CN100457116C - Cercopithecoidealin with anti-tumor function, its preparation and use - Google Patents
Cercopithecoidealin with anti-tumor function, its preparation and use Download PDFInfo
- Publication number
- CN100457116C CN100457116C CNB2006100273269A CN200610027326A CN100457116C CN 100457116 C CN100457116 C CN 100457116C CN B2006100273269 A CNB2006100273269 A CN B2006100273269A CN 200610027326 A CN200610027326 A CN 200610027326A CN 100457116 C CN100457116 C CN 100457116C
- Authority
- CN
- China
- Prior art keywords
- actinidin
- antitumor action
- preparation
- ethanol
- chromatography
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
An antineoplastic 'actinidine' in the form of coated tablet, capsule, mixture, softgel, particle, injection, or liposome is prepared from the root of lianoid pear through decocting, concentrating, filtering, chromatography with macroreticular resin, eluting by organic solvent, chromatography, concentrating, absorbing by polyamide, eluting and concentrating.
Description
Technical field
The present invention relates to plant amedica and its production and application field, particularly a kind of " Actinidin " and its production and application with antitumor action.
Background technology
" science of identification of Chinese materia medica " described Radix Actinidiae Chinensis is the root of Fructus actinidiae chinensis " A.chinensis Planch. Actinidia chinensisPlanch. " or Radix actinidiae argutae " Actinidia arguta Sieb.et Zucc Actinidia arguta (Sieb.et Zucc) Planch. ", Radix Actinidiae Chinensis and Fructus actinidiae chinensis or Radix actinidiae argutae are all done medicinal, the traditional Chinese medical science has that detoxifcation is invigorated blood circulation, the effect of clearing away heat-damp and promoting diuresis, is usually used in antitumor clinically.ShanXi Chinese Medicine Academy journal (VOL:28.NO:32005.5), the report of people such as Wei Peifeng " Radix Actinidiae Chinensis Chinensis extrat is induced the experimentation of apoptosis in gastric cancer ": adopt the ethanol extraction of Actinidia arguta Sieb.et Zucc that stomach cancer cell is had tangible lethal effect.The Shaanxi traditional Chinese medical science (2005 the 26th the 8th phases of volume): " Radix Actinidiae Chinensis is to the inhibiting experimentation of experimental rat gastric cancer ": adopt the ethanol extraction of Actinidia arguta Sieb.et Zucc, rat gastric cancer is had significant inhibition, and the effect of anti-stomach cancer metastasis is arranged.Hunan medicine (1985 second the 5th phases of volume), Kan Ke waits " actinidia root antitumor effect and to Immune Effects " clearly, adopt the water extract of Actinidia arguta Sieb.et Zucc root to have the effect of anti-mice U14 solid tumor, and strengthen the effect of cellular immunization and inhibition humoral immunization.Hebei Journal of Traditional Chinese Medicine (the 26th the 4th phase of volume of April in 2004), people such as Li Hao " Radix Actinidiae Chinensis is to the inhibiting experimentation of stomach cancer cell " report adopts commercially available Radix Actinidiae Chinensis, water extract, the aqueous solution of ethyl acetate extraction; 1.3mg/ml concentration, in the time of 24 hours, suppress stomach cancer cell and reach 87.61%.But its anti-tumor effective component and method of separating component are not done further research.
In Chinese patents such as CN200410022919, CN02114318, CN03156252, CN02129513, CN01128730, CN1478495A, CN1093594A, all related to preparing in the compound Chinese patent medicine with antitumaous effect and used Radix Actinidiae Chinensis or Radix Actinidiae Chinensis, these patents relate to the prescription of compound Chinese patent medicine mostly and form, prepare and use, rather than emphasize that the folk prescription Radix Actinidiae Chinensis extracts the new drug preparation of effective site and component separating.
Application number 2005100326666 relates to treats or prevents colorectal cancer, gastric cancer etc. to contain compound preparation and the preparation method and the application of Radix Actinidiae Chinensis and polygoni cuspidati,radix, Radix Gei japonici, preparation tablet, capsule, drop pill, and preparation such as freeze-dried powder.But this patent is not used the polyamide exquisiteness, and to macroporous resin not quantitatively, Radix Actinidiae Chinensis effective site does not have qualitative examination to determine; Lack dosage forms such as dispersible tablet, soft capsule, granule, liposome.
Application number 2005100326685 discloses anti-cancer Tengli root extract and its production and use, and this anti-cancer Tengli root extract can the lixiviate under 100 ℃ of temperature of 70 ∽ by water or organic solvent, concentrating under reduced pressure, acid adjustment, ethyl acetate extraction, polyamide chromatography, solvent elution, concentrate drying make then.But do not use macroporous resin to separate in the above-mentioned patent, and use ethyl acetate extraction, the method is brought problems such as equipment, labor are prevented, environmental protection when big commercial production, and the unresolved extract of the method prepares the preparation of new drug.
Summary of the invention
The objective of the invention is to overcome defective and the deficiency that prior art exists, a kind of " Actinidin " with antitumor action and its production and application is provided.
Purpose of the present invention is achieved through the following technical solutions: Actinidin is the total effective parts of a kind of compound anthraquinone glycoside etc., use 75% dissolve with ethanol, making becomes 1-10mg/ml alcohol liquid, point sample is (110 ℃ activate 1 hour) on high-efficient silica gel G thin layer chromatography (tlc) plate, the configuration chloroform: methanol: water: formic acid=3: 2: 0.2: 0.04 ratio is done developing solvent, the exhibition that launches is apart from 17cm, and taking-up is dried, and fluorescence down or sulphuric acid vanillin liquid spraying colour developing.Speckle (Fig. 1) with the Rf value in 0.00 to 0.100,0.167 to 0.300,0.400 to 0.533,0.700 to 0.833 or 0.900 to 0.967 scope shows that with methanol anion scanning the typical ion peak spectrum of Actinidin is 245,289,357,577,865amu (Fig. 2).
Actinidin preparation method and application comprise the steps:
1) Radix Actinidiae Chinensis decocting, decocting liquid is got supernatant concentration, adds chitosan clarification after-filtration, gets clear paste liquid;
2) clear paste liquid is through macroporous adsorption resin chromatography, and water is eluted to colourless, reuse organic solvent chromatography, and a stream part collection concentrates, and gets concentrated solution;
3) concentrated solution is adsorbed with polyamide, water is eluted to colourless, reuse organic solvent eluting, and eluent concentrates, and drying gets Actinidin.
4) Actinidin is added suitable adjuvant and additive after, make medicaments such as thin membrane coated tablet, hard capsule, mixture, soft capsule, granule, injection, liposome.
Water in the step 1) is deionized water or distilled water, and the 6-12 that adds water and be medical material weight doubly measures, and decocts each 1-3 hour 1-3 time; Be concentrated into 1: 1 or relative density 1.06-1.10 of medical material amount; Used chitosan be chitin take off the acetyl product, consumption is the 0.05-5% of medical material; Filter with filter cloth, filter paper, filter membrane.
Step 2) macroporous adsorbent resin in is D type or HPD type series; Granularity 20-60 order or 0.3mm-1.2mm; The aperture
Consumption is 0.5-2 a times of medical material amount; Flow velocity 0.5-10ml/g/ minute; Eluting water is deionized water or distilled water, and chromatography adopts organic solvent: 15%-30% ethanol, acetone or 50-85% ethanol, acetone; Collect the eluent of 50-85% ethanol or acetone, being concentrated into relative density is 1.100-1.350.
The polyamide consumption that adsorbs in the step 3) is 0.5-50 a times of concentrated solution; Granularity 100-120 order; The eluting organic solvent is ethanol or the acetone of 50-85%, preferred 50-75%; The concentrated solution relative density is 1.15-1.35; Dry vacuum available oven dry, vacuum-0.06-0.1Mpa, temperature≤80 ℃; Or centrifugal spray drying, centrifugal degree α=20000-50000; Inlet temperature 120-220 ℃, medicinal liquid flow velocity 10-200ml/ branch; Or vacuum lyophilization, vacuum-0.06-0.1Mpa, temperature-40 ℃-25 ℃, time 5-30 hour.
Adjuvant and additive that Actinidin in step 3) and the step 4) adds comprise cellulose family, stearic acid lipid, sodium benzoate, sucrose, starch, dextrin, cyclodextrin, lactose, the pre-powder process of Opadry film coating, sweet lactonaphthol, glycerol, sodium sulfite, PVP, PEG200-5000, two palmityl phosphatidyl alkali, two palmityl PHOSPHATIDYL ETHANOLAMINE, cholesterol, lecithin.Make medicaments such as thin membrane coated tablet, hard capsule, soft capsule, granule, injection, liposome.
Thin membrane coated tablet:
Actinidin 30-80%
Dextrin 1-20%
Lactose 1-20%
The pre-powder process 0.5-5% of Opadry film coating
PVP ethanol liquid 0.5-40%
Hard capsule:
Actinidin 30-90%
Dextrin 1-20%
Lactose 1-20%
PVP ethanol liquid 0.5-40%
Granule:
Actinidin 30-90%
Dextrin 1-20%
Lactose 1-20%
PVP ethanol liquid 0.5-40%
Mixture:
Actinidin 30-90%
Sucrose 1-20%
PEG400 1-10％
Antiseptic 0.5-1
Soft capsule:
Actinidin 30-90%
PEG4000 1-20％
Lactose 1-20%
Gelatin 1-20%
Glycerol 1-5%
Injection:
Actinidin 1-30%
Sodium sulfite 0.1-2%
Propylene glycol 0.1-10%
Water for injection 65-98%
Liposome:
Actinidin 1-30%
PEG200-5000 1-20％
Two palmityl phosphatidyl alkali 1-30%
Cholesterol 1-5%
Water for injection 0-96%
Actinidin of the present invention adopts mass spectral analysis, has determined the ion characteristic collection of illustrative plates, for further separation and purification provides definite target.Actinidin can be used for digestive tract tumor such as anti-treatment of esophagus cancer, gastric cancer, colorectal cancer, cancer of pancreas, hepatocarcinoma.
The invention provides the method for extracting Actinidin from Radix Actinidiae Chinensis, use this decocting method with deionized water or distilled water, impurity is few, effective ingredient rate of transform height; Chitosan clarification medicinal liquid is easy, economical, efficient, and loss of effective components is few; And chitosan is nontoxic, can not cause any bad side reaction to extract; Biggest advantage can be adsorbed objectionable impurities such as removing heavy metal when handling medicinal liquid.
Because macroporous adsorbent resin good separating effect, cost is low, easily produce, the present invention prepares in the method for Actinidin mainly the calculating according to D, DA series, HPD series plastics adsorbance, determine that the macroporous adsorbent resin consumption is the 0.5-2 optimum amount doubly of medical material amount, fully guarantee to adsorb effective ingredient, reduce the waste of resin;
The polyamide process for purification is that commercially available polyamide is milled into 100-120 purpose fine grained, more reasonably uses the polyamide consumption, improves the purity of raw material Actinidin, makes production operation easy, process stabilizing.
Figure of description
Fig. 1 Actinidin Actinidin thin-layer chromatogram
Fig. 2, Fig. 3, Fig. 4, Fig. 5 Actinidin mass spectral analysis collection of illustrative plates.
Specific implementation method
Concrete preparation method of the present invention illustrated by following routine embodiment, but protection scope of the present invention is not limited to this.
Embodiment 1
Get Radix Actinidiae Chinensis 100kg, add the 1200L deionized water, soaked 30 minutes, heated and boiled keeps boiling 3 hours, the leaching medicinal liquid; Decoction for the second time adds the 1000L deionized water, decocts 2 hours; Decoction for the third time adds the 800L deionized water, decocts 1 hour; Merge three times decocting liquid, standing over night, the leaching supernatant, vacuum concentration is to relative density 1.08; Clarifier chitosan 200g (with 1% acetum 20L dissolving), add in the water extracting liquid, stirring was left standstill 12 hours, and the leaching supernatant is by the pillar of D101-I type macroporous adsorbent resin 200kg, flow velocity 1L/ minute, be eluted to deionized water colourless, reuse 15% ethanol 200L eluting, and with 75% ethanol 500L eluting, collect 75% ethanol elution, reclaim ethanol, be concentrated into relative density 1.25 (70 ℃), admix the 25kg polyamide, the polyamide column of packing into, behind water and 15% ethanol elution,, collect 75% eluent with 75% ethanol 50L eluting, reclaim ethanol, be concentrated into relative density 1.35, at vacuum-0.06-0.1Mpa, 60 ℃ of following vacuum dryings of temperature, pulverize, obtain the Actinidin of 3.1kg.
Embodiment 2
Get Radix Actinidiae Chinensis 100kg, add the 1000L deionized water, soaked 45 minutes, heated and boiled keeps boiling 3 hours, the leaching medicinal liquid; Decoction for the second time adds the 800L deionized water, decocts 2 hours; Decoction for the third time adds the 800L deionized water, decocts 1 hour; Merge three times decocting liquid, standing over night, the leaching supernatant, vacuum concentration is to relative density 1.06; Clarifier chitosan 500g (with 1% acetum 50L dissolving) adds in the water extracting liquid, stirs and leaves standstill 12 hours, the leaching supernatant, by the pillar of HPD300-type macroporous adsorbent resin 200kg, flow velocity 1.2L/ minute, be eluted to colourless with deionized water, reuse 15% acetone 200L eluting, and, collect 75% acetone eluent with 75% acetone 500L eluting, reclaim acetone, be concentrated into relative density 1.30 (70 ℃), admix the 30kg polyamide, the polyamide column of packing into is behind water and the 15% acetone eluting, with 75% acetone 50L eluting, collect 75% eluent, reclaim acetone, be concentrated into relative density 1.15, centrifugal spray drying, centrifugal degree-20000-50000; Inlet temperature 120-220 ℃, medicinal liquid flow velocity 10-200ml/ branch; Obtain the Actinidin of 2.8kg.
Embodiment 3
Get Radix Actinidiae Chinensis 100kg, add the 1000L deionized water, soaked 45 minutes, heated and boiled keeps boiling 3 hours, the leaching medicinal liquid; Decoction for the second time adds the 800L deionized water, decocts 2 hours; Decoction for the third time adds the 800L deionized water, decocts 1 hour; Merge three times decocting liquid, standing over night, the leaching supernatant, vacuum concentration is to relative density 1.06; Clarifier chitosan 750g (with 1% acetum 75L dissolving), add in the water extracting liquid, stirring was left standstill 24 hours, and the leaching supernatant is by the pillar of DA201-type macroporous adsorbent resin 200kg, flow velocity 1.0L/ minute, be eluted to deionized water colourless, reuse 15% acetone 200L eluting, and with 75% acetone 500L eluting, collect 75% acetone eluent, reclaim acetone, be concentrated into relative density 1.30 (70 ℃), admix the 25kg polyamide, the polyamide column of packing into, behind water and the 15% acetone eluting,, collect 75% eluent with 75% acetone 50L eluting, reclaim acetone, be concentrated into relative density 1.15, vacuum lyophilization, vacuum-0.1Mpa, temperature-40 ℃ rises to 25 ℃, 25 hours time.Obtain the Actinidin of 3.8kg.
Embodiment 4
Actinidin, use 70% dissolve with ethanol, making becomes 1-10mg/ml alcohol liquid, point sample is (110 ℃ activate 1 hour) on high-efficient silica gel G thin layer chromatography (tlc) plate, the configuration chloroform: methanol: water: formic acid=3: 2: 0.2: 0.04 ratio is done developing solvent, the exhibition that launches is apart from 17cm, and taking-up is dried, and fluorescence down or sulphuric acid vanillin liquid spraying colour developing.Speckle with the Rf value in 0.00 to 0.100,0.167 to 0.300,0.400 to 0.533,0.700 to 0.833 or 0.900 to 0.967 scope.
Actinidin 400g
Dextrin 40g
Lactose 60g
The pre-powder process 10g of Opadry film coating
PVP ethanol liquid 0.5-40%
Embodiment 6
Actinidin 400g
Dextrin 40g
Lactose 60g
PVP ethanol liquid 0.5-40%
Make 1000 capsules
Embodiment 7
Actinidin 800g
Dextrin 1000g
Lactose 1200g
PVP ethanol liquid 0.5-40%
Make the 3g/1000 bag
Embodiment 8
Actinidin 100g
Sucrose 200g
PEG400 20g
Antiseptic 0.5-1%
Distilled water adds to full dose
Make 1000ml
Embodiment 9
Actinidin 400g
PEG4000 200g
Lactose 100g
Gelatin 100g
Glycerol 50g
Embodiment 10
Actinidin 10g
Sodium sulfite 2g
Propylene glycol 100ml
Water for injection adds to full dose
Make 1000ml
Embodiment 11
Actinidin 50g
PEG200-5000 50g
Two palmityl phosphatidyl alkali 100g
Cholesterol 5g
Make pro-liposome
Embodiment 12
With pro-liposome, add the injection water, cross the sterilization of 0.22 microporous filter membrane, fill N2, embedding gets lipidosome injection.
Embodiment 13
With pro-liposome, add the mannitol of injection water and 10%, cross the sterilization of 0.22 μ microporous filter membrane, packing, lyophilization gets injectable powder.
Embodiment 14
Movable normal behind the oral Actinidin extractum of mice, the end sees that animal has overt toxicity reaction and death condition, growth promoter normal (seeing below the body weight gain table).Huge inspection is dissected in off-test, and obvious pathological change is seen at the main organs end.
Experiment conclusion:
When the disposable oral Actinidin of mice is equivalent to Radix Actinidiae Chinensis 750g crude drug/kg, do not see that this medicine has any toxic reaction, it is safe taking in the dosage more than 370 times according to one day clinical consumption conversion Actinidin of people.
Embodiment 15
The extracorporeal anti-tumor function of Actinidin
1, is subjected to the reagent thing
Actinidin sample preparation: after the DMSO dissolving, add solution or uniform suspension that PBS is made into 100 μ g/ml, dilute with PBS then.
2, cell strain
Human breast carcinoma (Bcap-37), the former leukemia of the chronic marrow of people (K562), human breast carcinoma (MCF-7), human colon carcinoma (HT-29), murine melanoma (K111), become human lung fibroblast (WI38), Chinese hamster pneumonocyte (CHL), l cell (3T3), people's hepatocarcinoma (SMMC7721), human leukemia cell (W256), the above-mentioned cell strain of Humanmachine tumour (SKMEL28) frozen and go down to posterity by this laboratory.
3, culture medium
(1) RPMI (1640) cell culture medium (GIBCO): contain 10% deactivation newborn calf serum (Saida biological Pharmaceutical Co., Ltd., Shanghai city); The L-glutaminase (the import packing, SANGON); Sodium Pyruvate; 1 * 10
5U.L
-1Penicillin, 100mg.L
-1Streptomycin; Aseptic filtration, 4 ℃ of preservations.
(2) 0.25% trypsin solutions (Trypsin): available from Invitrogen company ,-20 ℃ of preservations.
(3) phosphate buffer (PBS): NaCl8g, KCl0.2g, Na
2HPO
41.15g, KH
2PO
40.2g, be dissolved in the 1L distilled water, 121 ℃ of autoclave sterilization 20min, 4 ℃ of preservations.
(4) MTT (AMRESCO) solution: be made into 5mg/ml solution with PBS.
(5) lysate: every 100ml deionization distilled water contains SDS10g, isobutanol 5ml, concentrated sulphuric acid 0.12ml.
4, experimental technique
Cytotoxicity to above-mentioned tumor cell line records by mtt assay.Concrete steps are as follows:
(1) sample preparation: sample is made gradient dilution with PBS, obtain the dilute sample that concentration is respectively 100 μ g/ml, 10 μ g/ml, 1 μ g/ml, 0.1 μ g/ml, 0.01 μ g/ml, 0.001 μ g/ml.
(2) will dilute good sample and add in flat 96 orifice plates, every hole 10 μ l make two parallel testings at every.The corresponding work of DMSO added in the entering plate, behind the gradient dilution in contrast.
(3) get the cell that is in exponential phase, cell is suspended in the RPMI-640 culture medium that contains 10% calf serum after trypsinization and washing, through the blue dyeing of Placenta Hominis exclusive method meter viable count, and regulates cell suspending liquid density to 2 * 10
5Cell/ml.
(4) in flat 96 orifice plates, every hole adds 90 microlitre cells, overnight incubation in 37 ℃, 5%CO2 cell culture incubator.
(5) flat 96 orifice plates that will add cell insulation 48 hours in 37 ℃, 5%CO2 cell culture incubator.
(6) add 20 μ l 5mg/mlMTT solution in every hole, continue in incubator, to be incubated 3～4 hours.
(7) every hole adds 100 μ l DMSO, continues incubated overnight in incubator, next day, surveys the optical density (can adopt the DG3022 enzyme-linked immunosorbent assay instrument) in each hole with the A570 wavelength.Each grade test solution hole average and negative control value relatively get the percent inhibition (establishing certain density standard antineoplastic agent simultaneously as positive control) of each grade concentration.Computing formula is:
IC％＝(C-T)/Cx100％
T is the negative matched group OD of a test solution group OD value C value
According to each IC% that detects each grade of liquid concentration, try to achieve IC then with regression curve
50Value.
5, experimental result
The results are shown in following table 1, Actinidin has the good antitumor effect to the former leukemia of the chronic marrow of people (K562), human colon carcinoma (HT-29), people's hepatocarcinoma (SMMC7721), Humanmachine tumour (SKMEL28), murine melanoma (K111), human breast carcinoma (MCF-7), murine melanoma (K111), human colon carcinoma (HT-29) cell strain.
Table .1 sample water is carried, ethanol extract is to the in-vitro multiplication inhibitory action of each cell strain:
Embodiment 16
The zoopery that Actinidin suppresses tumor sees Table 2
Method:
1, laboratory animal: Kunming mouse, ♀, 19-21g, by Shanghai Si Laike animal responsibility company limited.The quality certification number: SCXK (Shanghai) 2003-003
2, tumor source source: S180 tumor source has this laboratory to go down to posterity to protect kind
3, experimental technique:
Eugonic mice S180 ascites cells is diluted by 1: 5 with normal saline, mice one side oxter inoculation 0.2ml diluent, random packet, every group of 10 mices.Different extract medication groups, blank group are inoculated administration in the 2nd day, and continuous 7 days, put to death mice on the 9th day, take off the tumor piece and weigh, calculate tumour inhibiting rate.
4. experimental result: table 2
| Group | Sex | Quantity | Dosage (crude drug g/kg) | Tumour inhibiting rate (%) |
| 1L | ♀ | 10 | 4 | 16.36 |
| 1H | ♀ | 10 | 12 | 34.71 |
| 2L | ♀ | 10 | 8 | 16.48 |
| 2H | ♀ | 10 | 24 | 30.87 |
| 3L | ♀ | 10 | 20 | 34.71 |
| 3H | ♀ | 10 | 50 | 44.61 |
| 4L | ♀ | 10 | 20 | 52.80 |
| 4H | ♀ | 10 | 50 | 60.98 |
Annotate: (1) water extract 10g crude drug/ml; (2) ethanol extraction 10g crude drug/g; (3) macroporous resin eluate 15g crude drug/g; (4) Actinidin.
Embodiment 17
1. test objective
The test Actinidin is to the preliminary curative effect (table 3) of rat liver cancer (HAC) solid tumor
2. be subjected to the reagent thing
Actinidin; Compound method:, now with the current with normal saline dilution (0.1g/ml).
3. animal
Source and kind: Kunming mouse is provided by the court's Animal House
The quality certification number: Shanghai is moving closes the card word No. 107
Body weight: 19-21 gram
Sex: female
Each treated animal number: 10
4. transplanted tumor
Rat liver cancer (HAC cell) is that pharmacological room of Shanghai Institute of Pharmaceutical Industry goes down to posterity and keeps
5. Shi Yan key step and method
Get well-grown rat liver cancer (HAC cell), after normal saline dilution in 1: 4, every mice axil subcutaneous vaccination 0.2ml, random packet is established normal saline matched group and cyclophosphamide positive controls (100mg/kg, ip * 3 (1,3,5)), lot number 980401, sample sets (6.25,12.5,25ml/kg).Work giving different pharmaceutical next day, 0.5ml/ Mus, continuous irrigation stomach 7 days.Inoculate back 10 days and take off neck execution animal, dissect and get the tumor piece, relatively the size of each dosage group tumor weight.The result judges according to following formula:
6. result
Table 3 Actinidin is to the tumor-inhibiting action of rat liver cancer (HAC)
| Sample | Dosage (ml/kg) | Dosage regimen | Number of animals all the time | Heavy (g) x ± SD of tumor | Tumour inhibiting rate % |
| Blank (normal saline) | 25 | po×7 | 1010 | 3.18±0.77 | |
| Actinidin | 6.25 12.5 25 | po×7 po×7 po×7 | 1010 1010 1010 | 2.13±0.33 1.93±0.73 1.50±0.32 | 33.02 ** 39.31 ** 52.83 ** |
| CTX | 100mg/kg | ip×3 | 1010 | 0.17±0.07 | 94.66 ** |
Compare with the blank group:
*P＜0.01.
From last table 3 as seen, Actinidin has certain inhibitory action to rat liver cancer (HAC) when 25ml/kg dosage, and tumour inhibiting rate is 52.83%, has compared significant difference (P＜0.01=with the blank group.
Attached: test report
One. test apparatus:
The ABI 3200Q Trap level Four bar mass spectrograph (u.s.a. applied biosystem company) of connecting
Two. test procedure:
1. sample configuration: take by weighing sample 1.0mg, be dissolved in 1mL methanol, be made into 1mg/mL solution; Being diluted to concentration with methanol is 5ug/mL solution.
2. get methanol and do positive and negative ion scanning, sweep limits: 100～1000amu respectively;
3. get above-mentioned 5ug/mL sample solution and do positive and negative ion scanning, sweep limits: 100～1000amu respectively.
Three. test results and analysis:
1. result of the test is seen accompanying drawing 2-5.
2. interpretation of result
Compare with methanol cation scintigram, the scanning of sample cation does not see that strong characteristic ion peak is arranged.
With methanol anion scintigram relatively, in the scanning of sample anion occurs, occur 289, the strong quasi-molecular ions of 577amu, have characteristic.Simultaneously, because therefore 289=(577+1)/2 infers that 289 and 577 structures have certain dependency.In addition 245,357,865amu also has the characteristic ion peak
The typical ion peak spectrum of Actinidin is 245,289,357,577,865amu.
For further separation and purification provides definite target to Actinidin.
3. fill up the shortage of pharmaceutical preparation of the effective site of Radix Actinidiae Chinensis, develop the novel form of single medicinal material, reach the purpose of declaring the new drug dosage form.
Claims (15)
1. " Actinidin " with antitumor action is characterized in that:
Actinidin is a kind of compound anthraquinone glycoside, be to utilize Radix Actinidiae Chinensis decocting supernatant to add the chitosan clarification filtration, gained clear paste liquid is through macroporous adsorption resin chromatography, washing, the organic solvent chromatography, part concentrated solution reuse polyamide absorption of gained stream, washing, the organic solvent eluting obtains Actinidin, this Actinidin 75% dissolve with ethanol, making becomes 1-10mg/ml alcohol liquid, on the high-efficient silica gel G thin layer chromatography (tlc) plate, have 0.00 to 0.100,0.167 to 0.300,0.400 to 0.533,0.700 the speckle of the Rf value in 0.833 or 0.900 to 0.967 scope, typical ion peak spectrum is 245,289,357,577,865amu.
2. preparation method with antitumor action " Actinidin ", it is characterized in that: the Actinidin preparation method comprises the steps:
1) Radix Actinidiae Chinensis decocting, decocting liquid is got supernatant concentration, adds chitosan clarification after-filtration, gets clear paste liquid;
2) clear paste liquid is through macroporous adsorption resin chromatography, and water is eluted to colourless, reuse organic solvent chromatography, and a stream part collection concentrates, and gets concentrated solution;
3) concentrated solution is adsorbed with polyamide, water is eluted to colourless, reuse organic solvent eluting, and eluent concentrates, and drying gets Actinidin.
3. a kind of preparation method as claimed in claim 2 with antitumor action " Actinidin ", it is characterized in that: described water is deionized water or distilled water, the 6-12 that adds water and be medical material weight doubly measures, and decocts each 1-3 hour 1-3 time; Be concentrated into 1: 1 or relative density 1.06-1.10 of medical material amount; Used chitosan be chitin take off the acetyl product, consumption is the 0.05-5% of medical material; Filter with filter cloth, paper, film.
4. a kind of preparation method with antitumor action " Actinidin " as claimed in claim 2 is characterized in that: described macroporous adsorbent resin is D type or HPD type series; Granularity 20-60 order or 0.3mm-1.2mm; The aperture
Flow velocity 0.5-10ml/g/ minute.
5. a kind of preparation method with antitumor action " Actinidin " as claimed in claim 4 is characterized in that: it is 15%-30% ethanol, acetone or 50-85% ethanol, acetone that described chromatography adopts organic solvent.
6. a kind of preparation method with antitumor action " Actinidin " as claimed in claim 4 is characterized in that: the 0.5-2 that described macroporous adsorbent resin consumption is the medical material amount times, it is 75% ethanol or 75% acetone that chromatography adopts organic solvent.
7. a kind of preparation method as claimed in claim 2 with antitumor action " Actinidin ", it is characterized in that: the 0.5-50 that described polyamide consumption is a concentrated solution doubly, the chromatography organic solvent is ethanol or the 50-75% acetone of 50-75%, vacuum lyophilization, vacuum-0.06-0.1Mpa, temperature-40 ℃-25 ℃, time 5-30 hour.
8. a kind of preparation method with antitumor action " Actinidin " as claimed in claim 7 is characterized in that: described polyamide granularity 100-120 order, the chromatography organic solvent is 75% ethanol or 75% acetone.
9. a kind of application as claimed in claim 1 with antitumor action " Actinidin ", it is characterized in that: described pharmaceutical preparation is thin membrane coated tablet, comprising:
Actinidin 30-80%
Dextrin 1-20%
Lactose 1-20%
The pre-powder process 0.5-5% of Opadry film coating
PVP ethanol liquid 0.5-40%
10. a kind of application as claimed in claim 1 with antitumor action " Actinidin ", it is characterized in that: described pharmaceutical preparation is hard capsule, comprising:
Actinidin 30-90%
Dextrin 1-20%
Lactose 1-20%
PVP ethanol liquid 0.5-40%
11. a kind of application with antitumor action " Actinidin " as claimed in claim 1, it is characterized in that: described pharmaceutical preparation is granule, comprising:
Actinidin 30-90%
Dextrin 1-20%
Lactose 1-20%
PVP ethanol liquid 0.5-40%
12. a kind of application with antitumor action " Actinidin " as claimed in claim 1, it is characterized in that: described pharmaceutical preparation is liposome, comprising:
Actinidin 1-30%
PEG200-5000 1-20％
Two palmityl phosphatidyl alkali 1-30%
Cholesterol 1-5%
Water for injection 0-96%
13. a kind of application with antitumor action " Actinidin " as claimed in claim 1, it is characterized in that: described pharmaceutical preparation is injection, comprising:
Actinidin 1-30%
Sodium sulfite 0.1-2%
Propylene glycol 0.1-10%
Water for injection 65-98%
14. a kind of application with antitumor action " Actinidin " as claimed in claim 1, it is characterized in that: described pharmaceutical preparation is mixture:
Actinidin 30-90%
Sucrose 1-20%
PEG400 1-10％
Antiseptic 0.5-1%
15. a kind of application with antitumor action " Actinidin " as claimed in claim 1, it is characterized in that: described pharmaceutical preparation is soft capsule, comprising:
Actinidin 30-90%
PEG4000 1-20％
Lactose 1-20%
Gelatin 1-20%
Glycerol 1-5%
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100273269A CN100457116C (en) | 2006-06-06 | 2006-06-06 | Cercopithecoidealin with anti-tumor function, its preparation and use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100273269A CN100457116C (en) | 2006-06-06 | 2006-06-06 | Cercopithecoidealin with anti-tumor function, its preparation and use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1895302A CN1895302A (en) | 2007-01-17 |
| CN100457116C true CN100457116C (en) | 2009-02-04 |
Family
ID=37608040
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006100273269A Expired - Fee Related CN100457116C (en) | 2006-06-06 | 2006-06-06 | Cercopithecoidealin with anti-tumor function, its preparation and use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100457116C (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101518556B (en) * | 2009-04-07 | 2012-06-20 | 浙江大学 | Polysaccharide with antitumor and immunologic adjuvant function as well as preparation method and purpose thereof |
| CN101810651B (en) * | 2010-03-25 | 2011-08-17 | 赵正福 | Preparation method of kiwifruit frozen and dried fruit vegetable product with function of tumor inhibition |
| CN101791325B (en) * | 2010-03-26 | 2012-06-06 | 香港晓源药业集团股份有限公司 | Preparation method of Chinese goosebeery capsule for preventing and treating lung cancer |
| CN103705550B (en) * | 2012-10-08 | 2018-03-02 | 上海医药工业研究院 | A kind of Actinidin and its preparation method and application |
| CN103301218A (en) * | 2012-12-29 | 2013-09-18 | 浙江省中医院 | Chinese herbal preparation for treating colon cancer and preventing cancer metastasis and preparation method thereof |
| CN103202810A (en) * | 2013-04-01 | 2013-07-17 | 浙江中医药大学 | Chinese actinidia root polysaccharide lipidosome and preparation method thereof |
| CN103709206B (en) * | 2013-12-26 | 2015-12-30 | 长沙市博泰生物科技有限公司 | A kind of preparation method of Actinidin |
| CN111089932B (en) * | 2020-01-08 | 2022-02-18 | 河北中医学院 | Rapid multi-information thin-layer identification method for Chinese actinidia root particles and target decoction dry powder |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1156624A (en) * | 1996-10-30 | 1997-08-13 | 浙江中医学院 | Medicinal preparation with extractive from yangtao root and preparing process thereof |
-
2006
- 2006-06-06 CN CNB2006100273269A patent/CN100457116C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1156624A (en) * | 1996-10-30 | 1997-08-13 | 浙江中医学院 | Medicinal preparation with extractive from yangtao root and preparing process thereof |
Non-Patent Citations (6)
| Title |
|---|
| 中国猕猴桃属植物的主要化学成分与综合利用. 端木.林产化工通讯，第4期. 1994 |
| 中国猕猴桃属植物的主要化学成分与综合利用. 端木.林产化工通讯，第4期. 1994 * |
| 猕猴桃根抑制肿瘤细胞的实验研究. 邹益友，谭桂山，谢兆霞.湖南中医药导报，第5卷第4期. 1999 |
| 猕猴桃根抑制肿瘤细胞的实验研究. 邹益友，谭桂山，谢兆霞.湖南中医药导报，第5卷第4期. 1999 * |
| 藤梨根提取物诱导胃癌细胞凋亡的实验研究. 卫培峰，焦晨莉，张三印.陕西中医学院学报，第28卷第3期. 2005 |
| 藤梨根提取物诱导胃癌细胞凋亡的实验研究. 卫培峰，焦晨莉，张三印.陕西中医学院学报，第28卷第3期. 2005 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1895302A (en) | 2007-01-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100457116C (en) | Cercopithecoidealin with anti-tumor function, its preparation and use | |
| CN105884621B (en) | A kind of sesquiterpenoids and its preparation method and application | |
| CN102504006A (en) | Steroid saponin compound in siberian fritillary bulb | |
| CN102133220B (en) | Preparation method of pulsatilla saponin A | |
| CN104945452B (en) | A kind of preparation method and applications of phillygenol glucuronic acid derivative | |
| CN101185661A (en) | Technique for preparing liverwort extract and application of it in preparing medicine for treating liver cancer and hepatitis B | |
| CN102875615B (en) | Extraction method and application of falcate dolichos root or leaf glucoside A and total saponins of falcate dolichos root or leaf | |
| CN101537036A (en) | Soap pod saponin extract as well as preparation method and application thereof | |
| CN107266516B (en) | Triterpenoid with anti-tumor activity and the preparation method and application thereof | |
| CN110922440A (en) | Indole nitrogen glycoside compound, extraction method and application in preparation of medicine for preventing and treating nervous system diseases | |
| CN1486712A (en) | Extractive of Chinese medicinal material rabdosia as antitumor medicine and its prepn process | |
| CN101721434B (en) | Active ingredients of fomes officinalis, preparation method thereof and use thereof | |
| CN103083388B (en) | Preparation method of fructus gleditsiae total saponins | |
| CN105380968A (en) | Nitraria-tangutorum-bobr total alkaloid extract, preparing method thereof and application thereof | |
| CN101879170B (en) | Anti-tumor application of malonyl astragaloside I and aglycon thereof | |
| CN1954839B (en) | Medical composition prepared by caulis Marsdeniae Tenacissimae, ginseng and astragalus root | |
| CN101550120A (en) | Effective component of artemisia leaves and preparation method thereof | |
| CN102295678B (en) | Purpose of triterpenoid-saponin compound extracted form anemone taipaiensis | |
| CN104961667B (en) | A kind of purification process of sulforaphen | |
| CN113336808A (en) | Glucoside compound extracted and separated from lily, and method and application thereof | |
| CN104961717B (en) | The enrichment method of a kind of Sophoraflavanone G and application | |
| CN103156891B (en) | 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 | |
| CN103800351A (en) | Pharmaceutical application of akebin E | |
| CN100546602C (en) | Anti-tumor compound polygoni cuspidati,radix, Radix Actinidiae Chinensis, Radix Gei japonici preparation and its production and application | |
| CN102229640B (en) | Application of triterpenoid saponins compound extracted from anemone taipaiensis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Anhui Kang Hui Pharmaceutical Co., Ltd. Assignor: Zhao Zhengfu Contract fulfillment period: 2009.6.4 to 2026.6.5 contract change Contract record no.: 2009340000251 Denomination of invention: Cercopithecoidealin with anti-tumor function, its preparation and use Granted publication date: 20090204 License type: Exclusive license Record date: 2009.9.2 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2009.6.4 TO 2026.6.5; CHANGE OF CONTRACT Name of requester: ANHUI KANGHUI PHARMACEUTICAL CO., LTD. Effective date: 20090902 |
|
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090204 Termination date: 20140606 |