CN105582000B

CN105582000B - Application of the terpene substances in preparing treatment senile dementia or Alzheimer disease drugs in Bark of Eucommia Ulmoides or folium cortex eucommiae

Info

Publication number: CN105582000B
Application number: CN201610016096.XA
Authority: CN
Inventors: 张忠立; 左月明
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-01-12
Filing date: 2016-01-12
Publication date: 2018-10-12
Anticipated expiration: 2036-01-12
Also published as: CN105582000A

Abstract

The invention discloses application of the terpene substances in Bark of Eucommia Ulmoides or folium cortex eucommiae in preparing treatment senile dementia or Alzheimer disease drugs.Terpene substances itself refer to that ulmoprenol in prepared Bark of Eucommia Ulmoides or folium cortex eucommiae,1 deoxidation ulmoprenol,1 deoxidation △ 4,5 ulmoprenols,2,3 dihydroxy △ 4,5 ulmoprenols,4 ' acetyl group ulmoprenols,Eucommioside A,Folium cortex eucommiae glycosides A,Folium cortex eucommiae glycosides B,Folium cortex eucommiae glycosides C,One or more terpene substances in folium cortex eucommiae glycosides D are used to prepare treatment senile dementia or Alzheimer disease drugs,Terpene substances have treatment by aging in human body due to possessing the structural units such as common or similar monoterpene Cortex Eucommiae alcohols or sequiterpene huge pillar alkanes in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae,Cardiovascular and cerebrovascular disease leads to the cerebral ischemia of Brain hypoperfusion,Amyloid A β storage disorders are because of the senile dementia of formation or the function curative effect of Alzheimer disease,While terpene substances have auxiliary treatment brain blood caused by diseases of cardiovascular and cerebrovascular systems scarce and its sequel resulted from cardio-cerebral blood-vessel diseases in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae.

Description

Terpene substances are preparing treatment senile dementia or A Erci in Bark of Eucommia Ulmoides or folium cortex eucommiae Application in the silent medicine in sea

Technical field

The present invention relates to terpene substances in Bark of Eucommia Ulmoides or folium cortex eucommiae to prepare treatment senile dementia or Alzheimer disease Application in drug, belongs to technical field of traditional Chinese medicine preparation.

Background technology

Bark of Eucommia Ulmoides be Eucommiaceae plant Cortex Eucommiae Eucommia ulmoides Oliv. dry bark, sweet in flavor, micro-pungent, It is warm-natured, return liver and kidney channel, many effects such as tool filling liver kidney, strengthening the bones and muscles, blood pressure lowering, tocolysis.Bark of Eucommia Ulmoides is modern study proves that Cortex Eucommiae has There are bidirectional modulation blood pressure, strengthen immunity, antitumor, anti-oxidant, antiviral, anti-aging, reduction blood glucose, adjusting blood fat etc. multi-party The effect of face.Folium cortex eucommiae is the dried leaf of Eucommiaceae Cortex Eucommiae Eucommia ulmoides Oliv., in 2005 as a kind of new Medicinal material takes in Chinese Pharmacopoeia, and warm-natured, taste micro-pungent has the effect of filling liver kidney, strengthening the bones and muscles, blood pressure lowering.Folium cortex eucommiae modern pharmacology is ground Studying carefully has the effects that anti-aging, anti-oxidant, lowering blood pressure and blood fat, strengthen immunity.Inventor's current research confirm Bark of Eucommia Ulmoides or Folium cortex eucommiae total extract, which treats H/MAD, has the pharmacological activity of anti-A β damages.

Consider aging, the vascular risk factor (hypertension, atherosclerosis and transient ischemic attack etc.) is led The cerebral ischemia and A β of the brain Low perfusion of cause act in the AD origin causes of formation, and Bark of Eucommia Ulmoides or folium cortex eucommiae exist to preventing above-mentioned three kinds of principal elements Generation, development and the symptomatic treatment of H/MAD has its unique advantage, inventor's current research to confirm Bark of Eucommia Ulmoides or folium cortex eucommiae 60% Ethyl alcohol totality object can make H/MAD rat model learning and memories improve significantly, and the vigor of T-AOC in serum can be made extensive It is multiple, CAT, SOD, GSH-P can be significantly improved_XActivity level, enhancing body remove free radical ability；Containing for TChE can be reduced Amount, the degree for preventing the reduction of neurotransmitter, while the content of MDA capable of being reduced again to reduce body by radical damage, significantly Improve the oxidation resistance of H/MAD rats.In inventor's cell culture experiments in vitro, PC12 cellular damages are caused using A β 25-35 Model has detected the vigor of each experimental group cell using mtt assay, to confirm Bark of Eucommia Ulmoides or folium cortex eucommiae treatment senile dementia or The active component and active constituent of Alzheimer disease (AD) are that Bark of Eucommia Ulmoides or folium cortex eucommiae treat senile dementia or alzheimer ' The new drug research based theoretical of silent disease (AD).The studies above is not yet reported that we are ground by the inside and outside experiment of system Study carefully, finds Bark of Eucommia Ulmoides or folium cortex eucommiae to by aging, cardiovascular and cerebrovascular disease (heart disease, hypertension, atherosclerosis and short first Temporary property cerebral ischemia attack etc.) cause the cerebral ischemia of Brain hypoperfusion, amyloid A β storage disorders because formation is senile silly Slow-witted or Alzheimer disease disease has good therapeutic effect.Terpene substances, which are used to prepare, in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae controls Senile dementia or Alzheimer disease drugs are treated, terpene substances are common or similar because possessing in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae The structural units such as monoterpene Cortex Eucommiae alcohols or sequiterpene huge pillar alkanes and in human body have treatment drawn by diseases of cardiovascular and cerebrovascular systems The function curative effect of the senile dementia or Alzheimer disease that rise, while there is auxiliary treatment to be caused by diseases of cardiovascular and cerebrovascular systems Brain blood lack and its sequel resulted from cardio-cerebral blood-vessel diseases.It can answering with partial alternative Bark of Eucommia Ulmoides in clinical research field folium cortex eucommiae to find With theoretical foundation is provided, there is good academic and clinical value.

Invention content

The invention discloses terpene substances in Bark of Eucommia Ulmoides or folium cortex eucommiae to prepare treatment senile dementia or Alzheimer Application in medicine.Terpene substances itself refer to that ulmoprenol, 1- deoxidations ulmoprenol, 1- in prepared Bark of Eucommia Ulmoides or folium cortex eucommiae Deoxidation-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, one or more terpene substances in folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D be used to prepare treatment senile dementia or Alzheimer disease drugs, in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances because possess common or similar monoterpene Cortex Eucommiae alcohols or The structural units such as sequiterpene huge pillar alkanes and in human body have treatment by aging, cardiovascular and cerebrovascular disease (heart disease, hypertension, Atherosclerosis and transient ischemic attack etc.) lead to cerebral ischemia, the amyloid A β storage disorders of Brain hypoperfusion Because of the senile dementia of formation or the function curative effect of Alzheimer disease disease, while having auxiliary treatment by cardio-cerebrovascular Brain blood caused by disease lacks and its sequel resulted from cardio-cerebral blood-vessel diseases.

Foregoing invention uses following technical proposals.

Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, The preparation method of folium cortex eucommiae glycosides D, it is characterised in that steps are as follows：

(1) is impregnated with 0-100% ethyl alcohol by Bark of Eucommia Ulmoides or folium cortex eucommiae pulverizing medicinal materials at coarse powder, is added 4-12 times and measures solvent Heating and refluxing extraction several times, filters, merging filtrate；

(2) the above-mentioned filtrate decompressions of are concentrated into 0.5-1.8g raw medicinal herbs/ml, refrigerated overnight, analyse glue, filtering, and filtrate is again with having Solvent extracts or the upper macroporous absorbent resin of filtrate carries out gradient elution, recycles organic solvent, obtains silicagel column on medicinal extract；

(3) chloroform is used successively after the upper silicagel columns of：Methanol or dichloromethane：Methanol or chloroform：Methanol：Water or Dichloromethane：Methanol：Water gradient elution, with ulmoprenol, 1- deoxidations ulmoprenol, 1- in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Deoxidation-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D product TLC or HPLC detections as a contrast, merge corresponding fraction；

(4) shuts out ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Secondary alcohol, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, The corresponding fraction reduced pressure of folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D are evaporated, and are dissolved with ethanol in proper amount, are filtered, ethyl alcohol is recovered under reduced pressure extremely Alcohol content analyses glue between 10-60%, filters, and suitable polyamide is added in filtrate, stirs 0.5-2 hours, and it is small to stand 4-24 When, proper amount of active carbon is added in filtrate for filtering, stirs 0.5-2 hour, stands 4-24 hour, filters, filtrate be spray-dried or Freeze-drying is dried, and collects spray dried powder or freeze-dried powder, with 10-100% alcohol crystals and recrystallize to get.Or The reduced pressure of corresponding fraction is evaporated or crystallized product by person, and standby chromatography is suppressed in or high performance liquid preparative chromatography prepares purifying, Mobile phase be methanol-water 5-100% or acetonitrile water 5-100% isocratic or gradient elution, recycling eluent to get.

More specifically, in step (1), it is added 4-12 times and measures solvent heating and refluxing extraction 2-4 times, each 1-3 hours.

More specifically, in step (2), in 0-4 DEG C of refrigerated overnight.

More specifically, in step (2), it is 45-80 DEG C that temperature, which is concentrated under reduced pressure,.

More specifically, in step (2), filtrate is eluted with water, the washing of 25% ethyl alcohol successively through macroporous adsorbing resin for purification De-, 40% ethyl alcohol water elution, 60% ethyl alcohol water elution, 95% ethyl alcohol water elution, it is main to collect water elution position, 25% ethanol water Position, 40% ethyl alcohol water elution position are eluted, recycling eluent obtains silicagel column on medicinal extract.Wherein, water elution position is mainly single Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- Du in terpene Cortex Eucommiae alcohol compound Secondary alcohol, 4 '-acetyl group-ulmoprenol, eucommioside A, 25% ethyl alcohol water elution position is mainly (+) rosin in lignanoids substance Element -4,4'-O- β-D- double glucopyranoside, (+) 8- hydroxyls-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls Base-rosin element -4-O- β-D- glucopyranosides, (+) rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyranosides, (+) 8,9'- dihydroxy-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin Element -4,4'-O- β-D- double glucopyranoside, syringaresinol -4-O- β-D- glucopyranosides, middle fat element -4-O- β-D- pyrroles Mutter glucose glucosides, olivil -4'-O- β-D-Glucose glycosides, olivil -4-O- β-D- glucopyranosides, fallen leaves The double glucopyranosides of rosin spirit -4,4'-O- β-D-, 8- hydroxyls-lariciresinol -4'-O- β-D- glucopyranosides, fallen leaves Rosin spirit -4-O- β-D- glucopyranosides, olive element bioside, Auricled Hedyotis Herb element bioside, cloves glycerine-β-syringaresinol ether Bioside, 40% ethyl alcohol water elution position are mainly folium cortex eucommiae glycosides A in sequiterpene huge pillar alkyl compound, folium cortex eucommiae glycosides B, Cortex Eucommiae Leaf glycosides C, folium cortex eucommiae glycosides D.

Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidations-△ in terpene substances in Bark of Eucommia Ulmoides or folium cortex eucommiae prepared by preceding method 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, Cortex Eucommiae One or more in leaf glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D can be used for preparing treatment senile dementia or Alzheimer disease The preparation formulation of drug, the drug includes oral solution, granule, capsule, tablet, effervescent tablet, syrup, pill, cream taste Agent, soft capsule, ointment, emulsion, powder, sustained release agent, controlled release agent, targeting preparation, powder-injection, liquid drugs injection, injection, atomization Agent, microemulsion, gelling agent, nanometer formulation or various known formulations dosage forms or various acceptable preparation formulations, the drug also wrap Various folk prescriptions and compound preparation are included, said medicine and its preparation have auxiliary treatment brain blood caused by diseases of cardiovascular and cerebrovascular systems It lacks and its sequel resulted from cardio-cerebral blood-vessel diseases, said medicine and its preparation are lacked especially suitable for treatment brain blood and its cardiovascular and cerebrovascular is lost after being ill The senile dementia or Alzheimer disease disease that disease is formed.

Above-mentioned involved senile dementia or Alzheimer disease are by aging, cardiovascular and cerebrovascular disease (heart disease, high blood Pressure, atherosclerosis and transient ischemic attack etc.) cause the cerebral ischemia of Brain hypoperfusion, amyloid A β to deposit The cause of disease formed senile dementia or Alzheimer disease disease in any one.

It is an advantage of the invention that：It is de- to provide ulmoprenol in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances, 1- deoxidations ulmoprenol, 1- Oxygen-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, Folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, the extraction of folium cortex eucommiae glycosides D and preparation method, obtained material are Bark of Eucommia Ulmoides or folium cortex eucommiae terpene object Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl in matter It is one or more available in base-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D In the oral solution, granule, capsule, tablet, effervescent tablet, the syrup that prepare treatment senile dementia or Alzheimer disease drugs Agent, pill, paste nourishing agent, soft capsule, ointment, emulsion, powder, sustained release agent, controlled release agent, targeting preparation, powder-injection, liquid drugs injection, The drug of injection, Alevaire, microemulsion, gelling agent, nanometer formulation or various known dosage forms or various acceptable preparation formulations Bulk pharmaceutical chemicals.

Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, Folium cortex eucommiae glycosides D is in vivo and in vitro to by aging, cardiovascular and cerebrovascular disease, (heart disease, hypertension, atherosclerosis and transience brain lack Blood breaking-out etc.) cause the cerebral ischemia of Brain hypoperfusion, amyloid A β storage disorders because formation senile dementia or A Er Ci Haimo diseases have good therapeutic effect and stronger bioactivity, while having auxiliary treatment by diseases of cardiovascular and cerebrovascular systems Caused brain blood lacks and its sequel resulted from cardio-cerebral blood-vessel diseases.

Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, Folium cortex eucommiae glycosides D, mainly for the symptomatic treatment of etiology and pathogenesis, reaches sample when treating senile dementia or Alzheimer disease It is simultaneous to control.

Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- ulmoprenols, 2,3- in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, The preparation method of folium cortex eucommiae glycosides D mainly by being extracted from natural drug, is pure natural component and monomer, has good biology Compatibility.Wherein, 1- deoxidations-△ 4,5- ulmoprenol, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, Cortex Eucommiae Alcohol glycosides A, folium cortex eucommiae glycosides A, folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D etc. are noval chemical compound.

Description of the drawings

Fig. 1 is that the content of 25% alcohol elution Lignans is more than 90% chromatogram.

Fig. 2 is the UV absorption figure of Lignanoids compounds contained by 25% alcohol elution retention time 8-14 minutes Spectrum.

Fig. 3 is the UV absorption figure of Lignanoids compounds contained by 25% alcohol elution retention time 14-19 minutes Spectrum.

Fig. 4 is Cortex Eucommiae extract treatment senile dementia experiment A-F each group rat space search track.

Fig. 5 is elution position and monomeric compound experimental design and the screening process figure of Bark of Eucommia Ulmoides or folium cortex eucommiae.

Fig. 6 is influences of the A β 25-35 of various concentration to PC12 cell viabilities.

Fig. 7 is the A β of various concentration_25-35With PC12 cells incubate altogether for 24 hours after or 48h after cell viability figure.

The Cortex Eucommiae extract of Fig. 8 various concentrations is to impaired PC12 cytoprotections.

The eucommia leaf extract of Fig. 9 various concentrations is to impaired PC12 cytoprotections.

Protective effect of the monoterpene Cortex Eucommiae alcohols monomeric compound of Figure 10 various concentrations to impaired PC12 cells.

Protective effect of the sequiterpene huge pillar alkanes monomeric compound of Figure 11 various concentrations to impaired PC12 cells.

Protective effect of 1~7 monomer of Lignanoids compounds of Figure 12 various concentrations to impaired PC12 cells.

Protective effect of 9~14 monomer of Lignanoids compounds of Figure 13 various concentrations to impaired PC12 cells.

Specific implementation mode

With reference to embodiment, the invention will be further described：

Embodiment 1

Ulmoprenol (eucommiol), 1- deoxidations in monoterpene Cortex Eucommiae alcohol compound in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Ulmoprenol (1-deoxyeucommiol), 1- deoxidation-△ 4,5- ulmoprenols (1-deoxy- △ 4,5-eucommiol), 2,3- bis- Hydroxyl-△ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol), 4 '-acetyl group-ulmoprenol (4 '-acetyl- ) and eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2-hydroxyethyl-cyclohex- eucommiol 1-enone-2 "-O- β-D-glucopyranoside) preparation method, wherein：By Bark of Eucommia Ulmoides or folium cortex eucommiae pulverizing medicinal materials at thick Powder is impregnated 1 hour with 60% ethyl alcohol, and 8 times are measured solvent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate.It is above-mentioned Filtrate is concentrated under reduced pressure into 1.0g raw medicinal herbs/ml with 65 DEG C, 0-4 DEG C of refrigerated overnight, analyses glue, centrifugal filtration, AB-8 models on filtrate Macroporous adsorbing resin for purification is eluted with water, 25% ethanol elution, 40% ethanol elution, 60% ethanol elution and 95% second successively Alcohol elute, obtain each elution position, wherein water elution position mainly contains monoterpene Cortex Eucommiae alcohol compound, by water elution position into The centrifugation of row 60-80% ethanol precipitations removes impurity, and 80% ethanol extract of recycling obtains silicagel column on medicinal extract, used successively after loading Chloroform：Methanol elution gradient mainly collects chloroform：Methanol is 10：1、8：1、2：1 elution fraction, with above-mentioned Du Ulmoprenol, 1- deoxidations ulmoprenol, 1- deoxidation-△ 4,5- shut out in monoterpene Cortex Eucommiae alcohol compound in secondary skin or folium cortex eucommiae terpene substances Secondary alcohol, 2,3- dihydroxy-△ 4,5- ulmoprenols and 4 '-acetyl group-ulmoprenol and eucommioside A are reference substance TLC or HPLC inspection It surveys, merges corresponding fraction.Above-mentioned fraction is evaporated in 50 DEG C of reduced pressures, is dissolved with 95% ethyl alcohol, and filtering is crystallized and recrystallized, To obtain the final product.Or be evaporated the reduced pressure of above-mentioned fraction or crystallized product, standby chromatography or high performance liquid preparative chromatography system are suppressed in Standby purifying, mobile phase are the isocratic or gradient elution of methanol-water 5-20% or acetonitrile water 5-15%, and recycling eluent is concentrated under reduced pressure Be evaporated to get.Ulmoprenol in monoterpene Cortex Eucommiae alcohol compound is prepared in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances using the above method Yield is 1.6-13.5g/kg, is 99.2% through HPLC detection purity；1- deoxidation ulmoprenol yield is 0.9-10.2g/kg, warp It is 99.0% that HPLC, which detects purity,；, 1- deoxidation-△ 4,5- ulmoprenol yield be 0.6-9.4g/kg, through HPLC detection purity be 98.6%；2,3- dihydroxy-△ 4,5- ulmoprenol yield is 0.5-6.5g/kg, is 97.8% through HPLC detection purity；4 '-second Acyl group-ulmoprenol yield is 0.5-5.5g/kg, is 99.0% through HPLC detection purity；Eucommioside A yield is 0.5-8.8g/ Kg is 99.0% through HPLC detection purity；Chromatographic condition YMC-C18 chromatographic columns (4.6mm × 150mm, 3 μm), mobile phase is first Alcohol-water (10%-100% gradient elutions), Detection wavelength 210nm, flow velocity 1.0ml/min, 30 DEG C of column temperature, 10 μ of sample size l。

Embodiment 2

Ulmoprenol (eucommiol), 1- deoxidations in monoterpene Cortex Eucommiae alcohol compound in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances Ulmoprenol (1-deoxyeucommiol), 1- deoxidation-△ 4,5- ulmoprenols (1-deoxy- △ 4,5-eucommiol), 2,3- bis- Hydroxyl-△ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol), 4 '-acetyl group-ulmoprenol (4 '-acetyl- ) and eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2-hydroxyethyl-cyclohex- eucommiol 1-enone-2 "-O- β-D-glucopyranosi-de) structural identification.

Ulmoprenol (1-deoxyeucommiol) colorless oil, molecular formula C₉H₁₆O₄, ESI-MSm/z [M+H]⁺189。¹H-NMR(DMSO-d₆, 400MHz) and δ：3.88 (1H, m, H-1), 2.52 (1H, m, H-2), 2.15 (1H, dd, J=3.2, 16.4Hz, H-5a), 2.58 (1H, dd, J=6.8,16.4Hz, H-5b), 1.28 (1H, m, H-2 ' a), 1.77 (1H, m, H-2 ' 1H, d, J=12.5Hz, H-3 ' a), 4.06 b), 3.85 ((1H, d, J=12.5Hz, H-3 ' b), 3.97 (2H, t, H-2 "), 3.47 (2H, q, H-4 ')；¹³C-NMR(DMSO-d₆, 100MHz) and δ：74.8 (C-1), 52.6 (C-2), 135.4 (C-3), 137.5 (C- 4), 42.3 (C-5), 34.2 (C-2 '), 55.3 (C-3 '), 59.8 (C-4 '), 57.0 (C-2 ")；Ulmoprenol is known compound.

1- deoxidations ulmoprenol (1-deoxyeucommiol) colorless oil, molecular formula C₉H₁₆O₃, ESI-MS m/z [M+ H]⁺173。¹H-NMR(DMSO-d₆, 400MHz) and δ：1.90 (1H, m, H-1a), 1.40 (1H, m, H-1b), 2.75 (1H, m, H-2), 1.83 (1H, m, H-5a), 1.20 (1H, m, H-5b), 2.29 (2H, m, H-2 '), 3.86 (1H, d, J=12.5Hz, H-3 ' a), 4.08 (1H, d, J=12.5Hz, H-3 ' b), 3.99 (2H, dd, J=9.6,12.5Hz, H-2 "), 3.44 (2H, m, H-4 ')；¹³C- NMR(DMSO-d₆, 100MHz) and δ：28.0 (C-1), 43.0 (C-2), 137.8 (C-3), 139.7 (C-4), 36.2 (C-5), 32.7 (C-2 '), 55.2 (C-3 '), 59.7 (C-4 '), 57.1 (C-2 ")；1- deoxidation ulmoprenols are known compound.

1- deoxidations-△ 4,5- ulmoprenols (1-deoxy- △ 4,5-eucommiol) colorless oil, molecular formula are C₉H₁₆O₃, ESI-MS m/z [M+Na]⁺195.0993。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.20 (1H, m, H-1a), 1.95 (1H, m, H-1b), 2.30 (1H, m, H-2), 2.47 (1H, dd, J=9.6,13.2HZ, H-3), 5.56 (1H, s, H-5), 1.73 1H, m, H-3 ' a), 3.38 1H, m, H-2 ' b), 3.50 1H, m, H-2 ' a), 1.50 ((((1H, m, H-3 ' b), 3.45 (1H, m, H- A), 3.40 2 " (1H, m, H-2 " b), 3.97 (1H, m, H-4 ')；¹³C-NMR(DMSO-d₆, 100MHz) and δ：36.54 (C-1), 38.84 (C-2), 49.04 (C-3), 147.43 (C-4), 124.66 (C-5), 33.27 (C-2 '), 59.87 (C-3 '), 59.66 (C-4 '), 60.35 (C-2 ")；1- deoxidation-△ 4,5- ulmoprenols are noval chemical compound.

2,3- dihydroxy-△ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol) colorless oil, point Minor is C₉H₁₆O₆, ESI-MS m/z [M+Na]⁺243.0343。¹H-NMR(DMSO-d₆, 400MHz) and δ：4.16 (1H, m, H-1), 1H, m, H-2 ' a), 2.02 6.44 (1H, s, H-5), 2.48 ((1H, m, H-2 ' b), 3.35 (1H, m, H-3 '), 3.46 (1H, m, H- 4 '), 3.75 (1H, m, H-2 ")；¹³C-NMR(DMSO-d₆, 100MHz) and δ：65.77 (C-1), 63.23 (C-2), 51.33 (C-3), 174.10 (C-4), 133.51 (C-5), 31.76 (C-2 '), 72.43 (C-3 '), 71.54 (C-4 '), 66.79 (C-2 ")；2,3- Dihydroxy-△ 4,5- ulmoprenols are noval chemical compound.

4 '-acetyl group-ulmoprenol (4 '-acetyl-eucommiol) colorless oil, molecular formula C₁₁H₁₈O₅, ESI-MS m/z[M+Na]⁺253.0632。¹H-NMR(DMSO-d₆, 400MHz) and δ：3.92 (1H, m, H-1), 2.53 (1H, m, H-2), 2.58 1H, m, H-2 ' a), 1.27 (1H, m, H-5a), 2.14 (1H, m, H-5b), 1.76 ((1H, m, H-2 ' b), 4.08 (1H, d, J= 13.2, H-3 ' a), 3.89 (1H, d, J=13.2, H-3 ' b), 4.57 (1H, d, J=12.8, H-4 '), 3.46 (1H, m, H-2 "), 2.00 (3H, s, CH₃CO-)；¹³C-NMR(DMSO-d₆, 100MHz) and δ：74.33 (C-1), 52.63 (C-2), 129.23 (C-3), 142.13 (C-4), 42.42 (C-5), 34.00 (C-2 '), 55.60 (C-3 '), 59.95 (C-4 '), 62.43 (C-2 "), 20.65 (CH₃CO-), 170.28 (CH₃CO-)；4 '-acetyl group-ulmoprenol is noval chemical compound.

Eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2-hydroxyethyl-cyclohex-1- Enone-2 "-O- β-D-glucopyranoside) white solid, molecular formula C₁₆H₂₆O₁₀, ESI-MS m/z [M+Na]⁺401。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.71 (1H, m, H-4), 4.67 (1H, m, H-5), 2.64 (2H, dd, J=6.0,16.0, H-6), 1.13 (1H, t, J=6.8, H-2 '), 4.02 (1H, m, H-3 '), 3.63 (1H, m, H-4 '), 3.80 (1H, m, H-2 are " a), 3.48 (1H, m, H-2 " b), 4.10 (1H, d, J=8.0, H-1 " '), 2.92 (1H, m, H-2 " '), 3.05 (1H, m, H-3 " '), 3.02 (1H, m, H-4 " '), 3.03 (1H, m, H-5 " '), 3.64 (1H, m, H-6 " ' a), 3.42 (1H, m, H-6 " ' b)；¹³C-NMR (DMSO-d₆, 100MHz) and δ：204.54 (C-1), 140.45 (C-2), 173.97 (C-3), 31.04 (C-4), 68.43 (C-5), 44.20 (C-6), 15.12 (C-2 '), 51.69 (C-3 '), 58.81 (C-4 '), 63.78 (C-2 "), 102.58 (C-1 " '), 73.38 (C-2 " '), 76.72 (C-3 " '), 70.02 (C-4 " '), 76.79 (C-5 " '), 61.03 (C-6 " ')；Eucommioside A is Noval chemical compound.

Embodiment 3

Folium cortex eucommiae glycosides A is (6R, 7E, 9R)-in sequiterpene huge pillar alkyl compound in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances megastigma-4,7-dien-3-one-9-O-[β-D-xylopyranos-yl-(1″→6′)-β-D- Glucopyranoside], folium cortex eucommiae glycosides B be (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D- Xylopyranos-yl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides C be (6S, 9R)-megastigmane- 4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides D are (6S,9S)-megastigmane-4-ene-3-one-9-O-[β-D-xylopyranosyl-(1″→6′)-β-D- Glucopyranoside] preparation method, wherein：By Bark of Eucommia Ulmoides or folium cortex eucommiae pulverizing medicinal materials at coarse powder, impregnated with 60% ethyl alcohol 1 hour, 8 times were measured solvent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate.Above-mentioned filtrate and 65 DEG C of reduced pressures To 1.0g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, centrifugal filtration, AB-8 model macroporous adsorbing resin for purification on filtrate, according to It is secondary be eluted with water, 25% ethanol elution, 40% ethanol elution, 60% ethanol elution and 95% ethanol elution, obtain each elution portion Position, wherein 40% alcohol elution mainly contains sequiterpene huge pillar alkyl compound, 40% alcohol elution of recycling must soak Silicagel column on cream uses chloroform after loading successively：Methanol elution gradient mainly collects chloroform：Methanol is 5：1、3：1、 2：1 elution fraction, with folium cortex eucommiae glycosides A in sequiterpene huge pillar alkyl compound in above-mentioned Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances, Folium cortex eucommiae glycosides B, folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D are reference substance TLC or HPLC detection, merge corresponding fraction.Above-mentioned fraction is in 50 DEG C reduced pressure is evaporated, and is dissolved with 95% ethyl alcohol, filtering, crystallize and recrystallize to get.Or above-mentioned fraction is concentrated under reduced pressure and is steamed Dry or crystallized product, suppresses standby chromatography in or high performance liquid preparative chromatography prepares purifying, mobile phase be methanol-water 10-40% or The isocratic or gradient elution of acetonitrile water 5-40%, recycling eluent reduced pressure be evaporated to get.Cortex Eucommiae is prepared using the above method Folium cortex eucommiae glycosides A yield is 0.5-11.2g/kg in sequiterpene huge pillar alkyl compound in skin or folium cortex eucommiae terpene substances, through HPLC It is 99.0% to detect purity；Folium cortex eucommiae glycosides B yield is 0.2-3.8g/kg, is 98.6% through HPLC detection purity；Folium cortex eucommiae glycosides C Yield is 0.6-9.8g/kg, is 99.2% through HPLC detection purity；Folium cortex eucommiae glycosides D yield is 0.2-2.6g/kg, is examined through HPLC It is 97.5% to survey purity；Chromatographic condition YMC-C18 chromatographic columns (4.6mm × 150mm, 3 μm), mobile phase are methanol-water (10%- 100% gradient elution), Detection wavelength 235nm, flow velocity 1.0ml/min, 30 DEG C of column temperature, 10 μ l of sample size.

Embodiment 4

Folium cortex eucommiae glycosides A is (6R, 7E, 9R)-in sequiterpene huge pillar alkyl compound in Bark of Eucommia Ulmoides or folium cortex eucommiae terpene substances megastigma-4,7-dien-3-one-9-O-[β-D-xylopyranos-yl-(1″→6′)-β-D- Glucopyranoside], folium cortex eucommiae glycosides B be (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D- Xylopyranos-yl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides C be (6S, 9R)-megastigm- Ane-4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides D is (6S, 9S)-megastigmane-4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D- Glucopyranoside] structural identification.

Folium cortex eucommiae glycosides A is (6R, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D-xylopyranos- Yl- (1 " → 6 ')-β-D-glucopyranoside], white solid, molecular formula C₂₄H₃₈O₁₁, ESI-MS m/z [M+Na]⁺ 525.2308。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.35 (1H, d, J=16.4HZ, H-2a), 1.95 (1H, d, J= 16.4HZ, H-2b), 5.80 (1H, s, H-4), 2.64 (1H, m, H-6), 5.56 (1H, dd, J=6.4,15.2HZ, H-7), 5.68 (1H, dd, J=8.8,15.2HZ, H-8), 4.29 (1H, m, H-9), 1.18 (3H, d, J=6.0HZ, H-10), 0.95 (3H, s, H-11), 0.91 (3H, s, H-12), 1.85 (3H, s, H-13), 4.18 (1H, d, J=8.0, H-1 '), 2.94 (1H, m, H-2 '), 3.10 (1H, m, H-3 '), 3.08 (1H, m, H-4 '), 3.21 (1H, m, H-5 '), 3.87 (1H, dd, J=1.6,10.8HZ, H-6 ' A), 3.50 (1H, dd, J=6.4,10.8HZ, H-6 ' b), 4.19 (1H, d, J=5.6, H-1 "), 3.35 (1H, m, H-2 "), 3.32 (1H, m, H-3 "), 3.63 (1H, m, H-4 "), 3.66 (1H, dd, J=3.6,11.6HZ, H-5 " a), 3.32 (1H, dd, J =3.6,11.6HZ, H-5 are " b)；¹³C-NMR(DMSO-d₆, 100MHz) and δ：35.60 (C-1), 47.12 (C-2), 197.96 (C- 3), 124.81 (C-4), 162.13 (C-5), 54.42 (C-6), 127.24 (C-7), 136.51 (C-8), 74.41 (C-9), 20.61 (C-10), 27.35 (C-11), 26.72 (C-12), 22.91 (C-13), 100.71 (C-1 '), 73.46 (C-2 '), 76.59 (C-3 '), 69.82 (C-4 '), 75.43 (C-5 '), 67.71 (C-6 '), 103.25 (C-1 "), 70.48 (C-2 "), 72.52 (C-3 "), 67.19 (C-4 "), 64.75 (C-5 ")；Folium cortex eucommiae glycosides A is noval chemical compound.

Folium cortex eucommiae glycosides B is (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D-xylopyranos- Yl- (1 " → 6 ')-β-D-glucopyranoside], white solid, molecular formula C₂₄H₃₈O₁₁, ESI-MS m/z [M+Na]⁺ 525.2310。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.32 (1H, d, J=17.2HZ, H-2a), 1.86 (1H, d, J= 17.2HZ, H-2b), 5.80 (1H, s, H-4), 1.89 (1H, m, H-6), 5.75 (1H, dd, J=5.6,16.8HZ, H-7), 5.50 (1H, dd, J=6.4,16.8HZ, H-8), 3.75 (1H, m, H-9), 1.08 (3H, d, J=6.0HZ, H-10), 1.00 (3H, s, H-11), 0.93 (3H, s, H-12), 1.97 (3H, s, H-13), 4.16 (1H, d, J=8.0, H-1 '), 2.91 (1H, m, H-2 '), 3.12 (1H, m, H-3 '), 3.06 (1H, m, H-4 '), 3.13 (1H, m, H-5 '), 3.90 (1H, dd, J=1.6,11.2HZ, H-6 ' A), 3.50 (1H, dd, J=6.4,11.2HZ, H-6 ' b), 4.19 (1H, d, J=2.2, H-1 "), 3.36 (1H, m, H-2 "), 3.34 (1H, m, H-3 "), 3.62 (1H, m, H-4 "), 3.66 (1H, dd, J=3.6,12.0HZ, H-5 " a), 3.32 (1H, dd, J =3.6,12.0HZ, H-5 are " b)；¹³C-NMR(DMSO-d₆, 100MHz) and δ：35.85 (C-1), 46.92 (C-2), 198.05 (C- 3), 124.11 (C-4), 166.03 (C-5), 50.03 (C-6), 128.39 (C-7), 142.53 (C-8), 73.32 (C-9), 19.60 (C-10), 25.50 (C-11), 28.40 (C-12), 24.03 (C-13), 100.44 (C-1 '), 73.35 (C-2 '), 76.60 (C-3 '), 70.21 (C-4 '), 75.56 (C-5 '), 68.03 (C-6 '), 103.27 (C-1 "), 70.46 (C-2 "), 72.39 (C-3 "), 67.11 (C-4 "), 64.63 (C-5 ")；Folium cortex eucommiae glycosides B is noval chemical compound.

Folium cortex eucommiae glycosides C is (6S, 9R)-megastigmane-4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], white solid, molecular formula C₂₄H₄₀O₁₁, ESI-MS m/z [M+Na]⁺ 527.2465。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.32 (1H, d, J=16.8HZ, H-2a), 1.88 (1H, d, J= 16.8HZ, H-2b), 5.71 (1H, s, H-4), 1.90 (1H, m, H-6), 1.81 (1H, m, H-7a), 1.37 (1H, m, H-7b), 1.51 (1H, m, H-8), 3.74 (1H, m, H-9), 1.08 (3H, d, J=6.0HZ, H-10), 1.00 (3H, s, H-11), 0.93 (3H, s, H-12), 1.96 (3H, s, H-13), 4.16 (1H, d, J=7.6, H-1 '), 2.90 (1H, m, H-2 '), 3.11 (1H, m, H-3 '), 3.03 (1H, m, H-4 '), 3.28 (1H, m, H-5 '), 3.89 (1H, dd, J=1.6,10.8HZ, H-6 ' a), 3.47 (1H, dd, J=6.4,10.8HZ, H-6 ' b), 4.19 (1H, d, J=6.0, H-1 "), 3.33 (1H, m, H-2 "), 3.32 (1H, m, H-3 "), 3.61 (1H, m, H-4 "), 3.64 (1H, dd, J=3.6,12.0HZ, H-5 " a), 3.30 (1H, dd, J=3.6, 12.0HZ, H-5 are " b)；¹³C-NMR(DMSO-d₆, 100MHz) and δ：36.08 (C-1), 46.87 (C-2), 197.92 (C-3), 124.03 (C-4), 165.88 (C-5), 49.97 (C-6), 24.96 (C-7), 35.77 (C-8), 72.96 (C-9), 19.50 (C- 10), 26.21 (C-11), 28.32 (C-12), 24.96 (C-13), 100.37 (C-1 '), 73.28 (C-2 '), 76.62 (C-3 '), 70.16 (C-4 '), 75.48 (C-5 '), 67.97 (C-6 '), 103.17 (C-1 "), 70.36 (C-2 "), 72.38 (C-3 "), 67.00 (C-4 "), 64.50 (C-5 ")；Folium cortex eucommiae glycosides C is noval chemical compound.

Folium cortex eucommiae glycosides D is (6S, 9S)-megastigmane-4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], white solid, molecular formula C₂₄H₄₀O₁₁, ESI-MS m/z [M+Na]⁺ 527.2465。¹H-NMR(DMSO-d₆, 400MHz) and δ：2.37 (1H, d, J=16.8HZ, H-2a), 1.90 (1H, d, J= 16.8HZ, H-2b), 5.72 (1H, s, H-4), 1.88 (1H, m, H-6), 1.81 (1H, m, H-7a), 1.38 (1H, m, H-7b), 1.53 (1H, m, H-8), 3.65 (1H, m, H-9), 1.14 (3H, d, J=6.0HZ, H-10), 1.00 (3H, s, H-11), 0.94 (3H, s, H-12), 1.97 (3H, s, H-13), 4.17 (1H, d, J=7.6, H-1 '), 2.92 (1H, m, H-2 '), 3.13 (1H, m, H-3 '), 3.02 (1H, m, H-4 '), 3.05 (1H, m, H-5 '), 3.90 (1H, dd, J=1.6,10.8HZ, H-6 ' a), 3.50 (1H, dd, J=6.4,10.8HZ, H-6 ' b), 4.21 (1H, d, J=5.6, H-1 "), 3.35 (1H, m, H-2 "), 3.33 (1H, m, H-3 "), 3.63 (1H, m, H-4 "), 3.65 (1H, dd, J=3.6,12.0HZ, H-5 " a), 3.34 (1H, dd, J=3.6, 12.0HZ, H-5 are " b)；¹³C-NMR(DMSO-d₆, 100MHz) and δ：36.15 (C-1), 46.91 (C-2), 198.08 (C-3), 124.09 (C-4), 165.91 (C-5), 50.33 (C-6), 25.05 (C-7), 35.94 (C-8), 75.11 (C-9), 21.64 (C- 10), 24.12 (C-11), 24.67 (C-12), 24.02 (C-13), 102.57 (C-1 '), 76.57 (C-2 '), 73.48 (C-3 '), 70.02 (C-4 '), 75.41 (C-5 '), 68.02 (C-6 '), 103.28 (C-1 "), 70.43 (C-2 "), 72.46 (C-3 "), 67.20 (C-4 "), 64.71 (C-5 ")；Folium cortex eucommiae glycosides D is noval chemical compound.

Embodiment 5

(+) rosin element -4,4'-O- β-D- double glucopyranoside, (+) 8- in Bark of Eucommia Ulmoides or folium cortex eucommiae lignanoids substance Hydroxyl-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4-O- β-D- glucopyranosides, (+) Rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyranosides, (+) 8,9'- dihydroxy-pine Fat element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4,4'-O- β-D- double glucopyranoside, cloves fat Element -4-O- β-D- glucopyranosides, middle fat element -4-O- β-D- glucopyranoses glucosides, olivil -4'-O- β-D- grapes Glucosides, olivil -4-O- β-D- glucopyranosides, lariciresinol -4,4'-O- β-D- double glucopyranoside, 8- Hydroxyl-lariciresinol -4'-O- β-D- glucopyranosides, lariciresinol -4-O- β-D- glucopyranosides, olive element One or more total lignan substances contain in bioside, Auricled Hedyotis Herb element bioside, cloves glycerine-β-syringaresinol ether bioside Amount is more than 90% component or the preparation method of single component, wherein：By Bark of Eucommia Ulmoides or folium cortex eucommiae pulverizing medicinal materials at coarse powder, with 60% Ethyl alcohol impregnates 1 hour, and 8 times are measured solvent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate.Above-mentioned filtrate with 65 DEG C It is concentrated under reduced pressure into 1.0g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, centrifugal filtration, AB-8 model macroporous absorption trees on filtrate Fat is enriched with, and is eluted with water, 25% ethanol elution, 40% ethanol elution, 60% ethanol elution and 95% ethanol elution, is obtained successively 25% ethanol elution portion is recycled wherein content of 25% alcohol elution containing Lignans is more than 90% in each elution position Position obtains silicagel column on medicinal extract, and chloroform is used successively after loading：Methanol elution gradient mainly collects chloroform：Methanol is 6： 1、3：1、2：1、1:1 elution fraction, with (+) rosin element -4,4'-O- β-D- in above-mentioned Bark of Eucommia Ulmoides and its leaf lignanoids substance Double glucopyranosides, (+) 8- hydroxyls-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4-O- β-D- glucopyranosides, (+) rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyanosyls Glycosides, (+) 8,9'- dihydroxy-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4,4'-O- β-D- are double Glucopyranoside, syringaresinol -4-O- β-D- glucopyranosides, middle fat element -4-O- β-D- glucopyranoses glucosides, Chinese olive tree Fat element -4'-O- β-D-Glucose glycosides, olivil -4-O- β-D- glucopyranosides, lariciresinol -4,4'-O- β-D- Double glucopyranosides, 8- hydroxyls-lariciresinol -4'-O- β-D- glucopyranosides, the Portugals lariciresinol -4-O- β-D- Grape pyranoside, olive element bioside, Auricled Hedyotis Herb element bioside, cloves glycerine-β-syringaresinol ether bioside are reference substance TLC Or HPLC detections, merge corresponding fraction.Above-mentioned fraction is evaporated in 50 DEG C of reduced pressures, is dissolved with 95% ethyl alcohol, and filtering, crystallization is simultaneously Recrystallization to get.Or be evaporated the reduced pressure of above-mentioned fraction or crystallized product, standby chromatography is suppressed in or prepared by efficient liquid phase Chromatography prepares purifying, and mobile phase is the isocratic or gradient elution of methanol-water 5-40% or acetonitrile water 5-40%, and recycling eluent subtracts Pressure concentration be evaporated to get.Chromatographic condition：YMC-C18 chromatographic columns (4.6mm × 150mm, 3 μm), mobile phase is methanol-water (10%-100% gradient elutions), Detection wavelength are 205nm or 225nm, flow velocity 1.0ml/min, 30 DEG C of column temperature, sample size 10 μl.Wherein, the results are shown in Figure 1 more than 90% for the content of 25% alcohol elution Lignans.25% ethanol elution The ultraviolet absorpting spectrum of Lignanoids compounds contained by position retention time 8-14 minutes is as shown in Figure 2.25% ethanol elution portion The ultraviolet absorpting spectrum of Lignanoids compounds contained by position retention time 14-19 minutes is as shown in Figure 3.

Embodiment 6

(+) rosin element -4,4'-O- β-D- double glucopyranoside, (+) 8- in Bark of Eucommia Ulmoides or folium cortex eucommiae lignanoids substance Hydroxyl-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4-O- β-D- glucopyranosides, (+) Rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyranosides, (+) 8,9'- dihydroxy-pine Fat element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4,4'-O- β-D- double glucopyranoside, cloves fat Element -4-O- β-D- glucopyranosides, middle fat element -4-O- β-D- glucopyranoses glucosides, olivil -4'-O- β-D- grapes Glucosides, olivil -4-O- β-D- glucopyranosides, lariciresinol -4,4'-O- β-D- double glucopyranoside, 8- Hydroxyl-lariciresinol -4'-O- β-D- glucopyranosides, lariciresinol -4-O- β-D- glucopyranosides, olive element The structural identification of bioside, Auricled Hedyotis Herb element bioside, cloves glycerine-β-syringaresinol ether bioside.

1 dual-ring phase structure compound 1~7 of table¹³C-NMR data

2 monocycle oxygen lignan compound 10~14 of table¹³C-NMR data

Compound 1 is double glucopyranoside [(+) pinoresinol-4,4'-di- of (+) rosin element -4,4'-O- β-D- O-β-D-glucopyranoside]。

Compound 2 is (+) 8- hydroxyls-rosin element -4'-O- β-D- glucopyranosides [(+) 8-Hydroxypin- oresinol-4'-O-β-D-glucopyranoside]。

Compound 3 is (+) 8- hydroxyls-rosin element -4-O- β-D- glucopyranosides [(+) 8-Hydroxypin- oresinol-4-O-β-D-glucopyranoside]。

Compound 4 is (+) rosin element -8-O- β-D- glucopyranosides [(+) pinoresinol-8-O- β-D-gluco- pyranoside]。

Compound 5 is (+) rosin element -4-O- β-D- glucopyranosides [(+) pinoresinol-4-O- β-D-gluco- pyranoside]。

Compound 6 is (+) 8,9'- dihydroxy-rosin element -4'-O- β-D- glucopyranosides [(+) 8,9'- Dihydroxypino-resinol-4'-O-β-D-glucopyranoside]。

Compound 7 is double glucopyranoside [(+) 8- of (+) 8- hydroxyls-rosin element -4,4'-O- β-D- Hydroxypinoresinol-4,4'-di-O- β-D-glucopyranoside].

Compound 8 is (-)-syringaresinol -4-O- β-D- glucopyranosides [(-)-syringaresinol-4-O- β - D-glucopyranoside], white powder.¹H-NMR(DMSO-d₆, 400MHz) and δ：6.66 (2H, s, H-2,6), 6.60 (2H, S, H-2 ', 6 '), 4.89 (1H, d, J=5.2Hz, H-7), 4.66 (1H, d, J=4.0Hz, H-7 '), 3.11 (2H, m, H-8, 2H, m, H-9a, 9 ' a), 3.78,4.28 8 ') ((2H, m, H-9b, 9 ' b), 3.76 (6H, s, 2 × OCH₃), 3.75 (6H, s, 2 × OCH₃), 4.95 (1H, glc-H-1 "), 3.06~3.58 (4H, m, H-2 ", 3 ", 4 ", 5 ").

Compound 9 is (+)-middle fat element -4-O- β-D- glucopyranoses glucosides [(+)-medioresinol-4-O- β-D- Glucopyranoside], white powder.¹H-NMR(DMSO-d₆, 400MHz) and δ：6.95 (1H, d, J=1.6Hz, H-2 '), 7.04 (1H, d, J=8.4Hz, H-5 '), 6.84 (1H, dd, J=1.6,8.4Hz, H-6 '), 6.60 (2H, s, H-2,6), 4.67 (1H, d, J=3.6Hz, H-7), 4.60 (1H, d, J=4.0Hz, H-7 '), 3.04 (2H, m, H-8,8 '), 4.14 (2H, m, H- 9a, 9 ' a), 3.93 (2H, m, H-9b, 9 ' b), 3.77 (3H, s, OCH₃), 3.75 (6H, s, 2 × OCH₃), 4.87 (1H, d, J= 7.2Hz, glc-H-1 "), 3.04~3.51 (4H, m, H-2 ", 3 ", 4 ", 5 ").

Compound 10 is olivil -4'-O- β-D-Glucose glycosides [olivil-4'-O- β-D- glucopyranoside]。

Compound 11 is olivil -4-O- β-D- glucopyranosides [olivil-4-O- β-D- glucopyranoside]。

Compound 12 is double glucopyranoside [lariciresinol-4, the 4'-di- of lariciresinol -4,4'-O- β-D- O-β-D-glucopy-ranoside]。

Compound 13 is 8- hydroxyls-lariciresinol -4'-O- β-D- glucopyranosides [8- Hydroxylariciresinol-4'-O-β-D-glucopyranoside]。

Compound 14 is lariciresinol -4-O- β-D- glucopyranosides [lariciresinol-4-O- β-D- glucopyranoside]。

Embodiment 7

The heterogeneity and multi-cause that inventor falls ill from senile dementia or Alzheimer disease (AD), by brain aging And the cerebral ischemia of brain Low perfusion caused by the vascular risk factor (hypertension, atherosclerosis and transient ischemic attack etc.) It is combined with aβ protein encephalopathy reason lesion, promote aging aging using intraperitoneal injection galactolipin causes with ligation bilateral common carotid arteries repeatedly Brain Low perfusion and the mouse ventricles of the brain disposably inject amyloid beta and make heterogeneous and multi-cause senile dementia or Alzheimer Sick (Heterogeneitys/Multi-factors'Alzheimer ' s Disease, H/MAD) model, to inquire into Cortex Eucommiae extraction Influence of the object to H/MAD rat model Spatial memory abilities, while measuring total antioxidation energy in H/MAD rat model serum Power (T-AOC) and acetylcholinesterase (TChE) content, realize the combination of " illness-model-drug ", are further It studies effective substance and provides theoretical foundation with mechanism of action.

1. dry Bark of Eucommia Ulmoides 40kg is ground into coarse powder by medicinal material extract, 3 times are extracted with 60% alcohol reflux, every time 2.0 Hour, filtration merges extracting solution, is recovered under reduced pressure, gets dry extract.By 60% ethanol extract of Bark of Eucommia Ulmoides, (1g is equivalent to crude drug 11.76g), test liquid is prepared with the aqueous solution of 2% Tween-80.

2. 72 healthy SD male rats are randomly divided into blank group, false damage group, H/MAD models by the grouping of experimental animal Group, 60% ethanol extract high dose group of Cortex Eucommiae, 60% ethanol extract middle dose group of Cortex Eucommiae, Aricept group.It is every to test each group 12 rats of group.

3. replicating heterogeneous and multi-cause senile dementia and Alzheimer disease animal model：Each group experimental rat abdomen Chamber injection of d-galactose (50mg/kgd) causes subacute aging, first carried out after 4W bilateral common carotid arteries ligature repeatedly between section Property blocking blood flow, causing cerebral ischemia lesion, (rat chloraldurate 360mg/kg intraperitoneal injection of anesthesia, the positive middle part of neck is conventional to disappear Notch after poison, detaches bilateral common carotid arteries, and set " 4 " number silk thread is buckled.Screw thread is tensed, blocking blood flow 10min, unclamping screw thread makes blood After 10min is perfused, blocking blood flow 10min again observes 5min after the 2nd Reperfu- sion, skin suture, and the celebrating of notch local injection is big Mycin.) after the completion of operation, rat is fed 3 days, take not dead rat in good condition intracerebral injection condensed state A β again_1-40 (Aβ_1-40：It is diluted to 5 μ g/ μ l, 37 DEG C of incubation 1W with sterile saline), reference《Rat brain stereotaxic atlas》, determining to regard 3mm is hippocampus after intersection.Operating method is as follows：Rat chloraldurate 360mg/kg intraperitoneal injection of anesthesia is fixed on brain solid On position indicator, head skin preservation, routine disinfection field of operation skin, sterile lower operation does the notch of long 2~3cm, separation along cranium center line Periosteum exposes " people " word seam and " ten " word seam.The 3.0mm after bregma, center line right side is other to open 2.0mm, and cranium is opened with No. 7 pin drills Then bone, exposure endocranium, micro syringe slowly inject 5 μ 1A β from the vertical inserting needle 4.0mm in brain surface into brain tissue_1-40, Vertical inserting needle injects 5min, let the acupuncture needle remain at a certain point 5min, and skull is blocked up with dentistry mudding after injection.Antiphlogistic powder sterilizes, skin suture.

Blank group is disregarded；Model group is handled by above-mentioned modeling method respectively with administration group.Model group is in intracerebral Start 2% Tween-80 aqueous solution of gavage 6 days within the 3rd day after injection operation, 1 time a day；Start to connect within the 3rd day after each administration group operation Continuous gastric infusion 6 days, 1 time a day.

The injected clear water in pond makes the water surface be higher by 1~2cm of platform to 4.Morris water maze laboratories in advance, and water temperature control exists 24 2 DEG C of scholars or so.Pool inner wall be coated with it is pitch-dark, every time test before appropriate black ink must be added into pond, pond is divided into 4 A quadrant.Platform is placed among first quartile, rat is a little optionally put into water towards pool wall of pool in other three quadrants In, experiment lasts 5 days, and respectively test is primary for every morning each quadrant, tests 1min every time.If rat is not yet looked within 1min To platform, then rat is taken on platform and it is made to stand above 15s, if rat finds platform within lmin, also allow its Stand 15s on platform, just terminates primary training, so training 4 days.At the 5th day, platform is removed, every rat swims lmin, record Every rat passes through the time (incubation period) of original platform for the first time, every rat in lmin by the number of platform, swim across It total distance and the time stayed in quadrant where original platform and other three quadrants, calculates when first quartile is swum Between (t₁) account for entire swimming time (t_Always) percentage.

5. spectrophotometry measures the content of T-AOC and TChE after the detection of 2.3 rat behaviors, every group of rat hydration After chloral (300mg/kg) anesthesia, quickly open chest, row Culling heart blood, 3500r/min centrifugations 10min takes serum, be placed in -70 DEG C it is low Temperature preserves.By kit specification, ultraviolet spectrophotometry detects the content of T-AOC and TChE.

6. statistical method is handled with SAS software one-way analysis of variances, measurement data data are with mean ± standard deviationIt indicates.

7. result

7.1Morris water maze laboratories are compared with model group, and blank group rat incubation period is short, crosses platform often, first Quadrant residence time is long, significant difference (P ＜ 0.01)；Administration group can shorten incubation period, increase platform number, funny in first quartile Time lengthening is stayed, there is significant difference (P ＜ 0.05 or P ＜ 0.01) compared with model group, illustrates that administration group treatment is effective, prompts 60% ethanol extract group (especially middle dose group) of Cortex Eucommiae is with obvious effects.It the results are shown in Table 3 and Fig. 4.Cortex Eucommiae extract is treated It is that each group rat space search track is as shown in Figure 4 that senile dementia, which tests A-F,.

3 H/MAD rat model space exploration test results of table

Compared with model group,^*P ﹤ 0.05；Compared with model group,^**P ﹤ 0.01；N=10

Influence of 7.2 eucommia ulmoides extracts to T-AOC and TChE contents in H/MAD rat model serum

Compared with model group, T-AOC contents increase in blank group and false damage group rat blood serum, and TChE vigor reduces, poor Different notable (P ＜ 0.01)；Administration group can improve T-AOC contents, reduce TChE vigor, have significant difference (P compared with model group ＜ 0.05 or P ＜ 0.01), illustrate that 60% ethanol extract of Cortex Eucommiae (especially middle dose group) therapeutic effect is apparent, can make heterogeneous Property and multi-cause senile dementia and Alzheimer disease model rat brain in T-AOC vitality restoration, reduce TChE contents, increase The strong ability for removing free radical, antagonism oxidation reaction, while the reduction of neurotransmitter is prevented, to play the role of preventing AD. It the results are shown in Table 4.

The content of T-AOC and TChE in 4 H/MAD rat model serum of table

Embodiment 8

The heterogeneity and multi-cause that inventor falls ill from senile dementia and Alzheimer disease (AD), by cerebral senility Brain Low perfusion caused by the old and vascular risk factor and aβ protein encephalopathy reason lesion are combined, and are promoted using intraperitoneal injection D- galactolipins Aging and ligation bilateral common carotid arteries repeatedly cause brain Low perfusion and the mouse ventricles of the brain disposably inject amyloid beta make it is heterogeneous and Multi-cause senile dementia and Alzheimer disease (Heterogeneitys/Multi-factors'Alzheimer ' s Disease, H/MAD) model, inquire into eucommia leaf extract to H/MAD rat models learning and memory and oxidation resistance (T-AOC, TChE、CAT、SOD、MDA、GSH-P_X) influence, Primary Study folium cortex eucommiae is anti-oxidant, anti-aging effects mechanism, is folium cortex eucommiae Comprehensive development and utilization provides theoretical foundation.

1. dry folium cortex eucommiae 20kg is ground into coarse powder by medicinal material extract, 3 times are extracted with 60% alcohol reflux, every time 2.0 Hour, filtration merges extracting solution, is recovered under reduced pressure, gets dry extract.By 60% ethanol extract of folium cortex eucommiae, (1g is equivalent to crude drug 8.0g), test liquid is prepared with 2% polysorbate aqueous solution.

2. the grouping of animal and 72 healthy SD male rats are randomly divided into blank group, false damage group, H/MAD models Group, eucommia leaf extract high dose group, eucommia leaf extract middle dose group, Doneppezil Hydrochloride group, every group of 12 rats.

3. replicating heterogeneous and multi-cause senile dementia and Alzheimer disease model：The daily abdomen of each group experimental rat Chamber injection of d-galactose (50mgkg^-1), subacute aging is caused, first carry out bilateral common carotid arteries after 4W ligatures discontinuity repeatedly Blocking blood flow, causing cerebral ischemia lesion, (chloraldurate intraperitoneal injection of anesthesia, the positive middle part notch of neck, separation Bilateral Cervical always move Arteries and veins, set " 4 " number silk thread are buckled.Screw thread is tensed, blocking blood flow 10min blocks blood again after release screw thread makes hemoperfusion 10min 10min is flowed, 5min, skin suture are observed after the 2nd Reperfu- sion.) after the completion of operation, rat is fed 3 days, take not dead shape The good rat of state intracerebral injection condensed state A β again_1-40(chloral hydrate anesthesia is fixed on stereotaxic apparatus, head skin preservation, In sterile lower operation, the notch of long 2~3cm is done along cranium center line, detaches periosteum, expose " people " word seam and " ten " word seam.Preceding 3.0mm after fontanel, center line right side is other to open 2.0mm, opens skull, exposure endocranium with No. 7 pin drills, micro syringe hangs down from brain surface Then straight needle 4.0mm slowly injects 5 μ 1A β into brain tissue_1-40, 5min, let the acupuncture needle remain at a certain point 5min are injected, dentistry mudding is used after injection Stifled skull.Skin suture after anti-inflammatory.

Blank group is disregarded；Model group is handled by above-mentioned modeling method respectively with administration group；False damage group abdominal cavity First carry out separation arteria carotis communis after injecting normal saline 4W, 4W, only thread but do not ligature, then intracerebral injection equivalent physiology salt Water.False damage group starts 2% polysorbate aqueous solution of gavage 6 days in the 3rd day after intracerebral injection operation with model group, 1 time a day. Start within the 3rd day continuous gavage administration 6 days after each administration group operation, 1 time a day.

The injected clear water in pond makes the water surface be higher by 1~2cm of platform to 4.Morris water maze laboratories in advance, and water temperature control exists 24 2 DEG C of scholars or so.Pool inner wall be coated with it is pitch-dark, every time test before appropriate black ink must be added into pond, pond is divided into 4 A quadrant.Platform is placed among fourth quadrant, rat is a little optionally put into water towards pool wall of pool in other three quadrants In, experiment lasts 5 days, and respectively test is primary for every morning each quadrant, tests 1min every time.If rat is not yet looked within 1min To platform, then rat is taken on platform and it is made to stand above 15s, if rat finds platform within lmin, also allow its Stand 15s on platform, just terminates primary training, so training 4 days.At the 5th day, platform is removed, every rat swims lmin, record Every rat passes through the time (incubation period) of original platform for the first time, every rat in l min by the number of platform, swim across It total distance and the time stayed in quadrant where original platform and other three quadrants, calculates when fourth quadrant is swum Between (t₄) account for entire swimming time (t_Always) percentage.

5. spectrophotometry measures T-AOC, TChE, CAT, SOD, MDA, GSH-P_XContent rat behavior detect Afterwards, every group of rat chloraldurate (300mgkg^-1) after anesthesia, quickly open chest, row Culling heart blood, 3500r/min centrifugations 10min takes serum, is placed in -70 DEG C of Cord bloods.By kit specification, ultraviolet spectrophotometry detect T-AOC, TChE, CAT、SOD、MDA、GSH-P_XContent.

6. statistical method is analyzed using SAS softwares, experimental data is usedIt indicates, the number of one-way analysis of variance experiment According to.P ＜ 0.05 are statistically significant.

7. result

For 7.1Morris water maze laboratories compared with blank group and false damage group, platform is crossed in model group rats prolongation of latency It reduces, significant difference (P ＜ 0.01) short in fourth quadrant residence time；Compared with model group, administration Doneppezil Hydrochloride group, Eucommia leaf extract is high, middle dose group can shorten incubation period, increase platform number, extends (P ＜ in fourth quadrant residence time 0.05 or P ＜ 0.01), illustrate that each administration group treatment is effective, prompts eucommia leaf extract (especially middle dose group) with obvious effects. It is shown in Table 5.

5 eucommia leaf extract of table to H/MAD rat model space explorations test result (N=8)

Note：Compared with blank group and false damage group,^#P ﹤ 0.01, compared with model group,^*P ﹤ 0.05,^**P ﹤ 0.01

Influence of 7.2 eucommia leaf extracts to T-AOC and TChE contents in H/MAD rat model serum

Compared with blank group and false damage group, T-AOC contents reduce in model group rats serum, and the enhancing of TChE vigor is poor Different notable (P ＜ 0.01)；Administration each group can improve T-AOC contents, reduce TChE vigor, have significant difference compared with model group (P ＜ 0.05 or P ＜ 0.01) illustrates that eucommia leaf extract (especially middle dose group) therapeutic effect is apparent, can make H/MAD models The vitality restoration of T-AOC in rat brain reduces TChE contents, and the ability of free radical is removed in enhancing, and antagonism oxidation reaction is prevented simultaneously The only reduction of neurotransmitter is played the role of preventing AD.It is shown in Table 6.

6 eucommia leaf extract of table to T-AOC and TChE contents in H/MAD rat model serum influence (N=8)

Influence of 7.3 eucommia leaf extracts to CAT, SOD, MDA and GSH-PX content in H/MAD rat model serum

Compared with blank group and false damage group, model group rats CAT, SOD, GSH-P_XIt is substantially reduced (P ＜ 0.01), and MDA is horizontal significantly to increase (P ＜ 0.01)；Compared with model group, Doneppezil Hydrochloride group makes the unobvious that MDA levels decline, poor It is different not notable, but antioxidase CAT, SOD, GSH-P can be improved_XVigor (P ＜ 0.05)；Compared with model group, Cortex Eucommiae carries Each administration group MDA levels of object are taken to be decreased obviously (P ＜ 0.05 or P ＜ 0.01), and CAT, SOD, GSH-P_XIt is apparent to rise (P ＜ 0.05 or P ＜ 0.01), illustrate that administration group treatment is effective, and be better than Doneppezil Hydrochloride group.It the results are shown in Table 7.

7 eucommia leaf extract of table is to CAT, SOD, MDA, GSH-P in H/MAD rat model serum_XContent influence ( N=8)

Embodiment 9

The pathological characters of senile dementia and Alzheimer disease (AD) are that cerebral cortex and hippocampus appearance are a large amount of old Year spot (senile plaques, SP), neurofibrillary tangles (neurofibrillary tangles, NFT), neuron are a large amount of It loses and dementia shows, wherein A beta-aggregations are key link and the key factor and critical therapies target spot of AD morbidities, are done It is to prevent the effective way of AD to disturb the generation of A β and prevent its aggregation, and exploitation inhibits the new drug of the formation of A β fibrils and aggregation to preventing There is good academic and clinical value with symptomatic treatment AD.In inventor's cell culture experiments in vitro, A β 25-35 are utilized PC12 cellular damage models are caused, the vigor of each experimental group cell is had detected using mtt assay, is controlled to confirm Bark of Eucommia Ulmoides or folium cortex eucommiae The active component and active constituent of senile dementia and Alzheimer disease (AD) are treated, is that Bark of Eucommia Ulmoides or folium cortex eucommiae treatment are old Property dull-witted and Alzheimer disease (AD) new drug research based theoretical.

1. the elution position and monomeric compound experimental design of Bark of Eucommia Ulmoides or folium cortex eucommiae and screening process figure are as shown in Figure 5.

2. medicine preparation

(1) the A β 25-35 of 1mg add ultra-pure water 4.715ml to be made into 200 μm of ol/L, after 37 DEG C are incubated 24 hours, are placed in 2-8 It is stored in DEG C refrigerator, it is spare.

(2) 60% ethanol total extract solution of Bark of Eucommia Ulmoides：Precision weighs 60% ethanol total extract dry cream fine powder 2.51mg After adding 5.0ml to distill water dissolution, the stock solution of 0.502mg/ml is made into after 0.2 μm of filter degerming excessively.Serum-free when experiment DMEM culture solutions are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(3) 60% ethanol total extract solution of folium cortex eucommiae：Precision weighs 60% ethanol total extract dry cream fine powder 2.50mg After adding 5.0ml to distill water dissolution, the stock solution of 0.50mg/ml is made into after 0.2 μm of filter degerming excessively.Serum-free when experiment DMEM culture solutions are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(4) Bark of Eucommia Ulmoides water elution position solution：Precision weighs Bark of Eucommia Ulmoides water elution position dry cream fine powder 2.53mg and adds 5.0ml After distilling water dissolution, the stock solution of 0.506mg/ml is made into after 0.2 μm of filter degerming excessively.The DMEM cultures of serum-free when experiment Liquid is diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(5) 25% alcohol elution solution of Bark of Eucommia Ulmoides：Precision weighs 25% alcohol elution dry cream fine powder of Bark of Eucommia Ulmoides After 2.52mg adds 5.0ml to distill water dissolution, the stock solution of 0.504mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(6) 40% alcohol elution solution of Bark of Eucommia Ulmoides：Precision weighs 40% alcohol elution dry cream fine powder of Bark of Eucommia Ulmoides After 2.51mg adds 5.0ml to distill water dissolution, the stock solution of 0.502mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(7) 60% alcohol elution solution of Bark of Eucommia Ulmoides：Precision weighs 60% alcohol elution dry cream fine powder of Bark of Eucommia Ulmoides After 2.53mg adds 5.0ml to distill water dissolution, the stock solution of 0.506mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(8) 95% alcohol elution solution of Bark of Eucommia Ulmoides：Precision weighs 95% alcohol elution dry cream fine powder of Bark of Eucommia Ulmoides After 2.51mg adds 5.0ml to distill water dissolution, the stock solution of 0.502mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(9) folium cortex eucommiae water elution position solution：Precision weighs folium cortex eucommiae water elution position dry cream fine powder 2.54mg and adds 5.0ml After distilling water dissolution, the stock solution of 0.508mg/ml is made into after 0.2 μm of filter degerming excessively.The DMEM cultures of serum-free when experiment Liquid is diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(10) 25% alcohol elution solution of folium cortex eucommiae：Precision weighs 25% alcohol elution dry cream fine powder of folium cortex eucommiae After 2.51mg adds 5.0ml to distill water dissolution, the stock solution of 0.502mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(11) 40% alcohol elution solution of folium cortex eucommiae：Precision weighs 40% alcohol elution dry cream fine powder of folium cortex eucommiae After 2.52mg adds 5.0ml to distill water dissolution, the stock solution of 0.504mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(12) 60% alcohol elution solution of folium cortex eucommiae：Precision weighs 60% alcohol elution dry cream fine powder of folium cortex eucommiae After 2.53mg adds 5.0ml to distill water dissolution, the stock solution of 0.506mg/ml is made into after 0.2 μm of filter degerming excessively.Nothing when experiment The DMEM culture solutions of serum are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(13) 95% alcohol elution solution of folium cortex eucommiae：Precision weighs 95% alcohol elution dry cream fine powder of folium cortex eucommiae After 2.50mg adds 5.0ml to distill water dissolution, the stock solution of 0.50mg/ml is made into after 0.2 μm of filter degerming excessively.With no blood when experiment Clear DMEM culture solutions are diluted to the working solution of 500 μ g/ml, 100 μ g/ml, 20 μ g/ml successively.

(14) ulmoprenol (eucommiol), 1- deoxidation Cortex Eucommiaes in monoterpene Cortex Eucommiae alcohol compound in Bark of Eucommia Ulmoides or folium cortex eucommiae Alcohol (1-deoxyeucommiol), 1- deoxidation-△ 4,5- ulmoprenols (1-deoxy- △ 4,5-eucommiol), 2,3- dihydroxy- △ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol), 4 '-acetyl group-ulmoprenol (4 '-acetyl- ) and eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2-hydroxyethyl-cyclohex- eucommiol 1-enone-2 "-O- β-D-glucopyranoside) aqueous solution：Monoterpene Cortex Eucommiae alcohols compound monomer distilled water is matched At the stock solution of 5mmol/L, it is diluted to 8 μm of ol/L, 40 μm of ol/L, 200 μ successively with the DMEM culture solutions of serum-free when experiment The working solution of mol/L.

(15) folium cortex eucommiae glycosides A is (6R, 7E, 9R)-in sequiterpene huge pillar alkyl compound in Bark of Eucommia Ulmoides or folium cortex eucommiae megastigma-4,7-dien-3-one-9-O-[β-D-xylopyranos-yl-(1″→6′)-β-D- Glucopyranoside], folium cortex eucommiae glycosides B be (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D- Xylopyranos-yl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides C be (6S, 9R)-megastigmane- 4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides D are (6S,9S)-megastigmane-4-ene-3-one-9-O-[β-D-xylopyranosyl-(1″→6′)-β-D- Glucopyranoside] aqueous solution：Sequiterpene huge pillar alkyl compound monomer is made into the reserve of 5mmol/L with distilled water Liquid is diluted to the working solution of 20 μm of ol/L, 100 μm of ol/L, 500 μm of ol/L when experiment successively with the DMEM culture solutions of serum-free.

(16) double glucopyanosyls of (+) rosin element -4,4'-O- β-D- in Lignanoids compounds in Bark of Eucommia Ulmoides or folium cortex eucommiae Glycosides, (+) 8- hydroxyls-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4-O- β-D- grape pyrans Glucosides, (+) rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyranosides, (+) 8,9'- bis- The double glucopyranosides of hydroxyl-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4,4'-O- β-D-, Syringaresinol -4-O- β-D- glucopyranosides, middle fat element -4-O- β-D- glucopyranoses glucosides, olivil -4'-O- β - The double glucopyanosyls of D-Glucose glycosides, olivil -4-O- β-D- glucopyranosides, lariciresinol -4,4'-O- β-D- Glycosides, 8- hydroxyls-lariciresinol -4'-O- β-D- glucopyranosides, lariciresinol -4-O- β-D- glucopyranosides, olive The aqueous solution of olive element bioside, Auricled Hedyotis Herb element bioside, cloves glycerine-β-syringaresinol ether bioside：By lignanoids chemical combination Object monomer is made into the stock solution of 5mmol/L with distilled water, is diluted to 20 μm of ol/ successively with the DMEM culture solutions of serum-free when experiment L, the working solution of 100 μm of ol/L, 500 μm of ol/L or 8 μm of ol/L, 40 μm of ol/L, 200 μm of ol/L.

3. the A β 25-35 of various concentration determine modeling concentration and action time to the measurement of PC12 cell viabilities

The PC12 cells that exponential phase is in culture bottle are taken, after 0.25% trypsase EDTA digestion.With containing Cell is diluted to every milliliter of suspension and contains 2 × 10 by the DMEM high glucose mediums of 10% fetal calf serum⁵A cell is inoculated in the training of 96 holes Plate is supported, per 100 μ l (2 × 10 of hole⁴), it is put into CO₂Incubator, 37 DEG C, 5%CO₂Under the conditions of cultivate, after cell covers with bottom hole i.e. It can be used for testing.Experiment is divided into 4 groups, every group of 8 holes, and when experiment, which inhales, abandons old culture solution.Blank control group (Control)：Only per hole 100 μ l serum-free DMEM low sugar culture mediums are added；A β (25-35) model group：It is separately added into per hole and is trained with serum-free DMEM low sugar Support final concentration of 5 μM of basigamy, 10 μM, 20 μM of A β (25-35) or 10 μM, 20 μM, 40 μM of A β (25-35).37 DEG C of trainings After supporting for 24 hours or after 48h, final concentration of 0.5mg/ml MTT (10 μ l) are added per hole, continue to suck DMEM culture mediums after cultivating 4h, DMSO100 μ l are added per hole, shake mixing 10min, after endoparticle is completely dissolved after hole, extinction is measured at microplate reader 492nm Spend (A_492nm) value.

4. the Bark of Eucommia Ulmoides of various concentration or the total extract of folium cortex eucommiae and each elution position are to PC12 cells caused by A β 25-35 The protective effect of damage

Measurement of the A β 25-35 of same 3. various concentrations of cell culture to PC12 cell viabilities.Experiment is divided into 9 groups, every group 10 Hole inhales when experiment and abandons old culture solution.Blank control group (Control)：100 μ l serum-free DMEM low sugar cultures are only added per hole Base；A β (25-35) model group：Final concentration of 10 μM of the A β (25- prepared with serum-free DMEM low sugar culture mediums are added per hole 35)；It is each in (1)-(13) in 2. medicine preparations that final concentration of 10 μM of A β (25-35) and solution is added per hole for each administration group The drug of administration group.After 37 DEG C of cultures for 24 hours, final concentration of 0.5mg/ml MTT (10 μ l) are added per hole, continue to inhale after cultivating 4h DMEM culture mediums are removed, DMSO100 μ l are added per hole, mixing 10min are shaken, after endoparticle is completely dissolved after hole, in microplate reader Absorbance (A is measured at 492nm_492nm) value.

5. monoterpene Cortex Eucommiae alcohols monomeric compound, sequiterpene huge pillar alkanes monomeric compound, wood in Bark of Eucommia Ulmoides or folium cortex eucommiae Protective effect of the fat element class monomeric compound to PC12 cellular damages caused by A β 25-35

6. calculation formula

All data withIt indicates, comparison among groups are examined with t.The calculation formula of cell survival rate is：Experimental group A values/ Control group A value × 100%.

7 experimental results

Influences and determining modeling concentration and action time of the A β 25-35 of 7.1 various concentrations to PC12 cell viabilities

The A β of 5 μM, 10 μM, 20 μM various concentrations_25-35After being incubated for 24 hours altogether with PC12 cells, cell viability is remarkably decreased, performance For A_492nmValue is decreased obviously；And with A β_25-35The increase PC12 cell viabilities of dosage decline more obvious.It the results are shown in Table 8 and figure 6.Influences of the A β 25-35 of various concentration to PC12 cell viabilities is as shown in Figure 6.

Influences (n=8) of the A β 25-35 of various concentration to PC12 cell viabilities afterwards for 24 hours of table 8

Note：^**The p compared with blank control group<0.01

The A β of 10 μM, 20 μM, 40 μM various concentrations_25-35After being incubated altogether for 24 hours with PC12 cells or after 48h, cell viability is notable Decline, shows as A_492nmValue is decreased obviously；And with A β_25-35Dosage and the increase PC12 cell viabilities of time decline brighter It is aobvious.It the results are shown in Table 9,10 and Fig. 7.The A β 25-35 of various concentration incubate cell viability figure such as Fig. 7 institutes behind 24 or 48 altogether to PC12 cells Show.

Influences (n=10) of the A β 25-35 of various concentration to PC12 cell viabilities afterwards for 24 hours of table 9

Note：^**The p compared with blank control group<0.01

Influences (n=10) of the A β 25-35 of various concentration to PC12 cell viabilities after 10 48h of table

Note：^**The p compared with blank control group<0.01

Observe result：20 μM of A β 25-35 handled PC12 cells after 24 hours, and prodigious change has occurred in cellular morphology： The cilium of model group PC12 cell visible cell films periphery disappears, and vacuole occurs in cytoplasm, and cell aggregation is agglomerating, most of thin Born of the same parents become irregular shape and circle by polygon, and the refractivity of cell is deteriorated.The PC12 cells of Normal group, cell individual Full, protrusion is apparent, and polygon, diamond shape or fusiformis is presented in cell, and cell growing way is fine, and the refractivity of cell is normal.To say Bright A β_25-35It causes PC12 cellular damages model preferably to replicate the pathological change of AD, is the research preferable models of AD.Determine modeling Concentration and action time is：20 μM of A β 25-35 processing PC12 cells 24 hours.

The Bark of Eucommia Ulmoides or folium cortex eucommiae total extract of 7.2 various concentrations and each elution position are to A β_25-35Caused PC12 cells damage The protective effect of wound

The Bark of Eucommia Ulmoides or folium cortex eucommiae total extract of various concentration and each elution position are to A β_25-35Caused PC12 cellular damages There is protective effect, so that its cell viability is enhanced, show as A_492nmValue rises, and wherein Bark of Eucommia Ulmoides water elution position, 25% ethyl alcohol are washed The cytoprotection of de- position and 40% alcohol elution is relatively strong, folium cortex eucommiae water elution position, 25% ethanol elution Position and 40% alcohol elution are stronger to the protective effect of PC12 cells.Wherein, Bark of Eucommia Ulmoides or folium cortex eucommiae water elution position Predominantly monoterpene Cortex Eucommiae alcohol compound, Bark of Eucommia Ulmoides or 25% alcohol elution of folium cortex eucommiae are mainly Lignanoids compounds, Bark of Eucommia Ulmoides or 40% alcohol elution of folium cortex eucommiae are mainly sequiterpene huge pillar alkyl compound.The results are shown in Table 11,12 and Fig. 8, 9.The Cortex Eucommiae extract of various concentration is as shown in Figure 8 to impaired PC12 cytoprotections.The Zhong Ye of various concentration is extracted Object is as shown in Figure 9 to impaired PC12 cytoprotections.

The Cortex Eucommiae extract of 11 various concentration of table is to impaired PC12 cytoprotections (n=10)

Note：^{﹟ ﹟}The p compared with blank control group<0.01；^*The p compared with A β model groups<0.05；^**The p compared with A β model groups< 0.01

The eucommia leaf extract of 12 various concentration of table is to impaired PC12 cytoprotections (n=10)

Monoterpene Cortex Eucommiae alcohols monomeric compound is to PC12 caused by A β 25-35 in the Bark of Eucommia Ulmoides or folium cortex eucommiae of 7.3 various concentrations The protective effect of cellular damage

Ulmoprenol, 1- deoxidations ulmoprenol, 1- in monoterpene Cortex Eucommiae alcohol compound in the Bark of Eucommia Ulmoides or folium cortex eucommiae of various concentration The singulations such as deoxidation-△ 4,5- ulmoprenols, 2,3- dihydroxy-△ 4,5- ulmoprenols, 4 '-acetyl group-ulmoprenol, eucommioside A Object is closed to A β_25-35Caused PC12 cellular damages have protective effect, so that its cell viability is enhanced, show as A_492nmValue rises.As a result It is shown in Table 13 and Figure 10.The protective effect such as Figure 10 of the monoterpene Cortex Eucommiae alcohols monomeric compound of various concentration to impaired PC12 cells It is shown.

Protective effect (n=10) of the monoterpene Cortex Eucommiae alcohols monomeric compound of 13 various concentration of table to impaired PC12 cells

Sequiterpene huge pillar alkanes monomeric compound is to caused by A β 25-35 in the Bark of Eucommia Ulmoides or folium cortex eucommiae of 7.4 various concentrations The protective effect of PC12 cellular damages

Folium cortex eucommiae glycosides A in sequiterpene huge pillar alkyl compound in the Bark of Eucommia Ulmoides or folium cortex eucommiae of various concentration, folium cortex eucommiae glycosides B, The monomeric compounds such as folium cortex eucommiae glycosides C, folium cortex eucommiae glycosides D are to A β_25-35Caused PC12 cellular damages have protective effect, make its cell viability Enhancing, shows as A_492nmValue rises.It the results are shown in Table 14 and Figure 11.The sequiterpene huge pillar alkanes monomeric compound of various concentration to by The protection of the PC12 cells of damage is made as shown in figure 11.

Protective effect (n=of the sequiterpene huge pillar alkanes monomeric compound of 14 various concentration of table to impaired PC12 cells 10)

Lignanoids monomeric compound is to PC12 cells caused by A β 25-35 in the Bark of Eucommia Ulmoides or folium cortex eucommiae of 7.5 various concentrations The protective effect of damage

(+) rosin element -4,4'-O- β-D- in bis-epoxy lignin compound in the Bark of Eucommia Ulmoides or folium cortex eucommiae of various concentration Double glucopyranosides, (+) 8- hydroxyls-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4-O- β-D- glucopyranosides, (+) rosin element -8-O- β-D- glucopyranosides, (+) rosin element -4-O- β-D- glucopyanosyls Glycosides, (+) 8,9'- dihydroxy-rosin element -4'-O- β-D- glucopyranosides, (+) 8- hydroxyls-rosin element -4,4'-O- β-D- are double Olive resin in glucopyranoside, syringaresinol -4-O- β-D- glucopyranosides etc. and monocycle oxygen lignin compound Element -4'-O- β-D-Glucose glycosides, olivil -4-O- β-D- glucopyranosides, lariciresinol -4,4'-O- β-D- are double Glucopyranoside, 8- hydroxyls-lariciresinol -4'-O- β-D- glucopyranosides, lariciresinol -4-O- β-D- grapes The monomeric compounds such as pyranoside are to A β_25-35Caused PC12 cellular damages have protective effect, so that its cell viability is enhanced, show as A_492nmValue rises.It the results are shown in Table 15,16 and Figure 12,13.1~7 monomer of Lignanoids compounds of various concentration is to impaired The protective effect of PC12 cells is as shown in figure 12.9~14 monomer of Lignanoids compounds of various concentration is thin to impaired PC12 The protective effect of born of the same parents is as shown in figure 13.

Protective effect (n=10) of the lignanoids monomeric compound of 15 various concentration of table to impaired PC12 cells

Protective effect (n=10) of the lignanoids monomeric compound of 16 various concentration of table to impaired PC12 cells

Note：^{﹟ ﹟}The p compared with blank control group<0.01；^*The p compared with A β model groups<0.05；^**The p compared with A β model groups< 0.01。

Claims

1. terpene substances are preparing the application in treating senile dementia drug in Bark of Eucommia Ulmoides or folium cortex eucommiae, it is characterised in that：Du Terpene substances itself are washed with water after macroporous absorbent resin on the total extract of Bark of Eucommia Ulmoides or folium cortex eucommiae in secondary skin or folium cortex eucommiae De- position is prepared and ulmoprenol in monoterpene Cortex Eucommiae alcohol compound is prepared in Bark of Eucommia Ulmoides or folium cortex eucommiae (eucommiol), 1- deoxidations ulmoprenol (1-deoxyeucommiol), 1- deoxidation-△ 4,5- ulmoprenols (1-deoxy- △ 4,5- Eucommiol), 2,3- dihydroxy-△ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol), 4 '-acetyl group- Ulmoprenol (4 '-acetyl-eucommiol) and eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2- Hydroxyethyl-cyclohex-1-enone-2 "-O- β-D-glucopyranoside) in it is one or more,

2. terpene substances are preparing the application in treating Alzheimer disease drugs in Bark of Eucommia Ulmoides or folium cortex eucommiae, it is characterised in that： Terpene substances itself are to use water after macroporous absorbent resin on the total extract of Bark of Eucommia Ulmoides or folium cortex eucommiae in Bark of Eucommia Ulmoides or folium cortex eucommiae Elution position is prepared and ulmoprenol in monoterpene Cortex Eucommiae alcohol compound is prepared in Bark of Eucommia Ulmoides or folium cortex eucommiae (eucommiol), 1- deoxidations ulmoprenol (1-deoxyeucommiol), 1- deoxidation-△ 4,5- ulmoprenols (1-deoxy- △ 4,5- Eucommiol), 2,3- dihydroxy-△ 4,5- ulmoprenols (2,3-dihydroxyl- △ 4,5-eucommiol), 4 '-acetyl group- Ulmoprenol (4 '-acetyl-eucommiol) and eucommioside A (5-hydroxyl-3,4-dihydroxymethyl-2- Hydroxyethyl-cyclohex-1-enone-2 "-O- β-D-glucopyranoside) in it is one or more,

3. terpene substances are preparing the application in treating senile dementia drug in Bark of Eucommia Ulmoides or folium cortex eucommiae, it is characterised in that：Du Terpene substances itself are after macroreticular resin on the total extract of Bark of Eucommia Ulmoides or folium cortex eucommiae with 40% ethyl alcohol in secondary skin or folium cortex eucommiae Elution position be prepared with Bark of Eucommia Ulmoides or folium cortex eucommiae be prepared in sequiterpene huge pillar alkyl compound folium cortex eucommiae glycosides A be (6R, 7E,9R)-megastigma-4,7-dien-3-one-9-O-[β-D-xylopyranos-yl-(1″→6′)-β-D- Glucopyranoside], folium cortex eucommiae glycosides B be (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D- Xylopyranos-yl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides C be (6S, 9R)-megastigmane- 4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides D are (6S,9S)-megastigmane-4-ene-3-one-9-O-[β-D-xylopyranosyl-(1″→6′)-β-D- Glucopyranoside] in it is one or more,

4. terpene substances are preparing the application in treating Alzheimer disease drugs in Bark of Eucommia Ulmoides or folium cortex eucommiae, it is characterised in that： Terpene substances itself are after macroreticular resin on the total extract of Bark of Eucommia Ulmoides or folium cortex eucommiae with 40% second in Bark of Eucommia Ulmoides or folium cortex eucommiae Alcohol elution position, which is prepared, to be prepared folium cortex eucommiae glycosides A in sequiterpene huge pillar alkyl compound with Bark of Eucommia Ulmoides or folium cortex eucommiae and is (6R,7E,9R)-megastigma-4,7-dien-3-one-9-O-[β-D-xylopyranos-yl-(1″→6′)-β-D- Glucopyranoside], folium cortex eucommiae glycosides B be (6S, 7E, 9R)-megastigma-4,7-dien-3-one-9-O- [β-D- Xylopyranos-yl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides C be (6S, 9R)-megastigmane- 4-ene-3-one-9-O- [β-D-xylopyranosyl- (1 " → 6 ')-β-D-glucopyranoside], folium cortex eucommiae glycosides D are (6S,9S)-megastigmane-4-ene-3-one-9-O-[β-D-xylopyranosyl-(1″→6′)-β-D- Glucopyranoside] in it is one or more,

5. preparing treatment senile dementia according to terpene substances in claim 1, any one of 3 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in drug, it is characterised in that：Involved senile dementia is to lead to brain Low perfusion shape by aging, cardiovascular and cerebrovascular disease The cerebral ischemia of state, amyloid A β deposit any one in the senile dementia that etiology and pathogenesis is formed.

6. preparing treatment Alzheimer according to terpene substances in claim 2, any one of 4 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in medicine, it is characterised in that：Involved Alzheimer disease is to lead to the low filling of brain by aging, cardiovascular and cerebrovascular disease The cerebral ischemia of note state, amyloid A β deposit any one in the Alzheimer disease disease that etiology and pathogenesis is formed.

7. preparing treatment senile dementia according to terpene substances in claim 1, any one of 3 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in drug, it is characterised in that：Senile dementia brain blood caused by diseases of cardiovascular and cerebrovascular systems lacks and its heart and brain What angiosis sequelae was formed.

8. preparing treatment Alzheimer according to terpene substances in claim 2, any one of 4 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in medicine, it is characterised in that：Alzheimer disease brain blood caused by diseases of cardiovascular and cerebrovascular systems lack and its What sequel resulted from cardio-cerebral blood-vessel diseases was formed.

9. preparing treatment senile dementia according to terpene substances in claim 1, any one of 3 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in drug, it is characterised in that：The preparation formulation of the drug be oral solution, granule, capsule, tablet, effervescent tablet, Syrup, pill, paste nourishing agent, soft capsule, ointment, emulsion, powder, sustained release agent, controlled release agent, targeting preparation, powder-injection, liquid drugs injection Agent, injection, Alevaire, microemulsion, gelling agent, nanometer formulation.

10. preparing treatment alzheimer ' according to terpene substances in claim 2, any one of 4 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in silent medicine, it is characterised in that：The preparation formulation of the drug is oral solution, granule, capsule, tablet, bubble Rise piece, syrup, pill, paste nourishing agent, soft capsule, ointment, emulsion, powder, sustained release agent, controlled release agent, targeting preparation, powder needle Agent, liquid drugs injection, injection, Alevaire, microemulsion, gelling agent, nanometer formulation.

11. senile silly in preparation treatment according to terpene substances in claim 1, any one of 3 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in slow-witted drug, it is characterised in that：The drug further includes various folk prescriptions and compound preparation, and said medicine and its preparation are suitable For treating the senile dementia that brain blood lacks and its sequel resulted from cardio-cerebral blood-vessel diseases is formed.

12. preparing treatment alzheimer ' according to terpene substances in claim 2, any one of 4 Bark of Eucommia Ulmoides or folium cortex eucommiae Application in silent medicine, it is characterised in that：The drug further includes various folk prescriptions and compound preparation, said medicine and its preparation Suitable for treating the Alzheimer disease disease that brain blood lacks and its sequel resulted from cardio-cerebral blood-vessel diseases is formed.