CN103450323A - Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation - Google Patents
Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation Download PDFInfo
- Publication number
- CN103450323A CN103450323A CN2013103895413A CN201310389541A CN103450323A CN 103450323 A CN103450323 A CN 103450323A CN 2013103895413 A CN2013103895413 A CN 2013103895413A CN 201310389541 A CN201310389541 A CN 201310389541A CN 103450323 A CN103450323 A CN 103450323A
- Authority
- CN
- China
- Prior art keywords
- compound
- methyl alcohol
- preparation
- tuber
- dwarf lilyturf
- 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.)
- Pending
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Steroid Compounds (AREA)
Abstract
The invention provides a preparation method of an active component extracted from traditional Chinese medicine, namely, radix ophiopogonis, an active saponin compound contained in the component, and a separation and preparation method of the compound. The identification result shows that the active component of the radix ophiopogonis contains four active compounds. According to the experiment for an anti-senile dementia in vitro screening model, namely, rat pheochromocytoma cells, the active component of radix ophiopogonis and the four active components show remarkable nerve growth factor simulating activity, namely, NGFmimics, which can promote the outgrowth and protuberance of nerve cells. The preparation method has the advantages that the medical application of the active component of radix ophiopogonis and the active compound contained in the component in prevention of senile dementia is created; and a new treatment medicine is provided for preventing and treating senile dementia and other neurodegenerative diseases.
Description
Technical field
The invention belongs to medicine, field of food; be specifically related to from Chinese medicinal materials separation and Extraction and prepare the method for active ingredient the tuber of dwarf lilyturf; and relate to structure and the preparation method of contained saponins compound, and this active ingredient and the active compound application in preventing and treating the nerve degenerative diseases such as senile dementia and neurotrophy protection medicine, food.
Background technology
Senile dementia (Alzheimer ' s disease, alzheimer's disease) be a kind of lethality nerve degenerative diseases that carries out sexual development, clinical manifestation constantly worsens for cognitive and memory function, and carrying out property of activity of daily living goes down, and various neuropsychic symptoms and behavior disorder are arranged.At present, alzheimer's disease is only second to heart trouble, cancer, apoplexy, has become the fourth-largest cause of disease of mankind's death.The patient of China's alzheimer's disease surpasses 5,000,000, accounts for 1/4 of the total case load in the world, and along with the quickening of aging population, patient's quantity can be more huge, can have a strong impact on the stable and development of society.Therefore, the medicine of the nerve degenerative diseases such as the effective prevention of exploitation and treatment senile dementia has become whole world medical problem in the urgent need to address.
Medicine kind for senile dementia, mainly contain cholinergic agent, for example the medicine as the listing of acetylcholinesterase (Acetylcholinesterase, AChE) inhibitor has tacrine (tacrine), rivastigmine (rivastigmine), Huperzine A (huperzine A), E2020 (donepezil) etc.; β, gamma secretase Depressant; The brain metabolism regulators, as vincamine, nimodipine, cinnarizine; Affect the medicine of Radical Metabolism, as vitamins C in conjunction with vitamin-E etc.But these medicines can only delay the progress of the AD course of disease, and reduce gradually with the PD curative effect of medication, side effect is arranged, so find focus and difficult point that medicine and method for the AD cause of disease become the world of medicine.
Research shows, neurotrophic factor has important impact to the lysis of neurodevelopment and Adult Nervous System.Wherein nerve growth factor (nerve growth factor, NGF) is found to stop or to reduce neuronic regression in the nerve degenerative diseases animal model.First most important neurotrophic factor of finding as the mankind, NGF has the important regulating and controlling effect to the aspects such as growth, growth, differentiation and function maintenance of neurocyte.Further find that neurotrophic factor can stop or reduce neuratorphy, neurodegeneration, promote post-traumatic neural the reparation.But NGF contains more than 100 amino acid whose protein, and larger molecular weight and stronger polarity make it to pass through hemato encephalic barrier, have limited its application clinically.Therefore, find and there is the NGF of plan activity and can naturally become study hotspot by the micromolecular compound of hemato encephalic barrier.The micromolecular compound of intending the NGF activity should have neurotrophy and neuroprotective function.
Herbal medicine is the basic substance of Chinese materia medica, for the natural radioactivity organic compound provides huge resource.China's Development of Chinese Medicine history of several thousand, be distributed with valuable famous-region drug.The research that utilizes huge Chinese material medicine resource to carry out new composition and new activity is very important and significant.Some Chinese medicines have significant curative effect to senile dementia, and wherein foremost is the alkaloid extracted from Herba Lycopodii serrati---huperzine potassium (huperzine A).As the original new drug with independent intellectual property right of Chinese Academy of Sciences institute of materia medica exploitation, it is not only acetylcholinesterase (AChE) Depressant of efficient highly selective, and can also reduce the nerve cell death that L-glutamic acid brings out; There is the response to oxidative stress that obvious neuroprotective cell antagonism beta-amyloyd polypeptide (A β) produces; Can induce the nerve cell apoptosis effect to resistive to hydrogen peroxide, inhibitors of protein kinase C etc.
Having another name called dwarf lilyturf, book band grass, Radix Ophiopogonis the tuber of dwarf lilyturf, is Liliaceae Ophiopogon perennial evergreen herbaceous plant.Fibrous root is more sturdy, and often expand at the top of root or middle part becomes spindle shape meat fritter.As traditional Chinese medicine, can improve immunologic function the tuber of dwarf lilyturf; Various bacteria is had to restraining effect; Can strengthen the pituitary adrenal system function, improve the organism adaptation ability; Anti-arrhythmia and the peripheral vascular effect of expansion are arranged; Can improve hypoxia-bearing capability; Hypoglycemic activity is arranged.
For lung-dryness syndrome due to deficiency of YIN, cough with ropy sputum; Thermal burn stomach-Yin or deficiency of stomach-YIN, dry throat and mouth, constipation with dry stool; Deficiency of heart-yin or the heart channel of Hang-Shaoyin have heat, dysphoria and insomnia, red tongue with a little fluid.
Originate in Cixi Tianyuan town the tuber of dwarf lilyturf, after introduce a fine variety to Kan Dun gradually.Mainly be distributed in Cixi western part and northern Plain and Yuyao City east and northern Plain.The happiness of the property tuber of dwarf lilyturf is moistening.Cixi, Yuyao one band, affected by oceanic climate, warm moistening, the illumination aboundresources, and adding that soil is fertile contains sandyly, be the good living environment tuber of dwarf lilyturf, thereby output is high.The annual production of the nineties is stabilized in 200 tons of left and right, and part for export.
Summary of the invention
The preparation method who the purpose of this invention is to provide active ingredient in a kind of tuber of dwarf lilyturf.The preparation method that the present invention proposes, mainly realize by following steps:
(1) pulverizing and lixiviate:
Chinese medicine tuber of dwarf lilyturf
ophiopogon japonicus (Thunb.)ker-Gawl after crushed, with lixiviate under 50.0 L industrial grade benzenemethanol room temperatures 4 ~ 5 days, suction filtration is concentrated, obtain methyl alcohol extractive substance study, water and acetic acid ethyl dissolution study again, all be transferred to separating funnel and carry out the solvent distribution, upper strata ester layer and lower aqueous layer obtain ethyl acetate layer study and water layer study after concentrating respectively.
(2) separation and purifying:
The water layer study is first separated through octadecyl silane chromatographic column (ODS) opening column, and solvent system is methyl alcohol: water=50:50 by volume, 60:40,70:30,80:20,100:0, according to the TLC result, identical cut is merged, obtain 6 part A1-A6.Subsequently, by A2 part (14.2 g), (elution system is methyl alcohol: water=50:50) again with silica gel column chromatography, separate (silica gel 200-300 order, solvent system is methyl alcohol: chloroform=20:80 by volume, 50:50,100:0), according to the TLC result, identical cut is merged, obtain 4 part B1-B4, wherein (elution system is methyl alcohol: chloroform=20:80) quality is 1.1 g to the B2 part.By the B2 part, through the silica gel column chromatography separation, (solvent system is methyl alcohol: chloroform=10:90 by volume, 12:88,14:86,16:84 again, 18:82,20:80,30:70,50:50,100:0), according to the TLC result, identical cut is merged, obtain 8 cut C1-C8.Obtaining thus the C7(elution system is methyl alcohol: chloroform=30:70) for containing the active ingredient of saponin(e in the tuber of dwarf lilyturf.
Contained four kinds of saponins compounds of active ingredient its preparation method is provided in another object of the present invention has been isolation identification described tuber of dwarf lilyturf, wherein compound (
1) be 25 (
s)-ruscogenin 1-
o-β-d-xylopyranosyl-(1 → 2)-[
β-d-xylopyranosyl-(1 → 3)]-
β-d-fucopyranoside; Compound (
2) be neoruscogenin 1-
o-α-l-rhamnopyranosyl-(1 → 3)-
α-l-rhamnopyranosyl-(1 → 2)-
β-d-fucopyranoside; Compound (
3) be 25 (
s)-ruscogenin 1-
o-α-l-rhamnopyranosyl-(1 → 2)-[
β-d-xylopyranosyl-(1 → 3)]-4-
o-acetyl-
β-d-fucopyranoside; Compound (
4) be 25 (
s)-ruscogenin 1-
o-α-l-rhamnopyranosyl-(1 → 2)-[
β-d-xylopyranosyl-(1 → 3)]-
β-d-fucopyranoside. is compound wherein
1,
2,
3for new compound.Its structural formula is as follows:
Wherein:
Compound 1:R
1for
, R
2for
, R
3for H, R
4for ax Me;
Compound 2:R
1for
, R
2for H, R
3for H, R
4for=CH
2;
Compound 3:R
1for
, R
2for
, R
3for COCH
3, R
4for ax Me;
Compound 4:R
1for
, R
2for
, R
3for H, R
4for ax Me.
The preparation method of above compound, realize by following steps:
(1) pulverizing and lixiviate:
Chinese medicine tuber of dwarf lilyturf
ophiopogon japonicus (Thunb.)ker-Gawl after crushed, with lixiviate under 50.0 L industrial grade benzenemethanol room temperatures 4 ~ 5 days, suction filtration is concentrated, obtain methyl alcohol extractive substance study, water and acetic acid ethyl dissolution study again, all be transferred to separating funnel and carry out the solvent distribution, upper strata ester layer and lower aqueous layer obtain ethyl acetate layer study and water layer study after concentrating respectively.
(2) separation and purifying:
The water layer study is first separated through octadecyl silane chromatographic column (ODS) opening column, and solvent system is methyl alcohol: water=50:50 by volume, 60:40,70:30,80:20,100:0, according to the TLC result, identical cut is merged, obtain 6 part A1-A6.Subsequently, by A2 part (14.2 g), (elution system is methyl alcohol: water=50:50) again with silica gel column chromatography, separate (silica gel 200-300 order, solvent system is methyl alcohol: chloroform=20:80 by volume, 50:50,100:0), according to the TLC result, identical cut is merged, obtain 4 part B1-B4, wherein (elution system is methyl alcohol: chloroform=20:80) quality is 1.1 g to the B2 part.By the B2 part, through the silica gel column chromatography separation, (solvent system is methyl alcohol: chloroform=10:90 by volume, 12:88,14:86,16:84 again, 18:82,20:80,30:70,50:50,100:0), according to the TLC result, identical cut is merged, obtain 8 cut C1-C8.Obtaining thus the C7(elution system is methyl alcohol: chloroform=30:70) for containing the active ingredient of saponin(e in the tuber of dwarf lilyturf.Further with HPLC, purify that (flow velocity 3 mL/min, detect wavelength 210 nm for chromatographic condition: chromatographic column Develosil ODS-UG-5 (φ 20 * 250 mm), Nomura Chemical, and moving phase is methyl alcohol: water=80:20), obtain compound
1(t
r =33 min), compound
2(t
r =36 min) and compound
3(t
r =55 min), compound
4(t
r =38 min).
Another purpose of the present invention is to provide this active ingredient and contained saponin compound is prevented and treated the application in the disease medicaments such as senile dementia or food in preparation.Especially the application in preparing nerve degenerative diseases medicines such as preventing and treating Alzheimer's disease or food.Described medicine or food contain pharmacy or food acceptable carrier or thinner.
The present invention, in the guiding of PC12 cell bio-activity identification systems, separates through simple the extraction, from the tuber of dwarf lilyturf
ophiopogon japonicus(Thunb.) obtain active ingredient in the tuber of dwarf lilyturf and contained saponin compound in the Ker-Gawl extract, it has remarkable plan Nerve Growth Factor Activity.
Described pharmaceutically acceptable carrier refers to the pharmaceutical carrier of pharmaceutical field routine, and as thinner, vehicle wait in this way, weighting agent is as starch, sucrose, Microcrystalline Cellulose etc.; Tackiness agent is as starch slurry, hydroxypropylcellulose, gelatin, polyoxyethylene glycol etc.; Wetting agent is as Magnesium Stearate, differential silica gel, polyoxyethylene glycol etc.; The absorption enhancer polysorbate, Yelkin TTS etc., tensio-active agent poloxamer, lipid acid sorb are smooth, polysorbate etc., can also in composition, add in addition other assistant agent as flavouring agent, sweeting agent etc.
Saponins compound of the present invention can be with the unit dosage form administration, and route of administration can be enteron aisle or non-enteron aisle, comprises oral, muscle, subcutaneous and nasal cavity.
Active ingredient route of administration of the present invention can be intravenously administrable.Injection comprises intravenously administrable, intramuscular injection, subcutaneous injection and acupoint injection therapy.
The various formulations of pharmaceutical composition of the present invention can for example make active ingredient mix with one or more carriers according to the conventional production method preparation of pharmaceutical field, then are made into required formulation.
Form of administration can be solid preparation, capsule or liquid preparation, comprises tablet, capsule, dispersible tablet, oral liquid, infusion solutions, little pin, freeze-dried powder, ointment, liniment or suppository.
Described food formulation is solid or liquid.
Beneficial effect of the present invention is: (1) is separated and is obtained active ingredient in the tuber of dwarf lilyturf by organic solvent lixiviate, solvent distribution, chromatographic column, further high performance liquid chromatography (HPLC) purifying obtains active compound, the preparation method has simply, fast, the compound purity advantages of higher of acquisition.(2) active ingredient and compound have the activity of significant plan nerve growth factor, can in the nerve degenerative diseases such as prevention senile dementia, obtain application.(3), as micromolecular compound, the active compound of natural separation shows significant plan NGF activity in the in-vitro screening model PC12 of senile dementia cell.Using this compounds as primer, optimize structure, for the new drug development of prevention and nerve degenerative diseases such as treatment senile dementia etc. carries out basic research, will have important practical significance.
The accompanying drawing explanation
Fig. 1 adds the nervous process differentiation rate that the tuber of dwarf lilyturf, opening column was separated rear component C1-C8 PC12 cell after 48 hours.
Fig. 2 adds compound
1-4the nervous process differentiation rate result of PC12 cell after 48 hours.
Fig. 3 adds compound
1the Photomicrograph of PC12 cellular neural projection after 48 hours, a in figure, the negative contrast of 1% DMSO; B, NGF 40 ng/ml, positive contrast; C, compound
11
.
Embodiment
The present invention in conjunction with the accompanying drawings and embodiments, foregoing of the present invention is described in further detail, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to subordinate's example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
embodiment 1the extraction of the tuber of dwarf lilyturf with separate
(1) pulverizing and lixiviate:
By 10.0 kg Chinese medicine tubers of dwarf lilyturf (
ophiopogon japonicus (Thunb.)ker-Gawl) after pulverizing, with concussion lixiviate under 50.0 L methyl alcohol (technical grade) room temperatures 3 days.Suction filtration, concentrated after, obtain methyl alcohol extractive substance study.Then, with ethyl acetate and water dissolution study, all be transferred to separating funnel and carry out the solvent distribution, upper strata ester layer and lower aqueous layer obtain ethyl acetate layer study (15.9 g) and water layer study (2.9 kg) after concentrating respectively.
(2) separation of water layer study and purifying:
At first, water layer study (2.9 kg) evenly is divided into to 10 parts, every part all separates through reverse phase silica gel ODS chromatography column that (solvent system is methyl alcohol: water=50:50 by volume, 60:40,70:30,80:20,100:0), according to the TLC result, identical cut is merged, obtain 6 part A1-A6.Subsequently, by A2 part (14.2 g), (elution system is methyl alcohol: water=50:50) again with silica gel column chromatography, separate (silica gel 200-300 order, solvent system is methyl alcohol: chloroform=20:80 by volume, 50:50,100:0), obtain 4 part B1-B4, wherein (elution system is methyl alcohol: chloroform=20:80) quality is 1.1 g to the B2 part.By the B2 part, through the silica gel column chromatography separation, (solvent system is methyl alcohol: chloroform=10:90 by volume, 12:88,14:86,16:84 again, 18:82,20:80,30:70,50:50,100:0), according to the TLC result, identical cut is merged, obtain 8 cut C1-C8.Obtaining thus C7 is to contain the active ingredient of saponin(e in the tuber of dwarf lilyturf.Finally, by cut C7(107.0 mg, elution system is methyl alcohol: chloroform=30:70), with high-efficient liquid phase chromatogram purification (chromatographic condition: chromatographic column Develosil ODS-UG-5 (φ 20 * 250 mm), Nomura Chemical, flow velocity 3 mL/min, detect wavelength 210 nm, moving phase is methyl alcohol: water=80:20), obtain compound
1(t
r =33 min), compound
2(t
r =36 min) and compound
3(t
r =55 min), compound
4(t
r =38 min).
The physicochemical characteristics of 2 couples of resulting compound 1-4 of embodiment 1 of embodiment and the Qualitative Identification of compound structure:
Compound 1: colorless solid; It is C that high resolution mass spectrum demonstrates its molecular formula
43h
68o
16(HR ESI-TOF-MS m/z (M+H)
+calcd.for C
43h
69o
16: 841.4586, Found:841.4603).
Table 1 compound 1
1h with
13c NMR data (solvent: pyridine-d
5)
a500MHz, be coupling constant (Hz) in bracket,
b125MHz.
Compound 2: colorless solid; It is C that high resolution mass spectrum demonstrates its molecular formula
45h
70o
16(HR ESI-TOF-MS m/z (M+H)
+calcd.for C
45h
71o
16: 867.4742, Found:867.4736).
Table 2 compound 2
1h with
13c NMR data (solvent: pyridine-d
5)
a500MHz, be coupling constant (Hz) in bracket,
b125MHz.
Compound 3: colorless solid; It is C that high resolution mass spectrum demonstrates its molecular formula
46h
72o
17(HR ESI-TOF-MS m/z (M+Na)
+calcd.for C
46h
72o
17na:919.4667, Found:919.4600).
Table 3 compound 3
1h with
13c NMR data (solvent: pyridine-d
5)
a500MHz, be coupling constant (Hz) in bracket,
b125MHz.
Compound 4: colorless solid; It is C that high resolution mass spectrum demonstrates its molecular formula
44h
70o
16, (HR ESI-TOF-MS m/z (M+H)
+calcd.for C
44h
71o
16: 855.5737, Found:855.4728).
1h NMR (500MHz, in pyridine-d
5) δ: 1.72 (3H, d, J=6.0Hz, Me-6''), 1.49 (3H, d, J=6.5Hz, Me-6'), (1.40 3H, s, Me-19), 1.05 (3H, d, J=7.0Hz, Me-21), (1.03 3H, d, J=7.0Hz, Me-27), 0.83 (3H, s, Me-18).
embodiment 3in the tuber of dwarf lilyturf, active ingredient and contained compound thereof show significant plan NGF activity in the in-vitro screening model PC12 of senile dementia cell.
Test method:
1) cultivation of PC12 cell: inoculate 20 * 10
4individual cell, in the culture dish of 100 mm, containing 10 ml DMEM substratum (wherein containing 10% horse serum, 5% foetal calf serum), is often changed a subculture two days later, carries out succeeding transfer culture when cell density reaches requirement.First with PBS, cell is washed to twice, then add 10 ml PBS in culture dish, at 37 ℃, 5% CO
2interior standing 10 minutes of incubator, take out rear purge, the PBS that is suspended with cell transferred to the disposable centrifuge tube of 15 ml, centrifugal count with blood counting chamber afterwards, in 24 orifice plates, every hole first adds the DMEM substratum of 1 ml containing serum, and after cell counting, every hole connects 2 * 10
4individual cell, mix, at CO
2incubator is cultivated application of sample after 24 hours.
2) active testing: with the negative contrast of DMSO, the positive contrast of NGF 40 ng/mL, by the tuber of dwarf lilyturf separated portion or compound be configured to the DMSO solution of different concns, contain the DMEM solution (containing serum) of 1% DMSO and sample with 1 ml by after the former substratum replacement in every hole of 24 porocyte plates, put into 37 ℃, cultivate in the incubator of 5% CO2.Under inverted microscope every 24 hours, continuous 6 days observation of cell metamorphosis, record the nervous process differentiation rate (nervous process length is greater than the ratio of total cell number under the cell number of cell space diameter and the visual field) of cell, about 100 cells under each visual field, choose at random 3 places, and the statistics mapping analysis.
3) experimental result: referring to Fig. 1, Fig. 2, Fig. 3.
3
under the concentration of/ml, the active situation of each component C1-C8 on the PC12 cell after the opening column separation tuber of dwarf lilyturf after 48 hours.Wherein, C7 demonstrates the most significant active, C in Fig. 1: negative control: 1% DMSO; NGF (40 ng/ml): positive control).
1
concentration under, compound after 48 hours
1-4demonstrate significant plan NGF activity.C in Fig. 2: negative control: 1% DMSO; NGF (40 ng/ml): positive control).A in Fig. 3, the negative contrast of 1% DMSO; B, NGF 40 ng/ml, positive contrast; C, compound
11
.
Claims (10)
1. the preparation method of active ingredient in tuber of dwarf lilyturf is characterized in that:
(1) pulverizing and lixiviate:
Chinese medicine tuber of dwarf lilyturf after crushed, with lixiviate under the industrial grade benzenemethanol room temperature 4 ~ 5 days, suction filtration is concentrated, obtain methyl alcohol extractive substance study, water and acetic acid ethyl dissolution study again, all be transferred to separating funnel and carry out the solvent distribution, upper strata ester layer and lower aqueous layer obtain ethyl acetate layer study and water layer study after concentrating respectively;
(2) separation and purifying:
The water layer study is first separated through octadecyl silane chromatographic column opening column, solvent system is methyl alcohol: water=50:50 by volume, 60:40, 70:30, 80:20, 100:0, according to the TLC result, identical cut is merged, obtain 6 part A1-A6, subsequently, will be by methyl alcohol: water=50:50 wash-out and the A2 part come is separated with silica gel column chromatography again, used silica gel 200-300 order, solvent system is methyl alcohol: chloroform=20:80 by volume, 50:50, 100:0, merge identical cut, obtain 4 part B1-B4, will be by methyl alcohol: chloroform=20:80 wash-out and the B2 part come is separated through silica gel column chromatography again, solvent system is methyl alcohol: chloroform=10:90 by volume, 12:88, 14:86, 16:84, 18:82, 20:80, 30:70, 50:50, 100:0, according to the TLC result, identical cut is merged, obtain 8 cut C1-C8, wherein by methyl alcohol: the C7 that chloroform=the 30:70 wash-out obtains contains the active ingredient of saponin(e in the tuber of dwarf lilyturf.
2. the preparation method of active ingredient in a kind of tuber of dwarf lilyturf according to claim 1, is characterized in that, from the active ingredient obtained, prepares compound 1-compound 4 again, by following steps, realizes:
(1) pulverizing and lixiviate:
Chinese medicine tuber of dwarf lilyturf after crushed, with lixiviate under the industrial grade benzenemethanol room temperature 4 ~ 5 days, suction filtration is concentrated, obtain methyl alcohol extractive substance study, water and acetic acid ethyl dissolution study again, all be transferred to separating funnel and carry out the solvent distribution, upper strata ester layer and lower aqueous layer obtain ethyl acetate layer study and water layer study after concentrating respectively;
(2) separation and purifying:
The water layer study is first separated through octadecyl silane chromatographic column opening column, solvent system is methyl alcohol: water=50:50 by volume, 60:40, 70:30, 80:20, 100:0, according to the TLC result, identical cut is merged, obtain 6 part A1-A6, subsequently, will be by methyl alcohol: water=50:50 wash-out and the A2 part come is separated with silica gel column chromatography again, used silica gel 200-300 order, solvent system is methyl alcohol: chloroform=20:80 by volume, 50:50, 100:0, merge identical cut, obtain 4 part B1-B4, will be by methyl alcohol: chloroform=20:80 wash-out and the B2 part come is separated through silica gel column chromatography again, solvent system is methyl alcohol: chloroform=10:90 by volume, 12:88, 14:86, 16:84, 18:82, 20:80, 30:70, 50:50, 100:0, according to the TLC result, identical cut is merged, obtain 8 cut C1-C8, wherein by methyl alcohol: the C7 that chloroform=the 30:70 wash-out obtains contains the active ingredient of saponin(e in the tuber of dwarf lilyturf,
(3) obtain compound 1-4:
By elution system, be methyl alcohol: chloroform=30:70 wash-out cut C7 107.0 mg high-efficient liquid phase chromatogram purifications out, chromatographic condition is: chromatographic column Develosil ODS-UG-5 φ 20 * 250 mm, Nomura Chemical, flow velocity 3 mL/min, detect wavelength 210 nm, and moving phase is methyl alcohol: water=80:20, in retention time, it is 33 minutes, 36 minutes, 55 minutes, within 38 minutes, obtain respectively compound 1-4;
The structural formula of described compound is as follows:
Wherein:
Compound 1:R
1for
, R
2for
, R
3for H, R
4for ax Me;
Compound 2:R
1for
, R
2for H, R
3for H, R
4for=CH
2;
Compound 3:R
1for
, R
2for
, R
3for COCH
3, R
4for ax Me;
Compound 4:R
1for
, R
2for
, R
3for H, R
4for ax Me.
3. the application of active ingredient in preparation senile dementia prevention and cure medicine in the tuber of dwarf lilyturf that method according to claim 1 obtains.
4. the application of the compound 1-4 that wants 2 described methods to obtain according to right in preparation senile dementia prevention and cure medicine.
5. according to the described application of claim 3 or 4, it is characterized in that active ingredient or the compound 1-4 application in preparation control Alzheimer disease drug in the described tuber of dwarf lilyturf.
6. according to the arbitrary described application of claim 3-5, it is characterized in that, the route of administration of described medicine is enteron aisle and non-enteron aisle.
7. according to the arbitrary described application of claim 3-5, it is characterized in that, the formulation of described medicine is solid preparation or liquid preparation.
8. the application of active ingredient in preparation senile dementia prevention and cure food in the tuber of dwarf lilyturf that method obtains according to claim 1.
9. the application of the compound 1-4 that method obtains according to claim 2 in preparation senile dementia prevention and cure food.
10. application according to claim 8 or claim 9, is characterized in that, active ingredient or the compound 1-4 application in preparation control Alzheimer's disease food in the described tuber of dwarf lilyturf, and described food formulation is solid or liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013103895413A CN103450323A (en) | 2013-08-31 | 2013-08-31 | Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013103895413A CN103450323A (en) | 2013-08-31 | 2013-08-31 | Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103450323A true CN103450323A (en) | 2013-12-18 |
Family
ID=49733201
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2013103895413A Pending CN103450323A (en) | 2013-08-31 | 2013-08-31 | Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103450323A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383154A (en) * | 2016-05-17 | 2017-11-24 | 天士力制药集团股份有限公司 | A kind of new liriope muscari Baily Shandong this can steroid saponin compound and its preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102247524A (en) * | 2011-05-25 | 2011-11-23 | 浙江大学 | Preparation method of ophiopogon japonicus (Thunb.) Ker-Gawl total steroidal saponin extract, and application thereof in antisenescence pharmacy |
| CN102250197A (en) * | 2011-05-25 | 2011-11-23 | 浙江大学 | Preparation method and application of total steroidal saponin extracts of dwarf lilyturf roots |
-
2013
- 2013-08-31 CN CN2013103895413A patent/CN103450323A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102247524A (en) * | 2011-05-25 | 2011-11-23 | 浙江大学 | Preparation method of ophiopogon japonicus (Thunb.) Ker-Gawl total steroidal saponin extract, and application thereof in antisenescence pharmacy |
| CN102250197A (en) * | 2011-05-25 | 2011-11-23 | 浙江大学 | Preparation method and application of total steroidal saponin extracts of dwarf lilyturf roots |
Non-Patent Citations (2)
| Title |
|---|
| 叶英等: "麦冬来源的拟神经生长因子活性物质的化学及生物学研究", 《(2012)长三角药物化学研讨会》 * |
| 曲媛: "中药来源的拟神经生长因子活性物质的化学及生物学研究", 《中国博士学位论文全文数据库，医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383154A (en) * | 2016-05-17 | 2017-11-24 | 天士力制药集团股份有限公司 | A kind of new liriope muscari Baily Shandong this can steroid saponin compound and its preparation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2829275B1 (en) | Total flavone extract of abelmoschus manihot and preparation method thereof | |
| CN1977951B (en) | Gastrodia elata plant extract for preventing senile dementia and its preparing method | |
| JP5294509B2 (en) | Cycloastragenol monoglucoside, process for its production and use as a pharmaceutical composition | |
| CN102319275B (en) | Sunset abelmoschus flower extract, preparation and preparation method thereof | |
| CN105294623B (en) | A kind of Sesquiterpene lactones compound, its preparation method and application | |
| CN104910240B (en) | Triterpenoid saponin in Bougainvillea glabra, the hypoglycemic drug with it as active component, and its preparation method and application | |
| CN103360456B (en) | Triterpene compound and Synthesis and applications | |
| CN104262465A (en) | Rubiaceae cyclopeptides used as TAK1 inhibitor and preparation method thereof | |
| CN111647003B (en) | Triepoxyhexahydrochromone A, and pharmaceutical composition and application thereof | |
| CN103288914B (en) | Preparation method of traditional Chinese medicine manyflower tickclove herb extract and application in anti-senile dementia medicaments | |
| CN113754620B (en) | Lignan amide compound in fructus cannabis, and preparation method and application thereof | |
| CN102250197B (en) | Preparation method and application of total steroidal saponin extracts of dwarf lilyturf roots | |
| CN106674086B (en) | A kind of piperidones Alkaloid compound and its preparation method and application | |
| CN103450323A (en) | Radix ophiopogonis nerve growth factor activity simulating component and compound and preparation | |
| CN106236792B (en) | The preparation of Folum Ilicis extract and anti-alzheimer's disease purposes | |
| CN101774922B (en) | 2,3-dyhydroxyl parabens compound as well as preparation and application thereof | |
| CN111529515B (en) | Application of 12, 15-dioxo-alpha-cnidiene in pharmacy | |
| KR0183448B1 (en) | Anti-carcinogenic composition containing ginsenoside rg5 | |
| CN103342730B (en) | Preparation method of extract of traditional Chinese medicine herb of manyflower ticklover and use of the extract in anti-aging | |
| CN103204860B (en) | There is the amaryllidaceae alkaloid compounds of neuroprotective | |
| CN109206392B (en) | Coumarin compound and preparation method and application thereof | |
| CN115806490B (en) | Phenolic hetero-terpene compound with function of activating AMPK phosphorylation, pharmaceutical composition, preparation method and application | |
| CN113754622B (en) | Lipid lowering application of dibenzofuran compounds | |
| CN115806483B (en) | Phenolic hetercerpene (+) -Dayaolingzhiol M and its application in pharmacy and food | |
| CN115010689B (en) | Compound kurarinol A and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20131218 |