CN101537011B - Pharmaceutical composition for preventing or treating parkinsonism and preparation method thereof - Google Patents
Pharmaceutical composition for preventing or treating parkinsonism and preparation method thereof Download PDFInfo
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- CN101537011B CN101537011B CN200910135572XA CN200910135572A CN101537011B CN 101537011 B CN101537011 B CN 101537011B CN 200910135572X A CN200910135572X A CN 200910135572XA CN 200910135572 A CN200910135572 A CN 200910135572A CN 101537011 B CN101537011 B CN 101537011B
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- acanthopanacis senticosi
- caulis acanthopanacis
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- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 208000027089 Parkinsonian disease Diseases 0.000 title abstract 3
- 206010034010 Parkinsonism Diseases 0.000 title abstract 3
- NPJICTMALKLTFW-OFUAXYCQSA-N daucosterol Chemical compound O([C@@H]1CC2=CC[C@H]3[C@@H]4CC[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@H](C)CC[C@@H](CC)C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O NPJICTMALKLTFW-OFUAXYCQSA-N 0.000 claims abstract description 21
- QXMNTPFFZFYQAI-IMDKZJJXSA-N beta-sitosterol 3-O-beta-D-glucopyranoside Natural products CC[C@H](CC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3CC=C4C[C@H](CC[C@]4(C)[C@H]3CC[C@]12C)O[C@@H]5C[C@H](CO)[C@@H](O)[C@H](O)[C@H]5O)C(C)C QXMNTPFFZFYQAI-IMDKZJJXSA-N 0.000 claims abstract description 20
- QDFKFNAHVGPRBL-UHFFFAOYSA-N daucosterol Natural products CCC(CCC(C)C1CCC2C1CCC3C2(C)CC=C4CC(CCC34C)OC5OC(CO)C(O)C(O)C5O)C(C)C QDFKFNAHVGPRBL-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000003814 drug Substances 0.000 claims abstract description 15
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 229930182478 glucoside Natural products 0.000 claims description 59
- 150000008131 glucosides Chemical class 0.000 claims description 59
- KOWMJRJXZMEZLD-UHFFFAOYSA-N syringaresinol Chemical compound COC1=C(O)C(OC)=CC(C2C3C(C(OC3)C=3C=C(OC)C(O)=C(OC)C=3)CO2)=C1 KOWMJRJXZMEZLD-UHFFFAOYSA-N 0.000 claims description 18
- LVUPFEOCDSHRBL-UHFFFAOYSA-N syringaresinol Natural products COc1cccc(OC)c1C2OCC3C2COC3c4c(OC)cccc4OC LVUPFEOCDSHRBL-UHFFFAOYSA-N 0.000 claims description 18
- 239000003085 diluting agent Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 230000002265 prevention Effects 0.000 claims description 5
- 238000009472 formulation Methods 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 35
- 230000004083 survival effect Effects 0.000 abstract description 19
- 238000002474 experimental method Methods 0.000 abstract description 12
- 230000006378 damage Effects 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- 238000003908 quality control method Methods 0.000 abstract description 3
- 230000001681 protective effect Effects 0.000 abstract description 2
- 230000001225 therapeutic effect Effects 0.000 abstract description 2
- FFDULTAFAQRACT-OVUSVGQASA-N Eleutheroside D Chemical compound COC1=CC(C2C3C(C(OC3)C=3C=C(OC)C(O[C@@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)=C(OC)C=3)CO2)=CC(OC)=C1O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O FFDULTAFAQRACT-OVUSVGQASA-N 0.000 abstract 4
- QJVXKWHHAMZTBY-GCPOEHJPSA-N syringin Chemical compound COC1=CC(\C=C\CO)=CC(OC)=C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 QJVXKWHHAMZTBY-GCPOEHJPSA-N 0.000 abstract 2
- QJVXKWHHAMZTBY-KSXIZUIISA-N syringin Natural products COc1cc(C=CCO)cc(OC)c1O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O QJVXKWHHAMZTBY-KSXIZUIISA-N 0.000 abstract 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 210000005064 dopaminergic neuron Anatomy 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 238000012216 screening Methods 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 15
- 230000005779 cell damage Effects 0.000 description 13
- 208000037887 cell injury Diseases 0.000 description 13
- 230000001939 inductive effect Effects 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 239000004615 ingredient Substances 0.000 description 11
- 239000007788 liquid Substances 0.000 description 8
- FMGYKKMPNATWHP-UHFFFAOYSA-N Cyperquat Chemical compound C1=C[N+](C)=CC=C1C1=CC=CC=C1 FMGYKKMPNATWHP-UHFFFAOYSA-N 0.000 description 5
- 208000018737 Parkinson disease Diseases 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 239000006285 cell suspension Substances 0.000 description 5
- 238000002425 crystallisation Methods 0.000 description 5
- 230000008025 crystallization Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 238000012109 statistical procedure Methods 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- 230000000324 neuroprotective effect Effects 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- -1 1-methyl-4 phenylpyridine ion Chemical class 0.000 description 1
- 241000208340 Araliaceae Species 0.000 description 1
- 241001632410 Eleutherococcus senticosus Species 0.000 description 1
- 102100032352 Leukemia inhibitory factor Human genes 0.000 description 1
- 108090000581 Leukemia inhibitory factor Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000003291 dopaminomimetic effect Effects 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 125000004395 glucoside group Chemical group 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007937 lozenge Substances 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 239000000178 monomer Substances 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
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- 239000000829 suppository Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a pharmaceutical composition for preventing or treating parkinsonism and a preparation method thereof. The pharmaceutical composition consists of eleutheroside B, eleutheroside D, syringaresinol-di-glucoside and daucosterol. The invention determines the optimal dose level range of the four components and on the base, adopts an orthogonal test for screening out the optimal compatibility proportion of the four active components. Proved by experiment results, the pharmaceutical composition which is prepared by the compatibility of the eleutheroside B, the eleutheroside D, the syringaresinol-di-glucoside and the daucosterol according to a weight ratio of 1:2:8:8, can obviously improve PC12 cell survival rate and has obvious protective effects on dopaminergic neuron injuries. The active ingredients of the acanthopanax have simple and clear chemical compositions and quality control is easy to be conducted in production. The pharmaceutical composition can be used for preparing medicaments with good therapeutic effects for preventing and treating parkinsonism safely and reliably.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition, relate in particular to a kind of pharmaceutical composition of forming according to specific proportioning by the Radix Et Caulis Acanthopanacis Senticosi effective ingredient, the invention still further relates to the purposes of this pharmaceutical composition in prevention or treatment parkinson disease, belong to Parkinsonian treatment field.
Background technology
Parkinson disease (Parkinson ' s disease, be a kind of common central nervous system degenerative disease PD), its sickness rate increases with age growth.China has PD patient 1,700,000 now, and with newly-increased 100,000 people's speed increments in every year.Because this disease has a strong impact on patient's mobility and quality of life, the disability rate height, the course of disease is long, often needs lifelong treatment, thereby has brought serious society and financial burden to the mankind.So urgent needs is sought prevention and is treated Parkinsonian effective Chinese medicine, to delay Parkinsonian process and to prevent increasing the weight of of the state of an illness.In recent years, along with the application of modern biotechnology means at crude drug, the activated monomer in the Chinese medicine begins to become the focus that people pay close attention to, and the research aspect neurodegenerative diseases has obtained certain progress.
Radix Et Caulis Acanthopanacis Senticosi is dry root and the rhizome of Araliaceae Radix Et Caulis Acanthopanacis Senticosi (Acanthopanax senticosus (Rupr.et MaXim.) Harms.).Main product is in the mountain area, Heilongjiang Province.Acrid in the mouth, little hardship, warm in nature, nontoxic have the kidney tonifying spleen invigorating, the tonifying the kidney for tranquilization effect.Radix Et Caulis Acanthopanacis Senticosi compound recipe and Radix Et Caulis Acanthopanacis Senticosi injection have been used for Parkinsonian clinical treatment at present, and obtained certain curative effect, but up to now, Shang Weijian has Radix Et Caulis Acanthopanacis Senticosi effective ingredient proportioning is used for Parkinsonian treatment, so Radix Et Caulis Acanthopanacis Senticosi effective ingredient proportioning treatment parkinson disease have brilliant market developing prospect.
Summary of the invention
The purpose of this invention is to provide a kind of pharmaceutical composition that has significant curative effect for parkinson disease.
The objective of the invention is to be achieved through the following technical solutions:
A kind ofly prevent or treat Parkinsonian pharmaceutical composition, by Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, two glucosides of syringaresinol and daucosterol are formed;
In order to reach better therapeutic effect, preferred, the consumption proportion scope of each component is: Radix Et Caulis Acanthopanacis Senticosi glucoside B10～20 μ g/ml, Radix Et Caulis Acanthopanacis Senticosi glucoside D 20～40 μ g/ml, two glucoside 40～80 μ g/ml of syringaresinol, daucosterol 20～80 μ g/ml.
Preferred, by Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, two glucosides of syringaresinol and daucosterol were according to 1: 2: 8: 8 weight compatibility and forming.
Above-mentioned each chemical compound all can be bought by commercial sources and obtain, and also can prepare according to the disclosed method of existing literature.
The pharmaceutical composition of the present invention of pharmaceutically acceptable consumption with after pharmaceutically acceptable carrier or diluent cooperate, is prepared into any one suitable clinical preparation by the formulation method of this area routine with it.Usually the present composition is suitable for oral administration and drug administration by injection, also is fit to other medication.Said composition can be liquid preparation forms such as tablet, capsule, powder, granule, lozenge, suppository or oral liquid.
Determine the Parkinsonian active component of acanthopanax root for treating based on previous work, and isolation identification goes out on the basis of four chemical compounds in the active component, in depth study Radix Et Caulis Acanthopanacis Senticosi effective ingredient proportioning for system and treat Parkinsonian effect, the present invention adopts the method for cell culture, with 1-methyl-4 phenylpyridine ion (MPP
+) as the representative of external toxin, cause the PC12 cells injury, set up the cell model of PD.Adopt mtt assay to detect PC
12Cell survival rate is determined each dose of components horizontal extent in the acanthopanax root effective component.The consumption proportion scope of final definite each composition is: Radix Et Caulis Acanthopanacis Senticosi glucoside B 10～20 μ g/ml, Radix Et Caulis Acanthopanacis Senticosi glucoside D 20～40 μ g/ml, two glucoside 40～80 μ g/ml of syringaresinol, daucosterol 20～80 μ g/ml.
Dosage level scope according to Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, the two glucosides of syringaresinol and daucosterol, respectively at 3 dosage levels of its each composition design, the present invention as 4 factors investigating, presses L with Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, the two glucosides of syringaresinol and daucosterol
9(3
4) orthogonal array experimentizes, and adopts the MTT colorimetry, with the cell survival rate is to investigate index, adopts method of analysis of variance to determine the best proportion compatibility of Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, the two glucosides of syringaresinol and daucosterol.
Experimental result shows, 4 effective ingredient of Radix Et Caulis Acanthopanacis Senticosi are Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, and two glucosides of syringaresinol and daucosterol were according to 1: 2: 8: 8 weight proportion consists of best proportioning, the pharmaceutical composition formed of proportioning can significantly improve the PC12 cell survival rate thus, to MPP
+The PC12 dopaminergic nerve cell damage that causes has protective effect.
Radix Et Caulis Acanthopanacis Senticosi effective ingredient chemical constituent provided by the present invention is simply clear and definite, is easier to the quality control of medicine aborning.The Radix Et Caulis Acanthopanacis Senticosi effective ingredient steady quality that utilizes the present invention to obtain has the pharmacological action of neuroprotective unit, therefore can be used for preparing good effect, safe and reliable prevention and treat Parkinsonian medicine.
Each chemical constituent preparation technology in the pharmaceutical composition of the present invention is simple, and cost is low, and its toxic and side effects is little.Each chemical constituent is simply clear and definite, is easier to the quality control of medicine aborning.The Radix Et Caulis Acanthopanacis Senticosi effective ingredient proportioning steady quality that utilizes the present invention to obtain has the pharmacological action of neuroprotective unit, therefore can be used for preparing good effect, safe and reliable prevention and treat Parkinsonian medicine.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Experimental example 1
1, experimental cell strain: Shanghai cell institute of the Chinese Academy of Sciences
2, experiment medicine: MPP+SIGMA-RBI company
Radix Et Caulis Acanthopanacis Senticosi glucoside B standard substance
3, experimental technique: the PC12 cell is made into single cell suspension, with 5 * 10
4/ ml is inoculated in 96 well culture plates.Continuous culture was changed liquid after 24 hours, added the Radix Et Caulis Acanthopanacis Senticosi glucoside B diluent of variable concentrations in every hole respectively, made its final concentration be respectively 2.5,5, and 10,20,40,80 μ g/ml add final concentration 300 μ mol/L MPP simultaneously
+Solution.Continue to cultivate after 48 hours, every hole adds MTT solution (5mg/ml) 20 μ l, hatches 4 hours for 37 ℃ again.Stop cultivating the empty culture medium of the careful suction in back, every hole adds 150 μ l DMSO, vibrates 10 minutes, and crystallization is fully dissolved.Microplate reader detects each hole in 490nm place absorption value, calculates cell survival rate.Experimental result sees Table 1, table 2.(cell survival rate %=experimental group absorbance value/blank group absorbance value * 100%), (x ± s) expression carries out statistical procedures with the t check to data with mean ± standard deviation.
The Radix Et Caulis Acanthopanacis Senticosi glucoside B of table 1 variable concentrations is to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
The Radix Et Caulis Acanthopanacis Senticosi glucoside B of table 2 variable concentrations is to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
From experimental result as can be seen, the Radix Et Caulis Acanthopanacis Senticosi glucoside B of variable concentrations is to MPP
+There is evident difference in the influence of inductive cell injury.Compare with model group, 10,20 μ g/ml Radix Et Caulis Acanthopanacis Senticosi glucoside B group cell survival rate illustrates that apparently higher than model group (there is statistical significance P＜0.05) 10,20 μ g/ml Radix Et Caulis Acanthopanacis Senticosi glucoside B can significantly reduce MPP
+Damage effect, thereby make cell survival rate that by a relatively large margin raising be arranged.
Experimental example 2
1, experimental cell strain: Shanghai cell institute of the Chinese Academy of Sciences
2, experiment medicine: MPP+SIGMA-RBI company
Radix Et Caulis Acanthopanacis Senticosi glucoside D standard substance
3, experimental technique: the PC12 cell is made into single cell suspension, with 5 * 10
4/ ml is inoculated in 96 well culture plates.Continuous culture was changed liquid after 24 hours, added the Radix Et Caulis Acanthopanacis Senticosi glucoside D diluent of variable concentrations in every hole respectively, made its final concentration be respectively 2.5,5, and 10,20,40,80 μ g/ml add final concentration 300 μ mol/L MPP simultaneously
+Solution.Continue to cultivate after 48 hours, every hole adds MTT solution (5mg/ml) 20 μ l, hatches 4 hours for 37 ℃ again.Stop cultivating the empty culture medium of the careful suction in back, every hole adds 150 μ l DMSO, vibrates 10 minutes, and crystallization is fully dissolved.Microplate reader detects each hole in 490nm place absorption value, calculates cell survival rate.Experimental result sees Table 3, table 4.
(x ± s) expression carries out statistical procedures with the t check to the statistical procedures data with mean ± standard deviation.
The Radix Et Caulis Acanthopanacis Senticosi glucoside D of table 3 variable concentrations is to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
The Radix Et Caulis Acanthopanacis Senticosi glucoside D of table 4 variable concentrations is to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
From experimental result as can be seen, the Radix Et Caulis Acanthopanacis Senticosi glucoside D of variable concentrations is to MPP
+There is evident difference in the influence of inductive cell injury.Compare with model group, 20,40 μ g/ml Radix Et Caulis Acanthopanacis Senticosi glucoside D group cell survival rate illustrates that apparently higher than model group (there is statistical significance P＜0.05) 20,40 μ g/ml Radix Et Caulis Acanthopanacis Senticosi glucoside D can significantly reduce MPP
+Damage effect, thereby make cell survival rate that by a relatively large margin raising be arranged.
Experimental example 3
1, experimental cell strain: Shanghai cell institute of the Chinese Academy of Sciences
2, experiment medicine: MPP+SIGMA-RBI company
The two glucosides of syringaresinol
3, experimental technique: the PC12 cell is made into single cell suspension, with 5 * 10
4/ ml is inoculated in 96 well culture plates.Continuous culture was changed liquid after 24 hours, added the two glucoside diluents of syringaresinol of variable concentrations in every hole respectively, made its final concentration be respectively 2.5,5, and 10,20,40,80 μ g/ml add final concentration 300 μ mol/L MPP simultaneously
+Solution.Continue to cultivate after 48 hours, every hole adds MTT solution (5mg/ml) 20 μ l, hatches 4 hours for 37 ℃ again.Stop cultivating the empty culture medium of the careful suction in back, every hole adds 150 μ l DMSO, vibrates 10 minutes, and crystallization is fully dissolved.Microplate reader detects each hole in 490nm place absorption value, calculates cell survival rate.Experimental result sees Table 5, table 6.(x ± s) expression carries out statistical procedures with the t check to data with mean ± standard deviation.
The two glucosides of table 5 variable concentrations syringaresinol are to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
The two glucosides of table 6 variable concentrations syringaresinol are to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
From experimental result as can be seen, the two glucosides of the syringaresinol of variable concentrations are to MPP
+There is evident difference in the influence of inductive cell injury.Compare with model group, the two glucoside group cell survival rates of 40,80 μ g/ml syringaresinols illustrate that apparently higher than model group (there is statistical significance P＜0.05) the two glucosides of 40,80 μ g/ml syringaresinols can significantly reduce MPP
+Damage effect, thereby make cell survival rate that by a relatively large margin raising be arranged.
Experimental example 4
1, experimental cell strain: Shanghai cell institute of the Chinese Academy of Sciences
2, experiment medicine: MPP+SIGMA-RBI company
Daucosterol
3, experimental technique: the PC12 cell is made into single cell suspension, with 5 * 10
4/ ml is inoculated in 96 well culture plates.Continuous culture was changed liquid after 24 hours, added the daucosterol diluent of variable concentrations in every hole respectively, made its final concentration be respectively 2.5,5, and 10,20,40,80 μ g/ml add final concentration 300 μ mol/L MPP simultaneously
+Solution.Continue to cultivate after 48 hours, every hole adds MTT solution (5mg/ml) 20 μ l, hatches 4 hours for 37 ℃ again.Stop cultivating the empty culture medium of the careful suction in back, every hole adds 150 μ l DMSO, vibrates 10 minutes, and crystallization is fully dissolved.Microplate reader detects each hole in 490nm place absorption value, calculates cell survival rate.Experimental result sees Table 7, table 8.(x ± s) expression carries out statistical procedures with the t check to data with mean ± standard deviation.
The daucosterol of table 7 variable concentrations is to MPP
+The influence of inductive PC12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
The daucosterol of table 8 variable concentrations is to MPP
+The influence of inductive PC 12 cell injury (x ± s)
Annotate: compare with model group,
*P＜0.05
From experimental result as can be seen, the daucosterol of variable concentrations is to MPP
+There is evident difference in the influence of inductive cell injury.Compare with model group, 20,40,80 μ g/ml daucosterol group cell survival rates illustrate that apparently higher than model group (there is statistical significance P＜0.05) 20,40,80 μ g/ml daucosterols can significantly reduce MPP
+Damage effect, thereby make cell survival rate that by a relatively large margin raising be arranged.
Experimental example 5
1, experimental cell strain: Shanghai cell institute of the Chinese Academy of Sciences
2, experiment medicine: MPP+SIGMA-RBI company
Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, daucosterol
The two glucosides of syringaresinol
3, experimental technique: according to above-mentioned experimental result, obtain each dose of components horizontal extent (seeing Table 9), at 3 levels of each effective ingredient design, press L respectively
9(3
4) orthogonal array (seeing Table 10) experimentizes: the PC12 cell is made into single cell suspension, with 5 * 10
4/ ml is inoculated in 96 well culture plates.Continuous culture was changed liquid after 24 hours, added different sample diluting liquids in every hole respectively, and the final concentration that makes it is 250 μ g/ml, added final concentration 300 μ mol/L MPP simultaneously
+Solution.Continue to cultivate after 48 hours, every hole adds MTT solution (5mg/ml) 20 μ l, hatches 4 hours for 37 ℃ again.Stop cultivating the empty culture medium of the careful suction in back, every hole adds 150 μ l DMSO, vibrates 10 minutes, and crystallization is fully dissolved.Microplate reader detects each hole in 490nm place absorption value, calculates cell survival rate.(cell survival rate %=experimental group absorbance value/matched group absorbance value * 100%), experimental result sees Table 11, and The results of analysis of variance sees Table 12.
The dosage level scope of each effective ingredient of table 9
Table 10 L
9(3
4) orthogonal array
The proportion compatibility orthogonal experiments of each composition of table 11
Table 12 analysis of variance table
As seen from the above table, the D factor affecting is remarkable.A, B, C do not have marked difference, so selected best proportion compatibility is A
1B
1C
3D
3, promptly four kinds of effective ingredient proportionings are Radix Et Caulis Acanthopanacis Senticosi glucoside B: Radix Et Caulis Acanthopanacis Senticosi glucoside D: the two glucosides of syringaresinol: daucosterol is 1: 2: 8: 8.
Claims (6)
1. a prevention or treat Parkinsonian pharmaceutical composition is characterized in that: by Radix Et Caulis Acanthopanacis Senticosi glucoside B, and Radix Et Caulis Acanthopanacis Senticosi glucoside D, two glucosides of syringaresinol and daucosterol are formed.
2. according to the described pharmaceutical composition of claim 1, it is characterized in that: the consumption proportion scope of each component is: Radix Et Caulis Acanthopanacis Senticosi glucoside B10～20 μ g/ml, Radix Et Caulis Acanthopanacis Senticosi glucoside D20～40 μ g/ml, two glucoside 40～80 μ g/ml of syringaresinol, daucosterol 20～80 μ g/ml.
3. according to the described pharmaceutical composition of claim 1, it is characterized in that: this pharmaceutical composition is by Radix Et Caulis Acanthopanacis Senticosi glucoside B, Radix Et Caulis Acanthopanacis Senticosi glucoside D, and two glucosides of syringaresinol and daucosterol were according to 1: 2: 8: 8 weight compatibility and forming.
4. according to any one described pharmaceutical composition of claim 1-3, it is characterized in that: described pharmaceutical composition with after pharmaceutically acceptable carrier or diluent cooperate, is prepared into any one suitable clinical preparation by the formulation method of this area routine with it.
5. according to the described pharmaceutical composition of claim 4, it is characterized in that: described preparation is oral formulations or ejection preparation.
6. any one described pharmaceutical composition of claim 1-3 is preparing the purposes for the treatment of in the Parkinsonian medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910135572XA CN101537011B (en) | 2009-04-27 | 2009-04-27 | Pharmaceutical composition for preventing or treating parkinsonism and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910135572XA CN101537011B (en) | 2009-04-27 | 2009-04-27 | Pharmaceutical composition for preventing or treating parkinsonism and preparation method thereof |
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| Publication Number | Publication Date |
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| CN101537011A CN101537011A (en) | 2009-09-23 |
| CN101537011B true CN101537011B (en) | 2011-06-01 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN200910135572XA Expired - Fee Related CN101537011B (en) | 2009-04-27 | 2009-04-27 | Pharmaceutical composition for preventing or treating parkinsonism and preparation method thereof |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102755343A (en) * | 2012-08-08 | 2012-10-31 | 南京中医药大学 | Application of daucosterol in preparing medicines for promoting proliferation of neural stem cells |
| CN109223816B (en) * | 2018-11-20 | 2021-10-26 | 江苏省中国科学院植物研究所 | Application of eleutheroside D in preparation of anti-depression drug |
| KR102280202B1 (en) * | 2021-02-26 | 2021-07-22 | 옙바이오 주식회사 | Phamaceutical composition for treating neuroinflammation diseases comprising Eleutheroside B as an active ingredient |
| CN113209114B (en) * | 2021-05-18 | 2022-04-19 | 华中农业大学 | Application of PGC-1 alpha activator daucosterol in preparation of medicine for preventing and treating blood brain barrier injury |
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2009
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