CN101716262B - Extract from Iris tenuifolia, preparation method and application thereof - Google Patents
Extract from Iris tenuifolia, preparation method and application thereof Download PDFInfo
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- CN101716262B CN101716262B CN2009102438946A CN200910243894A CN101716262B CN 101716262 B CN101716262 B CN 101716262B CN 2009102438946 A CN2009102438946 A CN 2009102438946A CN 200910243894 A CN200910243894 A CN 200910243894A CN 101716262 B CN101716262 B CN 101716262B
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Abstract
The invention discloses an extract from Iris tenuifolia, a preparation method and application thereof. The method comprises the following steps of: 1) taking an underground part of the Iris tenuifolia, soaking the underground part in absolute methanol, collecting extracting solution, and then removing the absolute methanol of the extracting solution to obtain a total extract of the underground part of the Iris tenuifolia; and 2) dissolving the total extract of the underground part of the Iris tenuifolia obtained by the step 1) in water, then adding chloroform into the water for chloroform extraction, collecting a chloroform phase, and then removing the chloroform in the chloroform phase to obtain a chloroform extract. The chloroform extract has the effect of inhibiting the aggregation of amyloid beta, which indicates that the chloroform extract has certain effect of treating senile dementia.
Description
Technical field
The present invention relates to a kind of extract from Iris tenuifolia and preparation method thereof and application.
Background technology
Senile dementia (AD) is to cause dull-witted a kind of gradual neurodegenerative disease in elderly population.Different pathology signals and gradual senile plaque deposit and cause the deterioration of neurons neurofibrillary tangle to be accompanied by the decline of cognitive power and memory.The typical characteristic of AD is transmembrane glycoprotein β-amyloplaste predecessor albumen (β APP) hydrolysis and amyloid-β peptide (A β) (amyloid-β peptide chain (A the β)) accumulation that comes.Two kinds of main A β (Alzheimer ' s β-amyloid) variant is made of 40 and 42 aminoacid respectively, and they promote the proinflammatory reaction, activate the neuron excitotoxicity path, cause function of brain cell unusual even dead.Relevantly studies show that it all has the neuron excitotoxicity effect in cell and body as A β during with the polymerization of fibril form.Therefore, find the micromolecular compound that can the stop the A beta peptide aggregation very valuable (Ce ' line Rivie`re etc that in the AD treatment, becomes, Inhibitoryactivity of stilbenes on Alzheimer ' s β-amyloid fibrils in vitro, Bioorganic ﹠amp; MedicinalChemistry 15 (2007) 1160-1167).
Treatment to senile dementia at present mainly contains four big class medicines, be respectively cholinesterase inhibitor, antioxidant medicine, hormone replacement therapy medicine and NSAID (non-steroidal anti-inflammatory drug), but these medicines all exists risk or restriction to a certain extent.Screening study at present has the micromolecule lead compound that suppresses the A beta peptide aggregation, becomes one of focus of development treatment senile dementia.
The Iridaceae iris.Mainly be distributed in southwest, northwest and the Northeast in China.This genus has 300 kinds approximately, be distributed in widely all over the world, China has 60 kinds and 13 mutation approximately, (Mahmood U, KaulVK, Jirovetz L.Alkylated benzoquinones from Iris kumaonensis.Phytochemistry, 2002,2 (61): 923-926.Qin MJ, Xu LS, Tanaka T, et al.A preliminarystudy on thedistribution pattern of isoflavones in rhizomes of Iris from China and its systematicsignificance.Acta Phyto Sinica, 2000,38:343-349.) this platymiscium has been used as traditional folk medicine, be used for treating multiple disease, for example, cancer, inflammation, antibacterial and viral infection.At present, separate the chemical compound that obtains from Jris and have antitumor, antioxidation, effect such as malaria protozoon and tuberculosis (1.Atta-ur-RahmnM, et al.Anti-inflammatory isoflavonoids from the rhizomes of Iris germanica.Journalof Ethnopharmacology, 2003,86:177-180.2.Bonfils JP et al.Cytotoxicity of iridals, triterpenoids from Iris.on human tumor cell lines A2780 andK562.Planta Med, 2001,67:79-82).
Iris tenuifolia (Iris tenuifolia) is the Iridaceae iris, and its chemical constituent has certain report, (1.Kojima, K., Gombosurengyin, P., Ondogyni, P., Begzsurengyin, D., Zevgeegyin, O., Hatano, K., Ogihara, Y.Flavanones from Iris tenuifolia.Phytochemistry, 1997,44 (4): 711-714; 2.M.I.Choudhary, S.Hareem and H.Siddiqui, et al.A benzil andisoflavone from Iris tenuifolia, Phytochemistry, 2008,69,1880-1885).We continue to study the chemical constituent of this plant, have separated, identified a new chemical compound; Biological activity test shows: this chemical compound can stop the A beta peptide aggregation, and indication has certain curative effect to senile dementia.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of extract from Iris tenuifolia.
Preparation method provided by the invention may further comprise the steps:
1) gets the Iris tenuifolia under ground portion, extract, collect extracting solution, remove the absolute methanol of described extracting solution again, obtain Iris tenuifolia under ground portion total extract with absolute methanol;
2) the Iris tenuifolia under ground portion total extract that step 1) is obtained is soluble in water, adds chloroform again and carries out chloroform extraction, collects the chloroform phase, removes the chloroform of described chloroform in mutually again, obtains the extract of the present invention's protection.
Further, above-mentioned steps 1) in, described extraction repeats 3 times.
Above-mentioned steps 2) in, chloroform extraction can repeat 5 times.
The extract of method for preparing also belongs within protection scope of the present invention.
Above-mentioned extract stops the application in the A beta peptide aggregation product also to belong within protection scope of the present invention in preparation.
Further, the said goods can be a medicine.
The application of above-mentioned extract in the medicine of preparation treatment senile dementia also belongs within protection scope of the present invention.
Another purpose of the present invention is to provide a kind of medicine for the treatment of senile dementia.
The medicine of treatment senile dementia provided by the invention, its active component are above-mentioned extracts.
Experimental results show that: extract provided by the invention can stop A beta peptide aggregation (entanglement).Because the A beta peptide aggregation is a typical characteristic of senile dementia, therefore extract provided by the invention can be used to treat senile dementia.
Description of drawings
Fig. 1 is compound flow chart among the present invention.
The figure as a result that Fig. 2 descends A β entanglement degree for extract of the present invention, wherein aggregates represents not add the matched group of sample; 9 is extract of the present invention, and 10 is water-phase extract, and the vertical coordinate of Fig. 2 is that the fluorescence intensity note of matched group is done 1, and the fluorescence intensity of all the other groups is corresponding vertical coordinate with the ratio of the fluorescence intensity of matched group.
The specific embodiment
One, the preparation of chemical compound 1
Preparation flow as shown in Figure 1.
1) absolute methanol soaks
Get the fresh Iris tenuifolia under ground portion of 1661g, handle (each 5L absolute methanol/1kg material) three times with the absolute methanol soaking at room temperature, three times extracting solution merges, and reclaims methanol with Rotary Evaporators, obtains the total extract 241.8g of Iris tenuifolia under ground portion.
2) chloroform extraction
The extract that step 1) is obtained is dissolved in the 300mL distilled water, move in the 1000mL separatory funnel, adding isopyknic chloroform extracts, extraction is carried out 5 times altogether, the combined chloroform extract, with Rotary Evaporators the chloroform in the chloroform extraction liquid is reclaimed, obtain the chloroform extract 198.6g of Iris tenuifolia under ground portion, collect water-phase extract 23.2g simultaneously.This chloroform extract also is an extract provided by the invention.
Experimental example 2, extract provided by the invention suppress the activity rating of amyloid beta aggregation
Chloroform extract and water-phase extract that embodiment 1 is obtained carry out following activity analysis.
Reagent source: (1-40, M=4329.84) available from Switzerland Bachem company, thioflavin T (Tht) is available from Sigma company for a
Active appraisal experiment method reference literature Ce ' line Rivie`re etc.Inhibitory activity of stilbenes onAlzheimer ' s-amyloid fibrils in vitro, Bioorganic ﹠amp; Medicinal Chemistry15:1160-1167 (2007) carries out, and concrete steps are as follows:
1, A β tangles
(phosphate buffer PH=7.4), obtains the A β solution of 0.5mg/ml to 1mgA β albumen+2mlPBS, adds EDTA (1mM) and promotes to tangle, and at 37 ℃, 300rmp is hatched 72h, preserves in-80 ℃ of refrigerators.
2, the preparation of thioflavin T (Tht)
Be made into the Tht of 300uM, standby, in 4 ℃ of refrigerators, preserve, during fluorescence measurement, need be diluted to 3uM.
3, sample+A β
Add sample 1 or the sample 2 of A β solution that 20ul concentration is 20uM (the A β solution dilution of step 1 preservation becomes 20uM) and 2ul in the tube pipe, fully mixing, was hatched 30 minutes by 37 ℃.Sample 1 is that the chloroform extract that embodiment 1 obtains is dissolved in dimethyl sulfoxide (DMSO), forms the solution of 10ug/ml; Sample 2 is that the water-phase extract that embodiment 1 obtains is dissolved in dimethyl sulfoxide (DMSO), forms the solution of 10ug/ml.
4, fluorescence measurement
The Tht of 3uM that gets 1500ul adds the mixture of hatching in the 15ul step 3 and carries out fluorescence measurement, excitation wavelength in new tube pipe: 442nm, measure wavelength: 480nm.
Statistical method: with the variance analysis of SPSS statistical software, * * P＜0.001.
Experiment repeats 3 times, experimental result as shown in Figure 5, the entanglement degree of A β significantly descends in the experimental group of chloroform extract, and the water-phase extract effect is faint, illustrates that chloroform extract provided by the invention can stop the A beta peptide aggregation.Because the A beta peptide aggregation is a typical characteristic of senile dementia, therefore extract provided by the invention can be used to treat senile dementia.
Claims (8)
1. preparation method of extract from Iris tenuifolia comprises following step:
1) gets the Iris tenuifolia under ground portion, soak, collect extracting solution, remove the absolute methanol of described extracting solution again, obtain Iris tenuifolia under ground portion total extract with absolute methanol;
2) the Iris tenuifolia under ground portion total extract that step 1) is obtained is soluble in water, adds chloroform again and carries out chloroform extraction, collects the chloroform phase, removes the chloroform of described chloroform in mutually again, obtains extract.
2. method according to claim 1 is characterized in that: in the described step 1), absolute methanol soaks and repeats 3 times.
3. method according to claim 1 and 2 is characterized in that: described step 2), chloroform extraction repeats 5 times.
4. the extract of the arbitrary described method preparation of claim 1-3.
5. the application of the described extract of claim 4 in preparation prevention amyloid-β peptide chain aggregation products.
6. application according to claim 5 is characterized in that: described product is a medicine.
7. the application of the described extract of claim 4 in the medicine of preparation treatment senile dementia.
8. medicine for the treatment of senile dementia, its active component is the described extract of claim 4.
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| CN105030759A (en) * | 2015-08-27 | 2015-11-11 | 北京师范大学 | Novel application of 5,2',3'-trihydroxy-6,7-methylenedioxy-flavanone compound |
| CN111793050B (en) * | 2020-07-16 | 2023-12-01 | 中国药科大学 | New compounds in fine She Yuanwei and antioxidant activity thereof |
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