CN1775267A - Kaikoujian extract, Its preparing method and use - Google Patents
Kaikoujian extract, Its preparing method and use Download PDFInfo
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- CN1775267A CN1775267A CNA2005101008321A CN200510100832A CN1775267A CN 1775267 A CN1775267 A CN 1775267A CN A2005101008321 A CNA2005101008321 A CN A2005101008321A CN 200510100832 A CN200510100832 A CN 200510100832A CN 1775267 A CN1775267 A CN 1775267A
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Abstract
The present invention relates to a Chinese medicine tupistra chinensis extract, its preparation method and application. Said extract is obtained by extracting root, stem and leaf of tupistra chinensis, and is a brownish yellow powder. The characteristics of its chemical identification are as follows: Liebermann-Burchard reaction and Molish reaction are positive; the foam test is positive reaction, the foam height of alkaline liquor tube is higher than that of acid liquor tube; Rosen-Heimer reaction at 60deg.C can produce violet red color reaction, it is a steroid saponin compound. Said extract has the action of resisting tumor, resisting inflammation, inhibiting platelet agglutination and resisting thrombosis.
Description
Invention field
The present invention relates to the field of Chinese medicines, particularly a kind of Chinese medicine extract and its production and application is specifically related to Tupistra chinensis Bak. extract and its production and application.
Background technology
Tupistra chinensis Bak. belongs to (Tupistra) plant and is under the jurisdiction of Liliaceae (Liliaceae) Herba Convallariae family (Convallarieae).The Tupistra chinensis Bak. platymiscium has kind more than 20, and China is world's Tupistra chinensis Bak. platymiscium center of distribution, and kind accounts for 70% of the whole world.Tupistra chinensis Bak. is that the very high plant of Chinese endemic element belongs to, and mainly is distributed in southern areas such as China Sichuan, Hubei, Yunnan, Zhejiang, Guangxi, Jiangxi, Fujian.
The Tupistra chinensis Bak. platymiscium is China's medication among the people, how to be used as medicine with rhizome and Herb.Traditional Chinese medical science traditional theory is thought this platymiscium hardship, suffering, and is cold in nature, poisonous, has effects such as heat-clearing and toxic substances removing, expelling wind and removing dampness, eliminating stasis to stop pain, cures mainly diphtheria, laryngopharynx swelling and pain, rheumatic arthralgia, traumatic injury, stomachache, carbuncle sore tumefacting virus, poisonous snake lyssodexis etc.
The Tupistra chinensis Bak. platymiscium of existing at present research report has Tupistra chinensis Bak. (Tupistra chinensis Baker), curved stamen Tupistra chinensis Bak. (Tupistra wattii Hook.f.), tooth lobe Tupistra chinensis Bak. (Tupistra fimbriata Hand.-Mazz.) and Hemerocallis citrina Baroni Tupistra chinensis Bak. kinds more than 10 such as (Tupistra aurantiaca Wall.).Studies show that the contained chemical constituent of Tupistra chinensis Bak. platymiscium mostly is steroidal compounds, all contain steroid saponin (" Chinese wild plant resource ", 1999,19 (5): 59-62).Shen equality has been carried out separation, purification to the ethanol extraction of curved stamen Tupistra chinensis Bak., and measures the cytotoxic activity of the chemical compound that obtains with tumor cell K562 that cultivates and A2780a.Found that curved stamen ruscogenin B (wattigenin B), curved stamen ruscogenin C (Wattigenin C), steroid sapogenines (kitigenin) and convallagenin B (convallagenin B), and cardiotonic glycoside rhodexin A, wattoside F, wattoside E etc. have cytotoxic activity (" time precious traditional Chinese medical science traditional Chinese medicines " 2005,15 (12): 860-861).Taiwan's scholars Pan Wen refined (Pan WB) etc. produce Tupistra chinensis Bak. to the continent and have carried out similar research, find that steroid sapogenines Δ 25 (27)-pentrogenin, ranmogenin A etc. have cytotoxic activity (J.Nat.Prod. to the gastric carcinoma cells of cultivating (NUGC), 2003,66 (2): 161-168).Yet the antitumor action of the chemical compound of these Tupistra chinensis Bak. platymisciums is the result that the cell in vitro cytotoxic activity is measured, and the result who whether has antitumor action still not have experimentation at present in the body is confirmed.It once is principal agent with the Tupistra chinensis Bak. that old state just waits, and forms prescription QE-8407, has carried out anticancer pharmacology and clinical trial, and confirming has inhibitory action (" Yunnan University's journal " (natural science edition), 1989,1: 55) to tumor strains such as W256.But whether independent use has antitumor action for Tupistra chinensis Bak., and what chemical compound has problems such as antitumor action not do further research.In addition, there are some researches show also that Tupistra chinensis Bak. eliminates the phlegm, antiinflammatory and antibacterial (" folks of china medical magazine ", 2005,2:103-106), and effect (" Changjiang University's journal " (natural science edition), 2005,2 (2): 77-82) such as antifungal.These researchs are object with the methanol crude extract of Tupistra chinensis Bak. all, do not determine the compound property of Tupistra chinensis Bak. active component.
Summary of the invention
The object of the invention is to provide a kind of Tupistra chinensis Bak. extract.
Another purpose of the present invention provides a kind of Tupistra chinensis Bak. preparation method of extract.
Another purpose of the present invention provides a kind of pharmaceutical composition that contains the Tupistra chinensis Bak. extract.
Another purpose of the present invention provides the application of Tupistra chinensis Bak. extract in preparation treatment antitumor, antiinflammatory and anticoagulant medicine.
Tupistra chinensis Bak. extract of the present invention is that (Tupistra chinenese, Baker) extraction obtains from Liliaceae (Liliaceae) Tupistra chinensis Bak. genus (Tupistra) plant Tupistra chinensis Bak..
Tupistra chinensis Bak. of the present invention can be its rhizome, leaf or herb.
Tupistra chinensis Bak. extract of the present invention is a steroid oside compound, is pale brown toner powder.Being characterized as of its chemical identification: Liebermann-Burchard reaction, Molish reaction all are positive; Foam stability test is positive, and the foam height of alkali liquor pipe is than the foam height of acid solution pipe; Rosen-Heimer presents the aubergine chromogenic reaction when being reflected at 60 ℃.
Tupistra chinensis Bak. method for preparing extractive of the present invention can adopt oside compound separating and extracting process commonly used.
Tupistra chinensis Bak. method for preparing extractive of the present invention contains following steps: a prepares crude extract; The b purification.
Tupistra chinensis Bak. method for preparing extractive step a of the present invention can use Chinese medicine crude extract preparation method commonly used.For example the Tupistra chinensis Bak. water can be decocted, or obtain containing the Tupistra chinensis Bak. crude extract of Saponin with the 1-4 carbon alcoholic solution reflux Tupistra chinensis Bak. of variable concentrations.1-4 carbon alcohol of the present invention can be methanol, ethanol, propanol, isopropyl alcohol, n-butyl alcohol wherein one or more, preferably methanol and ethanol are wherein a kind of.
In the Tupistra chinensis Bak. method for preparing extractive of the present invention, Tupistra chinensis Bak. can use its rhizome, leaf or herb.
The Saponin purification process that Tupistra chinensis Bak. method for preparing extractive step b of the present invention can adopt those skilled in the art to use always.
Tupistra chinensis Bak. method for preparing extractive of the present invention, purification process can adopt extraction: the Tupistra chinensis Bak. crude extract is concentrated the extractum that obtains be dissolved in the water, extract with 4 or 5 carbon alcohol; Extract reclaims the Tupistra chinensis Bak. extract that the solvent after drying obtains containing total Saponin.Of the present invention 4 or 5 carbon alcohol can be n-butyl alcohol, isobutanol, n-amyl alcohol wherein one or more, n-butyl alcohol preferably.
In above-mentioned abstraction purification method, Tupistra chinensis Bak. crude extract extractum aqueous solution can be used organic solvent degreasing earlier.Described organic solvent can be chloroform, ethyl acetate, ether, petroleum ether wherein one or more.
In above-mentioned abstraction purification method, obtaining extract can precipitate with acetone or ether etc., reclaims precipitate, obtains the higher total Saponin of purity with this.
Tupistra chinensis Bak. method for preparing extractive of the present invention, purification process can also adopt chromatography.Chromatography of the present invention can be that macroporous resin chromatography, silica gel chromatography, alumina column chromatography method are wherein a kind of.Chromatography of the present invention is the macroporous resin chromatography preferably
Macroporous resin chromatography concrete steps of the present invention are: on the Tupistra chinensis Bak. crude extract behind the macroporous resin chromatographic column, after elder generation's water is eluted to the no polysaccharide chromogenic reaction of phenolsulfuric acid method mensuration, carry out eluting with ethanol then, collect the male position of Saponin chromogenic reaction, concentrate drying obtains the Tupistra chinensis Bak. extract.
Macroporous resin of the present invention is nonpolar or low pole polystyrene macroporous resin, for example HPD-100, D-101, AB-8, HP-20, HPD-300 etc.Macroporous resin of the present invention is the HPD-100 macroporous resin preferably.
The pharmaceutical composition that contains the Tupistra chinensis Bak. extract of the present invention contains the Tupistra chinensis Bak. extract and the adjuvant or the excipient commonly used for the treatment of effective dose.Can obtain the Tupistra chinensis Bak. extract according to method known to those skilled in the art and preferably use consumption and form.Those skilled in the art can obtain the type of service and the consumption of used adjuvant, excipient according to methods known in the art.
The pharmaceutical composition that contains the Tupistra chinensis Bak. extract of the present invention adopts this area preparation method commonly used.For example promptly with above-mentioned Tupistra chinensis Bak. extract and pharmaceutical adjuvant mix homogeneously.
The pharmaceutical composition that contains the Tupistra chinensis Bak. extract of the present invention, its dosage form can be any in injection, tablet, pill, capsule, electuary, oral liquid, suspension, spray, externally-applied liniment, the cutaneous permeable agent.Those skilled in the art can obtain used dosage form according to methods known in the art.
Tupistra chinensis Bak. extract of the present invention has antitumor action, antiinflammatory action and anticoagulant, anti thrombotic action.For a better understanding of the present invention, with relevant pharmacological testing of the present invention and result its purposes is described below.
One, antitumor action
1. the activity of the external tumor of the total Saponin of Tupistra chinensis Bak.
The total Saponin I of Tupistra chinensis Bak. of embodiment 1 preparation is mixed with 2mg/mL solution with phosphate buffer solution (PBS), and after the filtration sterilization, the aseptic PBS of reuse is diluted to 20 μ g/mL, 40 μ g/mL, 80 μ g/ μ L, 160 μ g/mL, 320 μ g/mL.
K562: human leukemia cell; HepG2: human liver cancer cell; LoVo: human large intestine cancer cell; C6: rat neuroglial cytoma; MCF7: human breast cancer cell; All cultivate, by 2.0 * 10 with the DMEM that adds 10% new-born calf serum
3Individual/hole is inoculated in 96 orifice plates, volume 90 μ L.Cultivate after 48 hours, every hole adds the extract 10 μ L of variable concentrations, and promptly final concentration is respectively 2 μ g/mL, 4 μ g/mL, 8 μ g/mL, 16 μ g/mL, 32 μ g/mL.Blank is PBS, and matched group and medicine group are all established 6 parallel holes.Continue to cultivate 48 hours, the adding volume ratio is 1: 10 a CCK-8 reagent, hatches 2.5 hours, measures 450nm absorbance (OD value) on the microplate reader, and reference wavelength is 630nm.Be calculated as follows suppression ratio: suppression ratio (%)=[1-(the average OD value of the medication group/average OD value of blank group)] * 100%, and according to the half-inhibition concentration (IC of the total Saponin of suppression ratio calculating Tupistra chinensis Bak. to various tumors
50), the results are shown in Table 1.
The total Saponin I of table 1. Tupistra chinensis Bak. is to the tumor cell proliferation inhibition effect
| Concentration (μ g/mL) | Suppression ratio (%) | ||||
| K562 | LoVo | HepG2 | C6 | MCF7 | |
| 2 4 8 16 32 | 27.4 47.9 61.2 71.9 82.2 | 28.6 50.9 63.0 71.7 84.9 | 41.4 51.8 66.5 81.6 92.3 | 20.1 38.1 56.3 61.4 79.0 | 44.8 57.9 73.6 87.9 93.2 |
| IC 50(μg/mL) | 5.22 | 4.72 | 3.37 | 7.68 | 2.65 |
Total Saponin III and separated part B, the C of the total Saponin II of Tupistra chinensis Bak. of embodiment 2 preparation and embodiment 3 preparations, (position A separates because of indissoluble D, and content is few, do not carry out this test) be mixed with 2mg/mL solution with phosphate buffer solution, after the filtration sterilization, the aseptic PBS of reuse is diluted to 20 μ g/mL, 40 μ g/mL, 80 μ g/mL, 160 μ g/mL.The same method is measured the activity that each sample suppresses the HepG2 cell proliferation.The results are shown in Table 2.The result shows that the total Saponin of Tupistra chinensis Bak. and each separated part thereof all have inhibitory action to the HepG2 tumor cell proliferation, and strong with acting as of polarity position B on the low side.
Total Saponin of table 2. Tupistra chinensis Bak. and separated part are to the effect of HepG2 tumor cell proliferation inhibition
| Concentration (μ g/mL) | Total Saponin II | Total Saponin III | Position B | Position C | Position D |
| 2 4 8 16 | 21.2 42.9 62.7 88.9 | 17.4 38.6 52.3 79.8 | 52.9 78.8 96.5 98.2 | 8.5 26.7 41.7 66.9 | 2.3 10.1 23.4 42.1 |
| IC 50(μg/mL) | 4.87 | 6.27 | ＜2.0 | 9.55 | 19.24 |
2, the activity of the total Saponin in-vivo tumour of Tupistra chinensis Bak.
The total Saponin I of the Tupistra chinensis Bak. physiological saline solution of embodiment 1 preparation is mixed with the solution of 0.1g/mL and 0.2g/mL, filtration sterilization, and-20 ℃ of preservations are faced with before thawing after the packing.
With the S that inoculates 7 days
180Ascites of sarcoma mice and normal saline dilute in 1: 1 ratio, the right groin subcutaneous injection of mice 0.2mL diluent, and press the body weight random packet: 11 of normal saline matched groups, two groups number of animals is 10 in addition.
The calculating of medication and tumour inhibiting rate: beginning administration in the 2nd day after the mouse inoculation tumor, once a day, continuous 8 days.Dosage is respectively: ring phosphamide (CTX) group 20mg/kg lumbar injection, and the total Saponin of Tupistra chinensis Bak. is pressed 0.2mL/10g body weight gastric infusion, and dosage is respectively 2.0g/kg and 4.0g/kg.Put to death mice, and peeled off the tumor piece and weigh in the 2nd day after the last administration.Calculate tumour inhibiting rate by " tumour inhibiting rate=(1-administration group tumor weight/normal saline matched group tumor is heavy) * 100% " formula.Experimental result sees Table 3.
The total Saponin of table 3. Tupistra chinensis Bak. is to the inhibitory action of S180 tumor-bearing mice tumor growth
| Grouping | Number of animals | Tumor heavy (g) | Suppression ratio |
| Normal saline contrast CTX (20mg/kg) Tupistra chinensis Bak. (2.0g/kg) Tupistra chinensis Bak. (4.0g/kg) | 11 10 10 10 | 2.14±0.61 0.81±0.41** 1.19±0.53** 1.02±0.58** | 62.1％ 44.4％ 52.3％ |
Annotate: compare * * p＜0.01 with the normal saline matched group.
Two, antiinflammatory action
The total Saponin III of the Tupistra chinensis Bak. physiological saline solution of embodiment 3 preparation is mixed with the solution of 0.1g/mL and 0.2g/mL, filtration sterilization, and-20 ℃ of preservations are faced with before thawing after the packing.
Bring out the antiinflammatory action of the total Saponin of inflammatory model check Tupistra chinensis Bak. of mice auricle swelling with dimethylbenzene: 48 of mices, male and female half and half are divided into 4 groups at random.Positive controls lumbar injection aspirin 150mg/kg, the total Saponin III of Tupistra chinensis Bak. is with 0.2ml/10g body weight gastric infusion, and negative control group is given with the volume normal saline and is irritated stomach.Every day 1 time, continuous 7 days.After the last administration 2 hours, each group mouse right ear upper and lower faces is smeared dimethylbenzene 20 μ l respectively, left ear is not coated with in contrast, puts to death mice after 1 hour, cuts ears, sweeps away auricle with the card punch of 8mm diameter, and balance is weighed.Deduct left ear weight as the swelling degree with auris dextra weight, calculate inhibitory rate of intumesce as follows: inhibitory rate of intumesce=(negative control group difference-administration group difference)/negative control group difference * 100%.The results are shown in Table 4.The result shows that the total Saponin of Tupistra chinensis Bak. can suppress the mice caused by dimethylbenzene xylene auricle edema, has significant antiinflammatory action.
The total Saponin III of table 4. Tupistra chinensis Bak. xylol causes the influence of mice auricle swelling
| Group | Number of animals | Swelling degree (mg) | Suppression ratio |
| Normal saline aspirin (150mg/kg) Tupistra chinensis Bak. (2.0g/kg) | 12 12 12 | 13.13±4.18 8.17±2.54** 10.14±3.61 | 37.8％ 22.8％ |
| Tupistra chinensis Bak. (4.0g/kg) | 12 | 9.10±3.04* | 30.7％ |
Annotate: compare * p＜0.05, * * p＜0.01 with the normal saline matched group.
Three, antiplatelet aggregative activity
The total Saponin I of the Tupistra chinensis Bak. physiological saline solution of embodiment 1 preparation is mixed with the solution of 5mg/mL, 10mg/mL, 20mg/mL, 40mg/mL.
External anti-platelet aggregation effect: the rabbit auricular vein is got blood, the anticoagulant in 1: 9 of 3.8% sodium citrate.900 rev/mins centrifugal 8 minutes, getting the upper strata, to be rich in platelet blood plasma standby.The blood plasma of drawing PRP is continued centrifugal, 3000 rev/mins centrifugal 20 minutes, get platelet poor plasma.Use the turbidimetry for Determination platelet aggregation rate.Get and be rich in platelet blood plasma 0.27mL and add in the test cup, add the total Saponin I of 30 μ L Tupistra chinensis Bak., matched group is with 10 μ L physiologic saline for substitute, 37 ℃ hatch 2 minutes after, add the gathering of Adenosine diphosphate (ADP) induced platelet.
The result shows that the total Saponin of Tupistra chinensis Bak. has tangible antiplatelet aggregative activity, and is dose-effect relationship.Calculate platelet aggregation inhibition rate with the platelet maximum agglutination rate, calculate as follows: suppression ratio=(matched group aggregation rate-administration group aggregation rate)/matched group aggregation rate * 100%.The results are shown in Table 5.The IC of the total Saponin anticoagulant of Tupistra chinensis Bak.
50Be 3.18mg/mL.
The total Saponin I of table 5. Tupistra chinensis Bak. antiplatelet aggregative activity
| Group | Number of times (n) | Maximum agglutination rate | Suppression ratio |
| Normal saline Tupistra chinensis Bak. (0.5mg/mL) Tupistra chinensis Bak. (1.5mg/mL) Tupistra chinensis Bak. (2.0mg/mL) | 5 5 5 5 | 41.79±7.26 36.45±7.36 31.52±5.83* 24.35±5.21** | 12.8％ 24.6％ 41.7％ |
| Tupistra chinensis Bak. (4.0mg/mL) | 5 | 11.87±4.42** | 53.6％ |
Annotate: compare * p＜0.05, * * p＜0.01 with the normal saline matched group.
Major advantage of the present invention has:
1. the total Saponin of Tupistra chinensis Bak. provided by the invention has effects such as definite antitumor, antiinflammatory and anticoagulant, and value for clinical application is preferably arranged.
2. the preparation of the total Saponin of Tupistra chinensis Bak. of the present invention can be adopted methods such as extraction and resin chromatography, and easy to operation, the reaction condition gentleness is suitable for and applies.
3. Tupistra chinensis Bak. raw medicinal material of the present invention is comparatively common in province, China south, and the medicine source is wider, is easy to realize industrialization.
The specific embodiment
Following examples make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Embodiment 1
Tupistra chinensis Bak. rhizome thin slice (2～4mm) 0.2 kilograms, add 1.5 premium on currency, to soak after 6 hours, heating decocted 2 hours, and the water outlet decocting liquid inclines; Add 1.5 liters in water then, continue to decoct 1.5 hours, the water outlet decocting liquid that inclines adds 1.5 liters in water again, decocts 1.5 hours with quadrat method; Merge 3 times decocting liquid, filter, standby.
The HPD-100 macroporous resin column chromatography: HPD-100 macroporous resin bed volume is 4.5cm (internal diameter) * 58cm (height), about 920mL.Sample on the above-mentioned Tupistra chinensis Bak. decocting liquid is used the distilled water eluting, about 1.8 liters/hour of flow velocity.Behind about 5 liters of distilled water flushings, effluent detects the polysaccharide reaction with sulfuric acid-phynol method and is light yellow; Then with 2.5 liters of eluting of 20% ethanol, 3 liter of 70% ethanol elution of reuse.Collect the position of 70% ethanol elution, detection Liebermann-Burchard reaction, Molish reaction all are positive, and are illustrated as Saponin class material.The position heat drying of 70% ethanol elution obtains total Saponin I, about 20.1 grams of weighing, and yield is 10.0%.
Contain drug regimen composition formula of Tupistra chinensis Bak. extract and preparation method thereof:
Tablet: 1000 amounts (200mg/ sheet)
(1) prescription:
Above-mentioned Tupistra chinensis Bak. extract 200g
Starch 40g
Dextrin 40g
Stearic acid 6.4g
(2) preparation method:
According to above-mentioned prescription mixing → every heavily about 300mg of an amount of alcohol granulation → 60-80 ℃ oven dry → tabletting sieves → add.
Embodiment 2
(2～4mm) 0.2 kilograms of Tupistra chinensis Bak. rhizome thin slices, use 1.5 liters 95% ethanol, 75% ethanol and 50% ethanol respectively successively, heating and refluxing extraction 2 hours, merge extractive liquid,, ethanol is reclaimed in distilling under reduced pressure, and resulting extractum adds the dissolving of 400 ml distilled waters, place in the separatory funnel, add 200 milliliters of petroleum ether extractiones 3 times, vibrated 5 minutes at every turn, left standstill 0.5 hour; Lower aqueous solution vibrated 5 minutes with the saturated n-butanol extraction of 200 ml waters 4 times at every turn, left standstill 1 hour.Merge n-butanol extraction solution, n-butyl alcohol is reclaimed in distilling under reduced pressure, and residue dissolves with 50 ml methanol, add 500 milliliters of acetone, standing over night, carefully inclining upper solution, precipitate Liebermann-Burchard reaction after testing, Molish reaction all are positive, and are Saponin class material.60 ℃ of vacuum dryings of precipitate obtain total Saponin II, 22.8 grams of weighing, and yield is 11.4%.
Contain drug regimen composition formula of Tupistra chinensis Bak. extract and preparation method thereof: 1000 bottles of amounts of freeze-dried powder (injection) (100mg/ props up)
(1) prescription:
Above-mentioned Tupistra chinensis Bak. extract 100g
Active carbon 25g
Water for injection is to 5000ml
(2) preparation method:
With water for injection the dissolving of total Saponin is added active carbon → mixing → be heated to 80 ℃ * 30min → buchner funnel to filter → 0.2 μ m membrane filtration degerming → packing, every bottle of 5ml → partly cover plug → lyophilization → gland.
Embodiment 3
(2～4mm) 0.2 kilograms of Tupistra chinensis Bak. rhizome thin slices, add 1.5 liters of methanol, heating and refluxing extraction 2 hours is extracted merge extractive liquid, repeatedly 4 times, methanol is reclaimed in distilling under reduced pressure, resulting extractum adds the dissolving of 400 ml distilled waters, places in the separatory funnel, adds 200 milliliters of petroleum ether extractiones 3 times, each vibration 5 minutes was left standstill 0.5 hour; Lower aqueous solution vibrated 5 minutes with the saturated n-butanol extraction of 200 ml waters 4 times at every turn, left standstill 1 hour.Merge n-butanol extraction solution, n-butyl alcohol is reclaimed in distilling under reduced pressure, and residue Liebermann-Burchard reaction after testing, Molish reaction all are positive, and for Saponin class material, promptly obtain total Saponin, and 25.4g weighs.Get the total Saponin dissolve with methanol of 12g, stir with H type tlc silica gel, air-dry standby.
Get the above-mentioned silica gel of mixing sample, place the dried post of H type tlc silica gel (2.6 * 73cm) tops, highly about 5.3cm.Analyse eluting with the water saturation n-butanol layer, thin layer chromatography result according to effluent, collected A, B, four positions of C, D by effusive order (polarity grows from weak to strong), the Liebermann-Burchard at four positions reaction after testing, Molish reaction all are positive, and are Saponin class material.N-butyl alcohol is reclaimed in distilling under reduced pressure, and residue is weighed, and A is 0.17g, and B is that 0.87g, C are that 5.29g, D are 2.60g.Gross weight is 8.93g.
Contain drug regimen composition formula of Tupistra chinensis Bak. extract and preparation method thereof:
Capsule: 1000 amounts (200mg/ grain)
(1) preparation method:
Above-mentioned Tupistra chinensis Bak. extract 200g
(2) method for making:
Raw material is distributed into capsule, every about 300mg.
Claims (11)
1. Tupistra chinensis Bak. extract, be to belong to (Tupistra) plant Tupistra chinensis Bak. (Tupistra chinensis from Liliaceae (Liliaceae) Tupistra chinensis Bak., Baker) Chinese medicine extract that obtains through extraction of rhizome, leaf or herb, it is characterized in that Chinese medicine extract is a steroid oside compound, described extract prepares by following steps: at first Tupistra chinensis Bak. rhizome, leaf or herb are obtained crude extract behind pure reflux or decocting; Then the Tupistra chinensis Bak. crude extract being concentrated the extractum that obtains is dissolved in the water, extract with 4 or 5 carbon alcohol, extract obtains the Tupistra chinensis Bak. extract after reclaiming solvent, or with behind the macroporous resin chromatographic column on the Tupistra chinensis Bak. crude extract, elder generation's water is eluted to the phenolsulfuric acid method and measures no polysaccharide chromogenic reaction, carry out eluting with ethanol then, obtain the Tupistra chinensis Bak. extract behind the concentrate drying of the male position of collection Saponin chromogenic reaction.
2. according to the described Tupistra chinensis Bak. extract of claim 1, it is characterized in that preparing the used alcohol of Tupistra chinensis Bak. crude extract is ethanol or methanol.
3. according to the described Tupistra chinensis Bak. extract of claim 1, it is characterized in that extracting used alcohol is n-butyl alcohol.
4. according to the described Tupistra chinensis Bak. extract of claim 1, it is characterized in that macroporous resin is the HPD-100 macroporous resin.
5. contain following steps according to one of them described Tupistra chinensis Bak. preparation method of extract of claim 1 to 4: water decocts Tupistra chinensis Bak., or obtains the Tupistra chinensis Bak. crude extract with the 1-4 carbon alcoholic solution reflux Tupistra chinensis Bak. of variable concentrations; Then with the separation and purification of Tupistra chinensis Bak. crude extract.
6. according to the described Tupistra chinensis Bak. preparation method of extract of claim 5, it is characterized in that the separation and purification of Tupistra chinensis Bak. crude extract is with behind the HPD-100 macroporous resin chromatographic column on the Tupistra chinensis Bak. crude extract, elder generation's water is eluted to the phenolsulfuric acid method and measures no polysaccharide chromogenic reaction, carry out eluting with ethanol then, obtain the Tupistra chinensis Bak. extract behind the concentrate drying of the male position of collection Saponin chromogenic reaction.
7. according to the described Tupistra chinensis Bak. preparation method of extract of claim 5, it is characterized in that the separation and purification of Tupistra chinensis Bak. crude extract is the Tupistra chinensis Bak. crude extract to be concentrated the extractum that obtains be dissolved in the water, use n-butanol extraction, extract obtains the Tupistra chinensis Bak. extract after reclaiming solvent.
8. pharmaceutical composition contains one of them described Tupistra chinensis Bak. extract of claim 1 to 4 for the treatment of effective dose and acceptable adjuvant pharmaceutically.
9. according to the application of one of them described Tupistra chinensis Bak. extract of claim 1 to 4 in preparation treatment antitumor, antiinflammatory and anticoagulant medicine.
10. according to the application of one of them described Tupistra chinensis Bak. extract of claim 1 to 4 in preparation treatment antitumor drug.
11. according to the application of the described pharmaceutical composition of claim 8 in preparation treatment antitumor, antiinflammatory and anticoagulant medicine.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103520445A (en) * | 2013-10-28 | 2014-01-22 | 武汉大学 | Application of Tupistra chinensis in preparation of antitumor drugs |
| CN103751486A (en) * | 2014-01-13 | 2014-04-30 | 姚娟 | Traditional Chinese medicine composition for treating lung cancer |
| CN105859825A (en) * | 2016-05-06 | 2016-08-17 | 广东药学院 | Spirostanol saponins and application thereof |
| CN105924494A (en) * | 2016-05-06 | 2016-09-07 | 广东药学院 | furostanol saponin compounds and application thereof |
| CN106509281A (en) * | 2016-12-21 | 2017-03-22 | 神农架林区中医医院 | campylandra chinensis sore-throat relieving tea and preparation method thereof |
| CN107361403A (en) * | 2017-08-07 | 2017-11-21 | 武汉纺织大学 | One kind contains Extracts of Tupistra chinensis additive in cigarette and preparation method thereof |
-
2005
- 2005-11-03 CN CNA2005101008321A patent/CN1775267A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103520445A (en) * | 2013-10-28 | 2014-01-22 | 武汉大学 | Application of Tupistra chinensis in preparation of antitumor drugs |
| CN103751486A (en) * | 2014-01-13 | 2014-04-30 | 姚娟 | Traditional Chinese medicine composition for treating lung cancer |
| CN105859825A (en) * | 2016-05-06 | 2016-08-17 | 广东药学院 | Spirostanol saponins and application thereof |
| CN105924494A (en) * | 2016-05-06 | 2016-09-07 | 广东药学院 | furostanol saponin compounds and application thereof |
| CN105924494B (en) * | 2016-05-06 | 2017-12-12 | 广东药科大学 | A kind of furostanol saponin class compound and its application |
| CN106509281A (en) * | 2016-12-21 | 2017-03-22 | 神农架林区中医医院 | campylandra chinensis sore-throat relieving tea and preparation method thereof |
| CN107361403A (en) * | 2017-08-07 | 2017-11-21 | 武汉纺织大学 | One kind contains Extracts of Tupistra chinensis additive in cigarette and preparation method thereof |
| CN107361403B (en) * | 2017-08-07 | 2018-09-18 | 武汉纺织大学 | A kind of cigarette of additive in cigarette and preparation method thereof containing Extracts of Tupistra chinensis |
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