CN101824067A - Barrigenol-type triterpenoid saponins compound, preparation method and application thereof - Google Patents
Barrigenol-type triterpenoid saponins compound, preparation method and application thereof Download PDFInfo
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- CN101824067A CN101824067A CN201010164035A CN201010164035A CN101824067A CN 101824067 A CN101824067 A CN 101824067A CN 201010164035 A CN201010164035 A CN 201010164035A CN 201010164035 A CN201010164035 A CN 201010164035A CN 101824067 A CN101824067 A CN 101824067A
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Abstract
The invention belongs to the technical field of medicine and provides a barrigenol-type triterpenoid saponins compound, a preparation method and an application thereof. The preparation method comprises the following steps that: obtaining extract solution or extract liquid by using the herbal blitzkrieg extraction technique or solvent extraction method; allowing the extract solution or extract liquid to pass through a non-polar resin column to obtain coarse total saponins; carrying out the rapid processing on the coarse total saponins by using the silica gel scintillation column chromatography; carrying out gradient elution on the solvent; and further obtaining barrigenol-type triterpenoid saponins compound by using the techniques of medium-pressure and low-pressure column chromatography and HPLC (high-performance liquid chromatography), wherein the barrigenol-type triterpenoid saponins is detected by using the techniques of thin-layer chromatography and HPLC. The prepared barrigenol-type triterpenoid saponins has better antitumor activity and improves the activity of learning and memory.
Description
Technical field
The invention belongs to medical technical field, relate to the saponins compound preparation, be specifically related to Barrigenol-type triterpenoid saponins compound, preparation method and application thereof.
Background technology
The basic parent nucleus of beautiful stamen alcohol type triterpene is the oleanane type triterpene that poly-hydroxy replaces.Beautiful stamen alcohol R
1(barrigenol R
1), beautiful stamen alcohol A
1(barrigenol A
1), beautiful stamen alcohol A
2(barrigenol A
2Orcamelliagenin A), barringtogenol C (barringtogenol C or theasapogenol B) and 16-desoxybarringtogenol C (16-deoxybarringtogenol C) all belong to beautiful stamen alcohol type triterpene.Up to the present, isolated beautiful stamen alcohol type triterpene is mainly derived from the vegitabilia, as the Barringtonia (Barringtonia) of Lecythidaceae, the Aesculus of Hippocastanaceae (Aesculus), the akeake of Sapindaceae belongs to (Dodonaea), Wood of Shinyleaf Yellowhorn genus (Xanthorcera) etc.
Beautiful stamen alcohol type triterpene has cytotoxic activity usually.Some plant milk extracts that contain beautiful stamen alcohol triterpene have a series of biological activitys, have the effect that the rat plasma tri-glyceride improves due to the sweet oil that suppresses as n-butyl alcohol extract in the bibliographical information Flower of Japanese Camellia; The total saponins that obtains from Dodonaea viscosa partly has the phagolysis of enhancing and activity against snails; There is the scholar from Aesculus hippocastanum, to get and has the beautiful stamen alcohol type triterpenoid that suppresses ethanol absorption and blood sugar reducing function; There is report from Careya arborea, to separate and has the saponin(e that anti-Leishmania acts on; From Barringtonia asiatica, separate and obtain having the triterpenoid saponin that makes ladybug larva antifeedant activity; Methanol extract and the dichloromethane extract of bibliographical information Myrtillocactus geometrizans have insecticidal action; Relevant bioactivity research to beautiful stamen alcohol type saponin monomer finds that Wood of Shinyleaf Yellowhorn total saponins and some monomer components have the remarkable effect that improves learning and memory of little mouse.
In order to study the pharmacologically active of this class natural product more fully, we sum up found a kind of fast, separate the preparation Barrigenol-type triterpenoid saponins compound and detect that it is antitumor and improve the active method of learning and memory from vegitabilia in a large number.
Summary of the invention
Barrigenol-type triterpenoid saponins compound, preparation method and application thereof have been the purpose of this invention is to provide.
The invention provides Barrigenol-type triterpenoid saponins compound, this saponin(e has following general structure:
R
1 R
2 R
3 R
4
1barrigenolR
1 OH OH OH OH
2barrigenolA
1 H OH OH OH
3barrigeno?A
2 H OH H OH
4barringtogenol?C OH OH H OH
5?16-deoxybarringtogenol?C OH OH H H
The basic parent nucleus of beautiful stamen alcohol type triterpene is five kinds (1-5) among the figure; C-16 wherein, C-21, C-22 can have acyl substituted on the C-28 position, and the kind of acyl substituted mainly comprises ethanoyl, angeloyl groups, cautious scrupulously and respectfully acyl group; At C-3, C-21, the C-28 position can connect bglii fragment, and the number of sugar is 1-5, and the kind of sugar is a furan sugar, glucuronic acid, glucose, pectinose, semi-lactosi, wood sugar, rhamnosyls etc., the mode of connection between the sugar have the 1-2 position to connect, and the 1-4 position connects, and the 1-3 position connects and is connected with the 1-6 position; One or two acyl substituted can be arranged on the furan sugar, and the acyl substituted type is mainly ethanoyl and angeloyl groups.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, can by in following two kinds of methods any one the preparation Barrigenol-type triterpenoid saponins compound,
Method one:
(1) pulverizes selected Chinese medicine (Chinese medicine and plants such as Sapindaceae akeake genus, Aesculus, Lecythidaceae Barringtonia, Theaceae Camellia), utilize sudden strain of a muscle formula extractive technique or solvent-extraction process then, adopt ethanol or the methanol extraction of 40%-100%, reclaim under reduced pressure extracting solution to determining alcohol is lower than 5%, gets supernatant liquor after centrifugal;
(2) supernatant liquor is through non-polar macroporous resin (nonpolar macroporous adsorption resin, as D101, HPD100, HPD400, models such as AB-8) column chromatography processing, water and pure gradient elution successively, each wash-out position utilizes thin-layer chromatography and HPLC technology (210nm detection), the efficient part of screening enrichment Barrigenol-type triterpenoid saponins composition, obtaining the flow point that 30%-80% alcohol elutes is the efficient part of Barrigenol-type triterpenoid saponins composition, promptly thick total saponins;
(3) thick total saponins is through quick mesolow reversed-phase column chromatography, and with methanol or ethanol/water gradient elution, decompression and solvent recovery, gained flow point utilize effective flow point of thin-layer chromatography and HPLC technology (210nm detection) screening enrichment Barrigenol-type triterpenoid saponins composition;
(4) the effective flow point of gained is through half preparation or preparation HPLC chromatographic separation in the above-mentioned steps (3), and with methanol, acetonitrile/water or methyl alcohol/acetonitrile/water are the moving phase wash-out, obtain Barrigenol-type triterpenoid saponins compound;
Method two:
(1) pulverizes selected Chinese medicine (Chinese medicine and plants such as Sapindaceae akeake genus, Aesculus, Lecythidaceae Barringtonia, Theaceae Camellia), utilize sudden strain of a muscle formula extractive technique or solvent-extraction process then, adopt ethanol or the methanol extraction of 40%-100%, reclaim under reduced pressure extracting solution to determining alcohol is lower than 5%, extracting solution is respectively with chloroform or methylene dichloride, ethyl acetate, n-butanol extraction 2 to 5 times, extraction solvent and extracting liquid volume were than 1: 3～3: 1, reclaim solvent, collect and obtain chloroform or dichloromethane extract, acetic acid ethyl ester extract, n-butyl alcohol extract;
(2) n-butyl alcohol extract is through non-polar macroporous resin post (nonpolar macroporous adsorption resin, as D101, HPD100, HPD400, models such as AB-8) chromatography, 4～8 retention volume of water and pure gradient elution successively, each wash-out position utilizes the efficient part of thin-layer chromatography and HPLC technology (210nm detection) screening enrichment Barrigenol-type triterpenoid saponins composition, obtaining the flow point that 30%-80% alcohol elutes is the efficient part of Barrigenol-type triterpenoid saponins composition, promptly thick total saponins;
(3) the middle thick total saponins of gained of acetic acid ethyl ester extract and step (2) is respectively through quick mesolow reversed-phase column chromatography, with methanol or ethanol/water gradient elution, decompression and solvent recovery, gained flow point utilize effective flow point of thin-layer chromatography and HPLC technology (210nm detection) screening enrichment Barrigenol-type triterpenoid saponins composition;
(4) the effective flow point of gained is through half preparation or preparation HPLC chromatographic separation in the above-mentioned steps (3), and with methanol, acetonitrile/water or methyl alcohol/acetonitrile/water are the moving phase wash-out, obtain Barrigenol-type triterpenoid saponins compound.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, the alcohol in the step (2) in described method one and the method two is ethanol, its gradient elution concentration is 10%-100%.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, step (3) in the described method one is that thick total saponins is through mesolow column chromatography processing fast, eluting solvent is the methanol or the ethanol/water of different ratios, and 1: 5～5: 1 flow point of blending ratio is the flow point of Barrigenol-type triterpenoid saponins composition.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, step in the described method two (3) is that the thick total saponins of gained is handled through quick mesolow column chromatography respectively in acetic acid ethyl ester extract and the step (2), eluting solvent be different ratios methanol or ethanol/water, 1: 5～5: 1 flow point of blending ratio is the flow point of Barrigenol-type triterpenoid saponins composition.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, step (4) in the described method one is that the middle gained flow point of step (3) is through further separating through high performance liquid chromatography, moving phase is methanol, acetonitrile/water or methyl alcohol/acetonitrile/water, is that 1: 5～4: 1, acetonitrile/water mixed solvent ratio are that 1: 10～3: 1, methyl alcohol/acetonitrile/water mixed solvent ratio are to obtain the Barrigenol-type triterpenoid saponins monomeric compound 1: 1: 10～4: 4: 2 the flow point from methanol mixed solvent ratio.
The preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention, step in the described method two (4) is that the gained flow point further separates through high performance liquid chromatography in the step (3), moving phase is methanol, acetonitrile/water or methyl alcohol/acetonitrile/water, is that 1: 5～4: 1, acetonitrile/water mixed solvent ratio are that 1: 10～3: 1, methyl alcohol/acetonitrile/water mixed solvent ratio are to obtain the Barrigenol-type triterpenoid saponins monomeric compound 1: 1: 10～4: 4: 2 the flow point from methanol mixed solvent ratio
The preparation method preparation method of Barrigenol-type triterpenoid saponins compound provided by the invention is simple and reliable, favorable reproducibility, and prepared Barrigenol-type triterpenoid saponins compound has anti-tumor activity and improves the activity of learning and memory.
Embodiment
The following examples will give further instruction to the present invention, but not thereby limiting the invention.
Embodiment 1 prepares beautiful stamen alcohol type triterpene compound from the Wood of Shinyleaf Yellowhorn carpopodium
(1) utilizes solvent-extraction process after the dry carpopodium of Wood of Shinyleaf Yellowhorn is pulverized, the alcohol reflux of employing 75% three times, reclaim under reduced pressure extracting solution to determining alcohol is lower than 5%, extracting solution is used chloroform, ethyl acetate, n-butanol extraction respectively, reclaims solvent collection and obtains chloroform, ethyl acetate, n-butyl alcohol extract;
(2) n-butyl alcohol extract is handled through non-polar macroporous resin (HPD-100) column chromatography, water and 30% successively, 60%, 80% ethanol gradient elution, each wash-out position utilizes the thin-layer chromatography technology, is developping agent with chloroform/methanol/water, is developer with sulfuric acid/ethanolic soln, the efficient part of 60% ethanol gradient elution position enrichment Barrigenol-type triterpenoid saponins composition is determined in screening, gets thick total saponins;
(3) the thick total saponins of gained is handled through quick mesolow column chromatography respectively in acetic acid ethyl ester extract and the step (2), use methyl alcohol successively: water is 2: 8,4: 6,5: 5,6: 4,9: 1 gradient elutions, each wash-out flow point utilizes the thin-layer chromatography technology screening to determine thick total saponins 4: 6,5: 5,6: 4 wash-out positions were the flow point of enrichment Barrigenol-type triterpenoid saponins composition;
(4) the gained flow point obtains 9 beautiful stamen alcohol saponin monomer compounds as follows through the preparation HPLC chromatographic separation in the step (3).
Embodiment 2 prepares beautiful stamen alcohol type triterpene compound from shinyleaf yellowhorn fruit shell
(1) utilize solvent-extraction process after the drying and ripening shinyleaf yellowhorn fruit shell is pulverized, adopt 90% alcohol reflux secondary, reclaim under reduced pressure extracting solution to determining alcohol is lower than 3%, leaves standstill 24 hours after centrifugal, gets supernatant liquor;
(2) supernatant liquor is handled through non-polar macroporous resin (D101) column chromatography, water and 40% successively, 70%, 90% ethanol gradient elution, each wash-out position utilizes HPLC technology (210nm detection) screening to determine the efficient part of 70% ethanol gradient elution position enrichment Barrigenol-type triterpenoid saponins composition, gets thick total saponins;
(3) the thick total saponins of gained is handled through quick mesolow column chromatography in the step (2), use methyl alcohol successively: water is 3: 7,4: 6,5: 5,6: 4,7: 3,9: 1 gradient elutions, each wash-out flow point utilizes the thin-layer chromatography technology screening to determine thick total saponins 5: 5, and 6: 4,7: 3 wash-out positions were the flow point of enrichment Barrigenol-type triterpenoid saponins composition;
(4) the gained flow point is through half preparation HPLC chromatographic separation in the step (3), and with methanol, acetonitrile/water obtains 13 beautiful stamen alcohol type triterpenoids as follows for the moving phase wash-out.
Embodiment 3 separates preparation cyclic-ahltin type triterpene compound from camellia
(1) the camellia dry seed utilizes solvent-extraction process after pulverizing, and adopts 100% methyl alcohol, 80 degree refluxing extraction secondaries, and reclaim under reduced pressure extracting solution to determining alcohol is lower than 3%, and the extracting solution n-butanol extraction reclaims solvent collection and obtains n-butyl alcohol extract;
(2) n-butyl alcohol extract is handled through non-polar macroporous resin (AB-8) column chromatography, water successively, 60%, 100% methyl alcohol gradient elution, each wash-out position utilizes the thin-layer chromatography technology, is developping agent with chloroform/methanol/water, is developer with sulfuric acid/ethanolic soln, the efficient part of 60% methyl alcohol gradient elution position enrichment Barrigenol-type triterpenoid saponins composition is determined in screening, gets thick total saponins;
(3) the thick total saponins of gained is handled through the mesolow column chromatography respectively in the step (2), use methyl alcohol successively: water is 3: 7,4: 6,6: 4,9: 1 gradient elutions, each wash-out flow point utilizes the thin-layer chromatography technology screening to determine thick total saponins 4: 6, and 6: 4 wash-out positions are the flow point of enrichment Barrigenol-type triterpenoid saponins composition;
(4) the gained flow point obtains 8 beautiful stamen alcohol saponin monomer compounds as follows through the preparation HPLC chromatographic separation in the step (3).
The mensuration of the anti tumor activity in vitro of the beautiful stamen alcohol of embodiment 4 separated portions from Wood of Shinyleaf Yellowhorn carpopodium type triterpene compound
Adopt mtt assay, by high flux screening, measured the anti tumor activity in vitro that institute separates the pure type triterpene compound of part jade stamen that obtains external.Experimental result is as shown in table 1.
Test operation
(1) cell cultures
The human body tumour cell of taking the logarithm vegetative period (human melanoma cell (A375-S2) and human cervical carcinoma cell (HeLa)) is 5 * 10 with the RPMI-1640 nutrient solution dilution that includes 10% (v/v) foetal calf serum (fetal bovine serum)
4The cell suspending liquid of individual/L is inoculated in 96 orifice plates, and 100 μ L/ holes place 37 ℃, saturated humidity, 5%CO
2Cultivate 24h in the incubator.
(2) add reagent
After sample dissolves with DMSO, with the RPMI-1640 nutrient solution dilution that contains 10% (v/v) foetal calf serum.Above-mentioned pastille nutrient solution is added in 96 orifice plates, be positioned in the incubator with cell cultures the same terms and cultivate 48h.
(3) result measures
The mtt assay operating process is as follows:
Mtt assay: inoculating cell is cultivated (24h) in advance, and drug treating (48h) adds MTT dyeing (4h), and the centrifugal supernatant liquor of removing adds the DMSO dissolving, and microplate reader 570nm measures the OD value down.
Data analysis is calculated in accordance with the following methods:
Inhibitory rate of cell growth=[1-administration group OD value/control group OD value] * 100%.
Adopt the LOGIT method to calculate medicine half-inhibition concentration (IC
50).
Test-results is as shown in table 1.
Table 1
1)“-”indicated?the?IC
50?value＞100μg/ml
The mensuration of the anti tumor activity in vitro of the beautiful stamen alcohol of embodiment 5 separated portions from shinyleaf yellowhorn fruit shell type triterpene compound
Adopt mtt assay, by high flux screening, measured the anti tumor activity in vitro that institute separates the pure type triterpene compound of part jade stamen that obtains external.Experimental result is as shown in table 2.
Test operation
(1) cell cultures
Six kinds of human body tumour cells of taking the logarithm vegetative period (people's acute myeloid leukaemia cell strain (HL-60), Human Prostate Cancer Cells strain (PC-3MIE8), human stomach cancer cell line (BGC-823), human breast cancer cell strain (MDA-MB-435), human hepatoma cell strain (Bel-7402), human cervical carcinoma cell strain (HeLa)) are 5 * 10 with the RPMI-1640 nutrient solution dilution that includes 10% (v/v) foetal calf serum (fetal bovine serum)
4The cell suspending liquid of individual/L is inoculated in 96 orifice plates, and 100 μ L/ holes place 37 ℃, saturated humidity, 5%CO
2Cultivate 24h in the incubator.
(2) add reagent
After sample dissolves with DMSO, with the RPMI-1640 nutrient solution dilution that contains 10% (v/v) foetal calf serum.Above-mentioned pastille nutrient solution is added in 96 orifice plates, be positioned in the incubator with cell cultures the same terms and cultivate 48h.
(3) result measures
The mtt assay operating process is as follows:
Mtt assay: inoculating cell is cultivated (24h) in advance, and drug treating (48h) adds MTT dyeing (4h), and the centrifugal supernatant liquor of removing adds the DMSO dissolving, and microplate reader 570nm measures the OD value down.
Data analysis is calculated in accordance with the following methods:
Inhibitory rate of cell growth=[1-administration group OD value/control group OD value] * 100%.
Adopt the LOGIT method to calculate medicine half-inhibition concentration (IC
50).
Test-results is as shown in table 2.
Table 2
The beautiful stamen alcohol of embodiment 5 separated portions from shinyleaf yellowhorn fruit shell type triterpene compound improve the active mensuration of learning and memory
Adopt external PC12D cytoactive screening system, the beautiful stamen alcohol of part type triterpene compound monomer has been carried out promoting the mensuration of the nerve synapse growth activity of NGF mediation the results are shown in Table 3.
Test operation
(1) cell cultures
The PC12D cell is stored in the DMEM substratum that includes 5% (v/v) foetal calf serum, 10% horse serum and 2mM glutamine (Dulbecco ' s Modified Eagle ' s medium, high glucose type), places 37 ℃, saturated humidity, 5%CO
2Cultivate in the incubator.Face the time spent and cultivate 1h, be inoculated in 24 well culture plates that cover with poly-L-Methionin (2 * 10 with the phosphate buffered saline buffer (PBS) that contains 1mM EGTA
4Individual/hole).
(2) add reagent
For test agent 1: be dissolved among the DMSO for the reagent thing, and to dilute with the substratum that contains 1% foetal calf serum and 2% horse serum be three gradients of 60,20,6 μ g/ml.
For test agent 2: be dissolved among the DMSO for the reagent thing, and to dilute with the substratum that contains 1% foetal calf serum and 2% horse serum be three gradients of 60,20,6 μ g/ml that add NGF (7S) simultaneously, making its concentration is 2ng/ml.
Negative control group: NGF (7S) adds the substratum that contains 1% foetal calf serum and 2% horse serum and is mixed with the solution that concentration is 2ng/ml.
Positive controls: NGF (7S), adding the substratum contain 1% foetal calf serum and 2% horse serum, to be mixed with concentration be 30ng/ml solution.
Cell after cultivating 24h, in 24 orifice plates, add above-mentioned prepare respectively organize sample solution, be positioned over and continue cultivation 48h the environment identical under with above-mentioned culture condition.
(3) result measures
In culture system, add the PBS buffered soln that contains 1% glutaraldehyde and fix, and be stored in the PBS solution.With length is to prolong between the cytolemma of cell dia more than one times to be defined as synapse cell, and adopts phase microscope to observe the generation of PC12D synapse cell.Each data point is observed the situation of 100 above cells at least, and record forms the number of the PC12D cell of cynapse.
Data processing:
Formation cynapse cell percentage [Neurite-bearing cells (%)]=
[for 2 groups of cell numbers of test agent/positive controls cell number] * 100%
Adopt aforesaid method, we have investigated the beautiful stamen alcohol of the part in shinyleaf yellowhorn fruit shell type triterpene in the external promoter action that the nerve synapse of NGF mediation is formed.It is compared with positive controls, and it is as shown in table 3 to form cynapse cell percentage result.
Table 3
1)NGF(2ng/ml，12％)was?used?as?the?negative?control.
2)Strong?activity：61-100％，moderate?activity：31-60％，mild?activity：10-30％.
3)“-”indicated?the?death?of?the?cells?for?the?cytotoxicity?of?the?fractions?or?compounds.
Claims (10)
1. Barrigenol-type triterpenoid saponins compound is characterized in that, this saponin(e has following general structure:
R
1 R
2 R
3 R
4
1barrigenol?R
1 OH OH OH OH
2barrigenol?A
1 H OH OH OH
3barrigeno?A
2 H OH H OH
4barringtogenol?C OH OH H OH
516-deoxybarringtogenol?C OH OH H H
The basic parent nucleus of beautiful stamen alcohol type triterpene is a 1-5 kind among the last figure, C-16 wherein, and C-21, C-22 can have acyl substituted on the C-28 position, and the kind of acyl substituted mainly comprises ethanoyl, angeloyl groups, cautious scrupulously and respectfully acyl group; At C-3, C-21, the C-28 position can connect bglii fragment, and the number of sugar is 1-5, and the kind of sugar is a furan sugar, glucuronic acid, glucose, pectinose, semi-lactosi, wood sugar, rhamnosyl, the mode of connection between the sugar have the 1-2 position to connect, and the 1-4 position connects, and the 1-3 position connects and is connected with the 1-6 position; One or two acyl substituted can be arranged on the furan sugar, and the acyl substituted type is mainly ethanoyl and/or angeloyl groups.
2. the preparation method of a Barrigenol-type triterpenoid saponins compound as claimed in claim 1 is characterized in that:
(1) pulverize selected Chinese medicine, utilize sudden strain of a muscle formula extractive technique or solvent-extraction process then, adopt ethanol or the methanol extraction of 40%-100%, reclaim under reduced pressure extracting solution to determining alcohol is lower than 5%, gets supernatant liquor after centrifugal;
(2) supernatant liquor is handled through the non-polar macroporous resin column chromatography, water and pure gradient elution successively, each wash-out position utilizes thin-layer chromatography and HPLC technology, the efficient part of screening enrichment Barrigenol-type triterpenoid saponins composition, obtaining the flow point that 30%-80% alcohol elutes is the efficient part of Barrigenol-type triterpenoid saponins composition, promptly thick total saponins;
(3) thick total saponins is through quick mesolow reversed-phase column chromatography, and with methanol or ethanol/water gradient elution, decompression and solvent recovery, gained flow point utilize effective flow point of thin-layer chromatography and HPLC technology screening enrichment Barrigenol-type triterpenoid saponins composition;
(4) the effective flow point of gained is through half preparation or preparation HPLC chromatographic separation in the above-mentioned steps (3), and with methanol, acetonitrile/water or methyl alcohol/acetonitrile/water are the moving phase wash-out, obtain Barrigenol-type triterpenoid saponins compound.
3. the preparation method of a Barrigenol-type triterpenoid saponins compound as claimed in claim 1 is characterized in that:
(1) pulverizes selected Chinese medicine, utilize sudden strain of a muscle formula extractive technique or solvent-extraction process then, adopt ethanol or the methanol extraction of 40%-100%, reclaim under reduced pressure extracting solution to determining alcohol is lower than 5%, extracting solution is respectively with chloroform or methylene dichloride, ethyl acetate, n-butanol extraction, reclaim solvent, collect and obtain chloroform or dichloromethane extract, acetic acid ethyl ester extract, n-butyl alcohol extract;
(2) n-butyl alcohol extract is handled through the non-polar macroporous resin column chromatography, water and pure gradient elution successively, each wash-out position utilizes the efficient part of thin-layer chromatography and HPLC technology screening enrichment Barrigenol-type triterpenoid saponins composition, obtaining the flow point that 30%-80% alcohol elutes is the efficient part of Barrigenol-type triterpenoid saponins composition, promptly thick total saponins;
(3) the middle thick total saponins of gained of acetic acid ethyl ester extract and step (2) is respectively through quick mesolow reversed-phase column chromatography, with methanol or ethanol/water gradient elution, decompression and solvent recovery, gained flow point utilize effective flow point of thin-layer chromatography and HPLC technology screening enrichment Barrigenol-type triterpenoid saponins composition;
(4) the effective flow point of gained is through half preparation or preparation HPLC chromatographic separation in the above-mentioned steps (3), and with methanol, acetonitrile/water or methyl alcohol/acetonitrile/water are the moving phase wash-out, obtain Barrigenol-type triterpenoid saponins compound.
4. according to the preparation method of claim 1 or 2 described Barrigenol-type triterpenoid saponins compounds, it is characterized in that: the alcohol of described step (2) is ethanol, and its gradient elution concentration is 10%-100%.
5. according to the preparation method of the described Barrigenol-type triterpenoid saponins compound of claim 1, it is characterized in that: described step (3) is handled through quick mesolow column chromatography for thick total saponins, eluting solvent is 1: 10～10: 1 methanol or an ethanol/water, and 1: 5～5: 1 flow point of blending ratio is the flow point of Barrigenol-type triterpenoid saponins composition.
6. according to the preparation method of the described Barrigenol-type triterpenoid saponins compound of claim 2, it is characterized in that: described step (3) is handled through quick mesolow column chromatography respectively for the thick total saponins of gained in acetic acid ethyl ester extract and the step (2), eluting solvent is 1: 10～10: 1 methanol or an ethanol/water, and 1: 5～5: 1 flow point of blending ratio is the flow point of Barrigenol-type triterpenoid saponins composition.
7. according to the preparation method of the described Barrigenol-type triterpenoid saponins compound of claim 1, it is characterized in that: described step (4) is that the middle gained flow point of step (3) is through further separating through high performance liquid chromatography, moving phase is methanol, acetonitrile/water or methyl alcohol/acetonitrile/water, is that 1: 5～4: 1, acetonitrile/water mixed solvent ratio are that 1: 10～3: 1, methyl alcohol/acetonitrile/water mixed solvent ratio are to obtain the Barrigenol-type triterpenoid saponins monomeric compound 1: 1: 10～4: 4: 2 the flow point from methanol mixed solvent ratio.
8. according to the preparation method of the described Barrigenol-type triterpenoid saponins compound of claim 2, it is characterized in that: described step (4) is further separated through high performance liquid chromatography for gained flow point in the step (3), moving phase is methanol, acetonitrile/water or methyl alcohol/acetonitrile/water, is that 1: 5～4: 1, acetonitrile/water mixed solvent ratio are that 1: 10～3: 1, methyl alcohol/acetonitrile/water mixed solvent ratio are to obtain the Barrigenol-type triterpenoid saponins monomeric compound 1: 1: 10～4: 4: 2 the flow point from methanol mixed solvent ratio.
9. the application of the described Barrigenol-type triterpenoid saponins compound of claim 1 in the preparation antitumor drug.
10. the application of the described Barrigenol-type triterpenoid saponins compound of claim 1 in the medicine of preparation raising learning and memory.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102180940A (en) * | 2011-04-02 | 2011-09-14 | 沈阳药科大学 | Preparation method of compound santhoceraside |
| CN103242412A (en) * | 2012-11-15 | 2013-08-14 | 沈阳药科大学 | Preparation method of shiny-leaved yellowhorn carpopodium and medical applications |
| CN103739658A (en) * | 2014-01-30 | 2014-04-23 | 苏州大学 | Compound, extraction method thereof, application thereof to preparation of antitumor drugs, and antitumor drugs prepared by using compound |
| CN103755769A (en) * | 2014-01-30 | 2014-04-30 | 苏州大学 | Camellia saponin compound, preparation method and application thereof and anti-tumour drug prepared from same |
| CN105061545A (en) * | 2015-08-17 | 2015-11-18 | 中国人民解放军第二一〇医院 | Triterpenoid saponin-type compounds of shinyleaf yellowhorn, as well as preparation method and application of compounds |
| CN105330717A (en) * | 2015-10-29 | 2016-02-17 | 淄博夸克医药技术有限公司 | Novel triterpenoid and preparation method and medical application thereof |
| CN108341849A (en) * | 2017-01-24 | 2018-07-31 | 复旦大学 | Beautiful stamen alcohols triterpenoid and preparation method thereof and the purposes in pharmacy |
| US20190256543A1 (en) * | 2004-09-07 | 2019-08-22 | Pacific Arrow Limited | Methods and compounds for modulating the secretion or expression of adhesion proteins or angiopoietins of cells |
| CN113072608A (en) * | 2020-01-03 | 2021-07-06 | 沈阳药科大学 | Triterpenoid saponin compound and application thereof |
-
2010
- 2010-05-06 CN CN201010164035A patent/CN101824067A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 李占林 等: "文冠果果壳化学成分及生物活性研究", 《2006第六届中国药学会学术年会论文集》 * |
| 李占林等: "文冠果化学成分及药理作用研究进展", 《沈阳药科大学学报》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190256543A1 (en) * | 2004-09-07 | 2019-08-22 | Pacific Arrow Limited | Methods and compounds for modulating the secretion or expression of adhesion proteins or angiopoietins of cells |
| US11046724B2 (en) * | 2004-09-07 | 2021-06-29 | Pacific Arrow Limited | Methods and compounds for modulating the secretion or expression of adhesion proteins or angiopoietins of cells |
| CN102180940A (en) * | 2011-04-02 | 2011-09-14 | 沈阳药科大学 | Preparation method of compound santhoceraside |
| CN103242412A (en) * | 2012-11-15 | 2013-08-14 | 沈阳药科大学 | Preparation method of shiny-leaved yellowhorn carpopodium and medical applications |
| CN103242412B (en) * | 2012-11-15 | 2016-03-30 | 沈阳药科大学 | A kind of preparation method of Wood of Shinyleaf Yellowhorn carpopodium glycosides and medicinal use |
| CN103739658A (en) * | 2014-01-30 | 2014-04-23 | 苏州大学 | Compound, extraction method thereof, application thereof to preparation of antitumor drugs, and antitumor drugs prepared by using compound |
| CN103755769A (en) * | 2014-01-30 | 2014-04-30 | 苏州大学 | Camellia saponin compound, preparation method and application thereof and anti-tumour drug prepared from same |
| CN103755769B (en) * | 2014-01-30 | 2016-01-20 | 苏州大学 | The antitumor drug of a kind of Sasanguasaponin compound, its preparation method, application and preparation thereof |
| CN103739658B (en) * | 2014-01-30 | 2016-05-25 | 苏州大学 | A kind of compound, its extracting method, it prepares the antineoplastic of application and the preparation thereof of antineoplastic |
| CN105061545A (en) * | 2015-08-17 | 2015-11-18 | 中国人民解放军第二一〇医院 | Triterpenoid saponin-type compounds of shinyleaf yellowhorn, as well as preparation method and application of compounds |
| CN105330717A (en) * | 2015-10-29 | 2016-02-17 | 淄博夸克医药技术有限公司 | Novel triterpenoid and preparation method and medical application thereof |
| CN108341849A (en) * | 2017-01-24 | 2018-07-31 | 复旦大学 | Beautiful stamen alcohols triterpenoid and preparation method thereof and the purposes in pharmacy |
| CN113072608A (en) * | 2020-01-03 | 2021-07-06 | 沈阳药科大学 | Triterpenoid saponin compound and application thereof |
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