 μ m) detect wavelength 350nm, 30 ℃ of column temperatures, and mobile phase is methanol (A)-0.1% glacial acetic acid gradient elution (0～10min, 25%～40% A; 10～60min, 40%～60% A; 60～70min, 60%～60%A), analysis time 70min, flow velocity 1.0mL/min.
Sample treatment: precision takes by weighing Spica Prunellae that Different Extraction Method obtains and/or french spinach extract 10.00mg in the 10mL volumetric flask, adds an amount of 60% dissolve with ethanol, ultrasonic dissolution 30min.The centrifugal 15min of 14000rpm gets supernatant, promptly gets.
The preparation of reference substance solution: the accurate respectively reference substance caffeic acid that takes by weighing constant weight; Rosmarinic acid; Rutin; Quercetin is an amount of, adds 60% alcoholic solution and processes every 1mL respectively and contain caffeic acid 77.28 μ g, rosmarinic acid 84.96 μ g, contains the reference substance solution of rutin 238.00 μ g, Quercetin 30.60 μ g.
Experimental technique and result: get reference substance and sample solution, press the analysis of chromatographic condition sample introduction, measure caffeic acid, rosmarinic acid, rutin and Quercetin peak area, calculate the content of caffeic acid, rosmarinic acid, rutin, Quercetin in the extract.
Ursolic acid, oleanolic acid assay
Instrument: Agilent 1200 high performance liquid chromatographs, chromatographic column: Alltima C
18(4.6 * 250mm, 5 μ m), wavelength 215nm, 30 ℃ of column temperatures, sample size 20 μ L.Mobile phase is methanol-0.1% phosphoric acid=85: 15, t=30min.
Sample treatment: precision takes by weighing Spica Prunellae that Different Extraction Method obtains and/or french spinach extract 10.00mg in the 10mL volumetric flask, adds an amount of 95% dissolve with ethanol, ultrasonic dissolution 30min.The centrifugal 15min of 14000rpm gets supernatant, promptly gets.
The preparation of reference substance solution: ursolic acid and the oleanolic acid reference substance of getting constant weight are an amount of, are made into every ml respectively and contain the 0.2250mg ursolic acid, the reference substance solution of 0.1790mg oleanolic acid.
Experimental technique and result: get reference substance and sample solution, press the analysis of chromatographic condition sample introduction, measure ursolic acid and oleanolic acid peak area, the content of ursolic acid and oleanolic acid in the calculating Spica Prunellae total triterpenes.
Other useful aspects of the present invention comprise:
Use avirulent, as to be applicable to further production technology and method, adopt simple ethanol stepwise gradient extraction step, obtain solvent position with biological effect.Through macroporous resin separation and concentration purification technique, adopt simple ethanol stepwise gradient separation component, obtain and the equal active component of crude drug prophylactic treatment pulmonary carcinoma.Whole processing step is simple, and yield is high, environmental friendliness.
It is not high that the present invention has overcome the prior art effective component extraction rate, and characteristics such as difficult quality control are used macroporous adsorbent resin active component is carried out separation and purification; Enrichment active component, preparation technology is easy, through compatibility; Obtain new compositions, made full use of herb resource; Prepared component mixture has stronger control activity to pulmonary carcinoma and relevant disease thereof; In addition, prepared component mixture can carry out large-scale production, the suitable new Chinese medicine of preventing and treating pulmonary carcinoma that is developed to.
For understand the present invention better, it implements advantage and specific objective that application reached thereof, should be with reference to the explanation of the attached drawings and the preferred embodiment of the invention.
Description of drawings
Fig. 1 is A549 cellular morphology figure behind the administration 24h;
Fig. 2 is HPLC figure: 1. caffeic acid; 2. rosmarinic acid; 3. rutin; 4. Quercetin; 5. oleanolic acid; 6. ursolic acid.
The specific embodiment
To combine embodiments of the invention further explain flesh and blood of the present invention below, but should notice that scope of the present invention does not receive any restriction of these instances.
Embodiment 1
Take by weighing Spica Prunellae 100g, measure 95% alcohol reflux 2 times, each 2 hours with 10 times.10 times of amounts of medicinal residues reuse, 75% alcohol reflux extracts each 2 hours 2 times.Medicinal residues are measured 40% alcohol reflux 2 times, each 2 hours with 10 times; 10 times of amounts of medicinal residues reuse, 10% alcohol reflux extracts each 2 hours 2 times; Filter respectively at four part ethanol extraction positions, and decompression recycling ethanol concentrates respectively as for getting extract.Be dissolved in water extract with appearance on the finite concentration on SP825 that handles well or HP20 macroporous adsorbent resin; Respectively with distilled water and 10% ethanol elution and discard eluent; With 50% ethanol elution of 8 times of resin bed volumes, collect 40%～60% ethanol elution, concentrating under reduced pressure; Vacuum drying gets extract I (flavone phenolic acid component); 95% ethanol elution of 10 times of resin bed volumes of reuse is collected 95% ethanol elution, concentrating under reduced pressure, and vacuum drying gets extract II (triterpene component); United extraction thing I and extract II, in the components composition of acquisition, its terpenoid, phenolic acid, the independent component ratio of flavone compatibility are all 1～100.Pulverize compositions, the conventional granulation, encapsulated, be prepared into 30 altogether.Instructions about how to take medicine: oral administration.One day 3 times, one time 3.
Phenolic acid described in this compositions: flavone: the weight ratio of triterpene is 15.8: 4.8: 7.4; Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 1.0: 14.8: 4.0: 0.8: 5.6: 1.8.
Embodiment 2, and is basic identical with embodiment 1, but wherein said phenolic acid: flavone: the weight ratio of triterpene is 9.5: 6.4: 5.0;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 1.0: 14.7: 3.9: 1.0: 4.4: 1.4.
Embodiment 3, and is basic identical with embodiment 1, but wherein said phenolic acid: flavone: the weight ratio of triterpene is or 14.4: 9.5: 3.6;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 1.0: 8.5: 5.8: 0.6: 4.1: 0.9.
Embodiment 4, and is basic identical with embodiment 1, but wherein said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 1.0: 13.4: 8.0: 1.5: 2.8: 0.8.
Embodiment 5, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 5: 2: 0.5.Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 0: 5: 2: 0: 0.25: 0.25.
Embodiment 6, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 70: 60: 40.Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 20: 50: 40: 20: 20: 20.
Embodiment 7, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 7: 2: 2;
Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 0: 7: 2: 0: 1.5: 0.5.
Embodiment 8, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 30: 40: 30;
Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 10: 20: 25: 15: 15: 15.
Embodiment 9, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 7: 2: 2;
Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 0: 7: 2: 0: 1.5: 0.5.
Embodiment 10, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 30: 40: 30;
Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 10: 20: 25: 15: 15: 15.
Embodiment 11, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 8: 3: 2.5;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 0: 8: 3: 0: 2.0: 0.5.
Embodiment 12, and is basic identical with embodiment 1, but the weight ratio of wherein said phenolic acid component, flavonoid component and triterpene component is 19: 15: 12;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 3: 16: 10: 5: 8: 4.
Claims (7)
1. an extraction prevents and treats the lung cancer drugs compositions from what Spica Prunellae belonged to the Spica Prunellae medical material, includes phenolic acid component, flavonoid component and triterpene component in the said composition, it is characterized in that,
The weight ratio of said phenolic acid component, flavonoid component and triterpene component is 5～70: 2～60: 0.5～40;
Described phenolic acid component is representative composition with caffeic acid and rosmarinic acid; Described flavonoid component is representative composition with rutin and Quercetin; Described triterpene component is representative composition with ursolic acid and oleanolic acid;
The weight ratio of described caffeic acid, rosmarinic acid, rutin, Quercetin, ursolic acid and oleanolic acid is 0～20: 5～50: 2～40: 0～20: 0.25～20: 0.25～20;
Described extraction is the compositions that obtains according to following method for preparing from the lung cancer drugs compositions of preventing and treating that Spica Prunellae belongs to the Spica Prunellae medical material:
Take by weighing Spica Prunellae 100g, measure 95% alcohol reflux 2 times, each 2 hours with 10 times; 10 times of amounts of medicinal residues reuse, 75% alcohol reflux extracts each 2 hours 2 times; Medicinal residues are measured 40% alcohol reflux 2 times, each 2 hours with 10 times; 10 times of amounts of medicinal residues reuse, 10% alcohol reflux extracts each 2 hours 2 times; Filter respectively at four part ethanol extraction positions, and decompression recycling ethanol is concentrated into dried extract respectively; Be dissolved in water extract with appearance on the finite concentration on SP825 that handles well or HP20 macroporous adsorbent resin; Respectively with distilled water and 10% ethanol elution and discard eluent; With 50% ethanol elution of 8 times of resin bed volumes, collect 40%～60% ethanol elution, concentrating under reduced pressure; Vacuum drying gets extract I--flavone phenolic acid component; 95% ethanol elution of 10 times of resin bed volumes of reuse is collected 95% ethanol elution, concentrating under reduced pressure, and vacuum drying gets extract II--triterpene component; United extraction thing I and extract II, in the components composition of acquisition, its terpenoid, phenolic acid, the independent component ratio of flavone compatibility are all 1～100.
2. extraction according to claim 1 prevents and treats the lung cancer drugs compositions from what Spica Prunellae belonged to the Spica Prunellae medical material, it is characterized in that,
The weight ratio of said phenolic acid component, flavonoid component and triterpene component is 7～30: 2～40: 2～30;
Described caffeic acid, rosmarinic acid, rutin, Quercetin, the weight ratio of ursolic acid and oleanolic acid is 0～10: 7～20: 2～25: 0～15: 2～15: 0.25～15.
3. extraction according to claim 2 prevents and treats the lung cancer drugs compositions from what Spica Prunellae belonged to the Spica Prunellae medical material, it is characterized in that,
The weight ratio of said phenolic acid component, flavonoid component and triterpene component is 8～19: 3～15: 2.5～12;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 0～3: 8～16: 3～10: 0～5: 2.5～8: 0.25～4.
4. extraction according to claim 3 prevents and treats the lung cancer drugs compositions from what Spica Prunellae belonged to the Spica Prunellae medical material, it is characterized in that,
Said phenolic acid: flavone: the weight ratio of triterpene is 15.8: 4.8: 7.4 or 15.7: 4.9: 5.8 or 9.5: 6.4: 5.0 or 14.4: 9.5: 3.6;
Said caffeic acid: rosmarinic acid: rutin: Quercetin: ursolic acid: the weight ratio of oleanolic acid is 1.0: 14.8: 4.0: 0.8: 5.6: 1.8 or 1.0: 14.7: 3.9: 1.0: 4.4: 1.4 or 1.0: 8.5: 5.8: 0.6: 4.1: 0.9 or 1.0: 13.4: 8.0: 1.5: 2.8: 0.8.
5. prevent and treat the lung cancer drugs compositions according to the described extraction of one of claim 1～4 from what Spica Prunellae belonged to the Spica Prunellae medical material, it is characterized in that described crude drug " Spica Prunellae " is meant that the Spica Prunellae that originates from Anhui, Guangxi belongs to the Spica Prunellae medical material.
6. the described extraction of claim 1 prevents and treats the application in the lung-cancer medicament from the lung cancer drugs compositions of preventing and treating that Spica Prunellae belongs to the Spica Prunellae medical material in preparation.
7. extraction according to claim 6 prevents and treats the application in the lung-cancer medicament from the lung cancer drugs compositions of preventing and treating that Spica Prunellae belongs to the Spica Prunellae medical material in preparation; It is characterized in that described dosage is: the extract that is equivalent to the Spica Prunellae of 50g crude drug amount is processed regular dosage form.
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| CN107441160B (en) * | 2017-08-22 | 2021-04-16 | 湖南中医药大学 | Pharmaceutical composition containing selfheal polysaccharide, triterpene and volatile oil and application thereof |
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| CN1965896A (en) * | 2006-11-03 | 2007-05-23 | 深圳清华大学研究院 | Prunella plant extract, preparation method, medical use and application thereof |
| CN101317883A (en) * | 2007-06-06 | 2008-12-10 | 上海中医药大学附属曙光医院 | Prunella spike active site and application of the same in preparing medicament composition |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1965896A (en) * | 2006-11-03 | 2007-05-23 | 深圳清华大学研究院 | Prunella plant extract, preparation method, medical use and application thereof |
| CN101317883A (en) * | 2007-06-06 | 2008-12-10 | 上海中医药大学附属曙光医院 | Prunella spike active site and application of the same in preparing medicament composition |
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