CN1303098C - Pseudo portulaca oleracea saponin compound, total sapon in and its application in medicine - Google Patents
Pseudo portulaca oleracea saponin compound, total sapon in and its application in medicine Download PDFInfo
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- CN1303098C CN1303098C CNB2004100893120A CN200410089312A CN1303098C CN 1303098 C CN1303098 C CN 1303098C CN B2004100893120 A CNB2004100893120 A CN B2004100893120A CN 200410089312 A CN200410089312 A CN 200410089312A CN 1303098 C CN1303098 C CN 1303098C
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Abstract
The present invention relates to three dammarane type triterpenoid saponins and a mixture with the three dammarane type triterpenoid saponins, which belongs to the technical field of medicine. The dammarane type triterpenoid saponins are extracted from bacopa monnieri herbs, and are used for treating tumor, depression and Alzheimer's disease. The three dammarane type triterpenoid saponins are respectively Bacopaside 1, Bacopasaponin C and Bacopasaponin D, and the mixture with the three dammarane type triterpenoid saponins (with the weight contents of the three dammarane type triterpenoid saponins of higher than or equal to 50%) is called Bacopa monnieri total saponins. Experimental researches prove that the three dammarane type triterpenoid saponins and the bacopa monnieri total saponins have inhibiting effects on tumor cell strains K-562, SHG-44, HCT-8, a-549, PC-3M, etc. of human bodies, have inhibiting effect on tumor tissues of S180 tumor-bearing mice, have prominent improvement effect on depression patients and scopolamine induced mouse brain memory damage, and are effective medicine for treating the tumor, the depression and the Alzheimer's disease.
Description
Technical field
The invention belongs to medical technical field, the mixture (Herba Bacopae monnieri total saponins) and the application in the medicine of diseases such as prevention and treatment tumour, spirit depressing and senile dementia thereof that are specifically related to a kind of saponins compound that extraction separation obtains from the herbal medicine Herba Bacopae monnieri and include saponins compound.
Background technology
Herba Bacopae monnieri is the herb of goatweed Herba Bacopae monnieri (Bacopa Monnieri (L.) Wettst), Herba Bacopae monnieri has clearing heat and cooling blood in the theory of traditional Chinese medical science, the effect of detoxify and promote the subsdence of swelling is used for the treatment of symptoms such as dysentery, conjunctival congestion with pain and swelling of the eye, erysipelas, swelling and pain of hemorrhoid, elephatiasis.
India's Ayurveda medicine theoretical (Ayurvedic system of medicine) thinks that Herba Bacopae monnieri has the effect of improving memory.
Summary of the invention
The objective of the invention is, according to the theoretical and treatment experience of India's Ayurveda medicine, in conjunction with modern chemistry and pharmacology means, extraction separation high-efficiency low-toxicity from Herba Bacopae monnieri can be used as the saponins compound of disease medicaments such as treatment tumour, spirit depressing and senile dementia and the mixture (Herba Bacopae monnieri total saponins) that contains this saponins compound.
The present invention's extraction separation from Herba Bacopae monnieri has the saponins compound of diseases such as treatment tumour, spirit depressing and senile dementia, obtain 3 kinds of dammarane type triterpenoid saponins, be respectively: Bacopaside I, Bacopasaponin D and Bacopasaponin C, and mixture---the Herba Bacopae monnieri total saponins that comprises this 1-3 kind dammarane type triterpenoid saponin.In this total saponins, saponin compound weight content 〉=50% usually.
Description of test, above-mentioned these dammarane type triterpenoid saponins, and the mixture Herba Bacopae monnieri total saponins that includes these dammarane type triterpenoid saponins, to human body tumour cell K-562, SHG-44, HCT-8, A-549, cell strains such as PC-3M all have restraining effect; The sarcoma of S180 tumor-bearing mice organized also restraining effect; Depressed patient, senile dementia and Scopolamine inductive mouse brain memory impairment are all improved significantly.They are the medicines of effectively treating tumour, depression and senile dementia.
3 kinds of dammarane type triterpenoid saponin chemical structures among the present invention are as follows:
Among the Bacopaside I, aglycon is pseudojujubogenin (a pseudo-Jujubogenin), and sugar chain is 3-O-{ α-L-arabinofuranosyl-(1 → 2)-[6-O-sulfonic group-β-D-glucopyranosyl-(1 → 3)] }-α-L-arabopyranose base.
Among the Bacopasaponin D, aglycon is jujubogenin (Jujubogenin), and sugar chain is 3-O-[β-D-glucopyranosyl (1 → 3) { α-L-arabinofuranosyl (1 → 2) }]-α-L-arabopyranose base.
Among the Bacopasaponin C, aglycon is pseudojujubogenin (a pseudo-Jujubogenin), and sugar chain is 3-O-[β-D-glucopyranosyl (1 → 3) { α-L-arabinofuranosyl (1 → 2) }]-α-L-arabopyranose base.
Dammarane type triterpenoid saponin provided by the invention and to include the preparation method of mixture Herba Bacopae monnieri total saponins of dammarane type triterpenoid saponin as follows: Herba Bacopae monnieri is extracted as extracting solvent with ethanol etc., with the concentrated medicinal extract that obtains of extracting solution, separate through macroporous resin column chromatography again, collect ethanol eluate, concentrate, promptly obtain the Herba Bacopae monnieri total saponins.To total saponins through silica gel column chromatography, detect the chromatography flow point with thin-layer chromatography, the flow point with identical single spot merges, and concentrates, separate and purifying through RP-18 lower pressure column or HPLC (high performance liquid chromatography) again, get Bacopaside I, BacopasaponinC and Bacopasaponin D.
By spectral analysis techniques such as acid hydrolysis and ultraviolet spectrometer, infrared spectrometer, high resolution fast atom bombardment MS instrument, hydrogen nuclear magnetic resonance spectrometer, carbon-13 nmr spectra instrument, the relevant spectrometer of heteronuclear volume, the relevant spectrometers of heteronuclear multikey, obtain above-mentioned 3 kinds of dammarane type triterpenoid saponin chemical structures, be followed successively by: the pseudo-Spina Date Seed glycosides of 3-O-α-L-arabinofuranosyl-(1 → 2)-[6-O-sulfonic group-β-D-glucopyranosyl-(1 → 3)]-α-L-arabopyranose base; 3-O-[β-D-glucopyranosyl (1 → 3) { α-L-arabinofuranosyl (1 → 2) } α-L-arabopyranose base] pseudo-Spina Date Seed glycosides; 3-O-[β-D-glucopyranosyl (1 → 3) { α-L-arabinofuranosyl (1 → 2) } α-L-arabopyranose base] the Spina Date Seed glycosides.
The present invention has carried out the evaluation experimental of antitumor, depression and senile dementia to Herba Bacopae monnieri total saponins and 3 compounds.
(1) pseudo-purslane saponin is to the restraining effect of vitro human tumour cell.
1. experiment purpose:
Estimate pseudo-purslane saponin to vitro human tumour cell K-562, SHG-44, HCT-8, A-549, the influence of cell strain growths such as PC-3M.
2. method:
Cytotoxic activity experiment: mtt assay screening sample.The concentration of three saponin(es and total saponins sample is respectively 100 μ g/ml, 10 μ g/ml, 1.0 μ g/ml, 0.1 μ g/ml and 0.01 μ g/ml.
Data processing: data are represented with inhibiting rate (%).According to every group of inhibition index of gained, the calculation formula of pressing inhibition rate of tumor cell: inhibiting rate=[necrocytosis mean-(cell survivaling number 1+ cell survivaling number 2)/2]/necrocytosis mean control group index] * 100%.
3. result: when 100 μ g/ml pseudo-purslane saponin to K-562, SHG-44, HCT-8, A-549, the growth of cell strains such as PC-3M has stronger restraining effect (table 1).
Table 1. pseudo-purslane saponin is to the restraining effect of human body tumour cell
Samples | Cell lines | Cytotoxic ratio(%) | ||||
100μg/ml | 10μg/ml | 1.0μg/ml | 0.1μg/ml | 0.01μg/ml | ||
Total saponins | A-549 | 94.36 | - | - | - | - |
HCT-8 | 90.92 | - | - | - | - | |
K-562 | 96.91 | 5.70 | - | - | - | |
PC-3M | 56.75 | - | - | - | - | |
SHG-44 | 94.43 | - | - | - | - | |
Bacopaside I | A-549 | 100.0 | 4.36 | - | - | 2.99 |
HCT-8 | 89.75 | - | - | - | - |
Samples | Cell lines | Cytotoxic ratio(%) | ||||
100μg/ml | 10μg/ml | 1.0μg/ml | 0.1μg/ml | 0.01μg/ml | ||
K-562 | 100.0 | 18.06 | 36.20 | 9.93 | - | |
PC-3M | 47.22 | 1.54 | 11.20 | 3.85 | 14.63 | |
SHG-44 | 100.0 | 7.39 | - | 11.94 | 9.96 | |
Bacopasaponin C | A-549 | 100.0 | - | - | - | - |
HCT-8 | 100.0 | - | - | - | - | |
K-562 | 100.0 | 64.02 | 34.50 | - | 61.69 | |
PC-3M | 100.0 | - | - | - | - | |
SHG-44 | 100.0 | 11.67 | 25.42 | 19.86 | 28.82 | |
Bacopasaponin D | A-549 | 55.28 | - | 32.66 | 2.47 | - |
HCT-8 | 34.80 | - | 2.96 | - | - | |
K-562 | 87.51 | - | - | 20.63 | 13.21 | |
PC-3M | 9.11 | 4.37 | 1.22 | 0.16 | 9.85 | |
SHG-44 | 42.50 | 11.67 | 9.31 | - | 10.07 |
4. conclusion: 3 dammarane type triterpenoid saponins and Herba Bacopae monnieri total saponins have stronger anti-tumor activity.
(2) pseudo-purslane saponin is to the restraining effect of S180 tumor-bearing mice tumour.
1. experiment purpose: estimate the influence of pseudo-purslane saponin to S180 tumor-bearing mice sarcoma tissue growth situation.
2. method
The S180 tumor-bearing mice, body weight 20g ± 5g, male and female half and half.Animal is divided into 4 experimental group at random, 1 cyclophosphamide-a control group and 1 blank group.Experimental group is 3 saponin(es of oral administration (50mg/kg) respectively, Herba Bacopae monnieri total saponins (100mg/kg), volume 0.5mL/kg (down together), famotidine group ig famotidine (30mg/kg), all (carboxymethyl cellulose, CMC) solution is made into desired concn to above medicine with the 5g/L carboxymethyl cellulose with preceding.The physiological saline of blank group ig respective volume.Above-mentioned respectively the group irritated stomach every day 1 time by measure, begins successive administration 7d from subcutaneous implantation tumour, and each group is all supplied with normal diet.Each organizes fasting 48h after the last administration, sarcoma is organized peeled off, and weighs, and calculates tumour inhibiting rate.
Data processing: data are with mean ± standard deviation (x ± SD) expression.According to every group of tumor tissue in vitro weight of gained, the calculation formula of pressing tumor control rate: tumor control rate=(the average knurl of the average knurl weight-administration of blank group group is heavy)/average knurl of control group heavy * 100%; Calculate tumor control rate.The relatively employing t check of each group difference.
3. result
The growth that 3 saponin(es of oral administration and Herba Bacopae monnieri total saponins (50-100mg/kg) can obviously suppress S180 tumor-bearing mice sarcoma tissue forms (table 2).
3 triterpenoid saponins of table 2 ig and Herba Bacopae monnieri total saponins are to the restraining effect of S180 tumor-bearing mice sarcoma tissue
samples | Dose (mg/kg) | Animal number Start/end | Animal weight(g) (cut of tumor) | Tumor weight(g) ±SD | Inhibition ratio(%) |
Blank | - | 20/20 | 28.82±2.36 | 2.00±0.76 | - |
CTX | 30 | 10/10 | 27.66±1.43 | 0.18±0.07 * | 91.12 |
Total saponins | 100 | 10/10 | 28.96±2.93 | 0.20±0.04 * | 90.18 |
Bacopaside I | 100 | 10/9 | 28.07±3.42 | 0.26±0.06 * | 87.03 |
Bacopasaponin C | 100 | 10/10 | 26.56±4.61 | 0.20±0.04 * | 90.18 |
Bacopasaponin D | 100 | 10/10 | 28.96±2.93 | 1.22±0.44 △ | 39.18 |
Compare with the blank group:
△P>0.05;
*P<0.01
4. conclusion: 3 dammarane type triterpenoid saponins and Herba Bacopae monnieri total saponins have stronger anti-tumor activity.
(3) pseudo-purslane saponin is to the influence of mouse non-swimming time.
1. experiment purpose: estimate the influence of pseudo-purslane saponin to the mouse forced swimming dead time.
2. method
Modeling and grouping: get the ICR mouse, male, body weight 20 ± 5g.Animal is divided into 4 experimental group at random, 1 extract with fluoxetine group and 1 blank group.The conventional raising, and irrigate stomach by 50mg/kg with the 3 kinds of triterpenoid saponins and the Herba Bacopae monnieri total saponins aqueous solution, 1 time/d, continuous 7 days.Carried out the forced swimming experiment after the last administration in one hour.
Data processing: write down each experimental group and the positive controls mouse average dead time in 6min.Each experimental group and positive group are checked and blank group comparing difference with t.
3. result
Pseudo-purslane saponin can significantly shorten the dead time (table 3) of mouse forced swimming.
Table 3 an ig3 triterpenoid saponin and Herba Bacopae monnieri total saponins are to the influence of mouse non-swimming time
The dosage group | Dead time | Per-cent | P |
Negative group | 124.4±27.0 | ||
Positive group | 24.9±30.8 | 80.0 | <0.01 |
Total saponins | 10.2±12.2 | 91.8 | <0.01 |
Bacopaside I | 27.3±16.4 | 77.2 | <0.01 |
Bacopasaponin C | 39.5±44.6 | 68.2 | <0.01 |
Bacopasaponin D | 25.9±30.6 | 79.2 | <0.01 |
4. conclusion
3 triterpenoid saponins of ig and Herba Bacopae monnieri total saponins (10-50mg/kg) have significant antidepressant effect.
(4) pseudo-purslane saponin is to the influence of mouse tail suspension dead time.
1. experiment purpose: estimate the influence of pseudo-purslane saponin to the mouse tail suspension dead time.
2 methods
Modeling and grouping: get the ICR mouse, male, body weight 20 ± 5g.Animal is divided into 4 experimental group at random, 1 extract with fluoxetine group and 1 blank group.The conventional raising, and irrigate stomach by 50mg/kg with the 3 kinds of triterpenoid saponins and the Herba Bacopae monnieri total saponins aqueous solution, 1 time/d, continuous 7 days.Hanged the tail experiment after the last administration in one hour.
Data processing: write down each experimental group and the positive controls mouse average dead time in 6min.Each experimental group and positive group are checked and blank group comparing difference with t.
3. result
3 triterpenoid saponins of ig and Herba Bacopae monnieri total saponins (10-50mg/kg) can significantly reduce the dead time (table 4) of outstanding tail mouse.
Table 4 an ig3 triterpenoid saponin and Herba Bacopae monnieri total saponins are to the influence of mouse tail suspension dead time
Group | Dead time | Per-cent | P |
Negative group | 102.3±29.4 | ||
Positive group | 28.3±23.6 | 72.3 | <0.01 |
Total saponins | 54.6±37.5 | 46.6 | <0.01 |
Bacopaside I | 40.2±40.8 | 60.7 | <0.01 |
Bacopasaponin C | 54.2±46.7 | 47.0 | <0.01 |
Bacopasaponin D | 56.2±34.2 | 45.1 | <0.01 |
4. conclusion:
3 Herba Bacopae monnieri triterpenoid saponins of ig and Herba Bacopae monnieri total saponins (10-50mg/kg) have significant antidepressant effect.
(5) nootropic effect of Herba Bacopae monnieri triterpenoid saponin and Herba Bacopae monnieri total saponins.
1. experiment purpose:
Estimate the therapeutic action of Herba Bacopae monnieri triterpenoid saponin and Herba Bacopae monnieri total saponins to senile dementia.
2. method:
Get the ICR mouse, male, body weight 20 ± 5g.Animal is divided into 6 groups at random, and negative control and positive controls are all irritated with isopyknic physiological saline at every turn, all the other four experimental group difference administration earlier 7 days, after the last administration, except that negative control group, all the other each group is all irritated stomach 10mg/kg with Scopolamine, carries out the diving tower experiment after the administration in one hour.
3. result: 3 triterpenoid saponins and Herba Bacopae monnieri total saponins (10-50mg/kg) can also make errors number obviously reduce (table 5) significant prolongation mouse diving tower latent period
Table 5 pseudo-purslane saponin is to the influence of Model of Dementia mouse anti dementia and nootropic effect
Group | Errors number (n) in the training 5min | Latent period behind the 24h (Tl)/s | Errors number (n) in the 3min | P |
Negative group | 5.77±3.83 | 73.23±49.84 | 0.47±0.23 | |
Positive group | 16.5±9.7 | 22.93±12.63 | 2.90±1.66 | <0.05 |
Total saponins | 9.38±5.75 | 41.46±14.45 | 1.00±1.00 | <0.01 |
Bacopaside I | 9.31±8.47 | 68.62±29.69 | 0.85±0.90 | <0.001 |
Bacopasaponin C | 5.17±4.32 | 54.75±18.88 | 0.67±0.25 | <0.001 |
Bacopasaponin D | 7.50±3.28 | 65.79±28.68 | 0.93±0.05 | <0.01 |
4. conclusion:
Pseudo-purslane saponin has effect of significant treatment senile dementia and short intelligence activity.
Embodiment
Embodiment 1:
Preparation Herba Bacopae monnieri total saponins
After Herba Bacopae monnieri herb 10kg pulverizes, with 90% alcohol reflux 3 times, each 2 hours, united extraction liquid, decompression and solvent recovery, get ethanol extraction, this extract with water-dispersion after, through the AB-8 macroporous resin adsorption, respectively with behind water and 40% ethanol elution, collect 70% ethanol eluate, solvent recuperation is to doing to such an extent that remaining solid is the Herba Bacopae monnieri total saponins.
The Herba Bacopae monnieri total saponins, light brown powder, Lieberman-Burchard and Keller-Killiani reaction all are positive.
Embodiment 2:
Preparation Bacopaside I
After Herba Bacopae monnieri herb 10kg pulverizes, with 90% alcohol reflux 3 times, each 2 hours, united extraction liquid, decompression and solvent recovery, get ethanol extraction, this extract with water-dispersion after, through the AB-8 macroporous resin adsorption, respectively with behind water and 40% ethanol elution, collect 70% ethanol eluate, solvent recuperation is to doing to such an extent that remaining solid is the Herba Bacopae monnieri total saponins.The Herba Bacopae monnieri total saponins is carried out silica gel column chromatography, carry out gradient elution, check the chromatography flow point with thin-layer chromatography with chloroform-methanol alcohol, flow point with identical single spot merges, concentrate, get flow point 1,2,3,4,5,6, wherein flow point 5 is through HPLC (MeOH:H
2O) separation, purifying obtain Bacopaside I.
Bacopaside I, white amorphous powder, mp260-261 ℃, [α]
D 28:-35.63 ° (c=0.20, MeOH), molecular formula: C
46H
72O
20S.
Embodiment 3:
Preparation Bacopasaponin C
After Herba Bacopae monnieri herb 10kg pulverizes, with 90% alcohol reflux 3 times, each 2 hours, united extraction liquid, decompression and solvent recovery, get ethanol extraction, this extract with water-dispersion after, through the AB-8 macroporous resin adsorption, respectively with behind water and 40% ethanol elution, collect 70% ethanol eluate, solvent recuperation is to doing to such an extent that remaining solid is the Herba Bacopae monnieri total saponins.The Herba Bacopae monnieri total saponins is carried out silica gel column chromatography, carry out gradient elution, check the chromatography flow point with thin-layer chromatography with chloroform-methanol, flow point with identical single spot merges, concentrate, get flow point 1,2,3,4,5,6, wherein flow point 4 is through HPLC (MeOH:H
2O) separation, purifying obtain Bacopasaponin C.
Bacopasaponin C, white amorphous powder, mp222-224 ℃, [α]
D 28:-47.49 ° (c=1.20, MeOH), molecular formula: C
46H
74O
17
Embodiment 4:
Preparation Bacopasaponin D
After Herba Bacopae monnieri herb 10kg pulverizes, with 90% alcohol reflux 3 times, each 2 hours, united extraction liquid, decompression and solvent recovery, get ethanol extraction, this extract with water-dispersion after, through the AB-8 macroporous resin adsorption, respectively with behind water and 40% ethanol elution, collect 70% ethanol eluate, solvent recuperation is to doing to such an extent that remaining solid is the Herba Bacopae monnieri total saponins.The Herba Bacopae monnieri total saponins is carried out silica gel column chromatography, carry out gradient elution, check the chromatography flow point with thin-layer chromatography with chloroform-methanol, flow point with identical single spot merges, concentrate, get flow point 1,2,3,4,5,6, wherein flow point 4 is through HPLC (MeOH:H
2O) separation, purifying obtain Bacopasaponin D.
Bacopasaponin D, white amorphous powder, mp234-241 ℃, [α]
D 28:-42.35 ° (c=0.14, MeOH), molecular formula: C
46H
74O
17
Embodiment 5:
Get Bacopaside I 100mg,,,, promptly get the injection lyophilized powder according to conventional freeze-dry process freeze-drying with N.F,USP MANNITOL 200mg with an amount of water for injection dissolving.Be used for the treatment of tumor disease.
Embodiment 6:
The Herba Bacopae monnieri total saponins is disperseed with an amount of water for injection, and spraying drying or vacuum-drying get Herba Bacopae monnieri total saponins dry powder, and taking polyethylene glycol 400 adds Herba Bacopae monnieri total saponins dry powder, and the limit edged is stirred to even, adopts pressing, makes soft capsule.Be used for the treatment of depressive illness.
Embodiment 7:
Get Herba Bacopae monnieri total saponins 100g, disperse with an amount of water for injection, add right amount of auxiliary materials, mixing is made particle, and drying is pressed into 1000, and sugar coating promptly gets Herba Bacopae monnieri total saponins sheet.Be used for the treatment of senile dementia.
Claims (4)
2. one kind is extracted the total saponins that obtains from Herba Bacopae monnieri, it is characterized in that for including the mixture of the arbitrary combination in pseudo-purslane saponin compounds Bacopaside I, 3 kinds of dammarane type triterpenoid saponins of Bacopasaponin D, Bacopasaponin C, wherein Bacopaside I and Bacopasaponin D according to claim 1, the structure of Bacopasaponin C is:
The weight content of pseudo-purslane saponin compounds 〉=50% wherein.
3. the application of the saponins compound that extraction obtains from Herba Bacopae monnieri as claimed in claim 1 in the medicine of preparation treatment tumour, dysthymia disorders and senile dementia.
4. the application of the total saponins that extraction obtains from Herba Bacopae monnieri as claimed in claim 2 in the medicine of preparation treatment tumour, dysthymia disorders and senile dementia.
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CN1833692B (en) * | 2005-03-15 | 2010-08-11 | 成都华高药业有限公司 | False portulaca oleracea extracts, its prepn. and usage |
CN102210692A (en) * | 2011-04-08 | 2011-10-12 | 中国人民解放军第二军医大学 | Application of pseudojujubogenin in preparation of antidepressant medicaments |
CN102389435A (en) * | 2011-09-22 | 2012-03-28 | 中国人民解放军第二军医大学 | Application of pseudojujubogenin to preparation of medicaments for resisting senile dementia |
CN103055007A (en) * | 2012-11-16 | 2013-04-24 | 中国人民解放军第二军医大学 | Application of total saponins of bacopa monnieri (L.) wettst. in preparation of medicaments for resisting cerebral ischemia |
CN105055625B (en) * | 2015-07-27 | 2018-07-31 | 中国人民解放军第二军医大学 | A kind of Chinese herbal medicine extract and application thereof |
CN107796882A (en) * | 2016-08-24 | 2018-03-13 | 山东颐正达医药科技有限公司 | The assay method of bacopa monnieri total saponin extracts active constituent content |
CN108003132B (en) * | 2017-11-10 | 2019-05-28 | 南阳师范学院 | Phloroglucinol derivatives compound and preparation and the application in preparation antidepressant |
CN109481435B (en) * | 2018-12-29 | 2021-04-23 | 湖北绿无界生物科技有限公司 | Compound composition for gynecology |
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CN1277555A (en) * | 1998-09-07 | 2000-12-20 | 马哈拉伊·克里森·潘迪塔 | Composition for improving mental capabilities in mammals |
WO2002003813A1 (en) * | 2000-07-12 | 2002-01-17 | Raj Kumar, Sujatha | Use of dammarane-type triterpenoid saponins |
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CN1277555A (en) * | 1998-09-07 | 2000-12-20 | 马哈拉伊·克里森·潘迪塔 | Composition for improving mental capabilities in mammals |
WO2002003813A1 (en) * | 2000-07-12 | 2002-01-17 | Raj Kumar, Sujatha | Use of dammarane-type triterpenoid saponins |
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