CN105294720B - A kind of dimerization chromone alkaloid compound and its preparation method and application - Google Patents
A kind of dimerization chromone alkaloid compound and its preparation method and application Download PDFInfo
- Publication number
- CN105294720B CN105294720B CN201510234377.8A CN201510234377A CN105294720B CN 105294720 B CN105294720 B CN 105294720B CN 201510234377 A CN201510234377 A CN 201510234377A CN 105294720 B CN105294720 B CN 105294720B
- Authority
- CN
- China
- Prior art keywords
- dimerization
- medicinal extract
- cassia
- chromone
- alkaloid compound
- 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.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/06—Peri-condensed systems
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Dentistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of dimerization chromone alkaloid compound and its preparation method and application, the dimerization chromone alkaloid compound is from leguminosae cassia medicinal plant purging cassia（Cassia fistula）Dry bark in it is isolated, its molecular formula be C26H21NO6, it is named as sausage element A, English entitled Fistulain A, with following structural formula：Preparation method is with leguminosae cassia medicinal plant purging cassia（Cassia fistula）Dry bark be raw material, extracted by medicinal extract, organic solvent extraction, MCI decolourings, silica gel column chromatography, high pressure liquid chromatography separating step and obtain.The compounds of this invention is the dimer chromone alkaloid of the first novel framework types by the-N connections of C 14 found from medicinal plant, and have preferable bioactivity, can be as cancer therapy drug and the lead compound of resisting tobacco mosaic virus, with preferable application prospect.
Description
Technical field
The invention belongs to the separation of the extracts active ingredients in national characters medicinal plant, Structural Identification and screening active ingredients
A kind of technical field, and in particular to dimerization chromone alkaloid compound and its preparation method and application.
Background technology
Legume purging cassia（Cassia fistula）, it is the national flower of Thailand, originates in South Asia south, be distributed in Burma
The ground such as pasture, Sri Lanka, south, the west and south of India and China's Mainland, is grown on the area of 1,000 meter of height above sea level.In China,
Purging cassia is a kind of conventional Chinese herbal medicine of the Dai nationality, and the Dai nationality is among the people to be widely used in skin infection, obesity, periodic fever and swell
The treatment of knurl disease etc..And purging cassia is called " pot holds together good " in Dai Nationality's language, its fruit, seed, tree root and bark can be used as medicine, wherein
Containing quinones, with discharge function；Fruit contains tannins compound, and bark can be done for manufacturing leather and red dye
Material etc..The Dai nationality is among the people to be used as medicinal plant and extensive use using fruit, root, branch, leaf and bark.Wherein, the leaf of purging cassia
It is bitter, puckery, it is cool in nature, there is swelling and pain relieving, fall fire toxin expelling, sharp water niter, scattered wind analgesic；The seed of purging cassia is used to treat
Disease in the liver and gallbladder, can also increase appetite；The root of purging cassia is mainly used to treat various skin diseases, leprosy, pulmonary tuberculosis and plum
Poison etc.；Its fruit has certain curative effect to inflammation, hepatopathy, sore-throat, bronchial astehma, rheumatism and rheumatoid arthritis.According to
Report, the different parts of the plant have anti-diabetic, antitumor, anti-inflammatory, antiviral, antibacterial, oxidation resistant activity.Early stage pair
In the chemical constitution study that the plant is carried out, what is mainly obtained is some conventional flavones, triterpene, naphthoquinones, carbohydrate and lignanoid
Class compound.From unlike early stage, isolated from purging cassia one of the present invention is novel passes through the two of C-14-N connections
Aggressiveness chromone alkaloid compound, and the compound has significant cytotoxic activity and antiviral activity.
The content of the invention
The first object of the present invention is to provide a kind of dimerization chromone alkaloid compound；Second purpose is to provide described
The preparation method of dimerization chromone alkaloid compound；3rd purpose is to provide the dimerization chromone alkaloid compound to exist
Prepare the application in anticancer and resisting tobacco mosaic virus medicine.
The first object of the present invention is achieved in that described dimerization chromone alkaloid compound is from dry beans
Section's Cassia medicinal plant purging cassia（Cassia fistula）Bark in it is isolated, its molecular formula be C26H21NO6, have
Following structures：
The compound is yellow jelly, name sausage element A, English entitled Fistulain A.
The second object of the present invention is achieved in that the preparation method of the dimerization chromone alkaloid compound, is
With legume purging cassia（Cassia fistula）Dry bark be raw material, extracted through medicinal extract, organic solvent extraction, MCI
Decolouring, silica gel column chromatography, high pressure liquid chromatography separating step, be specially：
A, medicinal extract are extracted：By legume purging cassia（Cassia fistula）Bark be crushed to 20 ~ 40 mesh, with organic
Solvent supersonic extract 2 ~ 5 times, 30 ~ 60 minutes every time, or cold soaking extract 2 ~ 5 times, 3 days every time, merge extract solution, filtering, subtract
Concentrated extracting solution is pressed, is stood, is filtered sediment, be condensed into medicinal extract a；
B, organic solvent extraction：Water of the weight than 1 ~ 2 times of amount is added in medicinal extract a, with the organic solvent isometric with water
Extraction 3 ~ 5 times, merges organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract b；
C, MCI decolourize：Medicinal extract b adds methanol water dissolves of the weight than 3 ~ 5 times of amounts, and upper MCI posts use 80%-95% methanol-waters
Elution, merges organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract c;
D, silica gel column chromatography：Silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 mesh, consumption be medicinal extract c weight 6 ~
10 times of amounts；Using volume proportion as 1:0~0:1 chloroform and acetone mixed organic solvents gradient elution, collects gradient eluent, dense
Contracting, is monitored through TLC, merges identical part；
E, high performance liquid chromatography separation：The elution that will be afforded with petroleum ether-acetone soln of volume content 50 ~ 90%
Liquid is purified through high performance liquid chromatography separation, produces described dimerization chromone alkaloid compound.
The structure of dimerization chromone compounds prepared by method described above is to identify to come by the following method：
The compounds of this invention is yellow jelly；（1）Ultraviolet spectra（Solvent is methanol）,λ max (log ε）：220
(4.46), 248 (3.68), 276 (3.84), 322 (3.48), 376 (2.92) nm；（2）Infrared spectrum（KBr
Compressing tablet）ν max： 3446, 3158, 2938, 1730, 1647, 1605, 1574, 1442, 1385, 1338, 1276,
1164, 1076, 862, 751 cm–1；（3）HRESIMS shows the compounds of this invention quasi-molecular ion peakm/z 466.1261
[M+Na]+（Calculated value is 466.1261）, with reference to13C and1H H NMR spectroscopies（Fig. 1 and Fig. 2, carbon spectrum hydrogen modal data ownership is shown in Table 1）Give
Go out its molecular formula for C26H21NO6, its degree of unsaturation is 17.1H NMR（CDCl3, 500 MHz）With13C NMR（CDCl3, 125
MHz）Data, are shown in Table 1.1H H NMR spectroscopies（Fig. 2）Show 3 methyl signals（δ H 2.26, s, H3-13; 2.38, s, H3-
9'; 2.51, s, H3-12'）, 2 methylene signals（δ H 3.45, s, H2-14; 4.24, s, H2-11）, 5 unimodal
Aromatic signal（δ H 6.21, s, H-3; 6.53, s, H-6'; 6.78, s, H-8'; 6.85, s, H-3';
6.93, s, H-10'）And a substituted benzene ring signal of 1,2,3,5- tetra-（δ H 6.59, d, J = 1.8 Hz, H-6;
6.66, d, J= 1.8 Hz, H-8）.In carbon spectrum and DEPT spectrums（Fig. 1）In observed 26 carbon atom signals, wherein 3
Methyl（Including two fragrant methyl）, 2 methylene, 7 fragrant methines and 14 quaternary carbon signals（Include 3 carbonyls and 5
Individual oxygen-containing quaternary carbon signal）.Wherein, 3 carbonyls and 18 double key carbons occupy 12 degrees of unsaturation, so the molecule is a height
The pentacyclic alkaloid of aromatization.Spectral data above compound isolated from Cassia before combining, initial guess,
Sausage element A is a heterodimer Alkaloid being made up of two different C13 skeletons.It is made up of A and B fragments.
According to A fragments（O-1 to C-14）Characteristic signal（δ C 166.4, 113.1, 180.1, 138.5,
121.1, 163.8, 103.6, 161.4, 115.9; δ H6.21, 6.59, 6.66）, thus it is speculated that it is a C13 skeleton
Chromone compounds（A and B rings）.This supposition is confirmed by HMBC correlations, i.e., H-3 is related to C-2/C-4/C-10, H-6
Related to C-8/C-10, H-8 is related to C-6/C-10（Fig. 3）.Secondly, also 1 acetonyl and 1 oh group are connected respectively
On C-5 and C-7, this H in being composed also by HMBC2- 11 is related to C-6/C-10, and 7-OH related to C-6/C-7/C-8
Confirmed.These data illustrate that the A fragments of compound are a 5- acetonyl -7- hydroxyl -2- chromone derivative, and its is main
Difference is C-14 and becomes methylene by original methyl, thus it is speculated that A fragments are connected by C-14 and B fragments.It is remaining
13 carbon atoms, include 2 methyl, and 4 methines, 7 quaternary carbon signals are under the jurisdiction of B fragments.HMBC correlations give B pieces
The general configuration of section.In HMBC spectrums, H-6' and C-5'/C-7'/C-8', H-8' and C-6'/C-7'/C-8a' have been observed,
H3- 9' and C-2'/C-3'/C-8a', H-3' and C-2'/C-4a'/C-9' and H-6' and C-4a'/C-5'/C-7'/C-8'/C-
10' correlation, so as to confirm the basic fragment of chromone in B fragments（C and D rings）.Meanwhile, also observe H- in HMBC spectrums
10' is related to C-4a'/C-5'/C-6'/C-11'/C-12''s and H-12' and C-14/C-4'/C-10'/C-11', this explanation
C-4', C-4a', C-5', C-10', C-11', C-12' and N atom together form a 2- pyridine derivatives piece
Section（Ring E）, it is combined by C-4', C-4a', and C-5' with C and D ring a pair of horses going side by sides.Finally, H is passed through2- 14 and C-4'/C-
11' and H3- 12' C-14 HMBC is related, it is determined that the order of connection of A and B fragments, so far, the dimerization chromone alkaloid
Class compound structure is determined.
The compound of table 11H and13C NMR datas（Solvent is CDCl3）(125 and 500 MHz)
| No. | 13C | 1H |
| 2 | 166.4 s | |
| 3 | 113.1 d | 6.21, s |
| 4 | 180.1 s | |
| 5 | 138.5 s | |
| 6 | 121.1 d | 6. 59 d (1.8) |
| 7 | 163.8 s | |
| 8 | 103.6 d | 6.66 d (1.8) |
| 9 | 161.4 s | |
| 10 | 115.9 s | |
| 11 | 50.5 t | 4.24 s |
| 12 | 207.9 s | |
| 13 | 30.8 q | 2.26 s |
| 14 | 58.2 t | 3.45 s |
| 2′ | 168.3 s | |
| 3′ | 98.2 d | 6.85s |
| 4′ | 146.3 s | |
| 4a′ | 110.2 s | |
| 5′ | 136.2 s | |
| 6′ | 108.3 d | 6.53s |
| 7′ | 175.1s | |
| 8′ | 105.3 d | 6.78 s |
| 8a′ | 155.9 s | |
| 9′ | 22.1q | 2.38 s |
| 10′ | 116.8 d | 6.93 s |
| 11′ | 141.6 s | |
| 12′ | 21.6 q | 2.51 s |
| 7-OH | 10.21 s |
The third object of the present invention be achieved in that described dimerization chromone alkaloid compound prepare anticancer and
Application in antiviral drugs.
The compounds of this invention is separated from purging cassia bark first, passes through nuclear magnetic resonance and measuring method of mass spectrum
It is defined as dimerization chromone alkaloid compound, and characterizes its concrete structure.Using the compounds of this invention as raw material, through dialogue blood
The acute early children grain NB4 cells of disease, Lung Adenocarcinoma A 549 Cell, people marrow neuroblastoma SHSY5Y cells, human prostata cancer PC3
Cell, the cytotoxic activity of human breast cancer MCF7 cell are respectively up to 5.5,4.6 and 8.8μM.The compounds of this invention is through resisting cigarette
The experiment of showy flowers of herbaceous plants mosaic virus, its relative inhibition is 20μM is issued to 32.8%, more than the relative suppression of positive reference substance Nanning mycin
Rate (30.5%) processed, its IC50It is worth for 43.8μM, activity is better than positive reference substance Nanning mycin (IC50=52.4μM), chemical combination is illustrated
Thing has good activity of resisting tobacco mosaic virus.
The compounds of this invention structure novel active is good, can be as anticancer and the guiding compound of antiviral drugs, in medicine
Thing is found and biological pesticide preparation aspect has good application prospect.
Brief description of the drawings
Fig. 1 is compound sausage element A carbon-13 nmr spectra（13C NMR）；
Fig. 2 is compound sausage element A proton nmr spectra（1H NMR）；
Fig. 3 compounds sausage element A main HMBC is related.
Embodiment
The present invention is further illustrated below in conjunction with the accompanying drawings, but the present invention is not any limitation as in any way, base
In present invention teach that any conversion or improvement made, each fall within protection scope of the present invention.
Dimerization chromone alkaloid compound of the present invention, is from legume Cassia purging cassia（Cassia fistula）Dry bark in it is isolated, its molecular formula be C26H21NO6, with following structures：
The compound is yellow jelly, is named as sausage element A, English entitled Fistulain A.
The preparation method of dimerization chromone alkaloid compound of the present invention, is with legume purging cassia（Cassia
fistula）Dry bark be raw material, extracted through medicinal extract, organic solvent extraction, MCI decolourings, silica gel column chromatography, efficient liquid phase
Chromatogram preparative separation step, be specially：
A, medicinal extract are extracted：By legume purging cassia（Cassia fistula）Bark be crushed to 20 ~ 40 mesh, with organic
Solvent supersonic extract 2 ~ 5 times, 30 ~ 60 minutes every time, or cold soaking extract 2 ~ 5 times, 3 days every time, merge extract solution, filtering, subtract
Concentrated extracting solution is pressed, is stood, is filtered sediment, be condensed into medicinal extract a；
B, organic solvent extraction：Water of the weight than 1 ~ 2 times of amount is added in medicinal extract a, with the organic solvent isometric with water
Extraction 3 ~ 5 times, merges organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract b；
C, MCI decolourize：Medicinal extract b adds methanol water dissolves of the weight than 3 ~ 5 times of amounts, and upper MCI posts use 80%-95% methanol-waters
Elution, merges organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract c;
D, silica gel column chromatography：Silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 mesh, consumption be medicinal extract c weight 6 ~
10 times of amounts；Using volume proportion as 1:0~0:1 chloroform and acetone mixed organic solvents gradient elution, collects gradient eluent, dense
Contracting, is monitored through TLC, merges identical part；
E, high performance liquid chromatography separation：The elution that will be afforded with petroleum ether-acetone soln of volume content 50 ~ 90%
Liquid is purified through high performance liquid chromatography separation, produces described dimerization chromone alkaloid compound.
The organic solvent of the step A is 90 ~ 100% methanol, 90 ~ 100% ethanol or 70 ~ 100% acetone.
The organic solvent of the step B is ethyl acetate, ether, petroleum ether, chloroform or dichloromethane.
The medicinal extract c of the D steps is before through silica gel column chromatography, methanol or acetone solution with weight than 1.5 ~ 3 times of amounts,
Then 0.8 ~ 1.2 times of 80 ~ 100 mesh silica gel mixed samples are weighed with medicinal extract,.
The chloroform of the D steps and the volume proportion of acetone mixed organic solvents are 20:1, 9:1, 8:2, 7:3, 6:4
With 1:1.
The high performance liquid chromatography separation purifying of the E steps is using 40 ~ 70% methanol or 20 ~ 60% acetonitrile as flowing
Phase, flow velocity 10 ~ 14ml/min, 21.2 × 250 mm, 5 μm of Zorbax PrepHT GF reverse phase preparative columns are stationary phase, purple
External detector Detection wavelength is 202 ~ 280nm, each μ L of sample introduction 10 ~ 100,10 ~ 40min chromatographic peak is collected, after repeatedly adding up
It is evaporated.
Application of the dimerization chromone alkaloid compound of the present invention in cancer therapy drug is prepared.
Application of the dimerization chromone compounds of the present invention in resisting tobacco mosaic virus medicine is prepared.
Cassia plant of the present invention is not limited by area and kind, can realize the present invention.
Embodiment 1
Take dry legume Cassia purging cassia（Cassia fistula）Bark 2.2kg, meal is broken to 40 mesh, uses
70% acetone ultrasonic extraction 4 times, 60 minutes every time, extract solution merged；Extract solution is filtered, and is concentrated under reduced pressure into the 1/4 of volume；It is quiet
Put, filter sediment, be condensed into 63.5g medicinal extract a；130g water is added in medicinal extract a, with the chloroform extraction 5 isometric with water
It is secondary, merge extraction phase, be concentrated under reduced pressure into 40g medicinal extract b；Medicinal extract b fills post with MCI, and 120g 80% methanol-water is added in medicinal extract b
Dissolving, then upper prop, with 90% methanol-water, 1 to 4 liter of elution, collects eluent, is concentrated under reduced pressure to give 31.2g medicinal extract c；Medicinal extract c exists
62g acetone solution is added in medicinal extract c, 100 mesh silica gel 32g is then added and mixes sample, mix after sample, post is filled with 200 mesh silica gel 200g；
It is respectively 20 with volume ratio:1, 9:1, 8:2, 7:3, 6:4 and 1:1 chloroform-acetone mixed organic solvents gradient elution,
Gradient eluent, concentration are collected, is monitored through TLC, merges identical part, obtains 6 part A-F, wherein, to the sample being collected into
Product part B 6.42g, repeats silica gel column chromatography, with volume ratio 9:1-1:2 petroleum ether-acetone mixed organic solvents gradient is washed
It is de-, gradient eluent, concentration are collected, is monitored through TLC, merges identical part, 6 part B1-B6 is obtained, wherein B3 portions
Point, i.e., 7:The mg of 3 parts about 650, then using 58% methanol as mobile phase, ml/min, 21.2 × 250mm, 5 μm of flow velocity 10
Zorbax PrepHT GF reverse phase preparative columns are stationary phase, and UV-detector Detection wavelength is 254 nm, each μ L of sample introduction 50, is received
Collect 25 min chromatographic peak, be evaporated after repeatedly adding up, produce described dimerization chromone alkaloid compound sausage element A.
Embodiment 2
Take dry legume Cassia purging cassia（Cassia fistula）The kg of bark 10.0, meal is broken to 40 mesh,
Extracted 4 times with 80% methanol cold soaking, 3 days every time, extract solution merged；Extract solution is filtered, and is concentrated under reduced pressure into the 1/4 of volume；It is quiet
Put, filter sediment, be condensed into 300g medicinal extract a；350g water is added in medicinal extract a, with the ethyl acetate extraction 5 isometric with water
It is secondary, merge extraction phase, be concentrated under reduced pressure into 210g medicinal extract b；Medicinal extract b fills post with MCI, and 600g 80% methanol-water is added in medicinal extract b
Dissolving, then upper prop, with 90% methanol-water, 5 to 15 liters of elutions, collects eluent, is concentrated under reduced pressure to give 150g medicinal extract c；In medicinal extract c
300g acetone solution is added, 100 mesh silica gel 150g is then added and mixes sample, post is filled with 200 mesh silica gel 1Kg, mixes upper prop after sample；With
Volume ratio is respectively 20:1, 9:1, 8:2, 7:3, 6:4 and 1:1 chloroform-acetone mixed organic solvents gradient elution, is received
Collect gradient eluent, concentration, monitored through TLC, merge identical part, obtain 6 part A-F, wherein, to the sample being collected into
Part B 32g, repeats silica gel column chromatography, with volume ratio 9:1-1:2 petroleum ether-acetone mixed organic solvents gradient elution, is received
Collect gradient eluent, concentration, monitored through TLC, merge identical part, 6 part B1-B6 are obtained, wherein B3 parts, i.e., 7:
The g of 3 parts about 2.8, then using 45% acetonitrile as mobile phase, the Zorbax that ml/min, 21.2 × 250mm, 5 μm of flow velocity 10
PrepHT GF reverse phase preparative columns are stationary phase, and UV-detector Detection wavelength is 254 nm, each μ L of sample introduction 80, collects 15
Min chromatographic peak, is evaporated after repeatedly adding up, and produces described dimerization chromone alkaloid compound sausage element A.
Embodiment 3
Compound sausage element A prepared by Example 1, is yellow jelly；
Assay method is：With nuclear magnetic resonance, structure is identified with reference to other spectroscopic techniques.
（1）Ultraviolet spectra（Solvent is methanol）,λ max (log ε）：220 (4.46), 248 (3.68), 276
(3.84), 322 (3.48), 376 (2.92) nm；
（2）Infrared spectrum（Pressing potassium bromide troche）ν max： 3446, 3158, 2938, 1730, 1647, 1605,
1574, 1442, 1385, 1338, 1276, 1164, 1076, 862, 751 cm–1；
（3）HRESIMS shows the compounds of this invention quasi-molecular ion peakm/z 466.1261 [M+Na]+（Calculated value is
466.1261）, with reference to13C and1H H NMR spectroscopies（Fig. 1 and Fig. 2, carbon spectrum hydrogen modal data ownership is shown in Table 1）Providing its molecular formula is
C26H21NO6, its degree of unsaturation is 17.1H NMR（CDCl3, 500 MHz）With13C NMR（CDCl3, 125 MHz）Data, are shown in Table
1。1H H NMR spectroscopies（Fig. 2）Show 3 methyl signals（δ H 2.26, s, H3-13; 2.38, s, H3-9'; 2.51, s,
H3-12'）, 2 methylene signals（δ H 3.45, s, H2-14; 4.24, s, H2-11）, 5 unimodal aromatic signals（δ H
6.21, s, H-3; 6.53, s, H-6'; 6.78, s, H-8'; 6.85, s, H-3'; 6.93, s, H-10'）With
And a substituted benzene ring signal of 1,2,3,5- tetra-（δ H 6.59, d, J = 1.8 Hz, H-6; 6.66, d, J = 1.8
Hz, H-8）.In carbon spectrum and DEPT spectrums（Fig. 1）In observed 26 carbon atom signals, wherein 3 methyl（Including two fragrance
Methyl）, 2 methylene, 7 fragrant methines and 14 quaternary carbon signals（Include 3 carbonyls and 5 oxygen-containing quaternary carbon signals）.
Wherein, 3 carbonyls and 18 double key carbons occupy 12 degrees of unsaturation, so the molecule is the five rings biology of a height aromatization
Alkali.More than spectral data combine before compound isolated from Cassia, initial guess, sausage element A be one by
The heterodimer Alkaloid of two different C13 skeletons compositions.It is made up of A and B fragments.
According to A fragments（O-1 to C-14）Characteristic signal（δ C 166.4, 113.1, 180.1, 138.5,
121.1, 163.8, 103.6, 161.4, 115.9; δ H6.21, 6.59, 6.66）, thus it is speculated that it is a C13 skeleton
Chromone compounds（A and B rings）.This supposition is confirmed by HMBC correlations, i.e., H-3 is related to C-2/C-4/C-10, H-6
Related to C-8/C-10, H-8 is related to C-6/C-10（Fig. 3）.Secondly, also 1 acetonyl and 1 oh group are connected respectively
On C-5 and C-7, this H in being composed also by HMBC2- 11 is related to C-6/C-10, and 7-OH related to C-6/C-7/C-8
Confirmed.These data illustrate that the A fragments of compound are a 5- acetonyl -7- hydroxyl -2- chromone derivative, and its is main
Difference is C-14 and becomes methylene by original methyl, thus it is speculated that A fragments are connected by C-14 and B fragments.It is remaining
13 carbon atoms, include 2 methyl, and 4 methines, 7 quaternary carbon signals are under the jurisdiction of B fragments.HMBC correlations give B pieces
The general configuration of section.In HMBC spectrums, H-6' and C-5'/C-7'/C-8', H-8' and C-6'/C-7'/C-8a' have been observed,
H3- 9' and C-2'/C-3'/C-8a', H-3' and C-2'/C-4a'/C-9' and H-6' and C-4a'/C-5'/C-7'/C-8'/C-
10' correlation, so as to confirm the basic fragment of chromone in B fragments（C and D rings）.Meanwhile, also observe H- in HMBC spectrums
10' is related to C-4a'/C-5'/C-6'/C-11'/C-12''s and H-12' and C-14/C-4'/C-10'/C-11', this explanation
C-4', C-4a', C-5', C-10', C-11', C-12' and N atom together form a 2- pyridine derivatives piece
Section（Ring E）, it is combined by C-4', C-4a', and C-5' with C and D ring a pair of horses going side by sides.
Finally, H is passed through2- 14 and C-4'/C-11' and H3- 12' C-14 HMBC is related, it is determined that A and B fragments
The order of connection, so far, the dimerization chromone alkaloid compound structure are determined, and are named as sausage element A.
The compound of table 11H and13C NMR datas（Solvent is CDCl3）(125 and 500 MHz)
| No. | 13C | 1H |
| 2 | 166.4 s | |
| 3 | 113.1 d | 6.21, s |
| 4 | 180.1 s | |
| 5 | 138.5 s | |
| 6 | 121.1 d | 6. 59 d (1.8) |
| 7 | 163.8 s | |
| 8 | 103.6 d | 6.66 d (1.8) |
| 9 | 161.4 s | |
| 10 | 115.9 s | |
| 11 | 50.5 t | 4.24 s |
| 12 | 207.9 s | |
| 13 | 30.8 q | 2.26 s |
| 14 | 58.2 t | 3.45 s |
| 2′ | 168.3 s | |
| 3′ | 98.2 d | 6.85s |
| 4′ | 146.3 s | |
| 4a′ | 110.2 s | |
| 5′ | 136.2 s | |
| 6′ | 108.3 d | 6.53s |
| 7′ | 175.1s | |
| 8′ | 105.3 d | 6.78 s |
| 8a′ | 155.9 s | |
| 9′ | 22.1q | 2.38 s |
| 10′ | 116.8 d | 6.93 s |
| 11′ | 141.6 s | |
| 12′ | 21.6 q | 2.51 s |
| 7-OH | 10.21 s |
Embodiment 4
Compound prepared by Example 2, is yellow jelly.Determine identical with implementing 3, confirm to implement 2 changes prepared
Compound is the dimerization chromone alkaloid compound --- sausage element A..
Embodiment 5
Any dimerization chromone alkaloid compound prepared by Example 1 and 2 carries out cytotoxicity assay experiment,
Test situation is as follows：
Cell line：Leukaemia (NB4), lung carcinoma cell (A549), human neuroblastoma cells (SHSY5Y), prostatitis
Adenocarcinoma cell (PC3), breast cancer cell (MCF7) are provided by Shanghai Pharmaceutical Inst., Chinese Academy of Sciences.
Experimental design：Above cell and various concentrations compound incubation 72 hours, the experiment of every plant of cell repeat one
It is secondary, data processing is carried out with the result tested twice, compound on intracellular propagation is evaluated using improvement MTT methods and SRB methods
Inhibition level, calculate inhibiting rate, according to inhibiting rate using Logit methods calculate IC50, comparative compound it is external anti-swollen
Tumor activity.
The proliferation inhibition rate of cell=(the OD values of blank control OD values-medicine feeding hole)/blank control OD value × 100%.
(a) MTT methods are improved
The suspension cell in exponential phase is taken, cell concentration is adjusted to 4 × 104/ ml, adds the culture of 96 holes
Plate, 90µL/ holes.Positive control is cis-platinum, uses physiological saline solution.The sample (1 of 10 μ l various concentrations is separately added into per hole
Number-No. 5 test solutions of test solution).Sample-adding group and control group are all provided with 4 multiple holes, and sample-adding group, the high concentration group of positive controls also set training
The dosing parallel hole of base is supported, every block of plate is equipped with 4 blank control wells (only plus culture medium).The final concentration of sample is respectively
10-2、10-1, 1,10 and 102 µG/mL, corresponding DMSO final concentration are respectively 0.1%, 0.01%, 0.001%, 0.0001%,
0.00001%.Sample is in final concentration 102 µDuring g/mL, with 0.1% DMSO as solvent control, remaining concentration uses physiology salt
Water makees negative control.Final concentration of the 10 of positive control drug cis-platinum-1、1、10 µg/mL.Cell is in 37 DEG C, 5% CO2Incubator
It is middle to be incubated respectively after 48h, add MTT (5 mg/ml, Sigma), 10µL/ holes.Continue to cultivate after 4 h, add three
Liquid [10% SDS -5% isobutanol -0.012mol/L HCL (w/v/v)], 100µL/ holes, use enzyme mark after standing overnight
Instrument determines the OD values in each hole under 570 nm, 630 nm dual wavelengths.
(b) SRB methods
The attached cell strain in exponential phase is taken, it is complete after 25% pancreatin conventional digestion, then with 15% calf serum
Cell concentration is adjusted to 5 × 10 by full RPMI-1640 culture mediums4/ mL, adds 96 well culture plates, 90μL/ holes.Cell exists
37℃, 5% CO2Addition positive control, negative control and given the test agent after 24h are incubated in incubator respectively (each tested dense
Spend ibid MTT methods, 10μL/ holes), the final concentration of sample is respectively 10-2、10-1、1、10、102 μG/mL, corresponding DMSO end
Concentration is respectively 0.1%, 0.01%, 0.001%, 0.0001%, 0.00001%.Sample is in final concentration 102 μ0.1%DMSO is used during g/mL
As solvent control, remaining concentration makees negative control with physiological saline.Final concentration of the 10 of positive control drug cis-platinum-1、1、10
μ g/mL, negative control is isometric physiological saline.Sample-adding group and control group are all provided with 4 multiple holes, sample-adding group, positive controls
High concentration group also set the dosing parallel hole of culture medium, every block of plate is equipped with 4 blank control wells (only plus culture medium).By 96
Well culture plate is placed in 37 DEG C, 5% CO2Be incubated in incubator after (cell and sample effect) 48h, add 4 DEG C, 50%
TCA (trichloroacetic acid) 50μL/ holes.Add after TCA, 96 well culture plates are placed in into 4 DEG C is incubated 1 hour, takes out culture plate,
Gently incline liquid in plate.5 times are gently rinsed with running water (by running water by gently being poured into beaker in plate, after light rolling again
Water is fallen), it is placed in air and air-dries to loseing washmarking.Then 0.4% SRB (being diluted with 1% acetic acid) prepared is added,
50 μL/ holes, at room temperature stand dyeing hypsokinesis in 30 minutes remove SRB solution, with 1% acetic acid rinse 4 times, with remove not with egg
The dyestuff that white matter is combined.It is placed in air and air-dries to after without washmarking, adds 10 mM and do not buffer Tris (slow blood ammonia acid) solution
150 μL/ holes (PH10 is prepared with tri-distilled water), after dyestuff is dissolved, in being vibrated 5 minutes on oscillator, are existed with ELIASA
Each hole OD values are read under 570nm wavelength.
(c) experimental result
Test result indicate that：Through female to the early children grain NB4 cells of Leukemia acute, Lung Adenocarcinoma A 549 Cell, people's marrow nerve
Cytoma SHSY5Y cells, human prostata cancer PC3 cells, the cytotoxic activity experiment of human breast cancer MCF7 cell, A pairs of sausage element
NB4, A549 and MCF7 cell line have preferable cytotoxic activity, IC50Value is respectively up to 5.5,4.6 and 8.8μM。
The compound sausage element A of table 2 cytotoxic activity
| Compounds | NB4 | A549 | SHSY5Y | PC3 | MCF7 |
| Sausage element A | 5.5 | 4.6 | >10 | >10 | 8.8 |
| Taxol | 0.02 | 0.02 | 0.1 | 0.1 | 0.05 |
Embodiment 6
Dimerization chromone alkaloid compound prepared by Example 1 and 2 carries out activity of resisting tobacco mosaic virus experiment,
Test situation is as follows：
Using half leaf method, anti-Tobacco mosaic is carried out to the compounds of this invention when the mass concentration of medicament is 50 mg/L
Virus activity is determined.On the plant of 5~6 age flue-cured tobaccos, the blade (leaf row is normal, disease-free without worm) suitable for test is chosen, first
Blade is uniformly sprinkled into fine emery powder, standby tobacco mosaic virus (TMV) source (3.0 × 10-3) is uniformly put on sprinkled with gold with writing brush
On the blade of emery, connect after the blade of all middle choosings after poison terminates, be immediately placed in the culture dish for fill decoction and handle 20 min,
Take out, wipe the globule and decoction on blade, the recovery of two and half leaves is emitted on and is covered with the glass jar of toilet paper moisturizing, and is covered
Glass cover, temperature control (23 ± 2) DEG C is placed on greenhouse natural light irradiation, and 2~3 d are that each processing sets second half to visible withered spot
Leaf is control, is additionally provided with 1 group of processing for commodity Ningnanmycin as a comparison, presses formula and calculates relative inhibition.
XI%=(CK-T)/CK × 100%
X：Relative inhibition (%), CK：The withered spot number (individual) that half in clear water connects malicious leaf is soaked in, T is soaked in decoction
Half connects the withered spot number (individual) of malicious leaf.
Through the experiment to resisting tobacco mosaic virus, its relative inhibition is issued to 32.8% at 20 μM, more than positive reference substance
The relative inhibition (30.5%) of Nanning mycin, its IC50It is worth for 43.8 μM, activity is better than positive reference substance Nanning mycin (IC50=
52.4 μM), illustrate that compound has good activity of resisting tobacco mosaic virus.
Claims (5)
1. a kind of dimerization chromone alkaloid compound, it is characterised in that the dimerization chromone compounds have following structures：
。
2. the preparation method of dimerization chromone alkaloid compound described in a kind of claim 1, it is characterised in that with legume
Purging cassia（Cassia fistula）Dry bark be raw material, extracted through medicinal extract, organic solvent extraction, MCI decolourize, silicagel column
Chromatography, high performance liquid chromatography preparative separation step, be specially：
A, medicinal extract are extracted：By legume purging cassia（Cassia fistula）Bark be crushed to 20 ~ 40 mesh, use 70% acetone
Or 80% methanol ultrasonic extraction 2 ~ 5 times, 30 ~ 60 minutes every time, or cold soaking extracts 2 ~ 5 times, 3 days every time, merges extract solution, mistake
Filter, be concentrated under reduced pressure extract solution, stands, filters sediment, be condensed into medicinal extract a；
B, organic solvent extraction：Water of the weight than 1 ~ 2 times of amount is added in medicinal extract a, with the ethyl acetate or chlorine isometric with water
It is imitative to extract 3 ~ 5 times, merge organic solvent extraction phase, be concentrated under reduced pressure into medicinal extract b；
C, MCI decolourize：Medicinal extract b adds the methanol water dissolves that weight is measured than 3 ~ 5 times, upper MCI posts, with 80%-95% methanol water elutions,
Merge organic solvent extraction phase, be concentrated under reduced pressure into medicinal extract c;
D, silica gel column chromatography：Silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 mesh, and consumption is 6 ~ 10 times of medicinal extract c weight
Amount；It is respectively 20 with volume ratio:1, 9:1, 8:2, 7:3, 6:4 and 1:1 chloroform-acetone mixed organic solvents gradient is washed
It is de-, gradient eluent, concentration are collected, is monitored through TLC, merges identical part, 6 part A-F is obtained, to the B portions being collected into
Divide and repeat silica gel column chromatography, with volume ratio 9:1-1:2 petroleum ether-acetone mixed organic solvents gradient elution, collects gradient and washes
De- liquid, concentration, are monitored through TLC, merge identical part；
E, high performance liquid chromatography separation：By the volume ratio 7 being collected into:3 petroleum ethers-acetone mixed organic solvents elution fraction is through height
Effect liquid phase chromatogram is isolated and purified, and high performance liquid chromatography separation purifying is using 40 ~ 70% methanol or 20 ~ 60% acetonitrile as flowing
Phase, the Zorbax PrepHT GF reverse phase preparative columns of 10 ~ 14ml/min, 21.2 × 250mm, 5 μm of flow velocity are stationary phase, ultraviolet inspection
It is 202 ~ 280nm, each μ L of sample introduction 10 ~ 100 to survey device Detection wavelength, collects 10 ~ 40min chromatographic peak, is evaporated after repeatedly adding up,
Produce the dimerization chromone alkaloid compound.
3. the preparation method of dimerization chromone alkaloid compound according to claim 2, it is characterised in that the D steps
Medicinal extract c is before through silica gel column chromatography, and then methanol or acetone solution with weight than 1.5 ~ 3 times of amounts weigh 0.8 ~ 1.2 with medicinal extract
80 ~ 100 mesh silica gel mixed samples again.
4. application of the dimerization chromone alkaloid compound in cancer therapy drug is prepared described in a kind of claim 1.
5. a kind of the answering in resisting tobacco mosaic virus medicine is prepared of dimerization chromone alkaloid compound described in claim 1
With.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510234377.8A CN105294720B (en) | 2015-05-08 | 2015-05-08 | A kind of dimerization chromone alkaloid compound and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510234377.8A CN105294720B (en) | 2015-05-08 | 2015-05-08 | A kind of dimerization chromone alkaloid compound and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105294720A CN105294720A (en) | 2016-02-03 |
| CN105294720B true CN105294720B (en) | 2017-07-18 |
Family
ID=55192625
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510234377.8A Expired - Fee Related CN105294720B (en) | 2015-05-08 | 2015-05-08 | A kind of dimerization chromone alkaloid compound and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105294720B (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106344577A (en) * | 2016-09-27 | 2017-01-25 | 全椒先奇医药科技有限公司 | Application of Fistulains A in anti-helicobacter pylori drugs |
| CN107865867A (en) * | 2016-09-27 | 2018-04-03 | 全椒先奇医药科技有限公司 | Applications of the Fistulains A in hypoglycemic medicament is reduced |
| CN106420746A (en) * | 2016-09-28 | 2017-02-22 | 南京康凯生物科技有限公司 | Application of Fistulains A in preparing medicine for treating nasopharynx cancer |
| CN106344578A (en) * | 2016-09-28 | 2017-01-25 | 南京康凯生物科技有限公司 | Application of Fistulains A in anti-hypoxic drugs |
| CN106344579A (en) * | 2016-09-29 | 2017-01-25 | 南京康凯生物科技有限公司 | Application of Fistulains A in drugs for myocardial ischemia treatment |
| CN106344580A (en) * | 2016-09-30 | 2017-01-25 | 南京康凯生物科技有限公司 | Application of Fistulains A in drugs for rheumatoid arthritis treatment |
| CN106309440A (en) * | 2016-09-30 | 2017-01-11 | 南京康凯生物科技有限公司 | Application of Fistulains A in preparation of antidepressants |
| CN106309441A (en) * | 2016-10-12 | 2017-01-11 | 南京康凯生物科技有限公司 | Application of Fistulains A in preparation of drugs for treating cervical cancer |
| CN107032867A (en) * | 2016-11-16 | 2017-08-11 | 徐州得铸生物科技有限公司 | A kind of composite biological fertilizer feed additives containing fistulainA |
| CN106551936A (en) * | 2016-11-18 | 2017-04-05 | 苏州枫灵医药科技有限公司 | One kind treats rhinitis medicine and its application |
| CN106580988A (en) * | 2016-11-24 | 2017-04-26 | 全椒先奇医药科技有限公司 | Drug for treating and preventing renal fibrosis and application thereof |
| CN106966888A (en) * | 2017-04-28 | 2017-07-21 | 云南民族大学 | A kind of preparation method and application of the novel anthraquinone analog compound with activity of resisting tobacco mosaic virus |
| CN107467712B (en) * | 2017-07-28 | 2019-05-21 | 云南中烟工业有限责任公司 | A kind of tobacco sauce additive and its application with antibacterial activity |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3870600A (en) * | 1971-03-29 | 1975-03-11 | Kamal Abdou Youssef | Selective and enrichment medium for the isolation identification and propagation of yeasts and fungi |
| CN104262316A (en) * | 2014-09-15 | 2015-01-07 | 云南民族大学 | Flavonoid compound as well as preparation method and application thereof |
| CN104292202A (en) * | 2014-09-15 | 2015-01-21 | 云南民族大学 | Flavonoid compound as well as preparation method and application of flavonoid compound |
-
2015
- 2015-05-08 CN CN201510234377.8A patent/CN105294720B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3870600A (en) * | 1971-03-29 | 1975-03-11 | Kamal Abdou Youssef | Selective and enrichment medium for the isolation identification and propagation of yeasts and fungi |
| CN104262316A (en) * | 2014-09-15 | 2015-01-07 | 云南民族大学 | Flavonoid compound as well as preparation method and application thereof |
| CN104292202A (en) * | 2014-09-15 | 2015-01-21 | 云南民族大学 | Flavonoid compound as well as preparation method and application of flavonoid compound |
Non-Patent Citations (2)
| Title |
|---|
| Cassiarins C-E, Antiplasmodial Alkaloids from the Flowers of Cassia siamea;Shiori Oshimi，等;《J. Nat. Prod.》;20090916;第72卷(第10期);第1899-1901页 * |
| Interactions of antiparasitic alkaloids with Leishmania protein targets: a molecular docking analysis;Ifedayo Victor Ogungbe，等;《Future Med. Chem.》;20131231;第5卷(第15期);第1777-1799页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105294720A (en) | 2016-02-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105294720B (en) | A kind of dimerization chromone alkaloid compound and its preparation method and application | |
| CN104860912B (en) | Dimer ketone compound and preparation method and application thereof | |
| CN105884621B (en) | A kind of sesquiterpenoids and its preparation method and application | |
| CN102796066B (en) | Flavone compound and preparation method and application thereof | |
| CN101123880A (en) | Anti-tumor compounds with angeloyl groups | |
| CN105348192B (en) | Isoquinoline alkaloids bases compound of antiviral activity and preparation method thereof in a kind of wing pod Cassia tora | |
| CN103130644A (en) | Method of obtaining and separating rosmarinic acid, apigenin and luteolin from elsholtzia haichowensis | |
| CN103665082B (en) | Hemsleya cucurbitane tetracyclic triterpenoid compound, pharmaceutical compositions containing same and application of compound and pharmaceutical composition | |
| CN107011143A (en) | The preparation method and application of two kinds of anthraquinone analog compounds | |
| CN103626824B (en) | Hemsleya amabilis cucurbitane tetracyclic triterpene compound, pharmaceutical composition comprising compound and application of pharmaceutical composition and compound | |
| CN104292203B (en) | A kind of Isocoumarin compounds and its preparation method and application | |
| CN104357505B (en) | Ketone alkaloid of a kind of indoles nafoxidine 2 and preparation method and application | |
| CN106966888A (en) | A kind of preparation method and application of the novel anthraquinone analog compound with activity of resisting tobacco mosaic virus | |
| CN109265423A (en) | A kind of chromone compounds and its preparation method and application | |
| CN107513049A (en) | Euphorbia diterpenoids moleplant seed diterpene A preparation method and application | |
| CN104262316B (en) | A kind of flavonoid compound and its preparation method and application | |
| CN105348247B (en) | A kind of isocoumarin class compound and its preparation method and application | |
| CN106986881A (en) | A kind of preparation method and application of isobenzofuran class compound | |
| CN105348193A (en) | Isoquinoline alkaloid compounds as well as preparation method and application of isoquinoline alkaloid compounds | |
| CN109535169A (en) | A kind of chromone derivative and its preparation method and application | |
| CN104829580A (en) | Isoflavone compound contained in tobacco and preparation method and application thereof | |
| CN102100692B (en) | Prenylflavanone compound and use thereof in preparation of anti-tumor medicaments | |
| CN103113439A (en) | Method for preparing kaempferol-3-O-Beta-D-glucuronide in euphorbia sororia | |
| CN106966952A (en) | The preparation method and application of two kinds of isobenzofuran class compounds | |
| CN105820044B (en) | A kind of novel C22Diterpene compound and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170718 Termination date: 20180508 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |