CN108451966A - It is a kind of to be used to substitute composition and its assay method that paris polyphylla prepares treatment of ovarian cancer drug - Google Patents
It is a kind of to be used to substitute composition and its assay method that paris polyphylla prepares treatment of ovarian cancer drug Download PDFInfo
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- CN108451966A CN108451966A CN201810458292.1A CN201810458292A CN108451966A CN 108451966 A CN108451966 A CN 108451966A CN 201810458292 A CN201810458292 A CN 201810458292A CN 108451966 A CN108451966 A CN 108451966A
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- G01N30/02—Column chromatography
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- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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Abstract
The composition and its assay method that the invention discloses a kind of to prepare treatment of ovarian cancer drug for substituting paris polyphylla.The composition being made of chonglou saponin VII, pennogenin 3 O α L Rha (1 → 2) [α L Rha (1 → 4)] β D Glc, chonglou saponin II, Gracillin and chonglou saponin I substantially can be with equivalent substitution paris polyphylla to LSD1 gene expressions depression effect, the two intensity is suitable；The composition can be used for equivalent substitution paris polyphylla and prepare LSD1 gene expression inhibitors；Those skilled in the art will know that, inhibiting LSD1 gene expressions that can effectively inhibit the proliferation of ovarian cancer cell, (expression of silence LSD1 genes can inhibit the proliferation of oophoroma HO8910 cells and induce its apoptosis, tumour the 5th phase of volume 36 in May, 2016), therefore, the composition can be used for equivalent substitution paris polyphylla and prepare treatment of ovarian cancer drug.
Description
Technical field
The present invention relates to a kind of compositions and its assay method preparing tumor therapeutic agent for substituting paris polyphylla.
Background technology
Paris Paris L. are broad sense liliaceous plant, and rhizome is used as medicine, and have clearing heat and detoxicating, swelling and pain relieving and cool liver
The effect of arresting convulsion, is mainly used for antitumor, hemostasis, immunological regulation, sedation and analgesia etc..The whole world has 24 kinds, and China has 19 kinds, master
It is distributed in the ground such as Yunnan, Sichuan, Guangxi, wherein paris polyphylla Parispolyphylla Smithvar.Yunnanensis
(Franch.) Hand.-Mazz. quilts《Chinese Pharmacopoeia》Version is included within 2010.
At present since medical industry production is continuously increased Paris polyphylla demand so that wild Paris polyphylla is fewer and fewer, resource
It is increasingly exhausted.If can search out can substitute the composition that Paris polyphylla is used as medicine, so that it may be obtained greatly in the method blent with chemistry
Paris polyphylla substitute is measured, the present situation of Paris polyphylla resource anxiety is alleviated.
Mainly contained in paris polyphylla chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 →
4)]-β-D-Glc (PGRR), chonglou saponin H, chonglou saponin II, Gracillin, chonglou saponin I etc..At present, in this case it is not apparent that
Which kind combination can substitute the drug effect of Paris polyphylla substantially in these ingredients.
Invention content
An object of the present disclosure is to provide a kind of composition preparing tumor therapeutic agent for substituting paris polyphylla；
The present invention second is designed to provide the assay method of above-mentioned composition.
The purpose of the present invention is realized by following technical solution：
First part：Cervical carcinoma equivalent composition
A kind of composition preparing p62 gene expression inhibitors for equivalent substitution paris polyphylla：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, Paris polyphylla
Saponin(e H, chonglou saponin II and Gracillin composition.
Application of the above-mentioned composition in terms of equivalent substitution paris polyphylla prepares p62 gene expression inhibitors.
A kind of composition preparing treatment of human cervical cancer drug for equivalent substitution paris polyphylla：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, Paris polyphylla
Saponin(e H, chonglou saponin II and Gracillin composition.
Application of the above-mentioned composition in terms of equivalent substitution paris polyphylla prepares treatment of human cervical cancer drug.
A kind of HPLC methods of each chemical composition content in paris polyphylla in measurement above-mentioned composition, including following chromatography ginseng
Number：
Chromatographic column：C18 filler chromatographic columns；
Mobile phase：Acetonitrile (A)-water (B)；
Elution program：0~10min, 20%~30%A；10~20min, 30%~40%A；20~35min, 40%~
50%A；35~45min, 50%~60%A；45~55min, 60%~80%A；55~60min, 80%~100%A.
Preferably, using 1100 liquid chromatographs of Agilent.
Preferably, using Agilent ZORBAX Extend-C18 liquid-phase chromatographic columns.
It is highly preferred that chromatographic column specification is 250mm × 4.6mm, 5 μm.
Preferably, column temperature is 30 DEG C.
Preferably, Detection wavelength 210nm.
Preferably, flow rate of mobile phase 1.0mL/min.
Preferably, sample size is 20 μ L.
Second part：Oophoroma equivalent composition
A kind of composition preparing LSD1 gene expression inhibitors for equivalent substitution paris polyphylla：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, Paris polyphylla
Saponin I I, Gracillin and chonglou saponin I compositions.
Application of the above-mentioned composition in terms of equivalent substitution paris polyphylla prepares LSD1 gene expression inhibitors.
A kind of composition preparing treatment of ovarian cancer drug for equivalent substitution paris polyphylla：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, Paris polyphylla
Saponin I I, Gracillin and chonglou saponin I compositions.
Application of the above-mentioned composition in terms of equivalent substitution paris polyphylla prepares treatment of ovarian cancer drug.
A kind of HPLC methods of each chemical composition content in paris polyphylla in measurement above-mentioned composition, including following chromatography ginseng
Number：
Chromatographic column：C18 filler chromatographic columns；
Mobile phase：Acetonitrile (A)-water (B)；
Elution program：0~10min, 20%~30%A；10~20min, 30%~40%A；20~35min, 40%~
50%A；35~45min, 50%~60%A；45~55min, 60%~80%A；55~60min, 80%~100%A.
Preferably, using 1100 liquid chromatographs of Agilent.
Preferably, using Agilent ZORBAX Extend-C18 liquid-phase chromatographic columns.
It is highly preferred that chromatographic column specification is 250mm × 4.6mm, 5 μm.
Preferably, column temperature is 30 DEG C.
Preferably, Detection wavelength 210nm.
Preferably, flow rate of mobile phase 1.0mL/min.
Preferably, sample size is 20 μ L.
Advantageous effect：
(1) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
The composition of building saponin(e H, chonglou saponin II and Gracillin composition substantially can be with equivalent substitution paris polyphylla to p62 gene tables
The depression effect reached, the two intensity are suitable；The composition can be used for equivalent substitution paris polyphylla and prepare p62 gene expression inhibitions
Agent；One skilled in the art will appreciate that inhibition p62 gene expressions can effectively inhibit proliferation (the RNA AF panels of cervical cancer cell
Influence of the p62 gene expressions to proliferation of cervical cancer HeLa cell, Bengbu Medical College's journal the 1st phase of volume 43 in January, 2018), because
This, the composition can be used for equivalent substitution paris polyphylla and prepare treatment of human cervical cancer drug；
(2) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
In the composition of building saponin(e H, chonglou saponin II and Gracillin composition, pennogenin -3-O- α-L-Rha (1 → 2) [α -
L-Rha (1 → 4)]-β-D-Glc are 11% to the independent inhibiting rate of p62 gene expressions, other 4 kinds of ingredients are to p62 gene expressions
Combination inhibiting rate be 19%, group of significantly less than the 5 kinds ingredients of sum of the two (11%+19%=30%) to p62 gene expressions
The phenomenon that conjunction inhibiting rate 53%, this 2 1+1 ＜, embodies pennogenin -3-O- α-L-Rha (1 → 2) [α-L- in the composition
Rha (1 → 4)] there are the effects for significantly cooperate with inhibition p62 gene expressions in-β-D-Glc and other 4 kinds of ingredients；
(3) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
The composition of building saponin I I, Gracillin and chonglou saponin I compositions substantially can be with equivalent substitution paris polyphyllas to LSD1 genes
The depression effect of expression, the two intensity are suitable；The composition can be used for equivalent substitution paris polyphylla and prepare LSD1 gene expressions suppression
Preparation；One skilled in the art will appreciate that inhibition LSD1 gene expressions can effectively inhibit proliferation (the silence LSD1 of ovarian cancer cell
The expression of gene can inhibit the proliferation of oophoroma HO8910 cells and induce its apoptosis, tumour the 5th phase of volume 36 in May, 2016),
Therefore, the composition can be used for equivalent substitution paris polyphylla and prepare treatment of ovarian cancer drug；
(4) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
In the composition of building saponin I I, Gracillin and chonglou saponin I compositions, pennogenin -3-O- α-L-Rha (1 → 2) [α -
L-Rha (1 → 4)]-β-D-Glc are 12% to the independent inhibiting rate of LSD1 gene expressions, other 4 kinds of ingredients are to LSD1 gene tables
The combination inhibiting rate reached is 22%, and significantly less than 5 kinds ingredients of sum of the two (12%+22%=34%) are to LSD1 gene expressions
The phenomenon that combination inhibiting rate 48%, this 2 1+1 ＜, embodies pennogenin -3-O- α-L-Rha (1 → 2) [α-in the composition
L-Rha (1 → 4)] there are the effects for significantly cooperate with inhibition LSD1 gene expressions in-β-D-Glc and other 4 kinds of ingredients.
Description of the drawings
Fig. 1 is the influence of paris polyphylla drug and composition to the expression of HeLa cell p62 genes；
Fig. 2 is the influence of paris polyphylla drug and composition to the expression of HO-8910 Proliferation of Human Ovarian Cell LSD1 genes.
Specific implementation mode
It is specific with reference to the accompanying drawings and examples to introduce essentiality content of the present invention, but protection of the present invention is not limited with this
Range.
One, experiment material
Paris polyphylla medicinal material is purchased from Bozhou medicinal material market, and the place of production is Lijiang, yunnan, is dried overnight for 55 DEG C after crushing, kept dry
It is spare.
1100 liquid chromatographs of Agilent；Sartorius BS210S type electronic balances, Beijing Sai Duolisi instruments system
Unite Co., Ltd；Sartorius CP225D type electronic balances, Beijing Sai Duolisi instrument systems Co., Ltd；Acetonitrile (chromatography
It is pure), methanol (chromatographically pure) be purchased from the U.S. world (TEDIA)；Water is Wahaha Pure Water.
RNAiso plus reagents, SYBRGreen PCRMaster Mix and PrimeScript R kits are purchased from
TaKaRa。
Two, experimental method
1, chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc
(PGRR), the assay of chonglou saponin H, chonglou saponin II, Gracillin, chonglou saponin I in paris polyphylla
HPLC methods include following chromatographic parameter：
Chromatograph：1100 liquid chromatographs of Agilent
Chromatographic column：Agilent ZORBAX Extend-C18 liquid-phase chromatographic columns (250mm × 4.6mm, 5 μm)；
Mobile phase：Acetonitrile (A)-water (B)；
Elution program：0~10min, 20%~30%A；10~20min, 30%~40%A；20~35min, 40%~
50%A；35~45min, 50%~60%A；45~55min, 60%~80%A；55~60min, 80%~100%A；
Column temperature：30℃.
Detection wavelength：210nm.
Flow rate of mobile phase：1.0mL/min.
Sample size：20μL.
The preparation of test solution：By paris polyphylla pulverizing medicinal materials, 40 mesh sieve is crossed, precision weighs 10g, sets in round-bottomed flask,
70% ethyl alcohol 40mL, water-bath reflux 2h is added, filtration takes filtrate, is repeated 3 times.Merging filtrate boils off drying after ethyl alcohol, obtains powder
End.Powder is dissolved in 250mL methanol, cross miillpore filter to get.
Precision measures 20 μ L of test solution and injects liquid chromatograph, with standard items external standard method chonglou saponin VII, partially
Promise saponin(e -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc (PGRR), chonglou saponin H, chonglou saponin II, fibre
The thin content of Dioscin, chonglou saponin I in paris polyphylla.
2, cell culture
Adenocarcinoma of the uterine cervix HeLa cells, HO-8910 Proliferation of Human Ovarian Cell are respectively in the DMEM in high glucose training containing 10% fetal calf serum
It supports in base, 37 DEG C, 5%CO2Under the conditions of secondary culture.Logarithmic growth phase cell is for testing.
3, medicine preparation
Paris polyphylla drug：By paris polyphylla pulverizing medicinal materials, 40 mesh sieve is crossed, precision weighs 10g, sets in round-bottomed flask, is added
70% ethyl alcohol 40mL, water-bath reflux 2h, filtration take filtrate, are repeated 3 times.Merging filtrate boils off drying after ethyl alcohol, obtains powder.
Composition medicine ingredient type such as following table, each ingredient are blent according to the content ratio in paris polyphylla with standard items
It is made.
| Serial number | Quantity | Ingredient type |
| Composition medicine 1 | 5 | Chonglou saponin VII, PGRR, chonglou saponin H, chonglou saponin II, Gracillin |
| Composition medicine 2 | 5 | Chonglou saponin VII, PGRR, chonglou saponin II, Gracillin, chonglou saponin I |
| Composition medicine 3 | 4 | Chonglou saponin VII, chonglou saponin H, chonglou saponin II, Gracillin |
| Composition medicine 4 | 4 | Chonglou saponin VII, chonglou saponin II, Gracillin, chonglou saponin I |
| Composition medicine 5 | 1 | PGRR |
4, it is grouped and is administered
By a concentration of 5.5 × 104The HeLa cell suspension inoculations of/mL after culture for 24 hours, are separately added into Yunnan in 96 orifice plates
Paris polyphylla drug, composition medicine 1, composition medicine 3, composition medicine 5 make its final concentration be equivalent to 250 μ g/ of paris polyphylla medicinal material
ML, while negative control group is set, every group of 6 parallel holes, 37 DEG C, 5%CO2After cultivating 48h in cell incubator, supernatant is abandoned in centrifugation
Cell is collected in liquid, PBS washings, spare.
By a concentration of 5.5 × 104The HO-8910 Proliferation of Human Ovarian Cell suspensions of/mL are inoculated in 96 orifice plates, after culture for 24 hours,
Being separately added into paris polyphylla drug, composition medicine 2, composition medicine 4, composition medicine 5 makes its final concentration be equivalent to paris polyphylla
250 μ g/mL of medicinal material, while negative control group is set, every group of 6 parallel holes, 37 DEG C, 5%CO2After 48h being cultivated in cell incubator,
Supernatant is abandoned in centrifugation, and cell is collected in PBS washings, spare.
5, qRT-PCR measures the expression of HeLa cell p62 genes and HO-8910 Proliferation of Human Ovarian Cell LSD1 genes
Above-mentioned cell is collected, cell is washed 3 times with PBS, it is total according to RNAiso Plus kit operation manuals extraction cell
Ultraviolet spectrophotometry row quantitative analysis is used in combination in RNA.Use PrimeScript RT kits by RNA reverse transcriptions for cDNA, with
CDNA is template, using the further row PCR amplification of SYBR-Green PCR Master Mix kits.
P62 upstream region of gene primer sequences are 5 '-TTTTCACTGTCAATTCACTGCA-3 '；
P62 downstream of gene primer sequences are 5 '-GGAGGCGAGGATGGAGGAA-3 '；
LSD1 upstream region of gene primer sequences are 5 '-CAAGTGTCAATTTGTTCGGG-3 '；
LSD1 downstream of gene primer sequences are 5 '-TTCTTTGGGCTGAGGTACTG-3 '；
Internal reference GAPDH upstream region of gene primer sequences are 5 '-GCAAATTCCATGGCACCGTC-3 '；
Internal reference GAPDH downstream of gene primer sequences are 5 '-TCGCCCCACTTGATTTTGG-3 '.
PCR reaction conditions：90 DEG C of 30s, 1 cycle；95 DEG C of 5s, 60 DEG C of 30s, totally 40 recycle.With 2-ΔΔCtIt is worth (Ct generations
Table cycle threshold) indicate p62mRNA, LSD1mRNA relative expression quantity.Experiment is repeated 3 times.
6, statistical method
The statistics and analysis of data is carried out using 5 softwares of Graph PadPrism.Measurement data is indicated with x ± s, multigroup
Between compare using one-way analysis of variance, compare examined using Tukey two-by-two.P ＜ 0.05 are that difference is statistically significant.
Three, experimental result
1, chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc
(PGRR), the assay result of chonglou saponin H, chonglou saponin II, Gracillin, chonglou saponin I in paris polyphylla
Chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc (PGRR),
The content of chonglou saponin H, chonglou saponin II, Gracillin, chonglou saponin I in paris polyphylla is as shown in the table.
| Compound name | Content (mg/g) | Compound name | Content (mg/g) |
| Chonglou saponin VII | 26.25 | Chonglou saponin II | 30.46 |
| PGRR | 10.50 | Gracillin | 1.95 |
| Chonglou saponin H | 4.18 | Chonglou saponin I | 96.52 |
2, the influence of paris polyphylla drug and composition to the expression of HeLa cell p62 genes
The expression of paris polyphylla drug, composition medicine 1, composition medicine 3, composition medicine 5 to HeLa cell p62 genes
Horizontal influence is as shown in following table and Fig. 1.
| Group | p62mRNA/GAPDHmRNA | Inhibiting rate |
| Negative control group | 1.00 | / |
| Paris polyphylla drug (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.45 | 55% |
| Composition medicine 1 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.47 | 53% |
| Composition medicine 3 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.81 | 19% |
| Composition medicine 5 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.89 | 11% |
Above-mentioned experimental result explanation：
(1) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
The composition of building saponin(e H, chonglou saponin II and Gracillin composition substantially can be with equivalent substitution paris polyphylla to p62 gene tables
The depression effect reached, the two intensity are suitable；The composition can be used for equivalent substitution paris polyphylla and prepare p62 gene expression inhibitions
Agent；One skilled in the art will appreciate that inhibition p62 gene expressions can effectively inhibit proliferation (the RNA AF panels of cervical cancer cell
Influence of the p62 gene expressions to proliferation of cervical cancer HeLa cell, Bengbu Medical College's journal the 1st phase of volume 43 in January, 2018), because
This, the composition can be used for equivalent substitution paris polyphylla and prepare treatment of human cervical cancer drug；
(2) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
In the composition of building saponin(e H, chonglou saponin II and Gracillin composition, pennogenin -3-O- α-L-Rha (1 → 2) [α -
L-Rha (1 → 4)]-β-D-Glc are 11% to the independent inhibiting rate of p62 gene expressions, other 4 kinds of ingredients are to p62 gene expressions
Combination inhibiting rate be 19%, group of significantly less than the 5 kinds ingredients of sum of the two (11%+19%=30%) to p62 gene expressions
The phenomenon that conjunction inhibiting rate 53%, this 2 1+1 ＜, embodies pennogenin -3-O- α-L-Rha (1 → 2) [α-L- in the composition
Rha (1 → 4)] there are the effects for significantly cooperate with inhibition p62 gene expressions in-β-D-Glc and other 4 kinds of ingredients.
3, the influence of paris polyphylla drug and composition to the expression of HO-8910 Proliferation of Human Ovarian Cell LSD1 genes
Paris polyphylla drug, composition medicine 2, composition medicine 4, composition medicine 5 are to HO-8910 Proliferation of Human Ovarian Cell
The influence of the expression of LSD1 genes is as shown in following table and Fig. 2.
| Group | LSD1mRNA/GAPDHmRNA | Inhibiting rate |
| Negative control group | 1.00 | / |
| Paris polyphylla drug (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.53 | 47% |
| Composition medicine 2 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.52 | 48% |
| Composition medicine 4 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.78 | 22% |
| Composition medicine 5 (is equivalent to 250 μ g/mL of paris polyphylla medicinal material) | 0.88 | 12% |
Above-mentioned experimental result explanation：
(1) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
The composition of building saponin I I, Gracillin and chonglou saponin I compositions substantially can be with equivalent substitution paris polyphyllas to LSD1 genes
The depression effect of expression, the two intensity are suitable；The composition can be used for equivalent substitution paris polyphylla and prepare LSD1 gene expressions suppression
Preparation；One skilled in the art will appreciate that inhibition LSD1 gene expressions can effectively inhibit proliferation (the silence LSD1 of ovarian cancer cell
The expression of gene can inhibit the proliferation of oophoroma HO8910 cells and induce its apoptosis, tumour the 5th phase of volume 36 in May, 2016),
Therefore, the composition can be used for equivalent substitution paris polyphylla and prepare treatment of ovarian cancer drug；
(2) by chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, again
In the composition of building saponin I I, Gracillin and chonglou saponin I compositions, pennogenin -3-O- α-L-Rha (1 → 2) [α -
L-Rha (1 → 4)]-β-D-Glc are 12% to the independent inhibiting rate of LSD1 gene expressions, other 4 kinds of ingredients are to LSD1 gene tables
The combination inhibiting rate reached is 22%, and significantly less than 5 kinds ingredients of sum of the two (12%+22%=34%) are to LSD1 gene expressions
The phenomenon that combination inhibiting rate 48%, this 2 1+1 ＜, embodies pennogenin -3-O- α-L-Rha (1 → 2) [α-in the composition
L-Rha (1 → 4)] there are the effects for significantly cooperate with inhibition LSD1 gene expressions in-β-D-Glc and other 4 kinds of ingredients.
Claims (10)
1. a kind of composition preparing LSD1 gene expression inhibitors for equivalent substitution paris polyphylla, it is characterised in that：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, chonglou saponin
II, Gracillin and chonglou saponin I compositions.
2. application of the composition described in claim 1 in terms of equivalent substitution paris polyphylla prepares LSD1 gene expression inhibitors.
3. a kind of composition preparing treatment of ovarian cancer drug for equivalent substitution paris polyphylla, it is characterised in that：
By chonglou saponin VII, pennogenin -3-O- α-L-Rha (1 → 2) [α-L-Rha (1 → 4)]-β-D-Glc, chonglou saponin
II, Gracillin and chonglou saponin I compositions.
4. application of the composition described in claim 1 in terms of equivalent substitution paris polyphylla prepares treatment of ovarian cancer drug.
5. a kind of HPLC methods measuring each chemical composition content in paris polyphylla in composition described in claim 1, special
Sign is, including following chromatographic parameter：
Chromatographic column：C18 filler chromatographic columns；
Mobile phase：Acetonitrile (A)-water (B)；
Elution program：0~10min, 20%~30%A；10~20min, 30%~40%A；20~35min, 40%~50%
A；35~45min, 50%~60%A；45~55min, 60%~80%A；55~60min, 80%~100%A.
6. HPLC methods according to claim 3, it is characterised in that：Using 1100 liquid chromatographs of Agilent.
7. HPLC methods according to claim 3, it is characterised in that：Using Agilent ZORBAX Extend-C18 liquid
Phase chromatographic column (250mm × 4.6mm, 5 μm).
8. HPLC methods according to claim 3, it is characterised in that：Column temperature is 30 DEG C.
9. HPLC methods according to claim 3, it is characterised in that：Detection wavelength is 210nm.
10. HPLC methods according to claim 3, it is characterised in that：Flow rate of mobile phase is 1.0mL/min.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810458292.1A CN108451966A (en) | 2018-05-14 | 2018-05-14 | It is a kind of to be used to substitute composition and its assay method that paris polyphylla prepares treatment of ovarian cancer drug |
| PCT/CN2018/103855 WO2019218543A1 (en) | 2018-05-14 | 2018-09-03 | Discovery and pharmaceutical use of equivalent component group of traditional chinese medicine rhizoma paridis yunnanensis, scutellaria indica, fructus xanthii sibirici, or bupleurum bicaule helm |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101693035A (en) * | 2009-10-15 | 2010-04-14 | 天津大学 | Medicinal preparation with inhibiting effect on tumor metastasis |
| CN101912514A (en) * | 2010-07-27 | 2010-12-15 | 天津大学 | Paris rhizome total saponin capsules with anti-tumor effect and preparation method thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101693035A (en) * | 2009-10-15 | 2010-04-14 | 天津大学 | Medicinal preparation with inhibiting effect on tumor metastasis |
| CN101912514A (en) * | 2010-07-27 | 2010-12-15 | 天津大学 | Paris rhizome total saponin capsules with anti-tumor effect and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 刘宗谕等: "重楼皂苷抑制卵巢癌细胞增殖和转移、诱导其凋亡分子机制研究", 《中国实验诊断学》 * |
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