CN103058970B - Inula wissmannian extracts, preparation for same, and application of extracts in preparation of anti-inflammatory medicine - Google Patents
Inula wissmannian extracts, preparation for same, and application of extracts in preparation of anti-inflammatory medicine Download PDFInfo
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Abstract
The invention provides a preparation for inula wissmannian extracts and an application of the extracts in preparation of an anti-inflammatory medicine. The extracts are germacrane sesquiterpene lactone inula wissmannian lactones A, B, C, D and E and inula cappa lactone with the following chemical structural formulas and absolute configurations shown in the specification, wherein the inula wissmannian lactones A, B, C, D and E and the inula cappa lactone generate an inhibition effect on the NO of the RAW264.7 macrophage induced by LPS (lipopolysaccharide); and in an xylene-induced ear swelling mouse model and a Freund complete adjuvant arthritis rat model, the inula wissmannian lactones A, B, C, D and E and the inula cappa lactone are found to be able to obviously inhibit the ear swelling and foot swelling of experimental animals, and the result indicates that the inula wissmannian lactones A, B, C, D and E and the inula cappa lactone are good in anti-inflammatory activity, capable of being used for preparing an anti-inflammatory medicine, and great in application value. The invention provides an extraction and separation method for the inula wissmannian lactones A, B, C, D and E and the inula cappa lactone.
Description
Technical field
The present invention relates to natural drug, be specifically related to the extraction and separation method of Inula wissmanniana effective constituent, purposes; Particularly relate to the extraction and separation method of Inula wissmanniana extract Germacrane Sesquiterpenoids lactone and preparing the application in anti-inflammatory drug.
Background technology
Composite family (Compositae) Inula (Inula L.) plant about has 100 kinds, is mainly distributed in Mediterranean Zone, also has distribution in other area that is European, African and Asia.This platymiscium in state-owned more than 20 plant and most mutation, wherein more than half plants has medicinal record (Chinese Plants will editorial board of the Chinese Academy of Sciences, Chinese Plants will [M], Beijing: Science Press, 1979,75:248-281).Chinese scholars is successively separated the compound (ZhaoYM obtaining the structure types such as flavones, sesquiterpene lactones, terpene from this platymiscium; Zhang ML; Shi QW, Kiyota H.Chemical constituents of plants from thegenus Inula.Chemistry & Biodiversity2006; 3:371-383).Sesquiterpene lactones is the characteristic chemical constituent of this platymiscium, is also main activeconstituents.There is the multiple effects such as anti-inflammatory, antitumor, antiviral, expelling parasite desinsection, immunosuppression.Sesquiterpene lactones also because its structure is simple, and has multiple mother nucleus structure type, can be used as the lead compound of research and development medicine.Plant Inula wissmanniana (Inulawissmanniana Hand.-Mazz.) is Inulaplants, and Chinese yunnan area is commonly called as " torch stalk ", is undershrub, produces south of Yunnan (Yuanjiang River, screen limit).Be born in Kai Kuang hillside fields, low mountain, height above sea level 1200 ~ 1650 meters.The present inventor has carried out the deep chemical constitution study of system to Inula wissmanniana, therefrom be separated and obtain 8 Germacrane Sesquiterpenoids constituents, find Inula wissmanniana extract and Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone there is significant anti-inflammatory activity.At present, there is no the bibliographical information of Inula wissmanniana chemical composition and pharmacologically active thereof, also have no the report relevant with the Inula wissmanniana method for preparing extractive in the present invention, purposes.
Summary of the invention
The object of technical problem to be solved by this invention is research and design Inula wissmanniana extract and extraction and separation method thereof and purposes.
The invention provides a kind of Inula wissmanniana extract, described extract be mainly Germacrane Sesquiterpenoids Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone, their structural formula and absolute configuration as follows respectively:
Inula wissmanniana lactone first:
Inula wissmanniana lactone second:
Inula wissmanniana lactone third:
Inula wissmanniana lactone fourth:
Inula wissmanniana lactone penta:
Wissmanniana lactone:
Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone be colourless lump shaped crystalline, be soluble in chloroform, acetone, methyl alcohol, ethanol, molecular formula is followed successively by C
22h
30o
6, C
20h
28o
5, C
20h
28o
4, C
19h
26o
4, C
15h
22o
3, C
22h
30o
7.Its chemical structure and absolute configuration are determined in conjunction with one dimension, two dimensional NMR wave spectrum and the analysis of X-ray single crystal diffraction by high resolution mass spectrum.
Another object of the present invention there is provided the preparation method of described Inula wissmanniana extract.
Preparation of the present invention comprises the preparation of extraction and isolation and monomeric compound.
Preparation method of the present invention comprises the following steps:
(1) Inula wissmanniana herb be dry, pulverize, extract 3 ~ 4 times with 1 ~ 10 times amount 80 ~ 95% ethanol soaking at room temperature, each 12 ~ 24 hours, merge to obtain extracting solution;
(2) concentrating under reduced pressure extracting solution is to without alcohol taste, obtains water suspendible fluid extract, and add 1 ~ 3 times of water dilution, use 2 ~ 6 times of sherwood oils, methylene dichloride, extraction into ethyl acetate successively, reclaim under reduced pressure respectively extracts part, obtains without aqueous extract;
(3) by above-mentioned methylene dichloride position application silica gel column chromatography, take volume ratio as the methylene chloride/methanol system gradient elution of 100:0 ~ 2:1, thin-layer chromatography detects, collect respectively containing Inula wissmanniana lactone first, second, third, fourth, penta and the flow point of wissmanniana lactone, again through dextran gel column chromatography, take volume ratio as the methylene chloride/methanol wash-out of 1:1, finally by liquid chromatography preparation, obtain Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone.
Another object of the present invention there is provided described Inula wissmanniana extract and is preparing the application in anti-inflammatory drug.
The present inventor by Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone to cell and animal experiment, result proves, Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone suppress scavenger cell RAW264.7 to produce the IC of NO
50value is respectively 4.90, and 2.17,5.09,1.09,1.75,10.28 μMs (aminoguanidine is positive drug IC
500.37 μM); In caused by dimethylbenzene xylene ear swelling mouse and Freund's complete adjuvant rat model of arthritis, find Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone can the obviously ear swelling of Inhibition test animal and foot swelling.Therefore, Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone there is good anti-inflammatory activity, can be used for preparing anti-inflammatory drug.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana extract.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone first.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone second.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone third.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone fourth.
Medicine of the present invention is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone penta.
The present invention makes public for the first time Inula wissmanniana extract, Inula wissmanniana lactone, Inula wissmanniana lactone first, second, third, fourth and penta; And provide their extraction and separation method and the application in pharmacy, provide new antiradiation drug for clinical, bring glad tidings to patient, have larger clinical value and social benefit.
Accompanying drawing explanation
Fig. 1 Inula wissmanniana lactone first X diffraction structure
Fig. 2 Inula wissmanniana lactone third X diffraction structure
Embodiment
The present embodiment is implemented under premised on technical solution of the present invention, gives detailed embodiment and process, but protection scope of the present invention is not limited to following embodiment.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.
Embodiment 1 Inula wissmanniana lactone first, second, third, fourth, penta and the preparation of wissmanniana lactone
After Inula wissmanniana herb (10.0kg) drying and crushing, extract 4 times (24,12,12,12h) with 95% ethanol (30L) cold soaking at room temperature 25 DEG C, recycling design, dryly without aqueous extract (631.4g) concentrated; Total medicinal extract is suspended in 1.5L water and extracts 3 times respectively by 5L sherwood oil, methylene dichloride, ethyl acetate successively, decompression and solvent recovery, obtains petroleum ether part (133.2g), methylene dichloride position (179.6g) and ethyl acetate extract (28.1g) respectively.
By normal phase silicagel column on the methylene dichloride position 120g of Inula wissmanniana, use methylene chloride-methanol (100% methylene dichloride successively, 100:1,50:1,20:1,10:1,5:1 and 100% methyl alcohol) gradient elution, elutriant detects through thin-layer chromatography, merges the elutriant of similar composition, and decompression and solvent recovery obtains 8 elution fraction.Get wherein the 1st elution fraction, upper silica gel column chromatography is with petroleum ether-ethyl acetate (20:1) wash-out, and through the elutriant of the similar composition of thin-layer chromatography combining data detection, decompression and solvent recovery obtains 6 eluted fraction.Wherein the 2nd eluted fraction (2.9g) is with the dissolving of the least possible methylene dichloride, standing at room temperature 25 DEG C, separate out lump shaped crystalline, be accredited as Inula wissmanniana lactone third (1.52g); 3rd flow point (3.3g), in the least possible dichloromethane solution, leaves standstill at room temperature 25 DEG C, separates out lump shaped crystalline Inula wissmanniana lactone first (1.23g); 4th flow point (0.9g) with Chromatographic Pure Methanol (3mL) dissolve, filter after through preparative chromatography method, with 70% methanol-eluted fractions, obtain compound Inula wissmanniana lactone fourth (t
r40.0min, 12.5mg).In addition, then by the 2nd elution fraction (12.4g), after methyl alcohol (30mL) dissolution filter, upper MCI column chromatography, with 80% methanol-eluted fractions, obtains 2 flow points.1st flow point (2.6g) upper Sephadex LH-20 gel column chromatography further, with methylene chloride-methanol 1:1 wash-out, elutriant detects through thin-layer chromatography, merge 8 elutriants of similar composition, recycling design obtains 8 eluted fraction, again the flow point (0.9g) of the 3rd (0.7g) and the 4th is wherein gone up preparative chromatography post respectively, with 55% methyl alcohol (10L) wash-out, obtain Inula wissmanniana lactone penta (t
r32.0min, 25.6mg), wissmanniana lactone (t
r58.2min, 31.1mg), and Inula wissmanniana lactone second (t
r75.0min, 30.0mg).
Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone
1h-NMR and
13c-NMR data are respectively in table 1 and table 2, and optical rotational activity spectrum, infrared spectra, mass spectrum, high resolution mass spectrum and X-ray single crystal diffraction data are as follows respectively:
Inula wissmanniana lactone first: brilliant without color lump; C
22h
30o
6;
(c0.10, CH
2cl
2); IR (KBr) ν
max2931,1768,1726,1661,1456,1382,1314,1245,1162,1134,1071,1019,975cm
-1; ESIMS m/z413.2 [M+Na]
+, m/z389.2 [M-H]
-; HRESIMS m/z413.1943 [M+Na]
+(calcd for C
22h
30o
6na, 413.1940); Crystal data: C
22h
30o
6, CH
2cl
2, M=390.46, orthorhombic, space group P2 (1) 2 (1) 2 (1), a=6.7751 (3)
α=90 °; B=8.9058 (4)
β=90 °; C=35.0890 (14)
γ=90 °; V=2117.19 (16)
z=4, D
calcd=1.225mg/m
3, crystallite size 0.20 × 0.15 × 0.10mm
3.Cu K α (λ=1.54178
), F (000)=840, T=133 (2) K.R value is respectively R
1=0.0317, and wR
2=0.0873, for22899 observed reflections [I>2 σ (I)] .Flack parameter is 0.05 (15).(storage number: CCDC835957) that related crystal data trnascription can obtain at Cambridge crystal data central toll-free, address: CCDC, 12Union Road, Cambridge CB21EZ, UK (fax:+441223336033or e-mail:data_requestccdc.cam.ac.uk).
Inula wissmanniana lactone second: brilliant without color lump; C
20h
28o
5;
(c0.10, CH
2cl
2); IR (KBr) ν
max3430,2928,2861,1763,1707,1650,1453,1381,1245,1150,1021,975cm
-1; ESIMS m/z371.1 [M+Na]
+, m/z347.1 [M-H]
-; HRESIMS m/z371.1836 [M+Na]
+(calcd for C
20h
28o
5na, 371.1834).
Inula wissmanniana lactone third: brilliant without color lump; C
20h
28o
4;
(c0.10, CH
2cl
2); IR (KBr) ν
max2937,2923,2856,1766,1705,1643,1450,1379,1234,1153,1144,1037,973cm
-1; ESIMS m/z355.2 [M+Na]
+, m/z331.2 [M-H]
-; HRESIMS m/z355.1856 [M+Na]
+(calcd for C
20h
28o
4na, 355.1855); Crystal data: C
20h
28o
4, CH
2cl
2, M=332.42, orthorhombic, space group P2 (1) 2 (1) 2 (1), a=11.9746 (5)
α=90 °; B=12.3428 (5)
β=90 °; C=12.4853 (5)
γ=90 °; V=1845.33 (13)
z=4, D
calcd=1.197mg/m
3, crystallite size 0.20 × 0.15 × 0.10mm
3.Cu K α (λ=1.54178
), F (000)=720, T=273 (2) K.R value is R respectively
1=0.0276, and wR
2=0.0767, for20005 observed reflections [I>2 σ (I)] .Flack parameter is 0.02 (15).(storage number: CCDC835958) that related crystal data trnascription can obtain at Cambridge crystal data central toll-free, address: CCDC, 12Union Road, Cambridge CB21EZ, UK (fax:+441223336033or e-mail:data_requestccdc.cam.ac.uk).
Inula wissmanniana lactone fourth: brilliant without color lump; C
22h
30o
6;
(c0.05, CH
2cl
2); IR (KBr) ν
max2930,2863,1770,1713,1633,1571,1454,1383,1317,1281,1250,1141,1072,1010,976cm
-1; ESIMS m/z341.2 [M+Na]
+, m/z317.2 [M-H]
-; HRESIMS:m/z341.1733 [M+Na]
+(calcd for C
19h
26o
4na, 341.1729).
Inula wissmanniana lactone penta: brilliant without color lump; C
15h
22o
3;
(c0.10, CH
2cl
2); IR (KBr) ν
max3441,2936,2865,1765,1663,1457,1382,1274,1249,1144,1079,1012,973cm
-1; ESIMS m/z273.1 [M+Na]
+, m/z249.1 [M-H]
-; HRESIMS:m/z273.1466 [M+Na]
+(calcd for C
15h
22o
3na, 273.1467).
Wissmanniana lactone: brilliant without color lump; C
22h
30o
7; ESIMS m/z429.2 [M+Na]
+, m/z405.2 [M-H]
-.
Table 1 Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone
1h-NMR data (CDCl
3, 400MHz)
Continued 1
Compound atom numbering is shown in structural formula of compound figure; Nuclear magnetisation displacement study unit is ppm; Be coupling constant values in bracket, unit is Hz.
Table 2 Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone
13c-NMR data (CDCl
3, 400MHz)
Compound atom numbering is shown in structural formula of compound figure; Nuclear magnetisation displacement study unit is ppm.
Embodiment 2 Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone to suppress LPS to induce the NO of RAW264.7 to generate active
(1) sample configuration
Obtained Inula wissmanniana lactone first, the second of embodiment 1, third, fourth, penta, wissmanniana lactone and positive control drug aminoguanidine (Sigma-Aldrich, purity >98.0%)) use DMSO(methyl-sulphoxide) (Merck) dissolve after, add the solution that PBS (-) (phosphate buffered saline buffer) is made into 10mM, further dilution is 0,0.1,0.5, the sample of 5,20 μMs of gradient concentrations.With the LPS of 10 μ g/mL (Lipopolysaccharides, lipopolysaccharides, sigma, Cat.L-2880) for inductor.
(2) experimental technique
Mouse macrophage RAW264.7(is purchased from Sheng Ke institute cellular resources center, Shanghai) at 37 ° of C, 5%CO
2in incubator, cellar culture is in DMEM nutrient solution.By 1 μ L/mL LPS(lipopolysaccharides during experiment) (being dissolved in distilled water) to add 100mL concentration be 2 × 10
6in the cell suspension of μ g/mL, indirectly reflect NO growing amount with Griess method by the content measuring cell conditioned medium liquid nitrite after 18h: get 100mL cell culture fluid, to add in equivalent Griess(lattice this) reagent, measure light absorption value.
(3) judgement criteria and statistical method
Optical density is surveyed, with NaNO in 570nm wavelength place
2standardized solution drawing standard curve, calculates the concentration of nitrite.Each group of experimental result carries out statistical study with SPSS software ONE WAY ANNOVA method (Chicago,U.S SPSS Inc.).
(4) experimental result
Experimental result show Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone induce the NO of RAW264.7 scavenger cell to generate to LPS to have obvious inhibit activities (the results are shown in Table 3), illustrate that it has anti-inflammatory activity.
Table 3 Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone to suppress LPS to induce the NO of RAW264.7 to generate active
Embodiment 3 Inula wissmanniana lactone first, second, third, fourth, penta and the activity of wissmanniana lactone antagonism caused by dimethylbenzene xylene inflammation
(1) sample configuration
The Inula wissmanniana total extract that embodiment 1 is obtained, petroleum ether part, methylene dichloride position, ethyl acetate extract and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone, use 0.5%Tween-80(tween-80 respectively) solution preparation becomes the suspension of desired concn, for subsequent use.Before use aspirin tablet (Sigma-Aldrich company) is mixed with the suspension of 200mg/kg as positive control.The administration volume of each group is 20mL/kg, oral administration.
(2) experimental technique
Kunming mouse 220, male, 18 ~ 22g, is divided into 22 groups at random, often organizes 10.If a group model contrast, one group of positive control and 20 groups of administration groups.After mouse is bought and conforms one week, start administration.Medication is: positive drug acetylsalicylic acid successive administration 5 days, and once a day, 0.5mL/ time, dosage is 200mg/kg; Each treatment group successive administration 5 days, once a day, 0.5mL/ time, Inula wissmanniana total extract group dosage is 400mg/kg, 800mg/kg, sherwood oil, methylene dichloride, ethyl acetate extract dosage are 200mg/kg, 400mg/kg, Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone dosage be 200mg/kg, 100mg/kg.Model group successive administration 5 days, once a day, 0.5mL/ time.30min after the administration of mouse last, auris dextra two sides is coated with dimethylbenzene 0.05mL/ and only causes inflammation, and left ear is not coated with, and puts to death after 60min, gets left and right ear same position disk with 6mm punch tool, precision weighing two ear weight.
(3) judgement criteria and statistical method
Swelling degree is represented with the difference of auris dextra and left ear weight.Each group of experimental result carries out statistical study with SPSS software ONEWAY ANNOVA method.
(4) experimental result
Positive drug (acetylsalicylic acid), Inula wissmanniana total extract, petroleum ether part, methylene dichloride position, ethyl acetate extract and Inula wissmanniana lactone first, second, third, fourth, penta, low, the high dose group of wissmanniana lactone, the ear swelling that p-Xylol causes has obvious restraining effect (P<0.05), wherein, with methylene dichloride position and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone high dose group effect is the most obvious.This experiment show Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone there is obvious anti-inflammatory activity (see table 4).
Table 4. Inula wissmanniana total extract, each extract part and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone is on the impact of mouse ear swelling test
Embodiment 4 Inula wissmanniana lactone first, second, third, fourth, penta and wissmanniana lactone antagonism Freund's complete adjuvant arthritic activity
(1) sample preparation
The Inula wissmanniana total extract that embodiment 1 is obtained, petroleum ether part, methylene dichloride position, ethyl acetate extract and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone, the suspension of desired concn is become respectively with 0.5%Tween-80 solution preparation, for subsequent use.Before use Indomethacin tablets is mixed with the suspension of 210mg/kg as positive control.The administration volume of each group is 10mL/kg, oral administration.
(2) experimental technique
Male rat, 190 ~ 200g, random point 22 groups, work, often organizes 10.If 1 group model, 1 group of positive control and 20 groups of administration groups.After rat is bought and conforms one week, start administration.Medication is: positive continuous oral administration 5 days, and once a day, dosage is 210mg/kg; Tested group of successive administration 5 days, once a day, abdominal injection, Inula wissmanniana total extract group dosage is 500mg/kg, 1000mg/kg, sherwood oil, methylene dichloride, ethyl acetate extract and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone dosage is 100mg/kg, 200mg/kg.Model group successive administration 5 days, once a day, oral injection blank solvent.Cause inflammation after last administration, survey Rat Right pawl solvent, after measuring, every Rat Right group injection Freund's complete adjuvant 0.05mL, surveys right sufficient volume again, tries to achieve Rat Right paw swelling after 24 hours.
(3) judgement criteria and statistical method
Each group of experimental result carries out statistical study with SPSS software ONE WAY ANNOVA method.
(4) experimental result
Positive drug (indomethacin), the high dose group at Inula wissmanniana total extract, methylene dichloride position, and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone is low, high dose group all obviously can suppress rat paw swelling, display Inula wissmanniana total extract, methylene dichloride position and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone all has certain anti-inflammatory activity (P<0.05, table 5).
Table 5. Inula wissmanniana total extract, each extract part and Inula wissmanniana lactone first, second, third, fourth, penta, wissmanniana lactone antagonism Freund's complete adjuvant arthritic activity
Claims (9)
1. Inula wissmanniana plant milk extract, it is characterized in that, described extract is Inula wissmanniana lactone first, Inula wissmanniana lactone second, Inula wissmanniana lactone third, Inula wissmanniana lactone fourth or Inula wissmanniana lactone penta, and their structural formula and absolute configuration are distinguished as follows:
Inula wissmanniana lactone first:
Inula wissmanniana lactone second:
Inula wissmanniana lactone third:
Inula wissmanniana lactone fourth:
Inula wissmanniana lactone penta:
2. prepare the method for Inula wissmanniana plant milk extract as claimed in claim 1, it is characterized in that,
The method comprises the following steps:
After Inula wissmanniana herb 10.0kg drying and crushing, at room temperature 25 DEG C, extract 4 times, 24,12,12,12h, recycling design with 95% ethanol 30L cold soaking, dryly without aqueous extract 631.4g concentrated; Total medicinal extract is suspended in 1.5L water and extracts 3 times respectively, decompression and solvent recovery by 5L sherwood oil, methylene dichloride, ethyl acetate successively, obtains petroleum ether part 133.2g, methylene dichloride position 179.6g and ethyl acetate extract 28.1g respectively;
By normal phase silicagel column on the methylene dichloride position 120g of Inula wissmanniana, use methylene chloride-methanol 100% methylene dichloride successively, 100:1,50:1,20:1,10:1,5:1 and 100% methanol elution gradient, elutriant detects through thin-layer chromatography, merges the elutriant of similar composition, and decompression and solvent recovery obtains 8 elution fraction; Get wherein the 1st elution fraction, upper silica gel column chromatography is with petroleum ether-ethyl acetate 20:1 wash-out, and through the elutriant of the similar composition of thin-layer chromatography combining data detection, decompression and solvent recovery obtains 6 eluted fraction; Wherein the 2nd eluted fraction 2.9g the least possible methylene dichloride dissolving, leave standstill at room temperature 25 DEG C, separate out lump shaped crystalline, be accredited as Inula wissmanniana lactone third 1.52g; 3rd flow point 3.3g, in the least possible dichloromethane solution, leaves standstill at room temperature 25 DEG C, separates out lump shaped crystalline Inula wissmanniana lactone first 1.23g; 4th flow point 0.9g Chromatographic Pure Methanol 3mL dissolves, filter after through preparative chromatography method, with 70% methanol-eluted fractions, obtain compound Inula wissmanniana lactone fourth t
r40.0min, 12.5mg; In addition, then by the 2nd elution fraction 12.4g, after methyl alcohol 30mL dissolution filter, upper MCI column chromatography, with 80% methanol-eluted fractions, obtains 2 flow points; 1st flow point 2.6g goes up Sephadex LH-20 gel column chromatography further, with methylene chloride-methanol 1:1 wash-out, elutriant detects through thin-layer chromatography, merge 8 elutriants of similar composition, recycling design obtains 8 eluted fraction, again 30.7g wherein and a 4th flow point 0.9g is gone up preparative chromatography post respectively, with 55% methyl alcohol 10L wash-out, obtain Inula wissmanniana lactone penta t
r32.0min, 25.6mg, wissmanniana lactone t
r58.2min, 31.1mg, and Inula wissmanniana lactone second t
r75.0min, 30.0mg.
3. Inula wissmanniana plant milk extract as claimed in claim 1 is preparing the application in anti-inflammatory drug.
4. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana plant milk extract.
5. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone first.
6. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone second.
7. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone third.
8. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone fourth.
9. apply as claimed in claim 3, it is characterized in that, described medicine is the pharmaceutical composition be made up of as activeconstituents and pharmaceutical excipient Inula wissmanniana lactone penta.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN103664839B (en) * | 2013-11-12 | 2016-01-13 | 中国人民解放军第二军医大学 | Inula wissmanniana lactone A and derivative thereof are preparing the application in antitumor drug |
| CN104945361B (en) * | 2015-06-19 | 2017-06-13 | 中国医学科学院药用植物研究所 | Germacrane Sesquiterpenoids derivative and preparation method and application |
| CN111714533A (en) * | 2020-07-21 | 2020-09-29 | 贵州光正制药有限责任公司 | Chrysanthemum indicum extract and preparation and application thereof |
| CN112402412B (en) * | 2020-11-18 | 2022-03-18 | 中国医学科学院药用植物研究所 | Application of inner ester compound of jingdao in preparing medicine for treating inflammation-caused diseases |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2005102311A1 (en) * | 2004-04-20 | 2005-11-03 | Deutsches Institut für Ernährungsforschung | Agonists of a bitter taste receptor and uses thereof |
| WO2008119527A2 (en) * | 2007-03-29 | 2008-10-09 | Deutsches Institut Für Ernährungsforschung Postdam-Rehbrücke Stiftung Des Öffentlichen Rechts | Agonists of bitter taste receptors and uses thereof |
| CN102274210A (en) * | 2011-06-17 | 2011-12-14 | 河北医科大学 | Applications of ineupatorolide B in preparation of antitumor drugs |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2005102311A1 (en) * | 2004-04-20 | 2005-11-03 | Deutsches Institut für Ernährungsforschung | Agonists of a bitter taste receptor and uses thereof |
| WO2008119527A2 (en) * | 2007-03-29 | 2008-10-09 | Deutsches Institut Für Ernährungsforschung Postdam-Rehbrücke Stiftung Des Öffentlichen Rechts | Agonists of bitter taste receptors and uses thereof |
| CN102274210A (en) * | 2011-06-17 | 2011-12-14 | 河北医科大学 | Applications of ineupatorolide B in preparation of antitumor drugs |
Non-Patent Citations (2)
| Title |
|---|
| Fang-Yuan Wang,等.Sesquiterpene lactones from inula cappa.《Phytochemistry Letters》.2012,第5卷(第3期),第639-642页. * |
| Hong-Gang Xie,等.Cytotoxic Germacranolide Sesquiterpene from Inula cappa.《Chem. Pharm. Bull.》.2007,第55卷(第8期),第1259-1260页. * |
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