CN109045078A - Heterostemonous Biebersteinia anticancer usage - Google Patents
Heterostemonous Biebersteinia anticancer usage Download PDFInfo
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- CN109045078A CN109045078A CN201811006093.3A CN201811006093A CN109045078A CN 109045078 A CN109045078 A CN 109045078A CN 201811006093 A CN201811006093 A CN 201811006093A CN 109045078 A CN109045078 A CN 109045078A
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- extract
- biebersteinia
- platymiscium
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- butanol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention discloses the purposes of Heterostemonous Biebersteinia platymiscium or its extract in the drug of preparation prevention or treatment liver cancer, and new possibility is provided for clinical application.
Description
Technical field
The present invention relates to the new applications of Heterostemonous Biebersteinia.
Background technique
Heterostemonous Biebersteinia platymiscium is distributed in Mediterranean sea region, West Asia, the Central Asia, northwest China and western Himalaya, belongs to Gu Dizhong
Sea distribution.About 5 kinds of this category, it is distributed in Mediterranean sea region, West Asia, the Central Asia, northwest China and western Himalaya, belongs to Tethys point
Cloth.China records 3 kinds, is distributed in the Northwest and western Tibet.This category is more generally as independent Heterostemonous Biebersteinia
Belong to Biebersteinia to handle.The domestic frequently seen plants in China have Heterostemonous Biebersteinia Biebersteinia heterostemon
Maxim., split Heterostemonous Biebersteinia Biebersteinia multifida DC., high mountain Heterostemonous Biebersteinia Biebersteinia odora more
Steph.ex Fisch。
Heterostemonous Biebersteinia, is Tibetan areas characteristic nationality drug, and multi-section Tibetan classics are all on the books.It is recorded according to " Jingzhubencao ",
" Mang Jian " is bitter, trembles with fear, controls ulcer boils, and dissipating swollen analgesic has special efficacy.Currently, studying both at home and abroad Heterostemonous Biebersteinia platymiscium medicinal material less.
Summary of the invention
The purpose of the present invention is to provide the antineoplastic new usages of Heterostemonous Biebersteinia platymiscium.
Specifically, the present invention provides Heterostemonous Biebersteinia platymisciums or its extract in the drug of preparation prevention or treatment liver cancer
Purposes.
Wherein, the drug is the drug for inhibiting liver cancer cell growth.
Wherein, the extract is selected from aqueous or not water-containing organic solvent extract.
Further, the polarity parameters of the organic solvent are selected from 3.4~4.4.Polarity parameters of the present invention are conventional
Polarity parameters, in the representative solvent of non-polar and polar, with n-hexane for 0.0, water 10.2.Other solvent polarity parameters
Have, methylene chloride: 3.4, n-butanol: 3.7, propyl alcohol: 4.0, butyl acetate 4.0, tetrahydrofuran: 4.2, chloroform: 4.1, acetic acid second
Ester: 4.3, isopropanol 4.3, chloroform 4.4 etc..
Certain organic solvents, when use, first can carry out saturated process, such as n-butanol etc. with water.
Wherein, the organic solvent is selected from the mixing of one or both of ethyl acetate, n-butanol.
In present invention experiment, on the basis of ethanol extract, petroleum ether, ethyl acetate, water-saturated n-butanol are successively used
It is extracted, obtained ligroin extraction, ethyl acetate extract, n-butanol extract and water extract all has certain
Resisting liver cancer activity, therefore, it can be stated that its bright ethanol extract also has certain resisting liver cancer activity.
If the organic solvent is selected from n-butanol, the preparation method of extract is selected from one of following:
Method one: Heterostemonous Biebersteinia platymiscium is taken, is directly extracted using n-butanol, extract is prepared；
Method two: taking the ethanol extract of Heterostemonous Biebersteinia platymiscium, then is successively using petroleum ether, ethyl acetate, water saturation just
Butanol extracts, and takes n-butanol extract that extract is prepared；
If the organic solvent is selected from ethyl acetate, the preparation method of extract is selected from one of following:
Method one: Heterostemonous Biebersteinia platymiscium is taken, is directly extracted using n-butanol, extract is prepared；
Method two: taking the ethanol extract of Heterostemonous Biebersteinia platymiscium, then is successively extracted using petroleum ether, ethyl acetate,
Take ethyl acetate extract that extract is prepared.
In the present invention, ethanol extract is the extract being prepared using conventional method using ethyl alcohol as solvent, at present
Also there is the preparation method of the ethanol extract.Ethyl alcohol generally selects concentration 80% or more, may further be 90% with
On ethyl alcohol, such as can be dehydrated alcohol, 95% ethyl alcohol.
In the present invention extracting method use conventional method, such as can be dipping, diacolation, reflux, ultrasound, microwave,
Flash, extraction etc..
When extraction, raw material can be crushed, it certainly, can also be without crushing.
The present invention also provides Heterostemonous Biebersteinia platymisciums or its extract in the drug that preparation inhibits liver cancer cell growth
Purposes.
Heretofore described Heterostemonous Biebersteinia platymiscium be selected from Heterostemonous Biebersteinia Biebersteinia heterostemon Maxim.,
Split Heterostemonous Biebersteinia Biebersteinia multifida DC., high mountain Heterostemonous Biebersteinia Biebersteinia odora Steph.ex more
One or both of Fisch.
The Heterostemonous Biebersteinia platymiscium that the present invention uses, can be used its all herbal medicine, can also use its aerial part.
Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically above content of the invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on above content of the present invention
The technology realized all belongs to the scope of the present invention.
Specific embodiment
Embodiment 1
Take 95% ethanol extract of Heterostemonous Biebersteinia Biebersteinia heterostemon Maxim., as far as possible recycling second
It after alcohol, then is successively extracted, is taken respectively above-mentioned organic using petroleum ether-water, ethyl acetate-water, water-saturated n-butanol-water
Layer, to get corresponding extract after recycling design.
Embodiment 2
95% ethanol extract of Heterostemonous Biebersteinia Biebersteinia heterostemon Maxim. is taken, recycling ethyl alcohol is simultaneously
It after extract drying, then is successively extracted, is taken respectively above-mentioned organic using petroleum ether, ethyl acetate, water-saturated n-butanol
Solvent extraction liquid, to get corresponding extract after recycling design.
Embodiment 3
1, culture solution is prepared
The culture of HepG2 cell selects RPMI-1640 culture medium, wherein containing 10% fetal calf serum and 1% culture
Base.
2, FBS is prepared
In the EP pipe that the FBS frozen is sub-packed in 15mL after melting at 4 DEG C, stored under the conditions of -20 DEG C.
3, HepG2 cell secondary culture
When the HepG2 cell of culture reached for 3~5 generation, the cell that upgrowth situation does not pollute well is chosen, is replaced fresh
Complete medium, remove culture medium within second day, with 3mL sterilizing 1 × PBS buffer solution rinse 3 times, 0.25% pancreatin digestion after
Add complete medium that cell suspension is made, then 1 000rpm is centrifuged 5min at room temperature, supernatant is removed, what addition was prepared in advance
Then frozen stock solution is divided in cryopreservation tube and seals, -80 DEG C of placements are placed on Liquid nitrogen storage for 24 hours.
4, cell cryopreservation and recovery
4.1 cell cryopreservations:
When the HepG2 cell of culture reached for 3~5 generation, the cell that upgrowth situation does not pollute well is chosen, is replaced fresh
Complete medium, outwell culture medium within second day, with 3mL sterilizing 1 × PBS buffer solution rinse several times, 0.25% pancreatin digestion
Afterwards plus cell suspension is made in complete medium, and then 1 000rpm is centrifuged 5min at room temperature, removes supernatant, is added and prepares in advance
Frozen stock solution, be then divided in cryopreservation tube and seal, -80 DEG C of placements are placed on Liquid nitrogen storage for 24 hours.
4.2 cell recoveries:
By the cryopreservation tube taken out in liquid nitrogen quickly be put into after sealing strip sealing orifice in 37 DEG C of water-baths shake back and forth with
Promote to melt, at this time it is noted that cell is not contaminated.After 75% cotton ball soaked in alcohol cleaning disinfection, 1 000rpm is centrifuged at room temperature
5min outwells the frozen stock solution in cryopreservation tube, and cell is transferred in culture bottle to (inoculating cell density is 4 × 105A/mL),
The complete medium for adding 10% is put into 37 DEG C of 5%CO2Incubator in cultivate, more renew immediately after second day cell is adherent
Culture medium.
5 cell proliferating determinings
With MTS method measurement Heterostemonous Biebersteinia different parts to the inhibiting effect of HepG2 cell.Choose the cell dissociation system in 3~7 generations
Obtain cell suspension (5 × 104A/mL), and every hole adds 100 μ L suspensions in 96 orifice plates, the sterilizing every hole 100 1 × PBS is added in surrounding
μL.Inoculated 96 orifice plate is put into 37 DEG C of 5%CO2Incubator in cultivate, after adherent, change the RPMI-1640 of serum-free
After culture medium synchronization process cell 12h, setting action time is for 24 hours respectively.Water extract, ethyl acetate extract, n-butanol portion
Position and petroleum ether part concentration gradient are respectively 10000,1000,100,10,0.1,0.01 μ g/mL, after drug-treated, to
The MTS solution of 10 μ L is added in every hole, and culture plate is continued to be incubated for 3 hours in incubator.With microplate reader measurement at 450nm
Absorbance, and returned to zero with blank well.The inhibiting rate of cell is calculated with following calculation formula:
Inhibiting rate (%)=[1- (ADrug-ABlank)/(AControl-ABlank)]
ADrug: various concentration treated absorbance
AControl: without the cell hole absorbance of drug-treated
ABlank: only add the absorbance of MTS and culture medium
6 experimental results
Each extract is shown in Table 1 to the inhibitory effect of HepG2 cell:
Table 1
As can be seen from the above table, using water extract, under the concentration of 1000 μ g/mL, certain inhibition makees liver cancer cells
With；And ethyl acetate extract, n-butanol extract, ligroin extraction are used, and under the concentration of 1000 μ g/mL, difference energy
Enough reach 100.89%, 70.99%, 96.05% inhibiting rate.
Claims (10)
1. the purposes of Heterostemonous Biebersteinia platymiscium or its extract in the drug of preparation prevention or treatment liver cancer.
2. purposes according to claim 1, it is characterised in that: the drug is the drug for inhibiting liver cancer cell growth.
3. purposes according to claim 1, it is characterised in that: the extract is selected from aqueous or not water-containing organic solvent
Extract.
4. purposes according to claim 3, it is characterised in that: the polarity parameters of the organic solvent are selected from 3.0~4.3.
5. purposes according to claim 3 or 4, it is characterised in that: the organic solvent is in ethyl acetate, n-butanol
One or two kinds of mixing.
6. purposes according to claim 3, it is characterised in that: the organic solvent is selected from ethyl alcohol；Further, ethyl alcohol is dense
Degree is 80% or more.
7. purposes according to claim 5, it is characterised in that: the organic solvent is selected from n-butanol, the system of extract
Preparation Method is selected from one of following:
Method one: Heterostemonous Biebersteinia platymiscium is taken, is directly extracted using n-butanol, extract is prepared；
Method two: taking the ethanol extract of Heterostemonous Biebersteinia platymiscium, then successively uses petroleum ether, ethyl acetate, water-saturated n-butanol
It extracts, takes n-butanol extract that extract is prepared.
8. purposes according to claim 5, it is characterised in that: the organic solvent is selected from ethyl acetate, extract
Preparation method is selected from one of following:
Method one: Heterostemonous Biebersteinia platymiscium is taken, is directly extracted using n-butanol, extract is prepared；
Method two: taking the ethanol extract of Heterostemonous Biebersteinia platymiscium, then is successively extracted using petroleum ether, ethyl acetate, takes second
Extract is prepared in acetoacetic ester extract.
9. the purposes of Heterostemonous Biebersteinia platymiscium or its extract in the drug that preparation inhibits liver cancer cell growth.
10. purposes described in any one according to claim 1~8, it is characterised in that: the Heterostemonous Biebersteinia platymiscium, which is selected from, to be smoked
Ox Biebersteinia heterostemon Maxim., split Heterostemonous Biebersteinia Biebersteinia multifida DC., height more
One or both of mountain Heterostemonous Biebersteinia Biebersteinia odora Steph.ex Fisch.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811006093.3A CN109045078B (en) | 2018-08-30 | 2018-08-30 | Anti-tumor application of smoked bull |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811006093.3A CN109045078B (en) | 2018-08-30 | 2018-08-30 | Anti-tumor application of smoked bull |
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| Publication Number | Publication Date |
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| CN109045078A true CN109045078A (en) | 2018-12-21 |
| CN109045078B CN109045078B (en) | 2021-07-30 |
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| CN201811006093.3A Active CN109045078B (en) | 2018-08-30 | 2018-08-30 | Anti-tumor application of smoked bull |
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