CN105368891A - Taxus chinensis cell culture medium for scale production of paclitaxel - Google Patents
Taxus chinensis cell culture medium for scale production of paclitaxel Download PDFInfo
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- CN105368891A CN105368891A CN201510960018.0A CN201510960018A CN105368891A CN 105368891 A CN105368891 A CN 105368891A CN 201510960018 A CN201510960018 A CN 201510960018A CN 105368891 A CN105368891 A CN 105368891A
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Abstract
The invention provides a taxus chinensis cell culture medium for scale production of paclitaxel. According to the culture medium, a nitrogen source and a phosphorus source are provided through a traditional autotrophy culture medium, certain calcium ions are added, and on this basis, it is unexpectedly discovered that the productive rate of the paclitaxel can be better increased by adding traditional Chinese medicinal materials of specific varieties and components. Based on the beneficial discovery, the traditional Chinese medicinal material components such as Chinese wax, fiveleaf akebia fruits, Chinese yam, orpiment, the tree of heaven ailanthus fruits, the bark of eucommia, radices rehmanniae, beewax and radix ophiopogonis which are used for treatment of human diseases originally are optimized through experiment means. The obtained culture medium achieves an outstanding culture effect unexpectedly, the productive rate of the paclitaxel of taxus chinensis cells is increased obviously, and meanwhile the production period is shorter. The splendid technical effect is achieved through the innovative technological means, meanwhile, cost is low, implementation is easy, and therefore good application prospects are achieved.
Description
Technical field
The present invention relates to technical field of cell culture, be specifically related to a kind of Taxus chinensis cell cultures base for large-scale production taxol.
Background technology
Ramulus et folium taxi cuspidatae, also known as Japanese yew, also claims red Bai Song, is China national one-level rare and protected trees, is also the endangered natural rare plants for anticancer of generally acknowledging in the world.
In the Taxus that taxol and analogue thereof exist only in gymnosperm taxaceae up to now and Austrotaxus, the Chinese yew genus plants whole world has 11 kinds, be distributed widely in the frigid zone of Eurasia and North America, temperate zone and subtropical zone, but the scattered and poor growth of distributing.Austrotaxus is only distributed in the Southern Hemisphere, rare numbers, and taxol content in Chinese yew is general very low, even if also less than 0.1% in the highest yewtree bark of current generally acknowledged content, but it is in great demand.
In order to solve the sharp contradiction between taxol supply and demand, correlative study person has carried out large quantity research in searching and expansion medicine source of Taxol approach for many years.Obtain successfully though the chemistry of taxol is complete synthesis, because the reasons such as route is long, yield is low cannot realize industrialization at present.In recent years, utilize yew cell isolated culture to obtain increasing concern to prepare taxol, because the training method of in vitro cell can obtain more various control, therefore its production efficiency is relatively high.But, be not easy by the cell cultures prepared for the purpose of taxol, first need to screen the strong cell strain of production performance, also to mate suitable culture condition simultaneously, the aspects such as this external nutrition supply need to design for cell strain feature and metabolic characteristic, thus ensure higher culture density and taxol yield.The substratum of prior art is still not ideal enough in promotion yew cell secretion taxol.
Summary of the invention
The present invention is intended to the technological deficiency for prior art; there is provided a kind of Taxus chinensis cell cultures base for large-scale production taxol, to solve technical problem lower for the promoter action of the Taxus chinensis cell cultures base of large-scale production taxol to its secretion taxol in prior art.
For realizing above technical purpose, the present invention by the following technical solutions:
A kind of Taxus chinensis cell cultures base for large-scale production taxol; comprise the composition of following weight part: SODIUMNITRATE 8 ~ 10 parts, dipotassium hydrogen phosphate 4 ~ 6 parts, 2 ~ 3 parts, magnesium sulfate; 1 ~ 2 part, calcium chloride; peptone 1 ~ 2 part, Insect Wax 1 ~ 2 part, Fructus Akebiae 1 ~ 2 part; Chinese yam 1 ~ 2 part; orpiment 1 ~ 2 part, Fruit of Tree-of-heaven Ailanthus 1 ~ 2 part, 50 ~ 60 parts, water.
As preferably, also comprise the bark of eucommia of 1 ~ 2 weight part.
As preferably, also comprise the glutinous rehmannia of 1 ~ 2 weight part.
As preferably, also comprise the beeswax of 1 ~ 2 weight part.
As preferably, also comprise the tuber of dwarf lilyturf of 1 ~ 2 weight part.
As preferably, described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.5 parts, magnesium sulfate, 1.5 parts, calcium chloride, peptone 1.5 parts, Insect Wax 1.5 parts, Fructus Akebiae 1.5 parts, Chinese yam 1.5 parts, orpiment 1.5 parts, Fruit of Tree-of-heaven Ailanthus 1.5 parts, 55 parts, water.
As preferably, described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts, 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts, Insect Wax 1.6 parts, Fructus Akebiae 1.2 parts, Chinese yam 1.1 parts, orpiment 1.4 parts, Fruit of Tree-of-heaven Ailanthus 1.5 parts, the bark of eucommia 1.4 parts, glutinous rehmannia 1.1 parts, 1.5 parts, beeswax, the tuber of dwarf lilyturf 1.8 parts, 57 parts, water.
In above technical scheme, described substratum is pulverizing after each composition mixing by formula ratio, is mixed to get with water, and described pulverizing can utilize all kinds of crushing material means well-known in the art to realize.Substratum of the present invention is particularly useful for the fluid suspension culture of yew cell, certainly also can be prepared as solid medium for seed culture or Secondary Culture.
The present invention by experiment means has extensively screened the nutritive substance as yew cell growth, determines formula of the present invention according to yew cell own metabolism characteristic and practical effect.This formula provides nitrogenous source and phosphorus source with traditional autotrophy substratum, and with the addition of certain calcium ion, and unexpected discovery helps lend some impetus to the productive rate of taxol by the Chinese medicinal materials adding particular types and composition on this basis.Based on discovery useful above, the present invention's means are preferred by experiment Insect Wax, Fructus Akebiae, Chinese yam, orpiment etc. are used for the Chinese medicinal materials composition of human disease treatment originally, gained substratum surprisingly obtains outstanding culture effect, the taxol yield of yew cell is significantly improved, and its production cycle is shorter simultaneously.The present invention obtains superior technique effect with the technique means of novelty, simultaneously cost lower, be easy to realize, therefore have a good application prospect.
Embodiment
Below will be described in detail the specific embodiment of the present invention.In order to avoid too much unnecessary details, in the examples below to belonging to known structure or function will not be described in detail.Apart from outside definition, technology used in following examples and scientific terminology have the identical meanings generally understood with those skilled in the art of the invention.
Embodiment 1
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 8 parts, dipotassium hydrogen phosphate 4 parts, 2 parts, magnesium sulfate; 1 part, calcium chloride, peptone 1 part, Insect Wax 1 part, Fructus Akebiae 1 part; Chinese yam 1 part, orpiment 1 part, Fruit of Tree-of-heaven Ailanthus 1 part, 50 parts, water.
Embodiment 2
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 10 parts, dipotassium hydrogen phosphate 6 parts, 3 parts, magnesium sulfate; 2 parts, calcium chloride, peptone 2 parts, Insect Wax 2 parts, Fructus Akebiae 2 parts; Chinese yam 2 parts, orpiment 2 parts, Fruit of Tree-of-heaven Ailanthus 2 parts, 60 parts, water.
Embodiment 3
A kind of Taxus chinensis cell cultures base for large-scale production taxol; be grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.3 parts, magnesium sulfate; 1.8 parts, calcium chloride; peptone 1.5 parts, Insect Wax 1.6 parts, Fructus Akebiae 1.7 parts; Chinese yam 1.8 parts; orpiment 1.2 parts, Fruit of Tree-of-heaven Ailanthus 1.8 parts, 55 parts, water.
Embodiment 4
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.8 parts, dipotassium hydrogen phosphate 5.4 parts; 2.7 parts, magnesium sulfate, 1.2 parts, calcium chloride, peptone 1.2 parts; Insect Wax 1.2 parts; Fructus Akebiae 1.5 parts, Chinese yam 1.6 parts, orpiment 1.4 parts; Fruit of Tree-of-heaven Ailanthus 1.7 parts; the bark of eucommia 1 part, the tuber of dwarf lilyturf 1.1 parts, 52 parts, water.
Embodiment 5
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 9.4 parts, dipotassium hydrogen phosphate 4.2 parts; 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.1 parts; Insect Wax 1 part, Fructus Akebiae 2 parts, Chinese yam 1.9 parts; orpiment 1.2 parts; Fruit of Tree-of-heaven Ailanthus 1.9 parts, glutinous rehmannia 1.3 parts, 1.2 parts, beeswax; the tuber of dwarf lilyturf 1.6 parts, 58 parts, water.
Embodiment 6
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.8 parts, dipotassium hydrogen phosphate 5.4 parts; 2.7 parts, magnesium sulfate; 1.2 parts, calcium chloride, peptone 1.2 parts, Insect Wax 1.2 parts; Fructus Akebiae 1.5 parts; Chinese yam 1.6 parts, orpiment 1.4 parts, Fruit of Tree-of-heaven Ailanthus 1.7 parts; the bark of eucommia 1.5 parts, 55 parts, water.
Embodiment 7
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 9.2 parts, dipotassium hydrogen phosphate 5.7 parts; 2.4 parts, magnesium sulfate; 1.5 parts, calcium chloride, peptone 1.5 parts, Insect Wax 1.7 parts; Fructus Akebiae 1.7 parts; Chinese yam 1.2 parts, orpiment 1.9 parts, Fruit of Tree-of-heaven Ailanthus 2 parts; 1.8 parts, beeswax, 59 parts, water.
Embodiment 8
For a Taxus chinensis cell cultures base for large-scale production taxol, be grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts; 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts; Insect Wax 1.6 parts, Fructus Akebiae 1.2 parts, Chinese yam 1.1 parts; orpiment 1.4 parts, Fruit of Tree-of-heaven Ailanthus 1.5 parts, the bark of eucommia 1.4 parts; glutinous rehmannia 1.1 parts; 1.5 parts, beeswax, the tuber of dwarf lilyturf 1.8 parts, 57 parts, water.
Above embodiments of the invention have been described in detail, but described content is only preferred embodiment of the present invention, not in order to limit the present invention.All make in application range of the present invention any amendment, equivalent to replace and improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. the Taxus chinensis cell cultures base for large-scale production taxol; it is characterized in that the composition comprising following weight part: SODIUMNITRATE 8 ~ 10 parts, dipotassium hydrogen phosphate 4 ~ 6 parts, 2 ~ 3 parts, magnesium sulfate; 1 ~ 2 part, calcium chloride; peptone 1 ~ 2 part, Insect Wax 1 ~ 2 part, Fructus Akebiae 1 ~ 2 part; Chinese yam 1 ~ 2 part; orpiment 1 ~ 2 part, Fruit of Tree-of-heaven Ailanthus 1 ~ 2 part, 50 ~ 60 parts, water.
2. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1, characterized by further comprising the bark of eucommia of 1 ~ 2 weight part.
3. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1, characterized by further comprising the glutinous rehmannia of 1 ~ 2 weight part.
4. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1, characterized by further comprising the beeswax of 1 ~ 2 weight part.
5. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1, characterized by further comprising the tuber of dwarf lilyturf of 1 ~ 2 weight part.
6. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1; it is characterized in that described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 9 parts, dipotassium hydrogen phosphate 5 parts, 2.5 parts, magnesium sulfate; 1.5 parts, calcium chloride; peptone 1.5 parts, Insect Wax 1.5 parts, Fructus Akebiae 1.5 parts; Chinese yam 1.5 parts; orpiment 1.5 parts, Fruit of Tree-of-heaven Ailanthus 1.5 parts, 55 parts, water.
7. the Taxus chinensis cell cultures base for large-scale production taxol according to claim 1, is characterized in that described substratum is grouped into by the one-tenth of following weight part: SODIUMNITRATE 8.5 parts, dipotassium hydrogen phosphate 5.5 parts; 2.1 parts, magnesium sulfate, 1.3 parts, calcium chloride, peptone 1.7 parts; Insect Wax 1.6 parts, Fructus Akebiae 1.2 parts, Chinese yam 1.1 parts; orpiment 1.4 parts, Fruit of Tree-of-heaven Ailanthus 1.5 parts, the bark of eucommia 1.4 parts; glutinous rehmannia 1.1 parts; 1.5 parts, beeswax, the tuber of dwarf lilyturf 1.8 parts, 57 parts, water.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510960018.0A CN105368891A (en) | 2015-12-18 | 2015-12-18 | Taxus chinensis cell culture medium for scale production of paclitaxel |
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| CN201510960018.0A CN105368891A (en) | 2015-12-18 | 2015-12-18 | Taxus chinensis cell culture medium for scale production of paclitaxel |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696436A (en) * | 2009-10-30 | 2010-04-21 | 梅州市杉维生物医药工程研究有限公司 | Special culture medium for culturing yew plant cells to produce taxol |
| CN102187782A (en) * | 2010-03-10 | 2011-09-21 | 德阳市食用菌专家大院 | Method for culturing lucid ganoderma by using culture medium containing non-medicinal parts of Chinese medicinal materials |
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2015
- 2015-12-18 CN CN201510960018.0A patent/CN105368891A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696436A (en) * | 2009-10-30 | 2010-04-21 | 梅州市杉维生物医药工程研究有限公司 | Special culture medium for culturing yew plant cells to produce taxol |
| CN102187782A (en) * | 2010-03-10 | 2011-09-21 | 德阳市食用菌专家大院 | Method for culturing lucid ganoderma by using culture medium containing non-medicinal parts of Chinese medicinal materials |
Non-Patent Citations (1)
| Title |
|---|
| 李永成: "东北红豆杉悬浮细胞与内生真菌在紫杉醇合成中相互关系的研", 《中国博士学位论文全文数据库，农业科技辑》 * |
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