CN105348193A - Isoquinoline alkaloid compounds as well as preparation method and application of isoquinoline alkaloid compounds - Google Patents
Isoquinoline alkaloid compounds as well as preparation method and application of isoquinoline alkaloid compounds Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
- C07D217/12—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with radicals, substituted by hetero atoms, attached to carbon atoms of the nitrogen-containing ring
- C07D217/14—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with radicals, substituted by hetero atoms, attached to carbon atoms of the nitrogen-containing ring other than aralkyl radicals
- C07D217/16—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with radicals, substituted by hetero atoms, attached to carbon atoms of the nitrogen-containing ring other than aralkyl radicals substituted by oxygen atoms
Abstract
The invention discloses isoquinoline alkaloid compounds as well as a preparation method and application of the isoquinoline alkaloid compounds. The isoquinoline alkaloid compounds are separated from dried barks of leguminosae Cassia fistula serving as a medicinal plant in Dai nationality and have the molecular formula as follows: Cl4H15NO4, are named as fistulalkaloid A and have the structure as the specification. The preparation method of the isoquinoline alkaloid compounds comprises the steps of extract extraction, organic solvent extraction, MCI decoloration, silica column chromatography and high-performance liquid chromatography separation realized by taking the leguminosae Cassia fistula serving as the medicinal plant in Dai nationality as a raw material. Cytotoxic activity experiments prove that the isoquinoline alkaloid compounds have relatively good cytotoxic activity for parts of tumor cell strains. The compounds disclosed by the invention are novel in structure and relatively good in biological activity and can be used as lead compounds of anticancer drugs.
Description
Technical field
The invention belongs to national characters active components in medicinal plant extracting and developing and Structural Identification technical field, be specifically related to a kind of isoquinoline alkaloids alkaloid compound and its preparation method and application.
Background technology
Species Cassia platymiscium Cassia fistula L. (
cassiafistula), be the national flower of Thailand, originate in south, South Asia, be distributed in the ground such as south, the west and south of Burma, Sri Lanka, India and China's Mainland, be grown on height above sea level 1, the area of 000 meter.In the Chinese Dai population treatment being widely used in skin infections, obesity, periodic fever and neoplastic disease etc. among the people.And Cassia fistula L. is named " pot holds together good " in Dai Nationality's language, cure mainly hemostasis at Xishuangbanna Prefecture, Yunnan Province, defaecation, bring down a fever.It is reported, the different sites of this plant has anti-diabetic, antitumor, anti-inflammatory, antiviral, antibacterial, oxidation resistant activity.The present invention is separated and obtains an isoquinoline alkaloids alkaloid compound from Cassia fistula L., and this compound has significant cytotoxic activity.
Summary of the invention
The first object of the present invention is to provide a kind of isoquinoline alkaloids alkaloid compound; Second object is the preparation method providing described isoquinoline alkaloids alkaloid compound; 3rd object is to provide described isoquinoline alkaloids alkaloid compound preparing the application in cancer therapy drug.
The first object of the present invention be achieved in that described isoquinoline alkaloids alkaloid compound be from leguminous plants Cassia fistula L. (
cassiafistula) dry bark in be separated and obtain, its molecular formula is C
14h
15nO
4, there is following structure:
This compound is yellow jelly, called after Cassia fistula L. alkali A, English fistulalkaloidA by name.
The second object of the present invention realizes like this, the preparation method of described isoquinoline alkaloids alkaloid compound, for raw material with the dry bark of leguminous plants Cassia fistula L. (Cassiafistula), through medicinal extract extraction, organic solvent extraction, MCI decolouring, silica gel column chromatography, high performance liquid chromatography preparative separation step, be specially:
A, medicinal extract extract: by leguminous plants Cassia fistula L. (
cassiafistula) bark be crushed to 20 ~ 40 orders, by organic solvent supersound extraction 2 ~ 5 times, each 30 ~ 60 minutes, united extraction liquid, filtration, concentrating under reduced pressure extracting solution, leave standstill, filtering throw out, is condensed into medicinal extract a;
B, organic solvent extraction: the water adding weight ratio 1 ~ 2 times amount in medicinal extract a, then with the isopyknic organic solvent extraction of water 3 ~ 5 times, merge organic solvent extraction phase, concentrating under reduced pressure becomes medicinal extract b;
C, MCI decolour: the methanol-water adding weight ratio 3 ~ 5 times amount at medicinal extract b dissolves, upper MCI post, and with 80%-90% methanol-water wash-out, merge organic phase, concentrating under reduced pressure becomes medicinal extract c;
D, silica gel column chromatography: silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 orders, and consumption is medicinal extract c weight 6 ~ 10 times amount; Take volume proportion as chloroform and the acetone mixed organic solvents gradient elution of 1:0 ~ 0:1, collect gradient eluent, concentrate, through TLC monitoring, merge identical part;
E, high performance liquid chromatography are separated: will with volume content be the elutriant that obtains of 50 ~ 90% sherwood oils-acetone soln wash-out through high performance liquid chromatography separation and purification, obtain described isoquinoline alkaloids alkaloid compound.
The structure of the isoquinoline alkaloids alkaloid compound prepared with aforesaid method identifies out by the following method:
The compounds of this invention is yellow jelly; UV spectrum (solvent is methyl alcohol),
λ max(log ε): 215 (4.28), 262 (3.58), 298 (3.13), 336 (3.42) nm; Infrared spectra (pressing potassium bromide troche) ν
max: 3418,3126,2953,1657,1622,1568,1455,1372,1236,1150,1046,858cm
– 1; High resolution mass spectrum (HRESIMS) shows the compounds of this invention quasi-molecular ion peak
m/z[284.0889 M+Na]
+(calculated value 284.0899), in conjunction with
1h and
13cNMR spectrum (table-1) determines that molecular formula is C
14h
15nO
4.Hydroxyl (3418cm is had in its infrared spectra display compound
-1), carbonyl (1657cm
-1), and aromatic ring (1622,1568,1455cm
-1) signal, UV spectrum 215,262,298,336nm has maximum absorption also to confirm to there is aromatic ring structure in compound.Compound
1h and
13c-NMR spectrum (table-1) shows it and contains 14 carbon and 15 hydrogen, comprises, one 1,6, the 7-isoquinoline 99.9 parent nucleus (C-1 ~ C-10 replaced; H-3, H-4, H-5, and H-8), a 3-pyruvic alcohol base (-CO-CH
2-CH
2-OH; C-1 ' ~ C-3 '; H
2-2 ' and H
2-3 ') and two methoxyl groups (
δ c56.0q and 56.2q;
δ h3.78s and 3.83s).H-3 and C-1, C-4, C-10 in compound; H-4 and C-3, C-9, C-10; H-5 and C-4, C-9, C-10; And H-8 with C-1, C-9, C-10 HMBC relevant (Fig. 3) also demonstrate that the existence of isoquinoline 99.9 parent nucleus.After the parent nucleus of compound is determined, remaining substituting group, the position of 3-pyruvic alcohol base, two methoxyl groups also can be determined by analyzing its HMBC Correlated Spectroscopy further.3-pyruvic alcohol base is substituted in C-1 position can by H
2-2 ' (
δ h3.24) and C-1 (
δ c156.8) HMBC is relevant to be determined; Two methoxy substitutions in C-6 and C-7 position can by methoxyl group hydrogen (
δ h3.78,3.83) and C-6 (
δ c152.3) and C-7 (
δ c154.6) HMBC is relevant to be determined.So far the structure of compound is confirmed, and this Compound nomenclature is: Cassia fistula L. alkali A, English fistulalkaloidA by name.
The third object of the present invention is achieved in that described isoquinoline alkaloids alkaloid compound is preparing the application in cancer therapy drug.
The compounds of this invention is separated first from Cassia fistula L. bark, is defined as isoquinoline alkaloids alkaloid compound by nucleus magnetic resonance and measuring method of mass spectrum, and characterizes its concrete structure.Take the compounds of this invention as raw material, to NB4, A549, SHSY5Y, PC3 and MCF7 cell strain, there is good cytotoxic activity, IC
50value reaches 0.61,0.52,1.2,0.83,0.47 respectively
μm.The compounds of this invention structure is simple better active, can be used as the guiding compound of cancer drug development.
Accompanying drawing explanation
Fig. 1 be compound Cassia fistula L. alkali A carbon-13 nmr spectra (
13cNMR);
Fig. 2 be compound Cassia fistula L. alkali A proton nmr spectra (
1hNMR);
The crucial HMBC of Fig. 3 compound Cassia fistula L. alkali A is correlated with.
Embodiment
Below in conjunction with accompanying drawing, the present invention is further illustrated, but limited the present invention never in any form, and any conversion done based on training centre of the present invention or improvement, all fall into protection scope of the present invention.
Isoquinoline alkaloids alkaloid compound of the present invention, be from leguminous plants Cassia fistula L. (
cassiafistula) dry bark in be separated and obtain, its molecular formula is C
14h
15nO
4,
Called after Cassia fistula L. alkali A, English fistulalkaloidA by name.
The preparation method of isoquinoline alkaloids alkaloid compound of the present invention, for raw material with the dry bark of leguminous plants Cassia fistula L. (Cassiafistula), through medicinal extract extraction, organic solvent extraction, MCI decolouring, silica gel column chromatography, high performance liquid chromatography preparative separation step, be specially:
A, medicinal extract extract: by leguminous plants Cassia fistula L. (
cassiafistula) bark be crushed to 20 ~ 40 orders, by organic solvent supersound extraction 2 ~ 5 times, each 30 ~ 60 minutes, united extraction liquid, filtration, concentrating under reduced pressure extracting solution, leave standstill, filtering throw out, is condensed into medicinal extract a;
B, organic solvent extraction: the water adding weight ratio 1 ~ 2 times amount in medicinal extract a, then with the isopyknic organic solvent extraction of water 3 ~ 5 times, merge organic solvent extraction phase, concentrating under reduced pressure becomes medicinal extract b;
C, MCI decolour: the methanol-water adding weight ratio 3 ~ 5 times amount at medicinal extract b dissolves, upper MCI post, and with 80%-90% methanol-water wash-out, merge organic phase, concentrating under reduced pressure becomes medicinal extract c;
D, silica gel column chromatography: silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 orders, and consumption is medicinal extract c weight 6 ~ 10 times amount; Take volume proportion as chloroform and the acetone mixed organic solvents gradient elution of 1:0 ~ 0:1, collect gradient eluent, concentrate, through TLC monitoring, merge identical part;
E, high performance liquid chromatography are separated: will with volume content be the elutriant that obtains of 50 ~ 90% sherwood oils-acetone soln wash-out through high performance liquid chromatography separation and purification, obtain described isoquinoline alkaloids alkaloid compound.
The organic solvent of described step A be the acetone of 70 ~ 100%, the ethanol of 90 ~ 100% or 90 ~ 100% methyl alcohol.
The organic solvent of described step B is methylene dichloride, chloroform, ethyl acetate ether or sherwood oil.
In described D step, medicinal extract c is before silica gel column chromatography, with acetone or the dissolve with methanol of weight ratio 1.5 ~ 3 times amount, then weighs 80 ~ 100 order silica gel mixed samples of 0.8 ~ 1.2 times with medicinal extract.
The chloroform of described D step and the volume proportion of acetone mixed organic solvents are 20:1,9:1,8:2,7:3,6:4 and 1:1.
The high performance liquid chromatography separation and purification of described E step is for moving phase with the methyl alcohol of 30 ~ 60%, flow velocity 10 ~ 14ml/min, with 21.2 × 250mm, the ZorbaxPrepHTGF reverse phase preparative column of 5 μm is stationary phase, UV-detector determined wavelength is 254nm, each sample introduction 10 ~ 100 μ L, collects the chromatographic peak of 10 ~ 40min, repeatedly cumulative rear evaporate to dryness.
Isoquinoline alkaloids alkaloid compound of the present invention is preparing the application in cancer therapy drug.
Cassia plant of the present invention does not limit by area and kind, all can realize the present invention.
Embodiment 1
Get dry leguminous plants Cassia Cassia fistula L. (
cassiafistula) bark 4.4kg, coarse reduction to 30 order, the acetone supersound extraction with 70% 4 times, each 60 minutes, extracting solution merge, extracting liquid filtering, is evaporated to 1/4 of volume, leave standstill, filtering throw out, is condensed into the medicinal extract a of 120g, in medicinal extract a, add 250g water, with the isopyknic chloroform extraction of water 5 times, merge extraction phase, concentrating under reduced pressure becomes 80g medicinal extract b, medicinal extract b MCI fills post, and 80% methanol-water adding 240g in medicinal extract b dissolves, and then upper prop, rises wash-out with 90% methanol-water 2 to 6, and collect elutriant, concentrating under reduced pressure obtains 62g medicinal extract c, medicinal extract c adds the acetone solution of 120g in medicinal extract c, then adds 100 order silica gel 62g and mixes sample, after mixing sample, fills post with 200 order silica gel 400g, 20:1 is respectively by volume ratio, 9:1, 8:2, 7:3, chloroform-acetone mixed organic solvents the gradient elution of 6:4 and 1:1, collect gradient eluent, concentrated, monitor through TLC, merge identical part, obtain 6 part A-F, wherein, to the sample part B 12g collected, repeat silica gel column chromatography again, with the sherwood oil-acetone mixed organic solvents gradient elution of volume ratio 9:1-1:2, collect gradient eluent, concentrated, monitor through TLC, merge identical part, obtain 6 part B1-B6, wherein B4 part, i.e. 6:4 part about 1.2g, again with 48% methyl alcohol for moving phase, flow velocity 15ml/min, 21.2 × 250mm, the ZorbaxPrepHTGF reverse phase preparative column of 5 μm is stationary phase, UV-detector determined wavelength is 254nm, each sample introduction 50 μ L, collect the chromatographic peak of 22.6min, repeatedly cumulative rear evaporate to dryness, obtain described new compound.
Embodiment 2
Get dry leguminous plants Cassia Cassia fistula L. (
cassiafistula) bark 10kg, coarse reduction to 40 order, extracts 4 times with the methyl alcohol cold soaking of 80%, each 3 days, extracting solution merge, extracting liquid filtering, is evaporated to 1/4 of volume, leave standstill, filtering throw out, is condensed into 300g medicinal extract a, in medicinal extract a, add 350g water, with the isopyknic extraction into ethyl acetate of water 5 times, merge extraction phase, concentrating under reduced pressure becomes 210g medicinal extract b, medicinal extract b MCI fills post, and 80% methanol-water adding 600g in medicinal extract b dissolves, and then upper prop, rises wash-out with 90% methanol-water 5 to 15, and collect elutriant, concentrating under reduced pressure obtains 150g medicinal extract c, add the acetone solution of 300g in medicinal extract c, then add 100 order silica gel 150g and mix sample, fill posts with 200 order silica gel 1Kg, mix upper prop after sample, 20:1 is respectively by volume ratio, 9:1, 8:2, 7:3, chloroform-acetone mixed organic solvents the gradient elution of 6:4 and 1:1, collect gradient eluent, concentrated, monitor through TLC, merge identical part, obtain 6 part A-F, wherein, to the sample part B 32g collected, repeat silica gel column chromatography again, with the sherwood oil-acetone mixed organic solvents gradient elution of volume ratio 9:1-1:2, collect gradient eluent, concentrated, monitor through TLC, merge identical part, obtain 6 part B1-B6, wherein B4 part, i.e. 6:4 part about 2.8g, again with 48% methyl alcohol for moving phase, flow velocity 15ml/min, 21.2 × 250mm, the ZorbaxPrepHTGF reverse phase preparative column of 5 μm is stationary phase, UV-detector determined wavelength is 254nm, each sample introduction 80 μ L, collect the chromatographic peak of 22.6min, repeatedly cumulative rear evaporate to dryness, obtain described new compound.
Embodiment 3
Compound prepared by Example 1 is yellow jelly;
Measuring method is: with nucleus magnetic resonance, in conjunction with other spectroscopic technique qualification structure.
The compounds of this invention is yellow jelly; UV spectrum (solvent is methyl alcohol),
λ max(log ε): 215 (4.28), 262 (3.58), 298 (3.13), 336 (3.42) nm; Infrared spectra (pressing potassium bromide troche) ν
max: 3418,3126,2953,1657,1622,1568,1455,1372,1236,1150,1046,858cm
– 1; High resolution mass spectrum (HRESIMS) shows the compounds of this invention quasi-molecular ion peak
m/z[284.0889 M+Na]
+(calculated value 284.0899), in conjunction with
1h and
13cNMR spectrum determines that molecular formula is C
14h
15nNaO
4.Hydroxyl (3418cm is had in its infrared spectra display compound
-1), carbonyl (1657cm
-1), and aromatic ring (1622,1568,1455cm
-1) signal, UV spectrum 215,262,298,336nm has maximum absorption also to confirm to there is aromatic ring structure in compound.Compound
1h and
13c-NMR spectrum (table-1) shows it and contains 14 carbon and 15 hydrogen, comprises, one 1,6, the 7-isoquinoline 99.9 parent nucleus (C-1 ~ C-10 replaced; H-3, H-4, H-5, and H-8), a 3-pyruvic alcohol base (-CO-CH
2-CH
2-OH; C-1 ' ~ C-3 '; H
2-2 ' and H
2-3 ') and two methoxyl groups (
δ c56.0q and 56.2q;
δ h3.78s and 3.83s).H-3 and C-1, C-4, C-10 in compound; H-4 and C-3, C-9, C-10; H-5 and C-4, C-9, C-10; And H-8 with C-1, C-9, C-10 HMBC relevant (Fig. 3) also demonstrate that the existence of isoquinoline 99.9 parent nucleus.After the parent nucleus of compound is determined, remaining substituting group, the position of 3-pyruvic alcohol base, two methoxyl groups also can be determined by analyzing its HMBC Correlated Spectroscopy further.3-pyruvic alcohol base is substituted in C-1 position can by H
2-2 ' (
δ h3.24) and C-1 (
δ c156.8) HMBC is relevant to be determined; Two methoxy substitutions in C-6 and C-7 position can by methoxyl group hydrogen (
δ h3.78,3.83) and C-6 (
δ c152.3) and C-7 (
δ c154.6) HMBC is relevant to be determined.So far the structure of compound is confirmed, and this Compound nomenclature is: Cassia fistula L. alkali A, English fistulalkaloidA by name.
Embodiment 4
Compound prepared by Example 2 is yellow jelly.Measure identical with enforcement 3, confirm that compound prepared by enforcement 2 is described isoquinoline alkaloids alkaloid compound---Cassia fistula L. alkali A.
Embodiment 5
Arbitrary isoquinoline alkaloids alkaloid compound Cassia fistula L. alkali A prepared by Example 1 and 2 carries out cytotoxicity assay test, and test situation is as follows:
Cell strain: leukemia cell (NB4), lung carcinoma cell (A549), human neuroblastoma cells (SHSY5Y), prostate cancer cell (PC3), breast cancer cell (MCF7) provide by Shanghai Pharmaceutical Inst., Chinese Academy of Sciences.
Experimental design: above cell and different concns compound incubation 72 hours, the experiment of every strain cell all repeats once, data processing is carried out by the result of twice experiment, adopt the suppression degree of improvement mtt assay and srb assay assessing compound on cell proliferation, calculate inhibiting rate, adopt Logit method to calculate IC according to inhibiting rate
50, the anti tumor activity in vitro of comparative compound.
Proliferation inhibition rate=(the OD value of blank OD value-medicine feeding hole)/blank OD value × 100% of cell.
(a) improvement mtt assay
Get the suspension cell being in logarithmic phase, cell concn is adjusted to 4 × 10
4/ ml, adds 96 well culture plates, 90
μl/ hole.Positive control is cis-platinum, uses physiological saline solution.Every hole adds the sample (No. 1 test solution-No. 5 test solutions) of 10 μ l different concns respectively.Application of sample group and control group all establish 4 multiple holes, and the high density group of application of sample group, positive controls also establishes the dosing parallel hole of substratum, and every block plate is equipped with 4 blank control wells (only adding substratum).The final concentration of sample is respectively 10
-2, 10
-1, 1,10 and 10
2 μthe final concentration of g/mL, corresponding DMSO is respectively 0.1%, 0.01%, 0.001%, 0.0001%, 0.00001%.Sample is at final concentration 10
2 μduring g/mL, with 0.1%DMSO as solvent control, all the other concentration all make negative control with physiological saline.The final concentration of positive control drug cis-platinum is 10
-1, 1,10
μg/mL.Cell at 37 DEG C, 5%CO
2after hatching 48h respectively in incubator, add MTT (5mg/ml, Sigma), 10
μl/ hole.After continuing to cultivate 4h, add three liquid [10%SDS – 5% Yi Ding Chun – 0.012mol/LHCL (w/v/v)], 100
μl/ hole, places the OD value measuring each hole after spending the night by microplate reader under 570nm, 630nm dual wavelength.
(b) srb assay
Get the attached cell strain being in logarithmic phase, after 25% pancreatin conventional digestion, then with 15% calf serum complete RPMI-1640 substratum, cell concn is adjusted to 5 × 10
4/ mL, adds 96 well culture plates, 90
μl/ hole.Cell at 37 DEG C, 5%CO
2positive control, negative control and given the test agent (the same mtt assay of each tested concentration, 10 are added after hatching 24h respectively in incubator
μl/ hole), the final concentration of sample is respectively 10
-2, 10
-1, 1,10,10
2 μthe final concentration of g/mL, corresponding DMSO is respectively 0.1%, 0.01%, 0.001%, 0.0001%, 0.00001%.Sample is at final concentration 10
2 μduring g/mL with 0.1%DMSO as solvent control, all the other concentration all make negative control with physiological saline.The final concentration of positive control drug cis-platinum is 10
-1, 1,10 μ g/mL, negative control is isopyknic physiological saline.Application of sample group and control group all establish 4 multiple holes, and the high density group of application of sample group, positive controls also establishes the dosing parallel hole of substratum, and every block plate is equipped with 4 blank control wells (only adding substratum).96 well culture plates are placed in 37 DEG C, 5%CO
2after hatching (cell and sample effect) 48h in incubator, add 4 DEG C, the TCA (trichoroacetic acid(TCA)) 50 of 50%
μl/ hole.After adding TCA, 96 well culture plates are placed in 4 DEG C and hatch 1 hour, take out culture plate, liquid in the plate that inclines gently.Rinsing gently 5 times (tap water is poured in plate gently by beaker, again water is gone after light rolling) with tap water, being placed in air air-dry to loseing washmarking.Then the 0.4%SRB (with 1% acetic acid dilution) prepared is added, 50
μl/ hole, removes SRB solution in left at room temperature hypsokinesis in 30 minutes of dyeing, rinses 4 times with 1% acetic acid, with remove not with the dyestuff of protein bound.Be placed in air air-dry to without after washmarking, add 10mM and do not cushion Tris (slow blood ammonia acid) solution 150
μl/ hole (PH10 prepares with tri-distilled water), after being dissolved by dyestuff, vibrates 5 minutes, reads each hole OD value by microplate reader under 570nm wavelength on vibrator.
(c) experimental result
Experimental result shows: through the cytotoxic activity experiment to Leukemia acute young grain NB4 cell morning, Lung Adenocarcinoma A 549 Cell, people's marrow neuroblastoma SHSY5Y cell, human prostata cancer PC3 cell, human breast cancer MCF7 cell, Cassia fistula L. alkali A is to NB4, A549 and PC3 cell strain has good cytotoxic activity, IC
50value reaches 8.2,5.6 and 6.8 respectively
μm.
The cytotoxic activity of table 2 compound Cassia fistula L. alkali A
| Compounds | NB4 | A549 | SHSY5Y | PC3 | MCF7 |
| Cassia fistula L. alkali A | 0.61 | 0.52 | 1.2 | 0.83 | 0.47 |
| Taxol | 0.02 | 0.02 | 0.1 | 0.1 | 0.05 |
Claims (8)
1. an isoquinoline alkaloids alkaloid compound, it is characterized in that described isoquinoline alkaloids alkaloid compound be from leguminous plants Cassia fistula L. (
cassiafistula) dry bark in be separated and obtain, called after Cassia fistula L. alkali A, English fistulalkaloidA by name, its molecular formula is C
14h
15nO
4, there is following structure:
。
2. a preparation method for isoquinoline alkaloids alkaloid compound described in claim 1, it is characterized in that with leguminous plants Cassia fistula L. (
cassiafistula) dry bark be raw material, through medicinal extract extraction, organic solvent extraction, MCI decolouring, silica gel column chromatography, high performance liquid chromatography preparative separation step, be specially:
A, medicinal extract extract: by leguminous plants Cassia fistula L. (
cassiafistula) bark be crushed to 20 ~ 40 orders, by organic solvent supersound extraction 2 ~ 5 times, each 30 ~ 60 minutes, united extraction liquid, filtration, concentrating under reduced pressure extracting solution, leave standstill, filtering throw out, is condensed into medicinal extract a;
B, organic solvent extraction: the water adding weight ratio 1 ~ 2 times amount in medicinal extract a, then with the isopyknic organic solvent extraction of water 3 ~ 5 times, merge organic solvent extraction phase, concentrating under reduced pressure becomes medicinal extract b;
C, MCI decolour: the methanol-water adding weight ratio 3 ~ 5 times amount at medicinal extract b dissolves, upper MCI post, and with 80%-90% methanol-water wash-out, merge organic phase, concentrating under reduced pressure becomes medicinal extract c;
D, silica gel column chromatography: silica gel column chromatography on medicinal extract c, dress post silica gel is 160 ~ 200 orders, and consumption is medicinal extract c weight 6 ~ 10 times amount; Take volume proportion as chloroform and the acetone mixed organic solvents gradient elution of 1:0 ~ 0:1, collect gradient eluent, concentrate, through TLC monitoring, merge identical part;
E, high performance liquid chromatography are separated: will with volume content be the elutriant that obtains of 50 ~ 90% sherwood oils-acetone soln wash-out through high performance liquid chromatography separation and purification, obtain described isoquinoline alkaloids alkaloid compound.
3. the preparation method of isoquinoline alkaloids alkaloid compound according to claim 2, it is characterized in that the organic solvent of described step A be the acetone of 70 ~ 100%, the ethanol of 90 ~ 100% or 90 ~ 100% methyl alcohol.
4. the preparation method of isoquinoline alkaloids alkaloid compound according to claim 2, is characterized in that the organic solvent of described step B is methylene dichloride, chloroform, ethyl acetate ether or sherwood oil.
5. the preparation method of isoquinoline alkaloids alkaloid compound according to claim 2, it is characterized in that in described D step, medicinal extract c is before silica gel column chromatography, with acetone or the dissolve with methanol of weight ratio 1.5 ~ 3 times amount, then weigh 80 ~ 100 order silica gel mixed samples of 0.8 ~ 1.2 times with medicinal extract.
6. the preparation method of isoquinoline alkaloids alkaloid compound according to claim 2, is characterized in that the chloroform of described D step and the volume proportion of acetone mixed organic solvents are 20:1,9:1,8:2,7:3,6:4 and 1:1.
7. the preparation method of isoquinoline alkaloids bases compounds according to claim 2, it is characterized in that the high performance liquid chromatography separation and purification of described E step is for moving phase with the methyl alcohol of 30 ~ 60%, flow velocity 10 ~ 14ml/min, with 21.2 × 250mm, the ZorbaxPrepHTGF reverse phase preparative column of 5 μm is stationary phase, and UV-detector determined wavelength is 254nm, each sample introduction 10 ~ 100 μ L, collect the chromatographic peak of 10 ~ 40min, repeatedly cumulative rear evaporate to dryness.
8. an isoquinoline alkaloids alkaloid compound according to claim 1 is preparing the application in cancer therapy drug.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105884621A (en) * | 2016-04-19 | 2016-08-24 | 秦瑞欣 | Sesquiterpenoids as well as preparation method and application thereof |
| CN106117138A (en) * | 2016-06-29 | 2016-11-16 | 云南中烟工业有限责任公司 | A kind of isoquinoline alkaloids alkaloid compound, its preparation method and application with antibacterial activity |
| CN110452170A (en) * | 2019-08-29 | 2019-11-15 | 云南中烟工业有限责任公司 | A kind of isoquinoline alkaloids bases compound and its preparation method and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104262316A (en) * | 2014-09-15 | 2015-01-07 | 云南民族大学 | Flavonoid compound as well as preparation method and application thereof |
| CN104292203A (en) * | 2014-09-16 | 2015-01-21 | 云南民族大学 | Isocoumarin compound and preparation method and application thereof |
| CN104610219A (en) * | 2015-01-14 | 2015-05-13 | 云南民族大学 | Xanthone compound with isopentene oxide group as well as preparation method and application of xanthone compound |
| CN104860912A (en) * | 2015-05-08 | 2015-08-26 | 云南民族大学 | Dimer ketone compound and preparation method and application thereof |
-
2015
- 2015-12-16 CN CN201510942660.6A patent/CN105348193A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104262316A (en) * | 2014-09-15 | 2015-01-07 | 云南民族大学 | Flavonoid compound as well as preparation method and application thereof |
| CN104292203A (en) * | 2014-09-16 | 2015-01-21 | 云南民族大学 | Isocoumarin compound and preparation method and application thereof |
| CN104610219A (en) * | 2015-01-14 | 2015-05-13 | 云南民族大学 | Xanthone compound with isopentene oxide group as well as preparation method and application of xanthone compound |
| CN104860912A (en) * | 2015-05-08 | 2015-08-26 | 云南民族大学 | Dimer ketone compound and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 于玲等: "傣药腊肠树有效成分的研究", 《河北化工》 * |
| 宫爽等: "天然药物中生物碱类成分抗肿瘤活性", 《吉林中医药》 * |
| 邓婕等: "综述异喹啉类生物碱分离与分析研究进展", 《化工时代》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105884621A (en) * | 2016-04-19 | 2016-08-24 | 秦瑞欣 | Sesquiterpenoids as well as preparation method and application thereof |
| CN106117138A (en) * | 2016-06-29 | 2016-11-16 | 云南中烟工业有限责任公司 | A kind of isoquinoline alkaloids alkaloid compound, its preparation method and application with antibacterial activity |
| CN106117138B (en) * | 2016-06-29 | 2018-06-15 | 云南中烟工业有限责任公司 | A kind of isoquinoline alkaloids bases compound, preparation method and application with antibacterial activity |
| CN110452170A (en) * | 2019-08-29 | 2019-11-15 | 云南中烟工业有限责任公司 | A kind of isoquinoline alkaloids bases compound and its preparation method and application |
| CN110452170B (en) * | 2019-08-29 | 2022-11-08 | 云南中烟工业有限责任公司 | Isoquinoline alkaloid compound and preparation method and application thereof |
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