CN103156891B - Preparation method of akebia fruit active components and preparation method of preparation of akebia fruit active components and application in preparation of antineoplastic medicines of preparation of foreknowledge sub-active components - Google Patents
Preparation method of akebia fruit active components and preparation method of preparation of akebia fruit active components and application in preparation of antineoplastic medicines of preparation of foreknowledge sub-active components Download PDFInfo
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Abstract
The invention belongs to the technical field of medicines, in particular to a preparation method of akebia fruit active components and a preparation method of the preparation of the akebia fruit active components and the application in the preparation of antineoplastic medicines of the akebia fruit active components. The preparation method comprises steps as below: smashing the akebia fruit medical materials into coarse powders, soaking the coarse powders in alcohol and heating, refluxing and extracting the akebia fruit medical materials for two to three times, filtering the akebia fruit medical materials, and merging filter liquor. Carrying out vacuum concentration, stewing and centrifugation, and therefore sediment I and supernatant fluid I are obtained. Adjusting the potential of hydrogen (PH) of the supernatant fluid I through alkali to 8 to 14, hydrolyzing the supernatant fluid I for 2 to 8 hours under the temperature of 60 Celsius degrees to 100 Celsius degrees, adding acid to adjust the PH of the supernatant fluid to 3 to 7 after the hydrolyzing, carrying out centrifugation, and therefore the sediment II and the supernatant fluid II after the hydrolyzing are obtained. Merging the sediment I and the sediment II, drying the sediment, and therefore the akebia fruit active components are obtained. The active components have significant antineoplastic activity, and can be used for manufacturing medicines of antineoplastic treatment or auxiliary antineoplastic treatment.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of preparation method of Fructus Akebiae active component and the preparation method of preparation thereof and its application in preparing antitumor drug, this active component has significant anti-tumor activity, can be used for preparing antitumor drug or adjuvant therapy medicaments of tumor.
Background technology
Fructus Akebiae is the dry almost ripe fruit of plants of Lardizabalaceae Caulis Akebiae [Akebia quinata (Thunb.) Decne.], threeleaf akebia [Akebia trifoliate (Thunb.) Koidz.] or Caulis Akebiae [Akebia trifoliate (Thunb.) Koidz. var. australis (Diels) Rehd.].Bitter in the mouth, cold.Return liver, gallbladder, stomach, urinary bladder channel.For liver-smoothing, qi-regulating, promoting blood circulation and stopping pain, diuresis, eliminating stagnation.For gastral cavity side of body distending pain, amenorrhea dysmenorrhea, dysuria, sucutaneous nodule mass in the abdomen.Bibliographical information: saponin component is the main component in Fructus Akebiae, have protect the liver, the multiple pharmacologically active such as relieving convulsion, pain relieving, antifungal, parasite killing, these effects have dependency with the clinical efficacy of Fructus Akebiae.Through research repeatedly, a few saponins in Fructus Akebiae have antineoplastic activity in addition, through systematic study, prepare exploitation and take the antitumor drug that this active component is main component.
Summary of the invention
The object of the present invention is to provide a kind of Fructus Akebiae extract, another object of the present invention is to provide the application in antitumor field of a kind of preparation method of Fructus Akebiae extract and this extract.
Fructus Akebiae active component of the present invention is achieved by the following technical solution:
Get Fructus Akebiae medical material, be ground into coarse powder, with 5-95% soak with ethanol 0.5-2h, 6-12 doubly measures heating and refluxing extraction 1-3 time, and each 1-3h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.2-1.0g raw medicinal herbs/mL in 50-70 ℃, static, centrifugal, must precipitate I and supernatant I; Supernatant I is adjusted PH to 8-14 with sodium hydroxide, and 60-100 ℃ is hydrolyzed 2-8 hour, adds hydrochloric acid and adjust PH to 3-7 after hydrolysis, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Merge precipitation I, precipitation II, dry, obtain Fructus Akebiae active component.
Through this technique, extract purification and saponin component content in Fructus Akebiae antitumor component can be greater than to 50%; The main saponin of measuring in precipitation I through HPLC method is triterpene saponin (3-O-β-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabopyranose Hederagenin, saponins B, saponins P
dwith saponins A) amount accounted for the more than 90% of total saponins amount in Fructus Akebiae, tool anti-tumor activity, and supernatant I anti-tumor activity is not obvious, but after basic hydrolysis, obtain precipitation II, by HPLC method, measure its main component identical with precipitation I, and have anti-tumor activity.
In initial preparation technology, supernatant II is adsorbed through macroporous adsorptive resins, water, 40% ethanol 70-90% ethanol elution successively after loading, collect 70-90% ethanol elution, concentrated, dry, obtains extract III.Research finds, the component of precipitation I, precipitation II and extract III is basic identical (mainly contains 3-O-β-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabopyranose Hederagenin, saponins B, saponins P
dwith saponins A), and in three part components, the content of total saponins is all greater than 50%.Yet the yield of precipitation I is 2-8%, the yield of precipitation II is 2-5%, and the yield of extract III is 0.1-0.3%, and from the viewpoint of economizing on resources and economic benefit, this part can be not considered macroporous resin, merges precipitation I and precipitation II in reality; Obtain Fructus Akebiae active component.In the present invention, precipitate I and precipitation II, using separately or merging uses (preferably merge and use) to may be used to prepare antitumor or for the medicine of tumor aid treatment, this active component can be used as active component and pharmaceutical carrier is mixed with the various preparations on pharmaceutics, the dispersity of Fructus Akebiae antitumor component in preparation can be liquid, also can be solid, the route of administration of said preparation can be oral, injection and topical.
in the present invention, by ultraviolet/visible spectrophotometry, measure saponin component content:
1, the preparation of oleanolic acid reference substance solution
Precision takes oleanolic acid reference substance 5mg, is transferred in 50mL volumetric flask, with dissolve with methanol, standardize solution, is made into the standard solution of 0.1mg/mL, and stored refrigerated is standby.
2, oleanolic acid maximum absorption wavelength determines
Accurate reference substance solution and the testing sample solution 1ml of drawing, each 2 parts (portion is done blank, portion is done and is developed the color), add in the dry tool plug test tube of label, in 60 ℃ of vacuum drying ovens, volatilize, blank group adds the methanol of 5ml, colour developing group adds the perchloric acid of 5ml, shake up, colour developing group is reacted 15min in 70 ℃ of water-baths, and jolting constantly, after reacting completely, put immediately cooling 15min in ice-water bath, retinue reagent is blank, in Shimadzu UV-2550 ultraviolet spectrophotometer, in 200～700nm wave-length coverage, scans.Result need testing solution and reference substance solution have absorption maximum at 310nm wavelength place, therefore determine that measuring wavelength is 310nm.
3, the drafting of standard curve
Precision pipettes reference substance solution 0.3,0.6,0.9,1.2,1.5,1.8 mL each 2 parts (portion is done blank, portion is done and is developed the color), be transferred in the tool plug test tube of 10mL, in 60 ℃ of vacuum drying ovens, volatilize, blank group adds the methanol of 5ml, colour developing group adds the perchloric acid of 5ml, shakes up, and colour developing group is reacted 15min in 70 ℃ of water-baths, and jolting constantly, after reacting completely, put immediately cooling 15min in ice-water bath, retinue reagent is blank, in Shimadzu UV-2550 ultraviolet spectrophotometer, at 200 ~ 500nm length scanning, at 310nm wavelength place, measure trap.The trap (A) of take is vertical coordinate, and oleanolic acid reference substance concentration (C) is abscissa, and drawing standard curve is made method of least square with trap A to concentration C and returned, and obtains standard curve equation to be: A=0.022C+0.045, R
2=0.9996(n=4), the range of linearity is 7.0～34.3 mg/mL, and in the range of linearity, the linear relationship of mass concentration and trap value is good.
4, saponin component content assaying method in sample
Get Fructus Akebiae medical material appropriate, pulverize, take medical material 34.90g, with 10 times of amount 70% ethanol (being 350ml), extract 2 times, each 1.5 hours, gained solution filters with Medium speed filter paper, is concentrated into below 100ml, is transferred to the volumetric flask of 100ml, be settled to scale, obtain the Fructus Akebiae extract of 0.349g/ml; Pipette Fructus Akebiae extracting solution 1ml, be transferred in the volumetric flask of 250ml, be settled to scale, obtain the Fructus Akebiae need testing solution of 1.396mg/ml.The accurate need testing solution 1ml that draws, 2 parts (portion is done blank, portion is done and is developed the color), add in the dry tool plug test tube of label, in 60 ℃ of vacuum drying ovens, volatilize, blank group adds the methanol of 5ml, colour developing group adds the perchloric acid of 5ml, shakes up, and colour developing group is reacted 15min in 70 ℃ of water-baths, and jolting constantly, after reacting completely, put immediately cooling 15min in ice-water bath, retinue reagent is blank, in Shimadzu UV-2550 ultraviolet spectrophotometer, at 200 ~ 500nm length scanning, at 310nm place, measure its trap.By standard curve, try to achieve in sample total saponins concentration and calculate content.
in the present invention, with HPLC high-efficient liquid phase technique, Fructus Akebiae active component is carried out to component analysis:
1, the preparation of need testing solution: get the about 50mg of Radix Pulsatillae antitumor component powder, accurately weighed, be placed in 50mL volumetric flask, dissolve with methanol is also settled to scale, filters, and gets subsequent filtrate 10mL as need testing solution.
2, chromatographic condition: chromatographic column: COSMOSIL(4.6*250mm, 5 μ m) C
18oDS post; Mobile phase: acetonitrile-water-phosphoric acid=45:55:0.1; Detect wavelength: 203nm; Column temperature: 25 ℃; Flow velocity: 1mL/min.Sample size 20 μ L.
Through component analysis, four main triterpene saponin in Fructus Akebiae active component are 3-O-β-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabopyranose Hederagenin, saponins B, saponins P
drefer to accompanying drawing with saponins A().
Accompanying drawing explanation
Fig. 1: the HPLC chromatogram of precipitation I
Fig. 2: the HPLC chromatogram of precipitation II
Fig. 3: the HPLC chromatogram of the Fructus Akebiae active component after precipitation I and precipitation II merge
In figure, 1 is that 3-O-β-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabopyranose Hederagenin, 2 is that saponins B, 3 is saponins P
d, 4 be saponins A.
the specific embodiment:
relevant Fructus Akebiae antitumor component preparation method specific embodiment:
Embodiment 1: the preparation method of Fructus Akebiae active component, wherein: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, with 20% soak with ethanol 0.5h, measure heating and refluxing extraction 2 times for 6 times, each 1h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.5g raw medicinal herbs/mL in 50 ℃, must precipitate I and supernatant I, and supernatant I hydro-oxidation sodium is adjusted PH to 9,70 ℃ of hydrolysis 7h, and after hydrolysis, hydrochloric acid is adjusted pH to 4, centrifugal, must precipitate II and hydrolysis and separate rear supernatant II; Dry, precipitation I yield is 5%, and precipitation II yield is 3.4%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 54%, and in precipitation II, total saponin content is 74%, and in the Fructus Akebiae active component after merging, the content of total saponins is 60%.
Embodiment 2: the preparation method of Fructus Akebiae active component, wherein: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, with 80% soak with ethanol 2h, measure heating and refluxing extraction 3 times for 12 times, each 3h, filters merging filtrate; Above-mentioned filtrate is evaporated to 1.0g raw medicinal herbs/mL in 70 ℃, centrifugal, must precipitate I and supernatant I, supernatant I hydro-oxidation sodium is adjusted PH to 10,80 ℃ of hydrolysis 6h, after hydrolysis, hydrochloric acid is adjusted pH to 5, centrifugal, must precipitate the rear supernatant II of II and hydrolysis, dry, precipitation I yield is 5.4%, and precipitation II yield is 3.8%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 66%, and in precipitation II, total saponin content is 85%, and in the Fructus Akebiae active component after merging, the content of total saponins is 74%.All the other are with embodiment 1.
Embodiment 3: the preparation method of Fructus Akebiae active component, wherein: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, with 50% soak with ethanol 1h, measure heating and refluxing extraction 2 times for 8 times, each 2h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.7g raw medicinal herbs/mL in 60 ℃, centrifugal, must precipitate I and supernatant I, supernatant I hydro-oxidation potassium is adjusted PH to 11,90 ℃ of hydrolysis 5h, after hydrolysis, hydrochloric acid is adjusted pH to 6, centrifugal, must precipitate the rear supernatant II of II and hydrolysis, dry, precipitation I yield is 4.6%, and precipitation II yield is 3.2%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 58%, and in precipitation II, total saponin content is 84%, and in the Fructus Akebiae active component after merging, the content of total saponins is 68%.All the other are with embodiment 1.
Embodiment 4: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, with 50% soak with ethanol 0.5h, measure heating and refluxing extraction 3 times for 12 times, each 1.5h, filters merging filtrate.Above-mentioned filtrate is evaporated to 0.5g raw medicinal herbs/mL in 60 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation sodium is adjusted PH to 12,90 ℃ of hydrolysis 4h, and after hydrolysis, sulphuric acid is adjusted pH to 7, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Dry, precipitation I yield is 2.4%, and precipitation II yield is 4.8%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 62%, and in precipitation II, total saponin content is 85%, and in the Fructus Akebiae active component after merging, the content of total saponins is 71%.All the other are with embodiment 1.
Embodiment 5: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, with 95% soak with ethanol 2h, measure heating and refluxing extraction 2 times for 6 times, each 3h, filters merging filtrate.Above-mentioned filtrate is evaporated to 0.5g raw medicinal herbs/mL in 50 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation potassium is adjusted PH to 13,100 ℃ of hydrolysis 3h, and after hydrolysis, hydrochloric acid is adjusted pH to 6, centrifugal, must precipitate II and hydrolysis and separate rear supernatant II; Dry, precipitation I yield is 4.4%, and precipitation II yield is 3.0%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 56%, and in precipitation II, total saponin content is 78%, and in the Fructus Akebiae active component after merging, the content of total saponins is 63%.All the other are with embodiment 1.
Embodiment 6: Fructus Akebiae pulverizing medicinal materials is become to coarse powder, and 10% soak with ethanol 1.5h, measures heating and refluxing extraction 1 time for 8 times, and each 3h, filters merging filtrate.Above-mentioned filtrate is evaporated to 0.5g raw medicinal herbs/mL in 50 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation sodium is adjusted PH to 14,60 ℃ of hydrolysis 8h, and after hydrolysis, sulphuric acid is adjusted pH to 6, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Dry, precipitation I yield is 5.0%, and precipitation II yield is 3.0%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 66%, and in precipitation II, total saponin content is 85%, and in the Fructus Akebiae active component after merging, the content of total saponins is 74%.All the other are with embodiment 1.
Embodiment 7: get Fructus Akebiae medical material, be ground into coarse powder, with 20% soak with ethanol 0.5h, measure heating and refluxing extraction 1 time for 6 times, each 1h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.2g raw medicinal herbs/mL in 50 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation sodium is adjusted PH to 12,90 ℃ of hydrolysis 6h, and after hydrolysis, hydrochloric acid is adjusted pH to 6, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Dry, precipitation I yield is 3.2%, and precipitation II yield is 4.2%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 55%, and in precipitation II, total saponin content is 75%, and in the Fructus Akebiae active component after merging, the content of total saponins is 67%.All the other are with embodiment 1.
Embodiment 8: get Fructus Akebiae medical material, be ground into coarse powder, with 80% soak with ethanol 2h, measure heating and refluxing extraction 3 times for 12 times, each 3h, filters merging filtrate; Above-mentioned filtrate is evaporated to 1.0g raw medicinal herbs/mL in 70 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation potassium is adjusted PH to 11,100 ℃ of hydrolysis 4h, and after hydrolysis, hydrochloric acid is adjusted pH to 5, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Dry, precipitation I yield is 7.4%, and precipitation II yield is 2.8%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 63%, and in precipitation II, total saponin content is 87%, and in the Fructus Akebiae active component after merging, the content of total saponins is 77%.All the other are with embodiment 1.
Embodiment 9: get Fructus Akebiae medical material, be ground into coarse powder, with 60% soak with ethanol 1h, 6-12 doubly measures heating and refluxing extraction 2 times, and each 2h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.7g raw medicinal herbs/mL in 60 ℃, centrifugal, must precipitate I and supernatant I, and supernatant I hydro-oxidation sodium is adjusted PH to 13,70 ℃ of hydrolysis 8h, and after hydrolysis, sulphuric acid is adjusted pH to 5, centrifugal, must precipitate the rear supernatant II of II and hydrolysis; Dry, precipitation I yield is 5.4%, and precipitation II yield is 3.8%; Merge precipitation I and precipitation II, obtain Fructus Akebiae active component.Through outer/visible spectrophotometry, detect, in precipitation I, total saponin content is 60%, and in precipitation II, total saponin content is 81%, and in the Fructus Akebiae active component after merging, the content of total saponins is 74%.All the other are with embodiment 1.
Embodiment 10: the preparation method of Fructus Akebiae active component, wherein: in Fructus Akebiae active component, saponin component content is more than 50%.All the other are with embodiment 1.
Embodiment 11: the preparation that described Fructus Akebiae active component is made, wherein: this Fructus Akebiae active component and excipient substance are mixed with the various preparations on pharmaceutics.All the other are with any one in embodiment 1,2,3,4,5,6,7,8,9 and 10.
Embodiment 12: the preparation that described Fructus Akebiae active component is made, wherein: the dispersity of this Fructus Akebiae active component in preparation can be liquid, can be also solid.All the other are with embodiment 11.
Embodiment 13: described Fructus Akebiae active component, wherein: the route of administration of said preparation can be oral, injection and topical.All the other are with embodiment 11.
Embodiment 14, by Fructus Akebiae active component prepared by the method for the embodiment of the present invention 1 or embodiment 2 or embodiment 3 or embodiment 4 or embodiment 5 or embodiment 6 or embodiment 7 or embodiment 8 or embodiment 9 or embodiment 10, be raw material, carry out lower series preparation preparation:
1, tablet preparation
Prescription (50):
Composition weight (g)
Active component 5.0 of the present invention
Lactose 4.8
Dextrin 9
Microcrystalline Cellulose 6
Magnesium stearate 0.2
According to the operational approach of conventional tablet, above mix homogeneously, wet granulation, finally adds magnesium stearate mix homogeneously to be pressed into every 500mg.
2, capsule preparation
Prescription (90):
Composition weight (g)
Active component 8 of the present invention
Lactose 5
Dextrin 9
Microcrystalline Cellulose 5
According to the operational approach of conventional tablet, above mix homogeneously, wet granulation, fill becomes capsule, every 300mg.
3, cataplasma preparation
Prescription:
Composition weight (g)
Active component 2 of the present invention
Polyacrylate 18
Laurocapram 0.6
Lauric acid 1.8
Propylene glycol 5.4
Ethyl acetate 15
According to the affected of common cataplasma, carry out, by above-mentioned, stir, coat on polyester adherent layer, 60 degree are dry after 2 minutes, cover poly-amino methyl film backing layer, section.
4, injectable powder preparation
Prescription:
Composition weight (g)
Active component 0.05 of the present invention
Hydrochloric acid is appropriate
Mannitol 50
According to the operation of conventional freeze-dried powder, carry out, active component of the present invention is added to appropriate water for injection and dissolve, add mannitol, be settled to 1000ml, regulate pH value between 4.0-5.5, aseptic filtration, lyophilization and get final product.
5, the preparation of aqueous injection
Fructus Akebiae antitumor component, be dissolved in appropriate water for injection, the injection water soluble adjuvant that adds metering, the active carbon that adds 0.1-0.5% is removed pyrogen, regulates PH to 6.0-7.5, after microporous filter membrane ultrafiltration, add water for injection to ormal weight, fill is in ampoule bottle or in infusion bottle, sealing by fusing/roll lid, through after the assay was approved, pack and get final product.
6, drop pill preparation
Prescription:
Composition weight (g)
Active component 0.1g of the present invention
Polyethylene glycol 6000 10g
Macrogol 4000 20g
Polyoxyethylene sorbitan monoleate 0.3g
Ethanol 1ml
Make 1000 balls
Method for making: Fructus Akebiae antitumor component of the present invention is dissolved in ethanol, become active component and disperse ethanol, add in the substrate that the polyethylene glycol 6000, Macrogol 4000, polyoxyethylene sorbitan monoleate of melting form, be stirred well to evenly, take dimethicone as coolant, and dripping becomes ball at 15 ℃, wipes ball, dry, obtain.
Embodiment 15: the active component of preparing by the method in the embodiment of the present invention one is raw material, carries out antitumor component pharmacodynamic study:
1, eliminating evil effect
The inhibitory action of 1.1 pairs of animal transplanting tumors
Adopt 7402 Liver Cancer Bearing Nude Mices, MFC TCM23 gastric cancer in nude mice and Mice Bearing Lewis lung cancer model, investigate the anti-tumor in vivo effect of Fructus Akebiae antitumor component (YZS).The nude mice of 7402 hepatocarcinoma, MFC TCM23 gastric cancer and A549 pulmonary carcinoma will be inoculated respectively, be divided at random positive controls (cyclophosphamide), solvent control group (blank), high, medium and low three the dosage groups (50 of Fructus Akebiae antitumor component of the present invention (YZS), 100,200mg/kg), gastric infusion is 10 days continuously, and last administration, after 24 hours, is put to death animal, separated oxter tumor is also weighed, and calculates tumor control rate.Be calculated as follows tumour inhibiting rate: tumour inhibiting rate (%)=[(the average tumor weight of the average tumor weight-administration of blank group group) the average tumor weight of/blank group] * 100%.Experimental result is as follows:
(1) impact on 7402 hepatocarcinoma: experimental result shows YZS(50,100,200mg/kg) can significantly suppress the tumor weight of nude mice, three dosage suppression ratio are respectively 16.55,44.82 and 57.24%.The results are shown in Table 1.
Table 1 Fructus Akebiae antitumor component (YZS) is right
7402the impact of Liver Cancer Bearing Nude Mice
* P<0.05, * * P<0.01 compares with blank group
(2) impact on MFC TCM23 gastric cancer in nude mice: YZS(50,100,200mg/kg) nude mice tumor weight average is had to certain inhibitory action.The results are shown in Table 2.
The impact of table 2 Fructus Akebiae antitumor component (YZS) on MFC TCM23 gastric cancer in nude mice
* P<0.05, * * P<0.01 compares with blank group
(3) impact on A549 pulmonary carcinoma: experimental result shows YZS(50,100,200mg/kg) the heavy suppression ratio of mice A549 lung cancer tumor is respectively to 14.21%, 33.73% and 47.34%.The results are shown in Table 3.
The impact of table 3 Fructus Akebiae antitumor component (YZS) on mice A549 pulmonary carcinoma
* P<0.05 compares with blank group
2, centralizing function
2.1 the impact of Fructus Akebiae antitumor component (YZS) on tumor animal immunologic function
Can obviously the raise killing activity of spleen index, splenocyte conversion ratio and NK cell of tumor-bearing mice of experimental result prompting Fructus Akebiae antitumor component (YZS).
Set up H
22tumor-bearing mice animal model
,fructus Akebiae antitumor component (YZS) (50,100,200mg/kg) gastric infusion, continuous 10 days, last administration was after 24 hours, by sacrifice of animal, get tumor-bearing mice Thymus and spleen, weigh, calculate respectively thymus index: [heavy (the mg)/body weight (g) of thymus] * 10, spleen index: [heavy (the mg)/body weight (g) of spleen] * 10.
(1) impact on mouse thymus exponential sum spleen index
Compare YZS(100,200 mg/kg with blank group) can obviously improve H
22the spleen index of tumor-bearing mice and thymus index, but cyclophosphamide obviously reduces spleen index and thymus index.Prompting, cyclophosphamide can cause tumor-bearing mice spleen and atrophy of thymus gland.Preliminary explanation YZS may improve the immunity of mice in performance antitumaous effect, the results are shown in Table 4.
Table 4 Fructus Akebiae antitumor component (YZS) is to H
22the immune impact of tumor-bearing mice (
± s)
| Group | Dosage (mg * kg -1×d) | Spleen index | Thymus index |
| YZS | 200 | 10.56±1.78* | ?3.78±1.89* |
| ? | 100 | 9.14±2.49 | 3.21±1.45 |
| ? | 50 | 8.45±2.89 | 2.79±1.34 |
| Cyclophosphamide | 30 | 7.02±2.10* | ? 1.02±0.49** |
| Blank group | ? | 8.70±2.89 | 2.56±1.23 |
* P<0.05, * * P<0.01, compares with blank group
(2) impact of human peripheral blood cell
Through analysis of accounts, with the comparison of blank group, cyclophosphamide group murine interleukin sum, lymphocyte and erythrocyte significantly reduce (
p<0.01), YZS respectively organizes murine interleukin sum, lymphocyte, mononuclear cell and neutrophilic granulocyte number and compares with blank group, do not have significant difference (
p>0.05).Prompting, cyclophosphamide obviously suppresses mouse immune cell, and each group of YZS is obviously different from the effect of cyclophosphamide, to mouse peripheral blood cell unrestraint effect.The results are shown in Table 5.
The impact of table 5 Fructus Akebiae antitumor component (YZS) on tumor-bearing mice peripheral blood immunocyte
* P<0.05, compares with blank group
3, median lethal dose(LD 50)
Adopt Fructus Akebiae antitumor component powder, with distilled water, be mixed with the solution of desired concn, adopt clean level mice, by Jiangxi College of Traditional Chinese Medicine Experimental Animal Center, provided.Median lethal dose(LD 50) when acute toxicity testing result shows Fructus Akebiae extract oral is 4.90g/kg.
Precipitation I prepared by the method for embodiment 16, employing embodiment 2 or embodiment 3 or embodiment 4 or embodiment 5 embodiment 6 or embodiment 7 or embodiment 8 or embodiment 9 or embodiment 10, precipitation II and Fructus Akebiae active component are carried out the research of antitumor drug effect, and effect is similar to Example 15.
Fructus Akebiae active component and excipient substance prepared by the method for embodiment 17, employing embodiment 2 or embodiment 3 or embodiment 4 or embodiment 5 embodiment 6 or embodiment 7 or embodiment 8 or embodiment 9 or embodiment 10 are mixed with the various preparations on pharmaceutics.
The preparation that Fructus Akebiae active component described in embodiment 18, employing embodiment 17 is made, wherein: as antitumor component, the dispersity in preparation can be liquid to Fructus Akebiae active component, can be also solid.
Embodiment 19, according to any one Fructus Akebiae active component described in the method for embodiment 2 or embodiment 3 or embodiment 4 or embodiment 5 embodiment 6 or embodiment 7 or embodiment 8 or embodiment 9 or embodiment 10 as antitumor component, wherein: Fructus Akebiae active component can be for the preparation of antitumor drug or adjuvant therapy medicaments of tumor as Studies on Anticancer Components.
Claims (1)
1. a preparation method for Fructus Akebiae active component, is characterized in that: get Fructus Akebiae medical material, be ground into coarse powder, with 5-95% soak with ethanol 0.5-2h, 6-12 doubly measures heating and refluxing extraction 1-3 time, and each 1-3h, filters merging filtrate; Above-mentioned filtrate is evaporated to 0.2-1.0g raw medicinal herbs/mL in 50-70 ℃, centrifugal, must precipitate I and supernatant I; Supernatant I is with adjusting PH with base to 8-14, and 60-100 ℃ is hydrolyzed 2-8 hour, and after being hydrolyzed, acid adding is adjusted pH to 3-7, centrifugal, must precipitate II and the rear supernatant II of hydrolysis, merges precipitation I and precipitation II, is dried, and obtains Fructus Akebiae active component; Described alkali is sodium hydroxide or potassium hydroxide; Described acid is hydrochloric acid or sulphuric acid.
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Non-Patent Citations (4)
| Title |
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| 《预知子中α2常春藤皂苷的HPLC 分析》;王家明;《中国药学杂志》;20060831;1212-1213 * |
| 王家明.《预知子中α2常春藤皂苷的HPLC 分析》.《中国药学杂志》.2006,1212-1213. * |
| 预知子的化学成分、药理作用与临床应用研究;高亚玲等;《河北化工》;20110531(第05期);35-36 * |
| 高亚玲等.预知子的化学成分、药理作用与临床应用研究.《河北化工》.2011,(第05期), * |
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