CN101265248A - Cembrene lactone type diterpenoid lobophilides A and B, and preparation method and use thereof - Google Patents
Cembrene lactone type diterpenoid lobophilides A and B, and preparation method and use thereof Download PDFInfo
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- CN101265248A CN101265248A CNA2007100380477A CN200710038047A CN101265248A CN 101265248 A CN101265248 A CN 101265248A CN A2007100380477 A CNA2007100380477 A CN A2007100380477A CN 200710038047 A CN200710038047 A CN 200710038047A CN 101265248 A CN101265248 A CN 101265248A
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Abstract
The invention relates to cembranolides with formulas of pod flexibilide A and pod flexibilide B extracted and separated from Sinularia polydactyla, preparation method and application thereof. A plurality of in vitro antitumor activity experiments indicate that the cembranolides have obvious tumor cell inhibiting activity, and prospect application in drug for treating caner/tumor. The invention also provides a precursor for the research of new drugs for treating various common multiple cancers. The cembranolides and the preparation method thereof are important to exploitation and utilization of the Chinese marine biological resources.
Description
Technical field
The present invention relates to medical technical field, specifically be a class from South China Sea beanpod soft coral (Lobophytum sp.), separate Xin Xisong alkane lactone type diterpenoid beanpod soft coral first, the second element that obtains (lobophilides A, B) and its production and use.This compounds has had strong inhibitory effects to various tumor cell strains, can be used as the lead compound of the new antitumor drug of class development, also can be used as the medicine of the various clinical common multiple cancers of treatment.
Background technology
Coral system is low to wait coelenteron door animal, and there is kind more than 6100 in such marine organisms whole world, wherein in state-owned 496 kinds.Soft coral is meant coelenteron door Anthozoa Alcyonaria soft coral suborder animal.The soft coral suborder is divided 6 sections, i.e. Paralcyoniidae, Alcyoniidae, Asterospiculariidae, Nephtheidae, Nidaliidae and Xeniidae again.
Be rich in the novel structure secondary metabolite in the soft coral, the structure type of its compound mainly comprises diterpene, sesquiterpene, polyhydroxy sterol and prostaglandin(PG) etc.The beanpod soft coral belongs to Alcyoniidae section, is that to receive publicity the earliest also be one of maximum soft coral of research, and research range relates to more than 20 kind.What its chemical ingredients report was maximum is western loose alkane type and Luo Bei type diterpene.
Living marine resources in development and use China are therefrom sought in the process of marine natural product of biologically active and prospect in medicine, and we find, the ethyl acetate crude extract of South China Sea beanpod soft coral in the extracorporeal anti-tumor screening experiment to A
549Human lung adenocarcinoma cell master strain shows good inhibition activity.Further biological activity follow-up study has caused the discovery of Xin Xisong alkane lactone type diterpenoid beanpod soft coral first, second element.
Literature search shows that beanpod soft coral first, second element is the western loose alkane lactone type diterpenoid of high oxidation, novel structure.The experiment of extracorporeal anti-tumor bioactivity screening shows that beanpod soft coral first element is to A
549Human lung adenocarcinoma cell line shows significant cytotoxicity; Beanpod soft coral second element is then to A
549Human lung adenocarcinoma cell line and P
388The mouse leukemia cell strain all has significant inhibitory effect.
Summary of the invention
The object of the present invention is to provide new western loose alkane lactone type diterpenoid beanpod soft coral first, the second element of a kind of extraction separation obtains from South China Sea beanpod soft coral two.
Another object of the present invention provides the preparation method of above-mentioned west loose alkane lactone type diterpenoid beanpod soft coral first, second element.
An also purpose of the present invention provides above-mentioned west loose alkane lactone type diterpenoid beanpod soft coral first, second element and uses in the medicine of preparation treatment cancer and/or tumour.
West of the present invention loose alkane lactone type diterpenoid beanpod soft coral first, second element has following chemical structural formula respectively:
These 2 compounds all are to produce from Lingshui, Chinese Sanya to extract the beanpod soft coral first to obtain, so difference called after beanpod soft coral first, second element.
The invention provides the method for extraction separation compound beanpod soft coral first, second element from South China Sea beanpod soft coral, step is as follows:
1) extracts: South China Sea beanpod soft coral is extracted with ketone solvent, get crude extract after the gained extracting solution concentrates; This crude extract is added in the NaCl solution, and suspendible is the back extracted with diethyl ether evenly, and the gained extraction liquid obtains ether medicinal extract after concentrating;
2) separate: the ether medicinal extract that obtains in the step 1) carries out silica gel column chromatography, with the petroleum ether/ethyl ether gradient elution; Wherein, 50: 50 wash-out parts of petroleum ether/ethyl ether volume ratio through the SephadexLH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out purifying, obtain beanpod soft coral second element; 20: 80 wash-out parts of petroleum ether/ethyl ether volume ratio through the SephadexLH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out purifying, obtain beanpod soft coral first element.
In step 1) was extracted, described ketone solvent was preferably acetone, and described NaCl solution is preferably 0.7～1.5N NaCl solution; And it is to adopt ultrasonic extraction that described ketone solvent extracts;
Described step 2) in, described petroleum ether/ethyl ether gradient elution is to carry out gradient elution in 100: 0 → 90: 10 → 80: 20 → 50: 50 → 20: 80 → 10: 90 → 0: 100 with the petroleum ether/ethyl ether volume ratio;
Described step 2) PetroChina Company Limited.'s ether/50: 50 wash-out parts of ether volume ratio, through the SephadexLH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, developping agent is 1: 8 a petroleum ether/ethyl ether of volume ratio, with R
fValue merges at the elutriant at 0.25～0.35 place, concentrating under reduced pressure, obtains beanpod soft coral second element;
Described step 2) 20: 80 wash-out parts of PetroChina Company Limited.'s ether/ether, through Sephadex LH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, developping agent is sherwood oil/acetone of 7: 3 of volume ratio, with R
fValue merges at the elutriant at 0.45～0.55 place, concentrating under reduced pressure, obtains beanpod soft coral first element.
The present invention has carried out the anti-tumor activity test to west loose alkane lactone type diterpenoid beanpod soft coral first, second element, shows that these compounds have obvious antineoplastic.Can be used for preparing the medicine of treatment cancer and/or tumour.
Embodiment
The present invention is further elaborated below in conjunction with embodiment, but the present invention does not limit this.
1H-NMR measures with Varian Inova 600 type instrument; MS (ESIMS and HRESIMS) measures with Q-TOF Micro LC-MS-MS type mass spectrograph; Employed silica gel is for Haiyang Chemical Plant, Qingdao produces; All kinds of SOLVENTS is produced by reagent company limited of traditional Chinese medicines group, is analytical pure.
If no special instructions, ratio is volume ratio between the liquid/liquid that relates in following examples.
Embodiment 1: the preparation of western loose alkane lactone type diterpenoid beanpod soft coral first, second element
(1) extracts: South China Sea beanpod soft coral dry weight 502g, with acetone 1000ml supersound extraction 3 times repeatedly, extracting solution merges the back concentrating under reduced pressure, the gained crude extract is suspended in the 500ml 1NNaCl solution, extract this suspension repeatedly 3 times with ether 500ml, the gained extraction liquid merges the back concentrating under reduced pressure and obtains ether medicinal extract 2.5g.
(2) separate: ether medicinal extract 2.5g is through 200-300 order silica gel column chromatography, with 100: 0 → 90: 10 → 80: 20 → 50: 50 → 20: 80 → 10: 90 → 0: 100 gradient elution of petroleum ether/ethyl ether, and each gradient consumption 1000ml; Wherein, 50: 50 elutriant enriched materials of petroleum ether/ethyl ether get 146.3mg, and this part is through Sephadex LH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, and developping agent is petroleum ether/ethyl ether 1: 8, with R
fValue merges at the elutriant at 0.25～0.35 place, concentrating under reduced pressure promptly obtains the plain 41.3mg of beanpod soft coral second; 20: 80 elutriant enriched materials of petroleum ether/ethyl ether get 520.0mg, this part is through Sephadex LH-20 gel filtration chromatography, and with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, developping agent sherwood oil/acetone 7: 3 is with R
fValue merges at the elutriant at 0.45～0.55 place, concentrating under reduced pressure promptly obtains the plain 136.8mg of beanpod soft coral first.
The physicochemical character of beanpod soft coral first element is as follows: white solid, optical activity [α]
D 22+ 33.8 ° of (c, 0.535, CHCl
3); 3411 (OH), 1735 (alpha, beta-unsaturated ketones), 1270,1020 (α, beta-unsaturated carboxylic acid ester) cm appear in the IR spectrum
-1Deng absorption peak; The positive ion mode electrospray ionization mass spectrum provides quasi-molecular ion peak m/z 355.2[M+Na]
+And 687.4[2M+Na]
+, high resolution positive ion mode electrospray ionization mass spectrum shows that its molecular formula is C
20H
28O
4M/z 355.1883[M+Na]
+, Δ=-0.2mmu}.
1H with
13C NMR data see Table 1.Simultaneously, by measuring the relevant spectrum of two-dimentional H-H (DQF-COSY), the relevant spectrum of H-C (HMQC), the long-range relevant spectrum of H-C (HMBC) and the relevant spectrum of H-H nuclear space dipole (ROESY), the signal ownership of all carbon atoms and hydrogen atom and the chemical structure of this compound have been determined.
The physicochemical character of beanpod soft coral second element is as follows: colorless oil, optical activity [α]
D 22+ 66.1 (c, 0.324, CHCl
3); 3479 (OH), 1732 (alpha, beta-unsaturated ketones), 1270,1027 absorption peaks such as (α, beta-unsaturated carboxylic acid esters) appear in the IR spectrum; The positive ion mode electrospray ionization mass spectrum provides quasi-molecular ion peak m/z 339.2[M+Na]
+And 655.1[2M+Na]
+, high resolution positive ion mode electrospray ionization mass spectrum shows that its molecular formula is C
20H
28O
3M/z 339.1955[M+Na]
+, Δ=+ 1.9mmu}.
1H with
13C NMR data see Table 2.Simultaneously, by measuring the relevant spectrum of two-dimentional H-H (DQF-COSY), the relevant spectrum of H-C (HMQC), the long-range relevant spectrum of H-C (HMBC) and the relevant spectrum of H-H nuclear space dipole (ROESY), the signal ownership of all carbon atoms and hydrogen atom and the chemical structure of this compound have been determined.
The nmr spectrum data table of table 1. beanpod soft coral first element
A, b
Annotate:
aVarian Inova 600MHz; Solvent C DCl
3, the ppm of δ chemical shift unit,
1H-NMR and
13C-NMR is respectively with chloroform (δ remaining in the solvent
H7.26ppm) and the interior mark of deuterochloroform (δ
C77.0);
bThe ownership of NMR signal is finished on two-dimensional spectrum bases such as HMQC, HMBC;
cFinish the signal ownership by the DEPT spectrum.
The nmr spectrum data table of table 2. beanpod soft coral second element
A, b
Annotate:
aVarian Inova 600MHz; Solvent C DCl
3, the ppm of δ chemical shift unit,
1H-NMR and
13C-NMR is respectively with chloroform (δ remaining in the solvent
H7.26ppm) and the interior mark of deuterochloroform (δ
C77.0);
bThe ownership of NMR signal is finished on two-dimensional spectrum bases such as HMQC, HMBC;
cFinish the signal ownership by the DEPT spectrum.
Experimental example 1
The anti-tumor activity test of loose alkane lactone type diterpenoid beanpod soft coral first, the second element in west
Test philosophy: have the desaturase relevant in the plastosome of mtt assay viable cell with NADP, can (3-(4 with xanchromatic MTT, 5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide) is reduced to insoluble bluish voilet Jia Za (Formazan), this enzyme of dead cell disappears, and MTT is not reduced.Separate with three liquid are molten that available microplate reader detects optical density value at the 570nm place behind the first Za.Optical density value is directly proportional with viable count.
Experimental technique: mtt assay is by different tumour generating rates, some amount is in the human lung adenocarcinoma cell line A-549 of logarithmic phase, mouse leukemia cell strain P-38890 μ l/ hole is inoculated in respectively in 96 well culture plates, cultivate to add after 24 hours and be subjected to reagent liquid (being the DMSO solution of beanpod soft coral first element, beanpod soft coral second element) 10 μ l/ holes, to each cell strain, each concentration is three multiple holes, the fresh culture 10 μ l/ holes of contrast.Tumour cell is at 37 ℃, 5%CO
2Cultivate after 48 hours under the condition, add the freshly prepared 5mg/mlMTT liquid of serum free medium 20 μ l/ holes; Continue to cultivate after 4 hours, (the 50 μ l/ holes of 10%SDS-5% isopropylcarbinol-0.01mol/LHCl) are in CO to add three liquid
2Spend the night in the incubator.Measure the OD value at 570nm with microplate reader then.
Experimental result is as follows:
Sum up: show through the extracorporeal anti-tumor pharmacology activity research, the loose alkane lactone type diterpenoid beanpod soft coral first element in west has significant inhibitory effect to the human lung adenocarcinoma cell line A-549 that is in logarithmic phase, and western loose alkane lactone type diterpenoid beanpod soft coral second element has significant inhibitory effect to human lung adenocarcinoma cell line A-549 and the mouse leukemia cell strain P-388 that is in logarithmic phase.Therefore, western loose alkane lactone type diterpenoid beanpod soft coral first element, second element can be used for preparing the medicine of treatment cancer and/or tumour.
Claims (8)
1. following western loose alkane lactone type diterpenoid beanpod soft coral first element or the second element of a class formation formula:
The plain beanpod soft coral of beanpod soft coral first second element.
2. the preparation method of the described west of claim 1 loose alkane lactone type diterpenoid beanpod soft coral first element or second element is characterized in that this method may further comprise the steps:
1) extracts: South China Sea beanpod soft coral is extracted with ketone solvent, get crude extract after the gained extracting solution concentrates; This crude extract is added in the NaCl solution, and suspendible is the back extracted with diethyl ether evenly, and the gained extraction liquid obtains ether medicinal extract after concentrating;
2) separate: the ether medicinal extract that obtains in the step 1) carries out silica gel column chromatography, with the petroleum ether/ethyl ether gradient elution; Wherein, 50: 50 wash-out parts of petroleum ether/ethyl ether volume ratio through the SephadexLH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out purifying, obtain beanpod soft coral second element; 20: 80 wash-out parts of petroleum ether/ethyl ether volume ratio through the SephadexLH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out purifying, obtain beanpod soft coral first element.
3. preparation method as claimed in claim 2 is characterized in that, in the described step 1), described ketone solvent is an acetone.
4. preparation method as claimed in claim 2 is characterized in that, in the described step 1), described NaCl solution is 0.7～1.5 N NaCl solution.
5. preparation method as claimed in claim 2 is characterized in that, in the described step 1), it is to adopt ultrasonic extraction that described ketone solvent extracts.
6. preparation method as claimed in claim 2, it is characterized in that, described step 2) in, described petroleum ether/ethyl ether gradient elution is to carry out gradient elution in 100: 0 → 90: 10 → 80: 20 → 50: 50 → 20: 80 → 10: 90 → 0: 100 with the petroleum ether/ethyl ether volume ratio.
7. preparation method as claimed in claim 2, it is characterized in that, described step 2) PetroChina Company Limited.'s ether/50: 50 wash-out parts of ether volume ratio, through Sephadex LH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, and developping agent is 1: 8 a petroleum ether/ethyl ether of volume ratio, with elutriant merging, the concentrating under reduced pressure of Rf value, obtain beanpod soft coral second element at 0.25～0.35 place;
Described step 2) 20: 80 wash-out parts of PetroChina Company Limited.'s ether/ether, through Sephadex LH-20 gel filtration chromatography, with 2: 1: 1 sherwood oil of volume ratio/chloroform/methanol wash-out, the gained elutriant detects through thin-layer chromatography, developping agent is sherwood oil/acetone of 7: 3 of volume ratio, with elutriant merging, the concentrating under reduced pressure of Rf value, obtain beanpod soft coral first element at 0.45～0.55 place.
8. the loose alkane lactone type diterpenoid in the described west of claim 1 beanpod soft coral first element or the second element application in preparation treatment cancer and/or tumour medicine.
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|---|---|---|---|
| CN200710038047A CN101265248B (en) | 2007-03-14 | 2007-03-14 | Cembrene lactone type diterpenoid lobophilides A and B, and preparation method and use thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109134184A (en) * | 2017-06-16 | 2019-01-04 | 中国科学院上海药物研究所 | Diterpene-kind compound, preparation method and the usage |
| CN111317748A (en) * | 2018-12-14 | 2020-06-23 | 中山大学 | Coral compound extract, composition containing the same and preparation method thereof |
| CN113402391A (en) * | 2021-05-07 | 2021-09-17 | 宁波大学 | Diterpenoid compound derived from Balanophora japonica, preparation method and application thereof |
-
2007
- 2007-03-14 CN CN200710038047A patent/CN101265248B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109134184A (en) * | 2017-06-16 | 2019-01-04 | 中国科学院上海药物研究所 | Diterpene-kind compound, preparation method and the usage |
| CN109134184B (en) * | 2017-06-16 | 2021-03-19 | 中国科学院上海药物研究所 | Diterpenoid compounds, preparation method and application thereof |
| CN111317748A (en) * | 2018-12-14 | 2020-06-23 | 中山大学 | Coral compound extract, composition containing the same and preparation method thereof |
| US11628191B2 (en) | 2018-12-14 | 2023-04-18 | National Sun Yat-Sen University | Coral composite extract, composition including the same and method of producing the same |
| US11738055B2 (en) | 2018-12-14 | 2023-08-29 | National Sun Yat-Sen University | Coral composite extract, composition including the same and method of producing the same |
| US11963985B2 (en) | 2018-12-14 | 2024-04-23 | National Sun Yat-Sen University | Coral composite extract, composition including the same and method of producing the same |
| CN113402391A (en) * | 2021-05-07 | 2021-09-17 | 宁波大学 | Diterpenoid compound derived from Balanophora japonica, preparation method and application thereof |
| CN113402391B (en) * | 2021-05-07 | 2022-04-26 | 宁波大学 | Diterpenoid compound derived from Balanophora japonica, preparation method and application thereof |
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