CN101525323A

CN101525323A - Novel styryl chromone type compound and preparation method and use thereof

Info

Publication number: CN101525323A
Application number: CN200910030308A
Authority: CN
Inventors: 杨春华; 刘静涵; 杨洋
Original assignee: Individual
Current assignee: Individual
Priority date: 2009-03-18
Filing date: 2009-03-18
Publication date: 2009-09-09
Anticipated expiration: 2029-03-18
Also published as: CN101525323B

Abstract

The invention discloses a novel styryl chromone type compound and a preparation method and the use thereof, the compound has the structure formula (1): wherein in a compound1, R1 is 3-methyl-2-butenyl, and R2 is OH; in a compound 2, R1 is OH, and R2 is 3-methyl-2-butenyl; in a compound 3, R1 is 3-methyl-3-butylene-2-sterol-1-base, and R2 is OH; and in a compound 4, R1 is OH, and R2 is 3-methyl-3-butylene-2-sterol-1-base. The styryl chromone type compound can be applied to prevent or treat tumours and infection and prepare health food.

Description

Novel styryl chromone type compound and its production and use

Technical field

The invention belongs to biomedicine field, be specifically related to a kind of styryl chromone type compound and its production and use.

Background technology

At present, natural product with 2-vinylbenzene chromone (2-styrylchromone) structure only obtains from the algae bio Chrysophaeum taylori of ocean, be respectively Hormothamnione and 6-Desmethoxyhormothamnione, these two kinds of natural products of bibliographical information all have antitumour activity (structure is as follows), but are trace ingredients.

Hormothamnione(R＝OMe)；6-Desmethoxyhormothamnione(R＝H)

Separate the 2-vinylbenzene chromone compounds that obtains and do not appear in the newspapers from the Lu Sheng higher plant, this patent discloses its discovery in the plane plant first, and the source is abundanter.

Tumour and bacterium and fungi infestation are modern common clinical, and sickness rate is ascendant trend year by year, and the present invention provides cheap, antitumor and anti-infectives, pharmaceutical intermediate or healthcare products safely and effectively for clinical, has good DEVELOPMENT PROSPECT.

Summary of the invention

The purpose of this invention is to provide a kind of novel styryl chromone type compound with pharmaceutical use.

Another object of the present invention provides a kind of preparation method of above-claimed cpd.

A further object of the invention provides a kind of purposes of above-claimed cpd.

Purpose of the present invention can reach by following measure:

A kind of 2-styryl chromone type compound has the structure as shown in the formula (I),

Wherein,

Compound 1:R ₁For

[3-methyl-2-butene base (isopentene group)], R ₂Be OH;

Compound 2:R ₁For

[3-methyl-2-butene base (isopentene group)];

Compound 3:R ₁For

[3-methyl-3-butene-2-alcohol-1-yl], R ₂Be OH;

Compound 4:R ₁Be OH, R ₂For

[3-methyl-3-butene-2-alcohol-1-yl].

Four compounds of the present invention, its structure is by ultraviolet, and is infrared, mass spectrum, the nuclear magnetic resonance spectrum integration analysis is determined, the nuclear magnetic resonance spectrum data such as the following table of compound:

Compound of the present invention is isolated from the bark of plane plant, the source comprises the plant of plane, mainly comprises a ball plane tree (Platanus occidentalis), two ball plane trees (Platanus acerifolia Willd) or sycamore (Platanus orientalis) that China introduces a fine variety.

The invention also discloses the preparation method of such styryl chromone:

With the meal of organic solvent extraction plane tree bark, reclaim solvent, get total medicinal extract, total medicinal extract is made saturated solution with dissolve with ethanol, adds water to and separates out a large amount of precipitations, the elimination precipitation, go up macroporous resin after filtrate concentrates and carry out wash-out, obtain each new compound of vinylbenzene chromone class again through chromatographic separation.

Concrete grammar is: the dry bark that comes off naturally of getting plane tree (Platanaceae) platymiscium plane tree, pulverize, use organic solvent extraction, extracting solution is made saturated solution, add water, to determining alcohol 30%～60%, separate out precipitation, separation of supernatant to an amount of volume (as medicinal extract), gets extract with supernatant concentration; Extract is carried out column chromatography with macroporous resin, with 30%～40%, 50%, 70%, 90% ethanol elution, collect 50%～70% ethanol eluate, concentrating under reduced pressure must contain total medicinal extract of vinylbenzene chromone.To total medicinal extract respectively through silicagel column, C ₁₈Anti-phase middle compression leg, sephadex lh-20 post, SephadexLH-20, preparative column chromatogram etc. are means such as separation and recrystallization repeatedly, promptly get new compound 1-4.

Raw material can be the bark that comes off naturally of plane shade trees such as a ball plane tree, two ball plane trees, sycamore.

The organic solvent that extracts usefulness can be ethanol, methyl alcohol, acetone etc.Wherein concentration of ethanol is 75%～95% (v/v).

Macroporous resin can be non-polar resins such as D101.

Cell experiment shows that compound 1-4 all has growth-inhibiting effect preferably to human tumor cell line HepG2, SMMC-7721, Hela, KB, K-562, Jurkat, MDA-MB-231, the U937 of growth in vitro.Bacteriostatic experiment shows that compound 1-4 all has certain growth-inhibiting effect to streptococcus aureus, gamboge look sarcina, Pseudomonas aeruginosa, dysentery bacterium and gypsum Yang Mao Xian bacterium.Animal acute toxicity experiment shows, this compounds mouse LD50 test dose is not seen animal dead greater than 1.0g/kg, therefore, compound 1-4 can be used as the potential new drug of safer prevention or treatment tumour or infection, also can be used as protective foods, comprise as prodrug or medicinal intermediates.

The present invention has good DEVELOPMENT PROSPECT for the treatment of tumour and infection provides potential safely and effectively drug candidate.

Description of drawings

Fig. 1 is a compound 1 ¹The H-NMR collection of illustrative plates;

Fig. 2 is a compound 1 ¹³The C-NMR collection of illustrative plates;

Fig. 3 is the HMBC collection of illustrative plates of compound 1;

Fig. 4 is the HSQC collection of illustrative plates of compound 1;

Fig. 5 is a compound 2 ¹The H-NMR collection of illustrative plates;

Fig. 6 is a compound 2 ¹³The C-NMR collection of illustrative plates;

Fig. 7 is the HMBC collection of illustrative plates of compound 2;

Fig. 8 is the HSQC collection of illustrative plates of compound 2;

Fig. 9 is a compound 3 ¹The H-NMR collection of illustrative plates;

Figure 10 is a compound 3 ¹³The C-NMR collection of illustrative plates;

Figure 11 is the HMBC collection of illustrative plates of compound 3;

Figure 12 is the HSQC collection of illustrative plates of compound 3;

Figure 13 is a compound 4 ¹The H-NMR collection of illustrative plates;

Figure 14 is a compound 4 ¹³The C-NMR collection of illustrative plates;

Figure 15 is the HMBC collection of illustrative plates of compound 4;

Figure 16 is the HSQC collection of illustrative plates of compound 4.

Embodiment

Embodiment 1

Get 5 kilograms on the dry bark that comes off naturally of two ball plane trees (Platanus acerifolia), pulverize, with 95% alcohol reflux 3-5 time, united extraction liquid, reclaim ethanol and be concentrated into solution and just become turbid, add isopyknic water precipitation, filter, get filtrate and be concentrated into medicinal extract (about 50g), medicinal extract is added on the D101 macroporous adsorptive resins of having handled well, use 30% successively, 50%, 70%, 90% ethanol elution, the ethanol eluate of collection 70% gets the about 10g of reactive site, this reactive site is separated with half preparative liquid chromatography, collects the position (about 4g) of 70% methanol-eluted fractions, with this 4g sample with 65% methanol-eluted fractions, get compound 1 (300mg), compound 2 (70mg), compound 3 (35mg), compound 4 (35mg).

Embodiment 2

Get 10 kilograms on the dry bark that comes off naturally of a ball plane tree (Platanus occidentali), pulverize, with 75% alcohol reflux 3-5 time, united extraction liquid, reclaim ethanol and be condensed into medicinal extract, add 95% ethanol and make saturated solution, it is about 40%～50% to determining alcohol to add suitable quantity of water, filters, and gets filtrate and is concentrated into medicinal extract (about 120g), medicinal extract is suspended in a spot of 40% the ethanol, be added on the D101 macroporous adsorptive resins of having handled well, use 40% successively, 50%, 70%, 90% ethanol elution, the ethanol eluate of collection 70% gets the about 25g of reactive site, this reactive site is separated with half preparative liquid chromatography, use the methanol-water wash-out, collect the position (about 8g) of 70% methanol-eluted fractions, with this 8g sample with 65% methanol-eluted fractions, collect different flow points, recrystallization gets compound 1 (700mg), compound 2 (130mg), compound 3 (60mg), compound 4 (50mg).

Effect experiment

1, cellulotoxic experiment:

HepG2 (human hepatoma cell strain)

HeLa (human cervical carcinoma cell strain)

KB (human nasopharyngeal carcinoma cell line)

K-562 (human erythroleukemia cell's strain)

Jurkat (people T lymphoma cell strain)

SMMC-7721 (human hepatoma cell strain)

MDA-MB-231 (human breast cancer cell)

U937 (people's monokaryon leukemia cell)

Compound 1-4 with the DMSO dissolving, is 5 concentration with containing the dilution of 10% calf serum RPMI-1640 respectively.HepG2, Hela, KB, K-562, Jurkat, SMMC-7721, MDA-MB-231, U937 cell are inoculated in 96 orifice plates with the density in 5000-10000/hole respectively; After 24 hours, when cell density reached 70～80%, dosing was on 96 orifice plates of the good human tumor cells of inoculation, and the final concentration of compound 1-4 is 0.5,2,5,20,50 μ g/ml, every kind of cell is provided with 3 parallel samples, with taxol and white birch acid is control group, and every kind of cell of control group is provided with 3 parallel samples, is according to group to feminine gender with the cell of handling through DMSO.Mixing is placed on 37 ℃, 5%CO ₂Continue to cultivate 24h in the incubator; To add 20 μ L concentration be 5mg/ml MTT in 4h every hole before finishing, centrifugal behind the continuation cultivation 4h, supernatant liquor is removed in suction, every hole adds dimethyl sulfoxide (DMSO) 200 μ L lysates, micro-mixer vibration 10～15min, fully dissolving is measured optical density(OD) OD value with enzyme-linked immunosorbent assay instrument in 570nm wavelength place, press the percentage that the inhibiting rate calculation formula is calculated anticancer.

Calculate growth inhibition ratio: after each hole count value deducts the blank well cell mean, average with each hole of concentration on the same group, calculate tumour cell survival rate (%), formula is as follows: (medication group OD value/control group OD value) * 100%

Growth of tumour cell inhibiting rate (%)=(1-medication group OD value/control group OD value) * 100%

Data are represented with x ± S, relatively check with t between group.

Logarithm with drug level is an X-coordinate, is that ordinate zou is drawn semilog plot with the cell inhibitory rate, carries out straight-line regression with SPSS software, calculates the drug level (IC that suppresses the growth of 50% cell ₅₀).

A, MTT detected result show that cell viability descends gradually along with drug effect dosage increases, and compound 1-4 handles the back cell viability and drops to 1/3 to 1/4 of DMSO control group, and for different clone, the effective dose of its cytotoxicity is also different.It seems that from this experimental result the effective dose of the cytotoxicity of compound 1 is at 0.5-10 μ g/ml.IC ₅₀Value is except that the Jurkat cell strain, and all the other are all less than 10, and the maximal percentage inhibition of compound pair cell strain all reaches more than 95%.The corresponding IC of 1 pair of selected JEG-3 of compound ₅₀Value sees Table 2.

Table 2, compound cell IC ₅₀(μ g/ml)

1

2

3

4

HepG2	2.46	2.68	3.68	3.57
HepG2	2.46	2.68	3.68	3.57	SMMC-7721	7.76	7.54	5.54	6.34
HeLa	7.78	3.66	4.65	3.67	SMMC-7721	7.76	7.54	5.54	6.34
HeLa	7.78	3.66	4.65	3.67	KB	1.55	1.98	1.15	1.58
Jurkat	15.12	10.69	13.12	11.98	KB	1.55	1.98	1.15	1.58
Jurkat	15.12	10.69	13.12	11.98	K562	9.95	7.32	4.33	5.54
MDA-MB-231	4.34	3.15	8.47	5.33	K562	9.95	7.32	4.33	5.54
MDA-MB-231	4.34	3.15	8.47	5.33	U937	＜0.5	＜0.5	＜0.5	＜0.5

B, the dyeing of Hoechst33258 nucleus detect cytotoxicity

Cell is observation of cell under fluorescent microscope after the Hoechst33258 dyeing, and with the negative control group of handling through DMSO of cell, the result shows that the negative control group cell membrane is complete, and kytoplasm enriches the full uniform blue-fluorescence of disperse that shows as of nuclear morphology.Dwindle fluorescent dye and strengthen chromatin and be fine and close dense bulk of dying or particulate state fluorescence and apoptotic body and form and present typical apoptosis morphological specificity and the visible nucleome of tumour cell group that compound 1-4 handles is long-pending.Experimental result explanation compound 1-4 has the intensive cytotoxicity to tumour cell, can cause apoptosis.

2, bacteriostatic experiment

Adopting broth culture to cultivate observation experiment result after 24 hours for 37 ℃, all is the minimum inhibitory concentration of this compound with asepsis growth.Experiment shows that compound 1-4 all has certain growth-inhibiting effect to streptococcus aureus, gamboge look sarcina, Pseudomonas aeruginosa, dysentery bacterium and gypsum Yang Mao Xian bacterium.Experimental result sees Table 3

Minimal inhibitory concentration (the MIC of table 3, compound 1-4; The μ g/ml of unit)

3, toxicity test

Animal acute toxicity experiment shows that the test dose of mice lavage compound is not seen animal dead all greater than 1.0g/kg.

Claims

1, a kind of novel styryl chromone type compound has the structure as shown in the formula (I),

Wherein,

Compound 1:R ₁For

R ₂Be OH;

Compound 2:R ₁Be OH, R ₂For

Compound 3:R ₁For

R ₂Be OH;

Compound 4:R ₁Be OH, R ₂For

2. the preparation method of the described styryl chromone type compound of claim 1 is characterized in that following these steps to carrying out:

A. get the dry bark of plane tree, pulverize,, extracting solution is made saturated solution, add water, remove precipitation with 75%～95% extraction using alcohol;

B. with supernatant concentration, adopt macroporous resin to carry out column chromatography,, concentrate with 30%～40%, 50%, 70%, 90% ethanol elution;

C. the part of 70% ethanol elution is wherein carried out separation and purification through column chromatography.

3, the preparation method of styryl chromone type compound according to claim 2 is characterized in that described plane tree is a ball plane tree (Platanus occidentalis), two ball plane trees (Platanusacerifolia Willd) or sycamore (Platanus orientalis).

4, the preparation method of styryl chromone type compound according to claim 2 is characterized in that described column chromatography is silicagel column, C ₁₈Compression leg, sephadex lh-20 post or high performance liquid preparative chromatography post in anti-phase.

5, the application of the described styryl chromone type compound of claim 1 aspect preparation prevention or treatment tumour, infection.

6, the application of the described styryl chromone type compound of claim 1 aspect the preparation protective foods.