CN106939031B - A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes - Google Patents
A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes Download PDFInfo
- Publication number
- CN106939031B CN106939031B CN201710086724.6A CN201710086724A CN106939031B CN 106939031 B CN106939031 B CN 106939031B CN 201710086724 A CN201710086724 A CN 201710086724A CN 106939031 B CN106939031 B CN 106939031B
- Authority
- CN
- China
- Prior art keywords
- spirosolane
- preparation
- added
- volume
- volumetric concentration
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J71/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton is condensed with a heterocyclic ring
- C07J71/0005—Oxygen-containing hetero ring
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides a kind of spirosolane type glycoalkaloid (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside and preparation method thereof and purposes, the spirosolane type glycoalkaloid is to human A549 cell lines, human large cell lung cancer cell H460 and people's Lung Squamous Carcinoma Cells SK-MES-1 all have inhibiting effect, it and is 19.22 μ g/ml to the 503nhibiting concentration of A549 cell, 503nhibiting concentration to H460 cell is 80.03 μ g/ml, 503nhibiting concentration to SK-MES-1 cell is 54.20 μ g/ml, it can be seen that (3 β that the present invention is prepared, 5 α , 22 α, 25R) and-spirosolane -3-O- β-D- glucopyranosyl-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside and its preparation can be used for treating non-small cell lung cancer.
Description
Technical field
The invention belongs to glycoalkaloid preparation method and medicinal usage technical fields, and in particular to a kind of spirosolane
Type glycoalkaloid and preparation method thereof and purposes.
Background technique
Glycoalkaloid is mainly distributed in Solanaceae and liliaceous plant, by hydrophobic aglycon and hydrophilic widow
Sugar chain is constituted, and aglycon includes three kinds of spirosolane, solanidane and other steroid derivatives, and oligonucleotide chain is by 3~4 monosaccharide
Composition, is a kind of steroid saponin.Induction biosynthesis glycoalkaloid primarily to defence microorganism, animal and insect invasion,
Therefore, glycoalkaloid has certain toxicity, but at the same time, it also has reduction cholesterol and hypertension, anti-inflammatory, promoting blood circulation
A variety of physiological activity such as analgesia, antiallergy, enhancing immunity of organisms, resisting pathogenic microbes, have potential medical value.Solanaceae
It is widespread in nature with liliaceous plant, such as bittersweet, potato and tomato plant, there is very strong economic valence
Value, studies of Solanum lyratum Thunb is sweet in flavor first recorded in " legendary god of farming hundred grass warp ", cold in nature, has heat-clearing, removing toxic substances, wind-dispelling, stagnation resolvation and other effects, research
The main chemical compositions for showing bittersweet are glycoalkaloid, and therefore, novel glycoalkaloid is extracted in research from studies of Solanum lyratum Thunb
It is of great significance.
In view of said medicine effect of glycoalkaloid, the world of medicine pays much attention to it, Chinese patent literature CN
101450144A discloses a kind of Solanum dulcamara total-saponin extract and its preparation method and application, which is dulcain B, another name for Sichuan Province
Sheep spring glycosides C, δ-one of solamarine and Dioscin (structural formula is as follows) or a variety of, above-mentioned Solanum dulcamara total-saponin extract are available
In diseases such as treatment gastritis, gastric duodenal ulcer, gastric cancer and psoriasis, and there is good therapeutic effect.It is well known that
Solanum dulcamara total-saponin extract is a kind of mixture comprising the compound of various structures, but not can determine that every kind of compound
Identical or corresponding effect is all had, i.e., can not include in Solanum dulcamara total-saponin extract disclosed in fully validated above-mentioned document
Compound good synergistic effect can be played to the diseases such as treatment gastritis, gastric duodenal ulcer, gastric cancer and psoriasis,
This is also that Chinese medical extract has the reason of unpredictalbe side effect, therefore, is carried out to medicinal plant extract further
Purification and research, the compound of Various Complex is separated and is extracted, and carries out confirmation to its effect and have great importance,
For this purpose, how to be efficiently separated and obtain having the compound of potential medical value to medicinal plant extract is all doctors
The project that medicine worker must face.
Summary of the invention
The technical problem to be solved by the present invention is to medicinal plant extract complicated component in the prior art, all the components
The effect of and influence each other cannot be fully validated defect, to provide a kind of compound spirosolane type sugar of one-component
Glycosides alkaloid, and then preparation method and its purposes in preparation treatment non-small cell lung cancer drug are provided.
For this purpose, the present invention realizes the technical solution of above-mentioned purpose are as follows:
The present invention provides a kind of spirosolane type glycoalkaloid, has structure shown in formula (I),
The present invention also provides a kind of preparation methods of above-mentioned spirosolane type glycoalkaloid, include the following steps:
(1) it takes bittersweet medicinal material to be added in alcoholic solution and carries out alcohol extracting, the alcohol extract of acquisition is refined total to get bittersweet
Alkali sample, it is spare；
(2) the bittersweet total alkali sample that step (1) is prepared is added in alcoholic solution and is dissolved by heating, with tlc silica gel
Sample is mixed, is then splined in silica gel column chromatography and carries out chromatography, with volume ratio for (2~3): 1 ethyl acetate and ethanol water
The eluant, eluent that solution is formed is eluted, and the volumetric concentration of the ethanol water is 93%~97%, is changed with aqueous hydrochloric acid solution
Good bismuth potassium iodide test solution is color developing agent, is detected with thin-layer chromatography, merges identical fraction, obtains two kinds of different groups of polarity
Point；
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 24%~26%:76%~74% → 29%~31%:71%~69%；Control flow rate of mobile phase is 25~35mL/
min；Controlling column temperature is 20-30 DEG C；Control sampling volume is 0.5~1.5mL；Ingredient is detected by mass detector, collects phase
The ingredient for being 901 to molecular mass, obtains the spirosolane type glycoalkaloid of the structure as shown in formula (I).
In the step (1), bittersweet total alkali sample is prepared method particularly includes: by the dry herb volumetric concentration of bittersweet
For 70% ethanol water refluxing extraction, filtering concentrates the filtrate to the medicinal extract that 50 DEG C of relative densities are 1.05, and distillation is added
Water dispersion, filtering, takes filtrate to add to D151 large pore resin absorption column, successively with the distilled water and 2~4 times of columns of 2~4 times of column volumes
The volumetric concentration of volume is the elution of 95% ethanol water, discards eluent, is then with the volumetric concentration of 3~5 times of column volumes
6 ‰ acidic alcohol aqueous solution elutes, and the volumetric concentration of ethyl alcohol is 95% in the acidic alcohol aqueous solution, collects acidic alcohol
Eluent is neutralized to neutrality with ammonium hydroxide, filtering, concentrate the filtrate to it is dry, with distillation water dispersion, the medical fluid after dispersion is added to
AB-8 macroporous absorbent resin discards eluent with the distillation water elution of 6~10 times of column volumes, then with 3~5 times of column volumes
Volumetric concentration is the elution of 95% ethanol water, collects ethanol eluate, is concentrated and dried to get bittersweet total alkali.
In the step (2), 30~40 parts by weight of bittersweet total alkali sample are weighed, the body of 300-1000 parts by volume is added
It is dissolved at being 80~100 DEG C in temperature in the ethanol water that product concentration is 93~97%, 10~20 parts by weight is then added
Tlc silica gel is uniformly mixed, and is splined in the silica gel column chromatography of the tlc silica gel of 780~820 parts by weight of addition and is carried out
Chromatography；
The relationship of the parts by weight and parts by volume is g/mL.
In the step (2), the eluant, eluent that ethyl acetate and ethanol water with volume ratio for 2.5:1 are formed is carried out
Elution, the volumetric concentration of the ethanol water are 95%.
In the step (2), the aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes:
0.8~0.9 parts by weight of basic bismuth nitrate are weighed, glacial acetic acid, 39~41 volumes of 9~11 parts by volume are sequentially added
The water of part and the liquor kalii iodide of 19~21 parts by volume, are uniformly mixed up to bismuth potassium iodide test solution；
The aqueous hydrochloric acid solution of 0.6mol/L, the bismuth potassium iodide test solution and the salt are added into the bismuth potassium iodide test solution
The volume ratio of aqueous acid is the bismuth potassium iodide test solution that 1:2 improves to get the aqueous hydrochloric acid solution；
The relationship of the parts by weight and parts by volume is g/mL.
In the step (3), mobile phase A: the volume ratio of Mobile phase B is 25%:75% → 30%:70%.
The present invention also provides the preparations including above-mentioned spirosolane type glycoalkaloid, or including above-mentioned preparation method
The preparation for the spirosolane type glycoalkaloid being prepared, with the spirosolane type glycoalkaloid be effectively at
Point, customary adjuvant is added, clinically acceptable preparation is made according to common process.
The preparation includes tablet, capsule, granule, syrup, powder, pill, tincture, vina, soft extract, pastille
Or mixture.
The present invention also provides the spiral steroid alkali that above-mentioned spirosolane type glycoalkaloid, above-mentioned preparation method are prepared
Alkane type glycoalkaloid, above-mentioned spirosolane type glycoalkaloid preparation preparation treatment non-small cell lung cancer drug in
Purposes.
Compared with prior art, above-mentioned technical proposal of the invention has the advantages that
1, spirosolane type glycoalkaloid of the present invention, be a kind of one-component, that is, chemical name be (3 β,
5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-glucopyranosyl-(1 → 4)-(1 → 2)-O- β-D-
β-D- galactopyranoside, to human A549 cell lines, human large cell lung cancer cell H460 and people's Lung Squamous Carcinoma Cells SK-
MES-1 all has inhibiting effect, and half suppression of the spirosolane type glycoalkaloid that is prepared of the present invention to A549 cell
Concentration processed is 19.22 μ g/ml, and the 503nhibiting concentration to H460 cell is 80.03 μ g/ml, to the half-suppressed of SK-MES-1 cell
Concentration is 54.20 μ g/ml, it is seen that spirosolane type glycoalkaloid component of the present invention is single, and function is definite, and can
For treating non-small cell lung cancer.
2, the preparation method of spirosolane type glycoalkaloid of the present invention, since glycoalkaloid contains polarized
Biggish sugar chain structure and the lesser steroidal unit of polarity, so that glycoalkaloid is not soluble in water, acetoneand ethyl acetate etc. is big
Most organic solvents can only be dissolved in a small number of organic solvents such as methanol, ethyl alcohol, contain nitrogen-atoms in glycoalkaloid structure, show
Certain alkalescent out, therefore can be dissolved in an acidic solution at salt, Precipitation, therefore present invention use under alkaline condition
Acidic alcohol-ammonia water precipitating refines bittersweet total alkali, then chromatographs using column, is detected with thin-layer chromatography, merges phase cocurrent flow
Part, obtain two kinds of different components of polarity；Pass through preparation after the small component of polarity in two kinds of components is dissolved with dimethylformamide
Chromatogram purification, mass detector detect ingredient, collect relative molecular mass be 901 ingredient, obtain it is of the present invention (3 β, 5
α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-glucopyranosyl-(1 → 4)-(1 → 2)-O- β-D-
β-D- galactopyranoside, the preparation method is easy to operate, scientific and reasonable.
3, the preparation method of spirosolane type glycoalkaloid of the present invention, with salt during column chromatography for separation
Aqueous acid improvement bismuth potassium iodide test solution be color developing agent, the color developing agent the preparation method comprises the following steps: into basic bismuth nitrate successively plus
Enter glacial acetic acid, water and liquor kalii iodide, then the aqueous hydrochloric acid solution of 0.6mol/L, the iodine are added into test solution after mixing
The volume ratio for changing bismuth potassium test solution and the aqueous hydrochloric acid solution is 1:2, obtains the bismuth potassium iodide examination of the aqueous hydrochloric acid solution improvement
Liquid, when the color developing agent which is prepared to be detected with thin-layer chromatography, for polar compound is with respect to background colour
It develops the color clear, easily determines.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art be briefly described, it should be apparent that, it is described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, is also possible to obtain other drawings based on these drawings.
Fig. 1 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans Portugal is prepared in the embodiment of the present invention 1
Grape glycosyl-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside structural formula；
Fig. 2 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside1HNMR spectrogram；
Fig. 3 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside13C NMR spectra；
Fig. 4 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside H-H cosy spectrogram；
Fig. 5 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside HMQC spectrogram；
Fig. 6 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside HMBC spectrogram；
Fig. 7 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside Tocsy spectrogram；
Fig. 8 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside Noesy spectrogram；
Fig. 9 is that (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- pyrans is prepared in the embodiment of the present invention 1
Glucosyl group-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside is to A549, H460 and SK-
The inhibitory effect figure of MES-1 cell.
Specific embodiment
Technical solution of the present invention will be clearly and completely described below, it is clear that described embodiment is this hair
Bright a part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not having
Every other embodiment obtained under the premise of creative work is made, shall fall within the protection scope of the present invention.In addition, below
Technical characteristic involved in described different embodiments of the present invention as long as they do not conflict with each other can be mutual
In conjunction with.
In following embodiment of the present invention and experimental example,
1, reagent and consumptive material:
Tissue Culture Flask (U.S. FALCON 353014)；
Penicillin/streptomycin solution (Jiangsu Province, China Kai Ji Biotechnology Ltd.
KGY002)；
0.25%Tripsin-EDTA (Jiangsu Province, China Kai Ji Biotechnology Ltd. KGY001)；
MTT (U.S. Amresco 0793)；
DMSO (U.S. SIGMA D2650)；
PBS (Jiangsu Province, China Kai Ji Biotechnology Ltd. KGB500)；
RPMI-1640 (U.S. GIBCO 31800-105)；
DMEM (U.S. GIBCO 12800-082)；
FBS (U.S. ExCell Biology FBS500)；
96wellcell culture plate (U.S. Corning Incorporated 3599)；
2, instrument and equipment
Superclean bench (Chinese Suzhou purifies SW-CJ-1FD)；
CO2Incubator (Japanese SANYO XD-101)；
Biological inverted microscope (Japanese OLYMPUS IX51)；
Table-type low-speed centrifuge (Chinese Shanghai Medical Treatment Equipment Co., Ltd Medical Equipment Plant 80-2)；
2.5uL, 10uL, 200uL, 1000uL pipettor (German eppendorf)；
Vertical pressure pot (the rich news of Chinese Shanghai, YXQ-LS-50)；
Electric drying oven with forced convection (Chinese Shanghai sage is glad, 101AS-3)；
Microplate reader (U.S. BioTek ELx800)；
Oscillator (Chinese Shanghai Hu Xi analysis instrument factory WH-2)；
Waters 2767/QDa prepares liquid phase mass spectrometry chromatograph: Masslynx4.1 chromatographic work station,
Waters2767 sample manager, Waters2489 ultraviolet-visible detector, Waters2545 binary high pressure chromatogram pump, QDa
Mass detector, chromatographic column are Waters SunFire Prep C18OBD (10 μm, 19 × 250mm) Column.
Embodiment 1
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-provided in this embodiment (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside preparation method the following steps are included:
(1) the dry herb of 10Kg bittersweet is taken, is added the ethanol water that 80L volumetric concentration is 70%, refluxing extraction 3 times,
2h, filtering, merging filtrate are extracted every time；Above-mentioned filtrate is concentrated into the medicinal extract for being 1.05 in 50 DEG C of relative densities, is added 10 times
The distillation water dispersion of medicinal extract quality, filtering, takes filtrate to add to D151 large pore resin absorption column, successively with the distillation of 3 times of column volumes
The ethanol water that the volumetric concentration of water and 3 times of column volumes is 95% elutes, and discards eluent, then containing with 4 times of column volumes
There is the ethanol water that volume fraction is 6 ‰ hydrochloric acid to elute, the volumetric concentration of ethyl alcohol is 95% in the acidic alcohol aqueous solution,
Collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry, with distillation water dispersion, after dispersion
Medical fluid add to AB-8 macroporous absorbent resin, with the distillation water elution of 8 times of column volumes, eluent is discarded, then with 4 times of column volumes
Volumetric concentration be 95% ethanol water elute, collect ethanol eluate, be concentrated and dried to get 35g bittersweet total alkali, yield
It is 3.5 ‰；
(2) take 35g bittersweet total alkali sample be added 95% ethanol water of 650mL volumetric concentration in temperature be 90 DEG C at it is molten
Then solution is added 15g tlc silica gel and is uniformly mixed, is splined on the silica gel column chromatography that the tlc silica gel of 800g is added
Middle carry out chromatography；The eluant, eluent that the ethyl acetate and ethanol water that are 2.5: 1 with volume ratio are formed is eluted, described
The volumetric concentration of ethanol water is 95%, using the bismuth potassium iodide test solution of aqueous hydrochloric acid solution improvement as color developing agent, with thin-layer chromatography
It is detected, merges identical fraction, obtain two kinds of different components of polarity；
The aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes: weigh basic bismuth nitrate 0.85g, successively
10mL glacial acetic acid, 40mL water and 20mL liquor kalii iodide is added, is uniformly mixed up to bismuth potassium iodide test solution；Take 1mL configured
The bismuth potassium iodide examination that the aqueous hydrochloric acid solution of 2mL0.6mol/L is improved to get the aqueous hydrochloric acid solution is added in bismuth potassium iodide test solution
Liquid.
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 25%:75% → 30%~70%；Control flow rate of mobile phase is 30mL/min；Controlling column temperature is 25 DEG C；Control sample introduction
Volume is 1mL；Ingredient is detected by mass detector, the ingredient that relative molecular mass is 901 is collected, obtains as shown in formula (I)
The spirosolane type glycoalkaloid of structure.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside1HNMR spectrogram,13C NMR spectra, H-H
Cosy spectrogram, HMQC spectrogram, HMBC spectrogram, Tocsy spectrogram and Noesy spectrogram are shown in Fig. 2-8 respectively.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside, structural formula is as shown in Fig. 1, wherein carbon atom
It is marked in structural formula number 1-27 shown in Fig. 1.Table 1 is the parsing result of Fig. 2-8.
The parsing result of 1 Fig. 2-8 of table
Wherein, in table 1 hydrogen spectrum and carbon modal data be1H-NMR (600MHz) and13C-NMR (150MHz), solvent are
It is obtained under conditions of deuterated methanol.
Embodiment 2
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-provided in this embodiment (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside preparation method the following steps are included:
(1) it refines bittersweet total alkali: taking the dry herb of 10Kg bittersweet, addition 60L volumetric concentration is 70% ethanol water, is returned
Stream extracts 4 times, extracts 2h every time, filters, merging filtrate；Above-mentioned filtrate is concentrated into the leaching for being 1.05 in 50 DEG C of relative densities
The distillation water dispersion of 8 times of medicinal extract quality is added in cream, and filtering takes filtrate to add to D151 large pore resin absorption column, successively with 2 times of columns
The volumetric concentration of the distilled water of volume and 4 times of column volumes is the elution of 95% ethanol water, eluent is discarded, then with 3 times of columns
Volume is eluted containing the ethanol water that volumetric concentration is 6 ‰ hydrochloric acid, and the volume of ethyl alcohol is dense in the acidic alcohol aqueous solution
Degree be 95%, collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry, with distillation moisture
It dissipates, the medical fluid after dispersion is added into AB-8 macroporous absorbent resin, with the distillation water elution of 6 times of column volumes, discards eluent, then
It is the elution of 95% ethanol water with the volumetric concentration of 5 times of column volumes, collects ethanol eluate, is concentrated and dried to get 35g bittersweet
Total alkali, yield are 3.5 ‰；
(2) it in temperature is 100 DEG C that take 30g bittersweet total alkali sample that 1000mL volumetric concentration is added, which be in 93% ethanol water,
Then lower dissolution is added 10g tlc silica gel and is uniformly mixed, is splined on the layer of silica gel that the tlc silica gel of 820g is added
Chromatography is carried out in analysis column；The eluant, eluent that the ethyl acetate and ethanol water that are 3: 1 with volume ratio are formed is eluted, institute
The volumetric concentration for stating ethanol water is 93%, using the bismuth potassium iodide test solution of aqueous hydrochloric acid solution improvement as color developing agent, with thin layer color
Spectrum is detected, and is merged identical fraction, is obtained two kinds of different components of polarity；
Aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes: weigh basic bismuth nitrate 0.8g, successively plus
Enter 11mL glacial acetic acid, 39mL water and 21mL liquor kalii iodide, is uniformly mixed up to bismuth potassium iodide test solution；Take the configured iodine of 1mL
Change bismuth potassium test solution, the bismuth potassium iodide test solution that the aqueous hydrochloric acid solution of 2mL0.6mol/L is improved to get the aqueous hydrochloric acid solution is added.
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 24%:76% → 31%~69%；Control flow rate of mobile phase is 25mL/min；Controlling column temperature is 30 DEG C；Control sample introduction
Volume is 0.5mL；Ingredient is detected by mass detector, the ingredient that relative molecular mass is 901 is collected, obtains such as formula (I) institute
Show the spirosolane type glycoalkaloid of structure.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2) customary adjuvant is added in-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside, is made according to common process
Tablet.
Embodiment 3
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-provided in this embodiment (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside preparation method the following steps are included:
(1) it refining bittersweet total alkali: taking the dry herb of 10Kg bittersweet, addition 100L volumetric concentration is 70% ethanol water,
Refluxing extraction 2 times, 2h is extracted every time, is filtered, merging filtrate；Above-mentioned filtrate is concentrated into the leaching for being 1.05 in 50 DEG C of relative densities
The distillation water dispersion of 12 times of medicinal extract quality is added in cream, and filtering takes filtrate to add to D151 large pore resin absorption column, successively with 4 times of columns
The volumetric concentration of the distilled water of volume and 2 times of column volumes is the elution of 95% ethanol water, eluent is discarded, then with 5 times of columns
Volume is eluted containing the ethanol water that volume fraction is 6 ‰ hydrochloric acid, and the volumetric concentration of ethyl alcohol is in acidic alcohol aqueous solution
95%, collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry, with distillation water dispersion, will
Medical fluid after dispersion adds to AB-8 macroporous absorbent resin, with the distillation water elution of 10 times of column volumes, eluent is discarded, then with 3
The volumetric concentration of times column volume is the elution of 95% ethanol water, collects ethanol eluate, is concentrated and dried total to get 35g bittersweet
Alkali, yield are 3.5 ‰；
(2) take 40g bittersweet total alkali sample be added 300mL volumetric concentration be in 97% ethanol water in the case where temperature is 80 DEG C
Then dissolution is added 20g tlc silica gel and is uniformly mixed, is splined on the silica gel column chromatography that the tlc silica gel of 780g is added
Chromatography is carried out in column；The eluant, eluent that ethyl acetate and ethanol water with volume ratio for 2:1 are formed is eluted, described
The volumetric concentration of ethanol water is 97%, using the bismuth potassium iodide test solution of aqueous hydrochloric acid solution improvement as color developing agent, with thin-layer chromatography
It is detected, merges identical fraction, obtain two kinds of different components of polarity；
Aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes: weigh basic bismuth nitrate 0.9g, successively plus
Enter 9mL glacial acetic acid, 41mL water and 19mL liquor kalii iodide, is uniformly mixed up to bismuth potassium iodide test solution；Take the configured iodate of 1mL
The bismuth potassium iodide test solution that the aqueous hydrochloric acid solution of 2mL0.6mol/L is improved to get the aqueous hydrochloric acid solution is added in bismuth potassium test solution.
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 26%:74% → 29%~71%；Control flow rate of mobile phase is 35mL/min；Controlling column temperature is 20 DEG C；Control sample introduction
Volume is 1.5mL；Ingredient is detected by mass detector, the ingredient that relative molecular mass is 901 is collected, obtains such as formula (I) institute
Show the spirosolane type glycoalkaloid of structure.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2) customary adjuvant is added in-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside, is made according to common process
Capsule.
Embodiment 4
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-provided in this embodiment (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside preparation method the following steps are included:
(1) it refining bittersweet total alkali: taking the dry herb of 10Kg bittersweet, addition 100L volumetric concentration is 70% ethanol water,
Refluxing extraction 2 times, 2h is extracted every time, is filtered, merging filtrate；Above-mentioned filtrate is concentrated into the leaching for being 1.05 in 50 DEG C of relative densities
The distillation water dispersion of 12 times of medicinal extract quality is added in cream, and filtering takes filtrate to add to D151 large pore resin absorption column, successively with 3 times of columns
The volumetric concentration of the distilled water of volume and 2 times of column volumes is the elution of 95% ethanol water, eluent is discarded, then with 4 times of columns
Volume is eluted containing the ethanol water that volume fraction is 6 ‰ hydrochloric acid, and the volume of ethyl alcohol is dense in the acidic alcohol aqueous solution
Degree be 95%, collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry, with distillation moisture
It dissipates, the medical fluid after dispersion is added into AB-8 macroporous absorbent resin, with the distillation water elution of 10 times of column volumes, discards eluent, so
It is afterwards the elution of 95% ethanol water with the volumetric concentration of 4 times of column volumes, collects ethanol eluate, be concentrated and dried white to get 35g
English total alkali, yield are 3.5 ‰；
(2) take 38g bittersweet total alkali sample be added 300mL volumetric concentration be in 94% ethanol water in the case where temperature is 80 DEG C
Then dissolution is added 16g tlc silica gel and is uniformly mixed, is splined on the silica gel column chromatography that the tlc silica gel of 780g is added
Chromatography is carried out in column；The eluant, eluent that ethyl acetate and ethanol water with volume ratio for 2.6:1 are formed is eluted, institute
The volumetric concentration for stating ethanol water is 94%, using the bismuth potassium iodide test solution of aqueous hydrochloric acid solution improvement as color developing agent, with thin layer color
Spectrum is detected, and is merged identical fraction, is obtained two kinds of different components of polarity；
The aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes: weigh basic bismuth nitrate 0.88g, successively
10.5mL glacial acetic acid, 40.5mL water and 19mL liquor kalii iodide is added, is uniformly mixed up to bismuth potassium iodide test solution；1mL is taken to configure
Good bismuth potassium iodide test solution, is added the bismuth iodide that the aqueous hydrochloric acid solution of 2mL0.6mol/L is improved to get the aqueous hydrochloric acid solution
Potassium test solution.
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 26%:74% → 29%~71%；Control flow rate of mobile phase is 35mL/min；Controlling column temperature is 20 DEG C；Control sample introduction
Volume is 1.5mL；Ingredient is detected by mass detector, the ingredient that relative molecular mass is 901 is collected, obtains such as formula (I) institute
Show the spirosolane type glycoalkaloid of structure.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2) customary adjuvant is added in-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside, is made according to common process
Syrup.
Embodiment 5
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-provided in this embodiment (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside preparation method the following steps are included:
(1) it refining bittersweet total alkali: taking the dry herb of 10Kg bittersweet, addition 100L volumetric concentration is 70% ethanol water,
Refluxing extraction 2 times, 2h is extracted every time, is filtered, merging filtrate；Above-mentioned filtrate is concentrated into the leaching for being 1.05 in 50 DEG C of relative densities
The distillation water dispersion of 12 times of medicinal extract quality is added in cream, and filtering takes filtrate to add to D151 large pore resin absorption column, successively with 2 times of columns
The volumetric concentration of the distilled water of volume and 3 times of column volumes is the elution of 95% ethanol water, eluent is discarded, then with 4 times of columns
Volume is eluted containing the ethanol water that volume fraction is 6 ‰ hydrochloric acid, and the volume of ethyl alcohol is dense in the acidic alcohol aqueous solution
Degree be 95%, collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry, with distillation moisture
It dissipates, the medical fluid after dispersion is added into AB-8 macroporous absorbent resin, with the distillation water elution of 8 times of column volumes, discards eluent, then
It is the elution of 95% ethanol water with the volumetric concentration of 5 times of column volumes, collects ethanol eluate, is concentrated and dried to get 35g bittersweet
Total alkali, yield are 3.5 ‰；
(2) it in temperature is 80 DEG C that take 32g bittersweet total alkali sample that 1000mL volumetric concentration is added, which be in 96% ethanol water,
Then lower dissolution is added 20g tlc silica gel and is uniformly mixed, is splined on the layer of silica gel that the tlc silica gel of 810g is added
Chromatography is carried out in analysis column；The eluant, eluent that ethyl acetate and ethanol water with volume ratio for 2.8:1 are formed is eluted,
The volumetric concentration of the ethanol water is 96%, using the bismuth potassium iodide test solution of aqueous hydrochloric acid solution improvement as color developing agent, with thin layer
Chromatography is detected, and identical fraction is merged, and obtains two kinds of different components of polarity；
The aqueous hydrochloric acid solution improvement bismuth potassium iodide test solution method particularly includes: weigh basic bismuth nitrate 0.81g, successively
9.6mL glacial acetic acid, 40mL water and 19.5mL liquor kalii iodide is added, is uniformly mixed up to bismuth potassium iodide test solution；1mL is taken to configure
Bismuth potassium iodide test solution, the bismuth potassium iodide examination that the aqueous hydrochloric acid solution of 2mL0.6mol/L is improved to get the aqueous hydrochloric acid solution is added
Liquid.
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um filter membrane
Filtering, is then isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18Liquid-phase chromatographic column；Using acetonitrile as mobile phase
A, 1%TFA aqueous solution are Mobile phase B, carry out gradient elution: 0~10min according to the procedure below, mobile phase A: the body of Mobile phase B
Product is than being 26%:74% → 29%~71%；Control flow rate of mobile phase is 35mL/min；Controlling column temperature is 20 DEG C；Control sample introduction
Volume is 1.5mL；Ingredient is detected by mass detector, the ingredient that relative molecular mass is 901 is collected, obtains such as formula (I) institute
Show the spirosolane type glycoalkaloid of structure.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-(1 that the present embodiment is prepared
→ 2) customary adjuvant is added in-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside, is made according to common process
Tincture.
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranose that 1 present invention of experimental example is prepared
Base-(1 → 2)-O- β-D- glucopyranosyl-inhibition of (1 → 4)-β-D- galactopyranoside to non-small cell lung cancer cell
Effect
1, experiment purpose
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-that the research present invention is prepared
(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside imitates the inhibition of non-small cell lung cancer cell
Fruit.
2, experimental method
2.1 cellular informatics
Three kinds of non-small cell lung cancer cell strains: human A549 cell lines, human large cell lung cancer cell H460, people's lung squama
Cancer cell SK-MES-1, cell are provided by Jiangsu Kai Ji Biotechnology Ltd.；A549/H460 complete medium is
90%RPMI640+10%FBS；SK-MES-1 complete medium is 90%DMEM+10%FBS；In 37 DEG C, 5%CO2, saturation it is wet
It is cultivated in the incubator of degree.
2.2 cell culture
(1) when the cell coverage rate in culture bottle reaches 80%~90%, original culture medium is sopped up；
(2) add trypsase (0.25%) appropriate, digest 1~2min；
(3) isometric culture medium containing serum is added after cell is all rounded and terminates digestion；
(4) cell is blown and beaten with liquid-transfering gun, cell all is suspended, then cell is drawn onto the centrifuge tube of 15ml,
1000rpm is centrifuged 5min；
(5) supernatant is outwelled, 1~2ml culture medium is added, cell is suspended to be transferred to again and continues to cultivate in culture bottle.
2.3MTT method detects cell Proliferation:
(1) respectively logarithmic growth phase A549, H460, SK-MES-1 cell dissociation, count, be configured to concentration be 5 × 104
The cell suspension of a/ml, 100 μ l cell suspensions are added in every hole in 96 porocyte culture plates；
(2) 96 porocyte culture plates are placed in 37 DEG C, 5%CO2It is cultivated for 24 hours in incubator；
(3) with the complete medium spirosolane type glycoalkaloid that is prepared of the dilution embodiment of the present invention 1 to table 2
In required drug concentration, 100 μ L above-mentioned pastille culture medium accordingly is added in every hole, while setting up blank control group and (being added androgynous
Product blank culture solution)；
(4) 96 porocyte culture plates are placed in 37 DEG C, 5%CO272h is cultivated in incubator；
(5) 96 orifice plates are subjected to MTT dyeing, λ=490nm measures OD value；
20 μ L MTT (5mg/ml) are added in the every hole A, continue to cultivate 4h in incubator；
B discards culture medium, and 150 μ LDMSO dissolution is added in every hole, and shaking table 10min is mixed gently；
C λ=490nm, microplate reader read the absorbance OD value in every hole, calculate inhibiting rate；
(6) each group inhibiting rate and drug 503nhibiting concentration (IC are counted50)
Inhibiting rate (%)=(ABlank control group-AMedicine group)/ABlank control group× 100%,
In formula, A is absorbance.
Pass through probit using SPSS (Staffstical Package for the Social Science) 17.0
Weighted regression method (Bliss method) calculates IC50。
3, experimental result
The spirosolane type glycoalkaloid that the present invention is prepared is to human A549 cell lines, human large cell lung cancer
The inhibitory effect of cell H460 and people's Lung Squamous Carcinoma Cells SK-MES-1 are as shown in table 2 and Fig. 9.
Suppression of the spirosolane type glycoalkaloid that 2 present invention of table is prepared to A549, H460, SK-MES-1 cell
Rate processed
By table 2 and Fig. 9 it is found that with spirosolane type glycoalkaloid concentration increase, the inhibition of cell is made
With gradually increasing, (3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-that the present invention is prepared
(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside is to the 503nhibiting concentration of A549 cell
19.22 μ g/ml, the 503nhibiting concentration to H460 cell is 80.03 μ g/ml, and the 503nhibiting concentration to SK-MES-1 cell is
54.20μg/ml。
4, experiment conclusion
(3 β, 5 α, 22 α, 25R)-spirosolane -3-O- β-D- glucopyranosyl-that the present invention is prepared (1 →
2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside is to human A549 cell lines, human large cell lung cancer
Cell H460 and people's Lung Squamous Carcinoma Cells SK-MES-1 has preferable inhibitory effect, and imitates to the inhibition of human A549 cell lines
Fruit is better than other two kinds of cancer cell effects, it is indicated above that (3 β, 5 α, 22 α, 25R)-spirosolane-that the present invention is prepared
3-O- β-D- glucopyranosyl-(1 → 2)-O- β-D- glucopyranosyl-(1 → 4)-β-D- galactopyranoside can be used for
Treat non-small cell lung cancer.
Obviously, the above embodiments are merely examples for clarifying the description, and does not limit the embodiments.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And it is extended from this it is obvious variation or
It changes still within the protection scope of the invention.
Claims (9)
1. a kind of spirosolane type glycoalkaloid, which is characterized in that there is structure shown in formula (I),
2. the preparation method of spirosolane type glycoalkaloid described in a kind of claim 1, which is characterized in that including walking as follows
It is rapid:
(1) the ethanol water refluxing extraction for being 70% by the dry herb volumetric concentration of bittersweet, filtering concentrate the filtrate to 50
The medicinal extract that DEG C relative density is 1.05, is added distillation water dispersion, and filtering takes filtrate to add to D151 large pore resin absorption column, successively
It is the elution of 95% ethanol water with the distilled water of 2~4 times of column volumes and the volumetric concentration of 2~4 times of column volumes, discards elution
Then liquid is eluted with the acidic alcohol aqueous solution that the volumetric concentration of 3~5 times of column volumes is 6 ‰, in the acidic alcohol aqueous solution
The volumetric concentration of ethyl alcohol be 95%, collect acidic alcohol eluent, be neutralized to neutrality with ammonium hydroxide, filter, concentrate the filtrate to it is dry,
With distillation water dispersion, the medical fluid after dispersion is added into AB-8 macroporous absorbent resin, with the distillation water elution of 6~10 times of column volumes,
Eluent is discarded, is then the elution of 95% ethanol water with the volumetric concentration of 3~5 times of column volumes, collects ethanol eluate, it is dense
Contracting is dry to get bittersweet total alkali sample, spare；
(2) the bittersweet total alkali sample that step (1) is prepared is added in alcoholic solution and is dissolved by heating, mixed with tlc silica gel
Then sample is splined in silica gel column chromatography and carries out chromatography, with volume ratio for (2~3): 1 ethyl acetate and ethyl alcohol is water-soluble
The eluant, eluent that liquid is formed is eluted, and the volumetric concentration of the ethanol water is 93%~97%, is improved with aqueous hydrochloric acid solution
Bismuth potassium iodide test solution be color developing agent, detected with thin-layer chromatography, merge identical fraction, obtain two kinds of different groups of polarity
Point；
(3) the small component of polarity in two kinds of components that step (2) obtains is dissolved with dimethylformamide, 0.45um membrane filtration,
Then it is isolated and purified using high performance liquid chromatography, chromatographic condition is as follows: C18 liquid-phase chromatographic column；Using acetonitrile as mobile phase A, 1%
TFA aqueous solution is Mobile phase B, carries out gradient elution: 0~10min according to the procedure below, mobile phase A: the volume ratio of Mobile phase B
For 24%~26%:76%~74% → 29%~31%:71%~69%；Control flow rate of mobile phase is 25~35mL/min；
Controlling column temperature is 20-30 DEG C；Control sampling volume is 0.5~1.5mL；Ingredient is detected by mass detector, collects opposite point
The ingredient that protonatomic mass is 901, obtains the spirosolane type glycoalkaloid of the structure as shown in formula (I).
3. preparation method according to claim 2, which is characterized in that in the step (2), weigh bittersweet total alkali sample
30~40 parts by weight, in the ethanol water that the volumetric concentration that 300-1000 parts by volume is added is 93~97% temperature be 80~
It is dissolved at 100 DEG C, the tlc silica gel that 10~20 parts by weight are then added is uniformly mixed, and is splined on 780~820 weight of addition
Chromatography is carried out in the silica gel column chromatography of the tlc silica gel of part；
The relationship of the parts by weight and parts by volume is g/mL.
4. preparation method according to claim 3, which is characterized in that be 2.5:1 with volume ratio in the step (2)
Ethyl acetate and the eluant, eluent that is formed of ethanol water eluted, the volumetric concentration of the ethanol water is 95%.
5. the preparation method according to claim 4, which is characterized in that in the step (2), the aqueous hydrochloric acid solution changes
Good bismuth potassium iodide test solution method particularly includes:
0.8~0.9 parts by weight of basic bismuth nitrate are weighed, the glacial acetic acid of 9~11 parts by volume, the water of 39~41 parts by volume are sequentially added
With the liquor kalii iodide of 19~21 parts by volume, it is uniformly mixed up to bismuth potassium iodide test solution；
The aqueous hydrochloric acid solution of 0.6mol/L, the bismuth potassium iodide test solution and the hydrochloric acid water are added into the bismuth potassium iodide test solution
The volume ratio of solution is the bismuth potassium iodide test solution that 1:2 improves to get the aqueous hydrochloric acid solution；
The relationship of the parts by weight and parts by volume is g/mL.
6. preparation method according to claim 2, which is characterized in that in the step (3), mobile phase A: Mobile phase B
Volume ratio be 25%:75% → 30%:70%.
7. including the preparation of spirosolane type glycoalkaloid described in claim 1, or any including claim 2-6
The preparation for the spirosolane type glycoalkaloid that preparation method described in is prepared, which is characterized in that with the spiral
Steroid alkali alkane type glycoalkaloid is effective component, and customary adjuvant is added, clinically acceptable preparation is made according to common process.
8. preparation according to claim 7, which is characterized in that the preparation includes tablet, capsule, granule, syrup
Agent, powder, pill, tincture, vina, soft extract, pastille or mixture.
9. spirosolane type glycoalkaloid described in claim 1, the described in any item preparation method systems of claim 2-6
The system of spirosolane type glycoalkaloid described in standby obtained spirosolane type glycoalkaloid or claim 7 or 8
Purposes of the agent in preparation treatment non-small cell lung cancer drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710086724.6A CN106939031B (en) | 2017-02-17 | 2017-02-17 | A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710086724.6A CN106939031B (en) | 2017-02-17 | 2017-02-17 | A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106939031A CN106939031A (en) | 2017-07-11 |
| CN106939031B true CN106939031B (en) | 2019-02-26 |
Family
ID=59468655
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710086724.6A Expired - Fee Related CN106939031B (en) | 2017-02-17 | 2017-02-17 | A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106939031B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111171107A (en) * | 2019-09-27 | 2020-05-19 | 中国中医科学院西苑医院 | Glucoside alkaloid, preparation method and application thereof |
| CN111440224A (en) * | 2019-11-26 | 2020-07-24 | 中国中医科学院西苑医院 | Glucoside alkaloid and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1634295A (en) * | 2004-09-27 | 2005-07-06 | 方芳 | Preparation method for climbing nightshade extract |
| CN102319319B (en) * | 2011-09-15 | 2013-05-08 | 中国中医科学院西苑医院 | Solanum lyratum extract, and anticancer medicine comprising solanum lyratum extract |
-
2017
- 2017-02-17 CN CN201710086724.6A patent/CN106939031B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN106939031A (en) | 2017-07-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101244129B (en) | Lhasa rhubarb extract, preparation method, and application in preparing preparation for treating cardiovascular and cerebrovascular diseases | |
| CN106674311B (en) | A kind of benzofuran glycosides compound and its preparation method and application | |
| Zhang et al. | Simultaneous quantification of seventeen bioactive components in rhizome and aerial parts of Alpinia officinarum Hance using LC-MS/MS | |
| CN106939031B (en) | A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes | |
| CN106928309B (en) | A kind of spirosolane type glycoalkaloid and preparation method thereof and purposes | |
| CN104262445B (en) | Camellia nitidissima saponin A, and preparation method and antitumor application thereof | |
| CN105998103B (en) | Chestnut flower activity extract and its preparation method and application | |
| CN107556325B (en) | The separation method of Alkaloid monomer in a kind of Diels Stephania Root | |
| CN105418726B (en) | A kind of marsdenia tenacissima total saponin extract and its extracting method | |
| QiN et al. | Composition analysis and structural identification of anthocyanins in fruit of waxberry. | |
| CN110204589B (en) | Effective component of feather cockscomb seed, extraction method and application thereof in preparing neuroprotective medicament | |
| CN106946973B (en) | A kind of compound and preparation method thereof, purposes | |
| CN107722087B (en) | Gynostemma pentaphylla flavonoid compound, preparation method thereof and application thereof in antitumor drugs | |
| CN108623645B (en) | Flavonoid compound and preparation method and application thereof | |
| US10874705B2 (en) | Extracts and isolated compounds from cakile arabica for treatment of ulcer | |
| CN105384786B (en) | A kind of compound and its preparation method and application | |
| CN106822167A (en) | A kind of new application of compound and preparation method thereof | |
| CN105294812B (en) | A kind of preparation method of Ardisia mamillata triterpenoid saponin reference substance | |
| KR100458003B1 (en) | New anticancer compound and its purification from Asparagus oligoclonos | |
| CN109369585A (en) | The method for extracting guainane type sesquiterpenoid | |
| CN116333029B (en) | Tea seed saponin compound and application thereof | |
| CN103739657B (en) | A kind of Sasanguasaponin compound, its preparation method, the antitumor drug applying and prepare | |
| CN108530505A (en) | A kind of flavonoid glycoside compound and its preparation method and application | |
| CN1095669C (en) | Medicine composition of saponin containing protopanaxyndiol component and preparing process and application thereof | |
| CN109694394B (en) | Preparation method and application of tannin extract in sea buckthorn |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190226 Termination date: 20220217 |