CN105503990A - Novel withanolides compound as well as preparation method and medical application thereof - Google Patents
Novel withanolides compound as well as preparation method and medical application thereof Download PDFInfo
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Abstract
The invention discloses a novel withanolides compound as well as a preparation method and medical application thereof, belongs to the field of drugs. The compound is firstly reported, is a withanolides compound of a novel structure, and can be obtained through extraction, separation and purification from the dried stem leaves of lophatherum gracile. In vitro tests prove that the toxicity of A beta 1-40 injures nerve cells and causes abundant apoptosis of PC12 and reduction of cell activity; and after the compound (I) is used for intervention, the apoptosis condition is improved, and thus the compound can be used for developing nerve protecting drugs.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to be separated from the dry cauline leaf of Herba Lophatheri the one obtained and sleep and add lactone compound, containing its pharmaceutical composition and its preparation method and application.
Background technology
Plant Herba Lophatheri (HerbaLoophatheri) has another name called Rhizome of Common Lophantherum, pheasant rice, Herba Lophatheri, for Gramineae Herba Lophatheri belongs to per nnial herb.Expand for fusiform meat block root in the middle part of Herba Lophatheri fibrous root, yellow-white; Leaf lanceolar, wide 2 ~ 3cm, vein is parallel, has obvious small crossvein, and blade face is without hair or have seta, and there is short bristle at edge; Panicle top is raw, and small ear is bordering on stockless, and arrangement is cob side slightly partially, wholely comes off; Caryopsis is oval.Herba Lophatheri is mainly born in the dark and damp place of hillside sylvan life, is distributed in each provinces and regions on the south the Changjiang river.Herba Lophatheri has effect of clearing heat and relieving fidgetness, diuresis, be mainly used in treating in the heart disease heat, cough with dyspnea, haematemesis, hot malicious wind, only quench one's thirst, press borax poison, dissolving phlegm, control fanatical unhappiness, apoplectic aphasia in silence, pain head wind, only palpitation with fear, pestilence fan be vexed, kill small worm, heat extraction delays the symptoms such as spleen.Medicinal material Herba Lophatheri (LophatherumgracileBrongn.) is the dry cauline leaf of Gramineae (Gramineae) herbaceous plant Herba Lophatheri (LophatherumgracileBrongn.), has heat-clearing and fire-purging, relieving restlessness is quenched the thirst, effect of inducing diuresis for treating stranguria syndrome.Ming Dynasty's LI Shi-Zhen claims it in Compendium of Material Medica: sweet, cold, nontoxic, remove dysphoria with smothery sensation, diuresis, clear away heart-fire.
Containing a large amount of flavonoid compounds and bioactive polysaccharide and other effective constituent in Herba Lophatheri, as phenolic acid compound, anthraquinone analog compound, terpene lactone, extraordinary amino acid and active skin, manganese, zinc, selenium and other trace elements.Functional factor mainly flavone glycoside and coumarins lactone contained in Herba Lophatheri.The content of its effective constituent and biological activity all have comparability with Ginkgo Leaf.
Modern pharmacological research shows, active oxygen radical in functional factor energy purged body contained in Herba Lophatheri; The activity of the Antioxidant Enzymes of induction organism inside; The anti-stress of enhancing body and anti-fatigue ability; Improve memory capability, the process etc. delayed senility.In addition, also abundant chlorophyll is contained in Herba Lophatheri.Research shows, chlorophyll is the important component of many vegetables anti-mutagenic activityes, has antitumor anti-canceration effect, and because chlorophyll has stronger anti-oxidant function, this is to prevention cardiovascular diseases and anti-agingly also have active effect.
Summary of the invention
The object of this invention is to provide a kind of one obtained that is separated from the dry cauline leaf of Herba Lophatheri to sleep and add lactone compound, containing its pharmaceutical composition and its preparation method and application.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
There is the compound (I) of following structural formula:
The preparation method of described compound (I), comprise following operation steps: the dry cauline leaf of Herba Lophatheri is pulverized by (a), extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 10% ethanol elution, 6 column volumes, then use 70% ethanol elution, 8 column volumes, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material; C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 90:1,60:1,35:1,15:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 20:1,15:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
Further, in step (a), extract with 75% alcohol heat reflux, united extraction liquid.
Further, described macroporous resin is D101 type macroporous adsorbent resin.
Pharmaceutical composition, the described compound (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
Described compound (I) application in the medicine preparing neuroprotective.
The application of described pharmaceutical composition in the medicine preparing neuroprotective.
When the compounds of this invention is used as medicine, directly can uses, or use with the form of pharmaceutical composition.
This pharmaceutical composition contains the compounds of this invention (I) for the treatment of significant quantity, and all the other are acceptable on pharmacology, nontoxic to humans and animals and pharmaceutically acceptable carrier of inertia and/or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, dripping pill, micropill, suspensoid, emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into water-based or oily solution, aseptic powder injection, liposome or the emulsion etc. of sterilizing.
Accompanying drawing explanation
Fig. 1 is compound (I) structural formula;
Fig. 2 is that compound (I) calculates ECD and experiment ECD figure.
Embodiment
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Main raw, reagent source and instrument type:
Ethanol, sherwood oil, ethyl acetate, propyl carbinol, methylene dichloride are analytical pure, and purchased from Shanghai Ling Feng chemical reagent company limited, methyl alcohol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.
Embodiment 1: compound (I) is separated preparation and structural identification
A the dry cauline leaf (10kg) of () Herba Lophatheri is pulverized, (30L × 3 time) are extracted with 75% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste (6L), use sherwood oil (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated propyl carbinol (6L × 3 time) to extract successively, obtain petroleum ether extract, acetic acid ethyl ester extract (381g) and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract D101 macroporous resin removal of impurities in (b) step (a), first use 10% ethanol elution, 6 column volumes, use 70% ethanol elution, 8 column volumes again, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material (125g); C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, successively with volume ratio be 90:1 (8 column volumes), the methylene chloride-methanol gradient elution of 60:1 (8 column volumes), 35:1 (8 column volumes), 15:1 (10 column volumes) and 1:1 (6 column volumes) obtains 5 components; D component 4 (27g) is separated further by purification on normal-phase silica gel in () step (c), successively with volume ratio be 20:1 (8 column volumes), the methylene chloride-methanol gradient elution of 15:1 (8 column volumes) and 5:1 (6 column volumes) obtains 3 components; E in () step (d), component 2 (15g) reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I) (151mg).
Structural identification: colorless amorphous powder; HR-ESIMS shows [M+Na]
+for m/z591.2610, can obtain molecular formula in conjunction with nuclear-magnetism feature is C
32h
40o
9, degree of unsaturation is 13.Hydrogen nuclear magnetic resonance modal data δ
h(ppm, DMSO-d
6, 400MHz): H-2 (5.99, dd, J=10.0, 2.0), H-3 (6.61, ddd, J=10.0, 4.8, 2.4), H-4 (2.67, m), H-4 (3.10, m), H-6 (5.47, d, J=6.0), H-7 (2.04, m), H-7 (2.78, m), H-8 (1.95, m), H-9 (2.84, m), H-11 (1.62, m), H-11 (2.63, m), H-12 (2.05, m), H-12 (2.76, m), H-15 (5.43, dd, J=9.9, 8.0), H-16 (2.78, dd, J=11.2, 9.9), H-16 (2.99, dd, J=11.2, 8.0), H-18 (4.65, d, J=11.6), H-18 (5.01, d, J=11.6), H-19 (1.20, s), H-21 (5.41, m), H-21 (5.59, m), H-22 (5.42, m), H-23 (2.58, m), H-23 (3.05, m), H-27 (2.04, s), H-28 (1.87, s), 15-OAc (2.17, s), 18-OAc (2.06, s), carbon-13 nmr spectra data δ
c(ppm, DMSO-d
6, 125MHz): 204.1 (C, 1-C), 128.0 (CH, 2-C), 145.7 (CH, 3-C), 33.6 (CH
2, 4-C), 135.1 (C, 5-C), 125.8 (CH, 6-C), 26.2 (CH
2, 7-C), 38.1 (CH, 8-C), 36.8 (CH, 9-C), 51.0 (C, 10-C), 23.5 (CH
2, 11-C), 26.2 (CH
2, 12-C), 57.8 (C, 13-C), 79.7 (C, 14-C), 76.9 (CH, 15-C), 43.1 (CH
2, 16-C), 61.8 (C, 17-C), 64.7 (CH
2, 18-C), 19.1 (CH
3, 19-C), 155.1 (C, 20-C), 109.6 (CH
2, 21-C), 71.3 (CH, 22-C), 33.9 (CH
2, 23-C), 150.7 (C, 24-C), 121.6 (C, 25-C), 166.7 (C, 26-C), 12.8 (CH
3, 27-C), 20.4 (CH
3, 28-C), 170.8 (C, 15-OAc), 21.5 (CH
3, 15-OAc), 171.1 (C, 18-OAc), 21.7 (CH
3, 18-OAc), carbon atom mark is see Fig. 1.IR spectrum shows that this compound contains hydroxyl (3262cm
-1), double bond (1638cm
-1) and ester group (1741cm
-1) group.
1hNMR composes display five methyl signals [δ H1.20 (s, H-19), 1.87 (s, H-28), 2.04 (s, H-27), 2.06 (s, 18-OAc), 2.17 (s, 15-OAc)], one containing Oxymethylene [δ H4.65 (d, J=11.6Hz, H-18), 5.01 (d, J=11.6Hz, H-18)], an olefinic methylene radical [δ H5.41 (m, H-21), 5.59 (m, H-21)], three olefinic methine proton signal [δ H5.47 (d, J=6.0Hz, H-6), 5.99 (dd, J=10.0, 2.0Hz, H-2), 6.61 (ddd, J=10.0, 4.8, 2.4Hz, H-3)], two containing oxygen methyne [δ H5.42 (m, H-22) and 5.44 (dd, J=9.9, 8.0Hz, H-15)].
13cNMR composes display 32 resonance carbon signal, five methyl, eight methylene radical (containing Oxymethylene δ C64.7, an olefinic methylene radical δ C109.6), seven methynes (three olefinic methyne δ C125.8,128.0 and 145.7; Two containing oxygen methyne δ C71.3 and 76.9), 12 quaternary carbons (four carbonyl δ C166.7,170.8,171.1,204.1; Four alkene quaternary carbon δ C121.6,135.1,150.7,155.1; Two contain oxygen quaternary carbon δ C61.8,79.7).Above-mentioned nuclear magnetic data shows that this compound is five-membered ring structure.Nuclear magnetic data shows that this compound contains one and comprises α, the nine carbon side chain unit [δ C12.8 (CH of β-unsaturated-gamma lactone
3, 27-C), 20.4 (CH
3, 28-C), 33.9 (CH
2, 23-C), 71.3 (CH, 22-C), 109.6 (CH
2, 21-C), 121.6 (C, 25-C), 150.7 (C, 24-C), 155.1 (C, 20-C), 166.7 (C, 26-C); δ H1.87 (s, H-28), 2.04 (s, H-27), 2.58 (m, H-23), 3.05 (m, H-23), 5.41 (m, H-21), 5.42 (m, H-22), 5.59 (m, H-21)], a 1-oxo-2,5 diene structures [δ C125.8 (CH, C-6), 128.0 (CH, C-2), 135.1 (C, C-5), 145.7 (CH, C-3), 204.1 (C, C-1)], two hydroxyl [δ C61.8 (CH, C-17), 79.7 (C, C-14)].In HMBC spectrum, H-15 (δ H5.43, dd, J=9.9,8.0Hz) and H
2with the dependency of corresponding acetyl carbon (δ C170.8 and 171.1) ,-18 (δ H4.65, d, J=11.6Hz and 5.01, d, J=11.6Hz) show that C-15 and C-18 position is respectively connected with an acetoxyl group.In addition, terminal ethylenic methylene signals [δ H5.41 (m, H-21), 5.59 (m, H-21)] and two carbon signal [δ C109.6 (CH
2, 21-C), 155.1 (C, 20-C)] show to there is double bond between C-20 and C-21.In ROESY spectrum, the dependency of H-15 and H-18 (δ H5.01, d, J=11.6Hz) shows that the acetoxyl group of C-15 position is α configuration.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and ROESY spectrum, and document is about correlation type nuclear magnetic data, can substantially determine this compound as shown in Figure 1, steric configuration is determined further by ECD test, theoretical value and experimental value basically identical (Fig. 2).
Embodiment 2: compound (I) pharmacological action is tested
One, material and instrument
PC12 cell strain, gifts in Anhui Chinese Medicine College experimental center.A β
1-40albumen is purchased from Sigma company.Compound (I) is self-control, and HPLC normalization method purity is greater than 98%.(import packing is purchased from Beijing match Mo Feishier biological chemistry Products Co., Ltd to DMEM/F12 substratum.NewbomCalfSerum is purchased from BeijingSolarbioScience & TechnologyCo., Ltd.MTT is purchased from Amresco company of the U.S..Triumphant base AnnexinV-FITC cell apoptosis detection kit is purchased from Nanjing Kai Ji Bioisystech Co., Ltd.Paraformaldehyde is purchased from chemical reagent company limited of Chinese Medicine group.Bcl-2 polyclonal antibody is purchased from Wuhan doctor's moral Bioisystech Co., Ltd.Bax polyclonal antibody is purchased from Beijing Bo Aosen Bioisystech Co., Ltd.Cyt-c polyclonal antibody is purchased from Wuhan doctor's moral Bioisystech Co., Ltd.Goat anti-rabbit igg/FITC is purchased from Beijing Bo Aosen Bioisystech Co., Ltd.Sheep anti-mouse igg/(H+L) is purchased from the green skies biotech firm in Jiangsu.Normal Goat Serum is purchased from Wuhan doctor's moral Bioisystech Co., Ltd.SABC immunohistochemical kit is purchased from Wuhan Boster Biological Technology Co., Ltd..DAB colouring reagents box is purchased from Wuhan doctor's moral Bioisystech Co., Ltd.
Electronic analytical balance (FA2004 type, Shanghai Lei Ci precision scientific instrument company), water-bath (HHSI1-4 type, Beijing medical apparatus and instruments factory), microplate reader (ELx-800 type, U.S. Bio-Tek), flow cytometer (EPICXL-MCL type, BeckmanCouiter company of the U.S.), high speed freezing centrifuge (3K30 type, Sigma Co., USA, CO2gas incubator (CO-150 type, NBS company limited of the U.S.), OLYMPUS fluorescent microscope (BX41 type, band DP70 camera system, Japanese Olympus Products).
Two, test method
1, A β
1-40hatch
A β
1-40be mixed with 1000 μm of ol/L storage liquid and packing with deionized water, be positioned over the storage of-20 DEG C, refrigerator, face and hatch 7d with taking out the last week at 37 DEG C of incubators, make it gathering aging.
2, the cultivation of cell
The DMEM/F12 substratum of PC12 cell strain containing 10% foetal calf serum, 100U/ and penicillin, 100U/mL Streptomycin sulphate is cultivated in glass culturing bottle, CO
2the culture condition of cell culture incubator is 37 DEG C, 5%CO
2concentration, saturated humidity, grows to after 80% abundance until cell and goes down to posterity 1 time (about 2-3d) with the trysinization of 0.25%, inverted microscope observation of cell upgrowth situation, and the vegetative period cell of taking the logarithm during experiment is tested.Seed cells into 96 well culture plate serum free mediums before carrying out MTT experiment to cultivate; Before flow cytomery, cell is seeded to 6 well culture plate serum-free culture both to cultivate; Immunohistochemical experiment all uses 24 well culture plate creep plate method culturing cells, and substratum is serum free medium.
3, cell administration process and grouping
Cell is divided into five groups, carries out drug intervention: 1. Normal group: normal PC12 cell after serum free medium inoculation 24h; 2. model group: 25 μm of ol/LA β
1-40; 3. compound (I) low dose group: 25 μm of ol/LA β
1-40+ 50mg/L compound (I); 4. dosage group in compound (I): 25 μm of ol/LA β
1-40+ 100mg/L compound (I); 5. compound (I) high dose group: 25 μm of ol/LA β
1-40+ 200mg/L compound (I).Detection indices is carried out after drug treating 24h.
4, MTT detects each group of cell viability
The cell of taking the logarithm vegetative period, with 1 × 10 after trysinization
5the cell density of individual/mL, every hole 100 μ L is inoculated in serum-free culture 24h in 96 orifice plates, then carries out drug intervention according to above-mentioned clustering method, and often group arranges 6 multiple holes, and after 24h, the MTT solution 20 μ L that every hole adds 5mg/mL continues at CO
2hatch 4h in cell culture incubator, then take out 96 well culture plates, abandoning supernatant, every hole adds the DMSO of 150 μ L, is placed on 10min that shaking table vibrates, detects the light absorption value in each hole after purple crystal dissolves completely by microplate reader at 490nm wavelength place.Only to add the hole of nutrient solution for zeroing reference, cell viability percentage expression, the cytoactive depending on control group is 100%.Result calculates: cell survival rate=(experimental group OD value-blank group OD value)/(control group OD value-blank group OD value) × 100%.
5, statistical study
Adopt SPSS17.0 analysis software to process statistics, data acquisition one-way analysis of variance, by mean scholar standard deviation
represent, adopt LSD to compare between group, P<0.05 is for having significant difference, and P<0.01 is extremely significant difference, and chart is drawn by Excel2003.
Three, result and conclusion
MTT test result shows, compare with normal group cell, model group cytoactive obviously reduces (P<0.01), compound (I) is intervened rear cytoactive and is increased, wherein the rising of high, middle dosage group cytoactive has statistical significance (P<0.01, P<0.05), low dose group cytoactive changes little (P>0.05).In table 1 (comparing * P<0.05 with model group, * * P<0.01).
Conclusion, this research confirms A β
1-40toxicity damage neurocyte, cause a large amount of apoptosis of PC12, cytoactive decline, compound (I) intervene after apoptosis situation improve.
Table 1MTT method compound (I) is to A β
1-40the impact of induction PC12 cytoactive (
n=6)
| Group | Treatment process | Cytoactive (%) |
| Normal group | – | 100±0.0 ** |
| Model group | 25μmol/LAβ 1-40 | 50.08±6.67 |
| Compound (I) low dose group | 25μmol/L Aβ 1-40+ 50mg/L compound (I) | 58.78±12.64 |
| Dosage group in compound (I) | 25μmol/L Aβ 1-40+ 100mg/L compound (I) | 63.77±6.81 * |
| Compound (I) high dose group | 25μmol/L Aβ 1-40+ 200mg/L compound (I) | 83.95±10.58 ** |
Embodiment 3
The preparation of tablet: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, vehicle is added, pelletizing press sheet than the ratio for 1:7 in itself and excipient weight.
Embodiment 4
The preparation of oral liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, oral liquid method for making makes oral liquid routinely.
Embodiment 5
The preparation of capsule or granule: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, add vehicle in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 6
The preparation of injection liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Embodiment 7
The preparation of aseptic powder injection: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, be dissolved in sterile water for injection, stirring makes molten, filter with aseptic suction funnel, aseptic essence filter again, be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains powder injection.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.Those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the protection domain of technical solution of the present invention.
Claims (7)
1. there is the compound (I) of following structural formula:
2. the preparation method of compound according to claim 1 (I), it is characterized in that comprising following operation steps: the dry cauline leaf of Herba Lophatheri is pulverized by (a), extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; B in () step (a), the removal of impurities of thing macroporous resin got by propyl carbinol, first use 10% ethanol elution, 6 column volumes, then use 70% ethanol elution, 8 column volumes, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material; C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 90:1,60:1,35:1,15:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 20:1,15:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
3. the preparation method of compound according to claim 2 (I), is characterized in that: in step (a), extracts, united extraction liquid with 75% alcohol heat reflux.
4. the preparation method of compound according to claim 2 (I), is characterized in that: described macroporous resin is D101 type macroporous adsorbent resin.
5. a pharmaceutical composition, is characterized in that: the compound according to claim 1 (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
6. compound according to claim 1 (I) application in the medicine preparing neuroprotective.
7. the application of pharmaceutical composition according to claim 5 in the medicine preparing neuroprotective.
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| CN201511017267.2A CN105503990A (en) | 2015-12-29 | 2015-12-29 | Novel withanolides compound as well as preparation method and medical application thereof |
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| CN201511017267.2A CN105503990A (en) | 2015-12-29 | 2015-12-29 | Novel withanolides compound as well as preparation method and medical application thereof |
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| CN201511017267.2A Withdrawn CN105503990A (en) | 2015-12-29 | 2015-12-29 | Novel withanolides compound as well as preparation method and medical application thereof |
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| CN105461782A (en) * | 2016-01-14 | 2016-04-06 | 郑平珍 | Novel triterpene compound and preparation and medical application thereof |
| CN105748866A (en) * | 2016-04-29 | 2016-07-13 | 陈芝维 | Traditional Chinese medicine compound with neuroprotective effect and preparation method thereof |
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| CN105748866A (en) * | 2016-04-29 | 2016-07-13 | 陈芝维 | Traditional Chinese medicine compound with neuroprotective effect and preparation method thereof |
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Address after: 810016 Qinghai University, 251 Ning Ning Road, Chengbei District, Qinghai, Xining Applicant after: Wu Jinfeng Address before: 213003 Jiangsu province Changzhou Yanling West Road Zhonglou District Mingjun homes 2 building room 2407 Applicant before: Wu Jinfeng |
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Application publication date: 20160420 |