CN108653209A - A kind of new type natural Tea Flower derives the preparation and its application of lipid nanometer vesica - Google Patents
A kind of new type natural Tea Flower derives the preparation and its application of lipid nanometer vesica Download PDFInfo
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- CN108653209A CN108653209A CN201810567200.3A CN201810567200A CN108653209A CN 108653209 A CN108653209 A CN 108653209A CN 201810567200 A CN201810567200 A CN 201810567200A CN 108653209 A CN108653209 A CN 108653209A
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- tea flower
- new type
- lipid nanometer
- natural tea
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- 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)
- A61K36/82—Theaceae (Tea family), e.g. camellia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- A61K2236/31—Extraction of the material involving untreated material, e.g. fruit juice or sap obtained from fresh plants
Abstract
The invention discloses preparations and its application that a kind of new type natural Tea Flower derives lipid nanometer vesica.The nano vesicle is to can be used as a kind of Nano medication of edible natural for treating disease from being extracted by way of repeatedly centrifuging in Tea Flower.It shows that the nano vesicle grain size is more uniform by carrying out a series of characterizations to nano vesicle, is 131.6 ± 1.6 nm, chromatography shows that the nano vesicle naturally loads a variety of catechin components.It is shown by co-culturing the nano vesicle of various concentration gradient and Several Kinds of Malignancy cell, the nano vesicle can effectively inhibit the proliferation of the cancer cells such as breast cancer, colon cancer, non-small cell lung cancer and cervical carcinoma, show good anticancer effect and broad spectrum anticancer performance.The extensive concern of scientific research personnel and pharmaceuticals is not caused for the drug derivative preparation and commercial development of Tea Flower at present, therefore the invention has convincingly demonstrated Tea Flower good medicinal application foreground.
Description
Technical field
The present invention relates to the fields such as Tea Science, pharmacy, pharmacology and Bio-Nano-Materials, and in particular to a kind of new type natural
Tea Flower derives the preparation of lipid nanometer vesica, characterizes its application in treating malignant tumor.
Background technology
Tea Flower, the component part important as tea tree, includes various active organic substance, be considered as it is a kind of it is natural in
Medicinal material.But since people do not know much have less understanding to Tea Flower, attention degree is far below tealeaves, and then causes it to be not yet received and effectively open
Hair and utilization.Since teacher Xu Jiying in 2002 is put forward for the first time " comprehensive utilization of Tea Flower resource " this theory, people by
Gradually start to carry out a series of research to Tea Flower.It is similar with tealeaves that current existing result of study shows that Tea Flower has
Biochemical composition, including tea polyphenols, protein, vitamins, caffeine, polysaccharide, flavonoids, superoxide dismutase and peroxidating
Hydrogen enzyme etc..Wherein, tea polyphenols are as a kind of common antioxidant, by many researchers have shown that it has good cancer resistance
Energy.Polyphenol content is more in Tea Flower, has accounted for prodigious ratio with catechin especially, has been up to about 70%, by EGCG, ECG, EC
Etc. multiple monomer compositions.In China's traditional medicine theory, " integration of drinking and medicinal herbs " theory is used till today by everybody always, camellia conduct
A kind of edibility, antioxidant activity show good natural material, there is the huge medicinal potential of clinic.
In recent years, it was discovered by researchers that in the prevalence of a kind of natural lipid nano vesicle, internal packet in fresh plant
Containing there are many different types of organic molecule, nucleic acid isoreactivity substance, a kind of good Nano medication can be used as to be used for portion
Divide the research of disease.For Nano medication compared to current hot topic, natural lipid nano vesicle has better bio-compatible
Property, the use of organic solvent in nano-particle preparation process is eliminated, the problems such as Nano medication can not be produced in enormous quantities is overcome.
2013, extraction obtained a kind of lipid nanometer vesica to Huang-ge Zhang seminars of the U.S. from grape fruit for the first time, this kind is received
Rice vesica can load treatment of the treatment molecule such as different antibodies or nucleic acid for diseases such as tumours（X. Houet al.Biomaterials2017: 124; 195-210）.Then, which confirms to equally exist similar knot in grape again
The substance of structure can effectively treat acute ulcer colitis disease (the S. Ju induced by dextran sulfate sodium et al.Mol. Ther.2013: 21; 1345-1357).Recently, Huang-ge Zhang seminars and U.S. Didier
Merlin seminars have obtained similar nano-lipid vesica from ginger, are respectively used to hepatopathy caused by alcohol, acute knot
The treatment of enteritis, chronic colitis and colon cancer has good therapeutic effect (M. Zhang et al.Biomaterials 2016: 101; 321-340. M. Zhang et al.Mol. Ther. 2016: 24; 1783-
1796. X. Zhuang et al.J. Extra. Ves. 2015: 4; 28713.)。
Invention content
In consideration of it, it is an object of the invention to extract a kind of natural lipid nano vesicle, the nanometer from fresh Tea Flower
Vesica naturally loads a variety of catechin components, shows the ability for preferably inhibiting tumor cell proliferation, can be used for a variety of pernicious
The chemotherapy of tumour.
Technical scheme of the present invention is specific as follows：
The preparation method that a kind of new type natural Tea Flower derives lipid nanometer vesica includes the following steps：
（1）Fresh Tea Flower tissue is put into juice extractor stirring to be crushed, obtains tissue magma；
（2）By step（1）Obtained tissue magma is 4oIt is centrifuged under the conditions of C, retains tissue supernatant；
（3）By step（2）Obtained tissue supernatant is collected 4oIt is centrifuged under the conditions of C, retains precipitation, phosphate buffer weight is used in combination
It is outstanding, obtain re-suspension liquid；
（4）By step（3）Collect obtained re-suspension liquid and further carry out sucrose gradient centrifugation, wherein sucrose concentration be respectively 8%,
30%, 45% and 60%；
（5）The natural tea tree collected between 30% and 45% spends lipid nanometer vesica（TFNPs）, it washs and is resuspended afterwards into PBS three times,
It puts to -80oC long-term preservations.
Further, the step（2）In, centrifugation rate is 6000 ~ 15000 rpm, and centrifugation time is 1 ~ 5h.
Preferably, the step（2）In, centrifugation rate is 20000 rpm, centrifugation time 3h.
Further, the step（3）In, centrifugation rate is 50000 ~ 80000 rpm, and centrifugation time is 1 ~ 5h.
Preferably, the step（3）In, centrifugation rate is 65000 rpm, centrifugation time 3h.
Further, the step（4）In, centrifugation rate is 50000 ~ 80000 rpm, and centrifugation time is 1 ~ 5h.
Preferably, the step（4）In, centrifugation rate is 65000 rpm, centrifugation time 3h.
Derive lipid nanometer vesica using new type natural Tea Flower is prepared by any of the above-described the method.
Lipid nanometer vesica is derived in malignant tumour using the new type natural Tea Flower prepared by any of the above-described the method
Application in treatment.
Compared with the prior art, the advantage of the invention is that：
（1）The present invention is extracted from fresh edible plants Tea Flower, and materials are simple and of low cost；
（2）The present invention is had found by spending lipid nanometer vesica to be co-cultured with Several Kinds of Malignancy cell natural tea tree, to more
Kind tumour cell shows the good ability for inhibiting its proliferation, has good broad spectrum anticancer performance；
（3）The it is proposed of the present invention can contribute to us and carry out further research and exploitation to Tea Flower, has prodigious
Economic and clinical conversion meaning.
Description of the drawings
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with attached drawing to the present invention make into
The detailed description of one step, wherein：
Fig. 1 is the preparation process and mechanism of action schematic diagram that natural tea tree spends lipid nanometer vesica in the embodiment of the present invention 1.
Fig. 2 is the Average Particle Diameters and particle diameter distribution that natural tea tree spends lipid nanometer vesica in the embodiment of the present invention 1；
Fig. 3 is the efficient liquid phase chromatographic analysis knot that natural tea tree spends lipid nanometer vesica content ingredient in the embodiment of the present invention 1
Fruit is schemed
Fig. 4 be the embodiment of the present invention 1 in natural tea tree spend lipid nanometer vesica to MCF-7 cells, CT-26 cells, A549 cells,
The in vitro toxicity result figure of Hela cells.
Specific implementation mode
Hereinafter reference will be made to the drawings, and the preferred embodiment of the present invention is described in detail, but is not limited to the present invention
Range.
Embodiment 1
（1）Fresh Tea Flower tissue is put into juice extractor stirring to be crushed, obtains tissue magma；
（2）By tissue magma 4 obtained aboveo20000 rpm centrifuge 3 h under the conditions of C, retain tissue supernatant；
（3）The tissue supernatant 4 that above-mentioned collection is obtainedo65000 rpm centrifuge 3 h under the conditions of C, retain precipitation, phosphoric acid is used in combination
Buffer solution is resuspended；
（4）The re-suspension liquid that above-mentioned collection is obtained further carries out sucrose gradient centrifugation, wherein sucrose concentration be respectively 8%,
30%, 45% and 60%, parameter of noncentricity 4o65000 rpm centrifuge 3 h under the conditions of C；
（5）The natural tea tree collected between 30% and 45% spends lipid nanometer vesica（TFNPs）, it washs and is resuspended afterwards into PBS three times,
It puts to -80oC long-term preservations.
Fig. 1 is the preparation process and mechanism of action schematic diagram that natural tea tree spends lipid nanometer vesica.
Experiment 1：It takes 50 μ L natural tea trees to spend lipid nanometer vesica diluting stock solutions to 1 mL, uses dynamic light scattering
Particle size analyzer（DLS）Analyze its Average Particle Diameters and particle diameter distribution.
Fig. 2 is that natural tea tree spends lipid nanometer vesica particle diameter distribution result figure, as a result shows that lipid nanometer vesica grain size is equal
One, average grain diameter is 131.6 ± 1.6nm.
Experiment 2：In order to probe into natural tea tree spend lipid nanometer Vesicle-Containing at being grouped as, we have carried out height to it
Effect liquid phase chromatogram is analyzed.
Fig. 3 is the constituent analysis figure that natural tea tree spends lipid nanometer Vesicle-Containing, as a result shows Tea Flower lipid nanometer
The catechin components such as EGCG, ECG, EC have been loaded in vesica.
Experiment 3：Natural tea tree spends the effect of lipid nanometer vesica treatment malignant tumour
Fig. 4 is that the natural tea tree of various concentration is spent lipid nanometer vesica and Several Kinds of Malignancy cell（MCF-7 cells, CT-26
Cell, A549 cells, Hela cells）Co-culture multiple time points（24h、48h、72h）, as a result show Tea Flower lipid nanometer
Vesica has the external ability for inhibiting malignant cell proliferation well, and independent with very strong time independence, concentration
Property and anticancer broad spectrum activity.
Finally illustrate, the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although passing through ginseng
According to the preferred embodiment of the present invention, invention has been described, but it will be understood by those of ordinary skill in the art that, it can
To make various changes to it in the form and details, without departing from of the invention defined by the appended claims
Spirit and scope.
Claims (9)
1. a kind of new type natural Tea Flower derives the preparation of lipid nanometer vesica, it is characterised in that：It comprises the steps of:
（1）Fresh Tea Flower tissue is put into juice extractor stirring to be crushed, obtains tissue magma；
（2）By step（1）Obtained tissue magma is 4oIt is centrifuged under the conditions of C, retains tissue supernatant；
（3）By step（2）Obtained tissue supernatant is collected 4oIt is centrifuged under the conditions of C, retains precipitation, phosphate buffer weight is used in combination
It is outstanding, obtain re-suspension liquid；
（4）By step（3）Collect obtained re-suspension liquid and further carry out sucrose gradient centrifugation, wherein sucrose concentration be respectively 8%,
30%, 45% and 60%；
（5）The natural tea tree collected between 30% and 45% spends lipid nanometer vesica（TFNPs）, it washs and is resuspended afterwards into PBS three times,
It puts to -80oC long-term preservations.
2. a kind of new type natural Tea Flower according to claim 1 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（2）Middle centrifugal rotational speed is 6000 ~ 25000 rpm, and centrifugation time is 1 ~ 5 h.
3. a kind of new type natural Tea Flower according to claim 1 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（3）Middle centrifugal rotational speed is 50000 ~ 80000 rpm, and centrifugation time is 1 ~ 5 h.
4. a kind of new type natural Tea Flower according to claim 1 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（4）Middle centrifugal rotational speed is 50000 ~ 80000 rpm, and centrifugation time is 1 ~ 5 h.
5. a kind of new type natural Tea Flower according to claim 2 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（2）Middle centrifugation rate is 20000 rpm, and centrifugation time is 3 h.
6. a kind of new type natural Tea Flower according to claim 3 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（3）Middle centrifugation rate is 65000 rpm, and centrifugation time is 3 h.
7. a kind of new type natural Tea Flower according to claim 4 derives the preparation of lipid nanometer vesica, it is characterised in that：
The step（4）Middle centrifugation rate is 65000 rpm, and centrifugation time is 3 h.
8. the preparation method according to claim 1-7, which obtains new type natural Tea Flower, derives lipid nanometer vesica.
9. deriving application of the lipid nanometer vesica in treating malignant tumor by new type natural Tea Flower according to any one of claims 8.
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| CN201810567200.3A CN108653209A (en) | 2018-06-05 | 2018-06-05 | A kind of new type natural Tea Flower derives the preparation and its application of lipid nanometer vesica |
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| CN201810567200.3A CN108653209A (en) | 2018-06-05 | 2018-06-05 | A kind of new type natural Tea Flower derives the preparation and its application of lipid nanometer vesica |
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| CN108653209A true CN108653209A (en) | 2018-10-16 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110227066A (en) * | 2019-07-03 | 2019-09-13 | 巫贵成 | Excretion body, pharmaceutical carrier and the extracting method of disease treatment, prevention and health care |
| CN113384705A (en) * | 2021-06-28 | 2021-09-14 | 西南大学 | Preparation of poloxamer modified liposome and application of poloxamer modified liposome in oral drug delivery |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04264027A (en) * | 1991-02-16 | 1992-09-18 | Taiyo Kagaku Co Ltd | Composition for preventing colon cancer |
| CN103479597A (en) * | 2012-06-14 | 2014-01-01 | 苏州恒宇生物科技有限公司 | Preparation method and use of grape source active component nano-scale membrane type vesicle |
| CN103479682A (en) * | 2012-06-14 | 2014-01-01 | 苏州恒宇生物科技有限公司 | Preparation method for plant source active component nano-scale membrane type vesicle |
-
2018
- 2018-06-05 CN CN201810567200.3A patent/CN108653209A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04264027A (en) * | 1991-02-16 | 1992-09-18 | Taiyo Kagaku Co Ltd | Composition for preventing colon cancer |
| CN103479597A (en) * | 2012-06-14 | 2014-01-01 | 苏州恒宇生物科技有限公司 | Preparation method and use of grape source active component nano-scale membrane type vesicle |
| CN103479682A (en) * | 2012-06-14 | 2014-01-01 | 苏州恒宇生物科技有限公司 | Preparation method for plant source active component nano-scale membrane type vesicle |
Non-Patent Citations (1)
| Title |
|---|
| 王伟伟等: "茶树花活性成分的提取、分离及生理功效研究进展", 《食品工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110227066A (en) * | 2019-07-03 | 2019-09-13 | 巫贵成 | Excretion body, pharmaceutical carrier and the extracting method of disease treatment, prevention and health care |
| CN113384705A (en) * | 2021-06-28 | 2021-09-14 | 西南大学 | Preparation of poloxamer modified liposome and application of poloxamer modified liposome in oral drug delivery |
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Application publication date: 20181016 |
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