CN103172555B - Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof - Google Patents
Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN103172555B CN103172555B CN201310095614.8A CN201310095614A CN103172555B CN 103172555 B CN103172555 B CN 103172555B CN 201310095614 A CN201310095614 A CN 201310095614A CN 103172555 B CN103172555 B CN 103172555B
- Authority
- CN
- China
- Prior art keywords
- compound
- acceptable salt
- preparation
- ratio
- gradient elution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides an indole alkaloid compound separated from rhizoma cimicifugae as well as a preparation method and an application thereof. The compound provided by the invention is a novel indole alkaloid compound which is separated from rhizoma cimicifugae. The invention further provides a pharmaceutical composition which contains the compound or pharmaceutically acceptable salts thereof, and an application of the compound in preparation of antitumor medicines.
Description
Technical field
The present invention relates to a kind of brand-new Benzazole compounds and its production and use, be specifically related to a kind of new indole alkaloid compound, preparation method and its usage of separating of extracting from rattletop.
Background technology
Rattletop is the famous conventional Chinese medicine of China, and underground rhizome is used as medicine, and its medicinal efficacy is clearing heat and detoxicating, elevate a turnable ladder yang-energy and delivers promoting eruption.The gynaecopathias such as headache, toothache, throat pain, measles and uterine prolapse that cure mainly among the people.In China, the pharmaceutical use of rattletop is just on the books in Shennong's Herbal, and is listed in " top grade " medicinal material.
The chemical composition of Rattleroot plant and pharmacologically active are domestic and international study hotspots always.Obtain more than 200 of triterpene and glycoside thereof, Phenylpropanoid Glycosides class, chromone and other type compounds from wherein separating nearly ten years.In the world the research of chemical composition in rattletop is mainly concentrated on its cyclic-ahltin alkane triterpenes composition, to its pharmacology activity research taking oestrogen-like hormone, osteoporosis as main.Treatment involution syndrome and osteoporotic medicine are all developed respectively in developed countries such as Germany, Japan, the U.S..Different from the focus of research in the world, in order to expand the frontier of compound research in rattletop, make full use of China's cimicifuga plant resource, extract the new compound of isolating construction novelty and develop its new purposes, contriver is through a large amount of experimental studies, successfully from rattletop, extract to separate and obtain a kind of brand-new compound, and be surprised to find this compound and there is anti-tumor activity well.
Summary of the invention
The object of the present invention is to provide a kind of new antitumor indole alkaloid compound that extraction is separated to from rattletop and extracting method and the application of this compound.
The invention provides new indole alkaloid compound or its pharmacy acceptable salt that a kind of structure is following.
The present invention also provides the preparation method of this compound to be:
(1) rattletop rhizome is raw material, and alcohol reflux reclaims solvent, obtains medicinal extract.
(2) medicinal extract water dissolution, uses sherwood oil, ethyl acetate, n-butanol extraction successively, obtains acetic acid ethyl ester extract.
(3) acetic acid ethyl ester extract silica gel low pressure column chromatography, with normal hexane acetone gradient elution, collects merging and obtains 6 streams part.
(4) wherein second part is used silica gel low pressure column chromatography, with chloroform-methanol system gradient elution, collects altogether 8 parts.
(5) carry out anti-phase C18 medium pressure column chromatography by the 2nd part in these 8 parts, with acetonitrile-water gradient elution, obtain this compound crude product, then obtain this compound by efficient preparation liquid phase purifying.
Ethanol in step (1) is more than 70% ethanol; Normal hexane-acetone gradient elution system in step (3), the ratio of eluent is: 8: 1-6: 1-4: 1-2: 1-1: 1-1: 2; Chloroform-methanol gradient elution system in step (4).The ratio of eluent is 100: 0-80: 20; Acetonitrile-water gradient elution system in step (5), the ratio of eluent is 70: 30-80: 20; In step (5), efficiently prepare liquid-phase condition, acetonitrile-water, 75: 25, flow velocity 5ml/min.
The present invention also provides this compound and pharmacy acceptable salt thereof in the application of preparing in medicine for treating tumor thing.
The acceptable salt of medicine of the compounds of this invention, particularly forms pharmaceutically acceptable non-toxic salt, comprises hydrochloride, vitriol, sulphite, mesylate, organic acid salt etc.Can prepare these salt according to conventional method for transformation.
The compounds of this invention is as the pharmaceutical composition of activeconstituents.Pharmaceutical composition of the present invention, this compound that contains physiology significant quantity and the acceptable salt of medicine thereof, also can comprise as required medicine acceptable carrier, said this compound and the acceptable salt of medicine thereof are 0.1-99.9% in the weight ratio of composition, and the weight ratio of medicine acceptable carrier in composition is 0.1-99.9%.Pharmaceutical composition of the present invention exists to be applicable to medicinal dosage form, and these dosage forms can be: tablet, capsule, oral liquid, mixture, suck agent, granule, electuary, pill, powder, paste, sublimed preparation, suspensoid, injection, suppository, creme, sprays, drops, pill, patch.
The compounds of this invention is white powder,
Accompanying drawing 1 is the compounds of this invention ir data: IR (KBr) vmax (cm-1): 3244,2957,1709,1619,1470,1227,1101,998em; Accompanying drawing 2 is the compounds of this invention high resolution mass spectrum (HRTOF-ESIMS) (m/z :) 407.2099[M+Na]+calculated value (C26H280N2Na, 407.2094); Accompanying drawing 3 is the compounds of this invention hydrogen spectrum spectrogram, and accompanying drawing 4 is the compounds of this invention carbon spectrum spectrogram; Accompanying drawing 5 is the compounds of this invention HMQC spectrogram; Accompanying drawing 6 is the compounds of this invention HMBC spectrogram; Accompanying drawing 7 is the compounds of this invention H-HCOSY spectrogram.
(see accompanying drawing 4 from carbon spectrum, table 1) and hydrogen spectrum (see accompanying drawing 3, table 1) information observe, in conjunction with HMQC (seeing accompanying drawing 5), this compound has 26 carbon, contains carbonyl signal and phenyl ring and two key signals, dragendorff reacting positive (bismuth potassium iodide reaction), the information providing according to high resolution mass spectrum, determines that molecular formula is C
26h
28n
2o, degree of unsaturation is that 14, IR composes at 3300-3200 and 1709,1619cm
-1there is strong absorption, in prompting molecule, contain amide segments, according to phenyl ring proton peak form characteristics in hydrogen spectrum, meet AABB spin coupling system.
1hNMR shows ,-CH (CH
3)
2structure fragment.In conjunction with H-HCOSY (seeing accompanying drawing 7), infer-CH
2-CH-CH
2-CH (CH
3)
2structure fragment, in conjunction with H-HCOSY and HMBC, infer CH by ethylene linkage proton signal
3-C (CH
2the structure fragment of)=CH-CH-, in conjunction with HMBC, determines this compound structure by these two structure fragments.
Table 1 compound
1h and
13c nuclear magnetic resonance data (deuterated DMSO, 400MHz)
By the compounds of this invention being carried out to antitumor activity in vitro (Alamar blue method), find that compound of the present invention has good broad-spectrum anti-tumor biological activity.
Brief description of the drawings
Fig. 1: the infrared spectra of formula I compound;
Fig. 2: the high resolution mass spectrum of formula I compound;
Fig. 3: the hydrogen spectrum of formula I compound;
Fig. 4: the carbon spectrum of formula I compound;
Fig. 5: the HMQC spectrum of formula I compound;
Fig. 6: the HMBC spectrum of formula I compound;
Fig. 7: the H-HCOSY spectrum of formula I compound.
Embodiment
Listed embodiment contributes to those skilled in the art to understand better the present invention below, but does not limit the present invention in any way.
Preparation example 1 is prepared compound
Rattletop (cimicifugae rhizoma) rhizome (5kg) is raw material, after raw material pulverizing, through 80% ethanol (according to the quantity of solvent of 8-10 times of volume) refluxing extraction 3 times, each 2 hours, merge the extracting solution of three times, reclaim solvent, obtain medicinal extract; By medicinal extract water suspendible, use successively sherwood oil, ethyl acetate extraction, obtains acetic acid ethyl ester extract 250g; 200~300 object silica gel column chromatographies for acetic acid ethyl ester extract, normal hexane-acetone gradient elution (8: 1-6: 1-4: 1-2: 1-1: 1-1: 2), by thin-layer chromatography, detect the wash-out situation of composition in stream part, collect the stream part containing this compound, merge concentrating under reduced pressure evaporate to dryness; The enriched material 40g of preparation is used to silica gel low pressure column chromatography again, with chloroform-methanol system gradient elution (100: 0~80: 20), detect by thin-layer chromatography, collect the composition that is rich in this compound, merge concentrating under reduced pressure, obtain enriched material 4g; 4g enriched material is carried out to anti-phase C
18medium pressure column chromatography, with acetonitrile-water gradient elution (70: 30~80: 20m), merges the concentrated crude product 90mg that obtains; By efficient preparation liquid phase acetonitrile-water (80: 20) purifying repeatedly, obtain compound, be 99.2% through HPLC detection level.
Preparation example 2 tablets
Take 2 grams of preparation example 1 gained compounds, 100 grams of starch, 40 grams of pregelatinized Starchs, 9 grams of sodium starch glycolatees, fully mix, and cross 80 mesh sieves, add 95% appropriate amount of ethanol to make softwood, 12 orders are granulated, and 60 DEG C are dried 3 hours, the whole grain of 12 mesh sieves, add the Magnesium Stearate of particle gross weight 0.3%, the heavily about 150mg of trim plate, compressing tablet, makes 1000 altogether.
Preparation example 3 capsules
Take 2 grams of preparation example 1 gained compounds, 50 grams of starch, 49 grams, dextrin, fully mixes, and crosses 80 mesh sieves, uses 95% alcohol granulation, crosses 12 mesh sieves, is filled in No. 1 capsule, makes altogether 1000.
Test example 1 antitumor activity in vitro
1. material:
RPMI1640 and DMEM nutrient solution, penicillin, Streptomycin sulphate, foetal calf serum are purchased from Gibco company, and pancreatin, MTT, DMSO are purchased from Amresco company.
Compound: preparation example 1 prepares.
2. process of the test:
Tumor cell culture: H460 human lung carcinoma cell, Hela human cervical carcinoma cell lines and MCF mono-7 human breast cancer cells are cultivated respectively in the RPMI1640 containing 10%FBS (containing the Streptomycin of 100u/ml Penicillin and 100 μ g/ml) substratum, the cell substratum of collecting well-grown and get in logarithmic phase is made into the cell suspension that density is 1 × 105 cell/ml, be inoculated in equably in 96 well culture plates, every hole 100ul (being that every porocyte number is 104), at 5%CO2, in 37 DEG C of incubators, cultivate 24 hours after cell is completely adherent, formula 1 compound treatment of different concns for administration group, compound final concentration is respectively 50 μ g/ml, 40 μ g/ml, 30 μ g/ml, 20 μ g/ml, 10 μ g/ml, 5 μ g/ml, 1 μ g/ml.Establish control group and blank group simultaneously, every group 6 multiple holes, at 5%CO2, in 37 DEG C of incubators, cultivate after 48hr, adding the 10mg/mlMTT of 10 μ l to make final concentration is 0.05%, hatches after 4hr, centrifugal supernatant discarded liquid culture medium, add 100 μ l DMSO, in microplate reader, vibrate after 60s, in 5: 70nm measures absorbance.
3. test-results:
Test-results shows, compound of the present invention all has more significant restraining effect (being shown in Table 2) to above three kinds of tumours.
The external IC50 value to three kinds of tumor cell lines of table 2 formula I compound
Claims (6)
2. a method of extracting compound claimed in claim 1, its step comprises:
(1) rattletop rhizome is raw material, and 80% alcohol reflux reclaims solvent, obtains medicinal extract;
(2) medicinal extract water dissolution, with sherwood oil, ethyl acetate extraction, obtains acetic acid ethyl ester extract successively;
(3) acetic acid ethyl ester extract silica gel low pressure column chromatography, normal hexane-acetone gradient elution, wherein the ratio of normal hexane-acetone eluent is: 8:1-6:1-4:1-2:1-1:1-1:2, collects the stream part containing this compound, merge concentrating under reduced pressure evaporate to dryness, obtain the enriched material containing this compound;
(4) enriched material of preparing in the 3rd step is used to silica gel low pressure column chromatography again, with chloroform-methanol system gradient elution, wherein the ratio of chloroform-methanol eluent is: 100:0-80:20, collects the composition that is rich in this compound, merge concentrating under reduced pressure, obtain the enriched material containing this compound;
(5) enriched material of preparing in the 4th step is carried out to anti-phase C18 medium pressure column chromatography, with acetonitrile-water gradient elution, wherein the ratio of acetonitrile-water eluent is 70:30-80:20, obtains this compound crude product;
(6) crude product is obtained to this compound by efficient preparation liquid phase purifying, efficiently preparing liquid-phase condition is that acetonitrile-water eluent ratio is 75:25, flow velocity 5ml/min.
3. compound pharmacy acceptable salt as claimed in claim 1, is characterized in that acceptable salt comprises: hydrochloride, vitriol, sulphite, mesylate, organic acid salt.
5. pharmaceutical composition described in claim 4, is characterized in that its formulation comprises: tablet, capsule, oral liquid, suck agent, granule, pill, powder, paste, sublimed preparation, suspensoid, injection, suppository, ointment, plaster, creme, sprays, drops, pill, patch.
6. compound as claimed in claim 1 or its pharmacy acceptable salt are in the application of preparing in antitumor drug.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310095614.8A CN103172555B (en) | 2012-12-07 | 2013-03-22 | Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210526127.8 | 2012-12-07 | ||
CN201210526127 | 2012-12-07 | ||
CN201310095614.8A CN103172555B (en) | 2012-12-07 | 2013-03-22 | Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103172555A CN103172555A (en) | 2013-06-26 |
CN103172555B true CN103172555B (en) | 2014-07-09 |
Family
ID=48632822
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310095614.8A Active CN103172555B (en) | 2012-12-07 | 2013-03-22 | Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103172555B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106860624B (en) * | 2017-01-25 | 2020-08-04 | 浙江大学 | Cimicifugae rhizoma extract, two cimicifugae flavone bases, and preparation method and application thereof |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7309787B2 (en) * | 2005-07-13 | 2007-12-18 | Allergan, Inc. | Kinase inhibitors |
US7723372B2 (en) * | 2008-03-19 | 2010-05-25 | Hoffman-La Roche Inc. | Spiroindolinone derivatives |
-
2013
- 2013-03-22 CN CN201310095614.8A patent/CN103172555B/en active Active
Also Published As
Publication number | Publication date |
---|---|
CN103172555A (en) | 2013-06-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109575099B (en) | Dammarane sapogenin derivative and preparation method and application thereof | |
CN104341430A (en) | 3-phenylcoumarin robustic acid as well as extraction method and application thereof | |
CN105153084A (en) | Novel diterpene compound as well as preparation method and medicinal application thereof | |
CN101824014B (en) | Compounds with anti-tumor activity in chloranthus japonicus and purpose thereof | |
CN103113218A (en) | Mapping-agathis dammara type diterpene compound, and preparation method and application thereof | |
CN103172555B (en) | Indole alkaloid compound separated from rhizoma cimicifugae as well as preparation method and application thereof | |
CN102267891B (en) | Novel triterpenoid compound and preparation method thereof | |
CN104892713A (en) | Preparation method and applications of cucurbitacin C and analogs thereof | |
CN102030800B (en) | Abies holophylla triterpenoid compound, extraction separation thereof and application thereof | |
CN115181156B (en) | Hederagenin derivative and application thereof in preparation of antitumor drugs | |
CN108796022B (en) | Preparation method and application of saikosaponin A and saikosaponin D | |
CN109879921A (en) | Compound with anti-tumor activity separated from rhizoma anemarrhenae and preparation method thereof | |
CN107266516B (en) | Triterpenoid with anti-tumor activity and the preparation method and application thereof | |
CN106749124B (en) | Neighbour's double tetrahydrofuran type Annonaceousacetogenicompounds compounds with anti-tumor activity and the preparation method and application thereof | |
CN113072526B (en) | Anthraquinone/coumarin dimer compound and preparation method and application thereof | |
CN112898263B (en) | Coumarin parallel lignan compound separated from fingered citron and liver protection application thereof | |
CN105801634B (en) | A kind of preparation method and application of straight chain alcohol glycoside compound in green peel of walnut | |
CN101245089A (en) | Process for producing a pair of novel ginsengenin and its compound body, and preparations thereof | |
CN101721434B (en) | Active ingredients of fomes officinalis, preparation method thereof and use thereof | |
CN102988525A (en) | Preparation method for total lignans in hawthorn seeds, and novel application | |
CN102675252B (en) | There is Cesong alkyl type diterpine compound and the application thereof of anti-tumor activity | |
CN101538259A (en) | Crystal form A of zedoary root cyclic diolefine, preparation method thereof and application thereof in preparing anti-tumor drugs | |
CN106810551A (en) | Two kinds of new carbon skeleton alkaloid compounds and its extraction separation method | |
CN101735189A (en) | Preparation method, preparation and application of tricin | |
CN101062045B (en) | Use of triterpene saponin compounds in preparing tumor medicine |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant |