CN104803838B - A kind of Herba Hedyotis Lanceae anthraquinone analog compound and preparation method thereof - Google Patents

Abstract

The invention discloses anthraquinone compound of saponifera and a preparation method thereof. The structural formula of the compound is: The preparation method comprises the following steps: (1) extracting the saphophylla in a solvent, concentrating the extract under reduced pressure, recovering the solvent until it has no alcohol smell; (2) diluting the obtained extract with water, and then using polar From small to large solvents, petroleum ether, chloroform, ethyl acetate, and n-butanol, extract in sequence to obtain the corresponding extracts, respectively concentrate the extracts under reduced pressure, and collect the dried extracts of the chloroform parts; (3) dry the obtained chloroform parts The extract is separated and purified by column chromatography to obtain the anthraquinone compounds of the saponifera. The anthraquinone compound of the present invention is a compound discovered for the first time, which enriches the types of existing compounds, has good pharmacological and physiological activities, has antitumor, hemostatic, antibacterial, purgative, and diuretic effects, and has suitable polarity in terms of drug absorption. To further promote the development and utilization of saber-leaf ear grass.

Description

Anthraquinone compound and preparation method thereof

technical field

The invention belongs to the field of preparation of pharmaceutical compounds, and in particular relates to anthraquinone compound of genus saponifera and a preparation method thereof.

Background technique

Sword-leaf eargrass (Hedyotis caudatifolia Merr.et Metcalf), is the whole herb of Rubiaceae eargrass plant Hedyotis caudatifolia Merr.et Metcalf, also known as juvenile red, long-tailed ear grass, millennium tea, postpartum tea, tuberculosis grass, is a Guangxi Folk herbal medicine has the functions of relieving cough and reducing phlegm, invigorating the spleen and eliminating accumulation, etc.

Currently, the researches involved in sword-leaf eargrass mainly include research on the microstructure of roots, stems and leaves, GC-MS analysis of volatile oil components and the protective effect of its extracts on the liver. Folks often have old Chinese medicine for the treatment of hepatitis B, and the effect is remarkable. Compared with its widespread use in the folk, there are very few researches on C. saponifera at home and abroad at present. The research on the chemical constituents of C. saponifera is almost blank except for the volatile oil components. This status quo has greatly affected the rational promotion and use of P. saponifera. Therefore, it is extremely important to carry out systematic research on the active ingredients of P. raptoria. The rational development and utilization of Auricularia fragrans resources provide a material basis for promoting the standardization and internationalization of Auricularia fragrans as a traditional Chinese medicine.

Anthraquinone compounds have a wide range of biological activities. Most of the anthraquinone compounds have certain antibacterial activity and laxative effect. Some anthraquinone compounds also have hemostatic, anti-tumor, anti-osteoporosis, diuretic and other effects.

Contents of the invention

The technical problem to be solved by the present invention is to provide a kind of anthraquinone compound and its preparation method. The anthraquinone compound is a compound discovered for the first time, has good pharmacological and physiological activities, and has antitumor, hemostatic, antibacterial, It has purgatory and diuretic effects, and has suitable polarity in terms of drug absorption, which further promotes the development and utilization of phobia.

In order to solve the problems of the technologies described above, the present invention adopts the following technical solutions:

A saponifera anthraquinone compound, the structural formula is:

The preparation method of the described saphophylla anthraquinone compound comprises the following steps:

(1) Pulverize the saphophylla, place it in a solvent for extraction, the weight of the solvent is 4 to 12 times the weight of the saphophylla, the solvent is one or a mixture of water and an organic solvent, and the organic solvent is methanol , ethanol, acetone, acetonitrile, chloroform, acetone, petroleum ether and ethyl acetate, or one or more of them are mixed; Concentrate the extract under reduced pressure, recover the solvent until it has no alcohol smell, and obtain the extract;

(2) Dilute the extract obtained in step (1) with water to an aqueous solution with a concentration of 0.02 to 1 g/ml, and then extract successively with solvents petroleum ether, chloroform, ethyl acetate, and n-butanol with increasing polarity to obtain For the corresponding extracts, respectively concentrate the extracts under reduced pressure, and collect the dry extracts from the chloroform parts;

(3) Separating and purifying the dried extract from the chloroform part obtained in step (2) by column chromatography to obtain the anthraquinone compound of the genus flavinus; the support of the column chromatography is silica gel, aluminum oxide, poly Amide or octadecylsilane, the eluent is a chloroform/acetone mixture with a volume ratio of 100:1 to 5:1 or a chloroform/methanol mixture with a volume ratio of 200:1 to 6:1.

The leaching of described step (1) is: the ethanol that is 95% by volume concentration, the ethanol that volume concentration is 80%, the ethanol that volume concentration is 60%, the ethanol that volume concentration is 40% and water are used as solvent to Fenugreek is subjected to percolation extraction, the mass ratio of Fenugreek to each solvent is 1:8, the extracts are combined, the obtained extracts are concentrated under reduced pressure, and the solvent is recovered until there is no alcohol smell to obtain the extract.

The column chromatography of the step (3) adopts normal phase silica gel column chromatography.

It also includes purifying the target fraction obtained through normal phase silica gel column chromatography in step (3) using Sephadex LH-20, reversed phase silica gel column chromatography or high performance liquid chromatography, to obtain the saponifera anthraquinone compound .

The particle size of the silica gel is 200-300 mesh.

The step (3) also includes: performing column chromatographic separation on the dry extract of the chloroform part obtained in step (2), and then using the gradient elution of the chloroform/acetone mixed solution, and the volume ratio of the chloroform/acetone in the chloroform/acetone mixed solution is sequentially : 100:1→80:1→50:1→20:1→10:1→5:1→2:1→1:1; the collected elution fraction is the chloroform/acetone volume ratio of 50:1 The solution eluted from the mixed solution was detected by thin-layer chromatography for color development, and the same fractions were combined, concentrated and dried under reduced pressure to obtain dark green feathery crystals; then separated by silica gel column chromatography, volume Gradient elution with chloroform/methanol at a ratio of 5:1 to 1:2 yielded yellow-green feathery crystals, which were repeatedly purified by Sephadex LH-20 column chromatography, and then washed with a mixture of chloroform/methanol at a volume ratio of 1:1 The light green feathery crystals are obtained, which are the said saponiferium anthraquinone compounds.

The adsorbent of the column chromatography is 100-200 mesh silica gel.

The whole herb of the saphophyllum plant is used for the saphophyllium.

Compared with prior art, the beneficial effect of the present invention is:

The anthraquinone compound of the saponifera of the present invention is a compound discovered for the first time, which enriches the types of existing compounds. The anthraquinone compound has good pharmacological and physiological activities, has anti-tumor, hemostatic, antibacterial, purgatory and diuretic effects, and has suitable polarity in terms of drug absorption, and further promotes the development and utilization of the saber ear grass.

Description of drawings

Fig. 1 is the mass spectrometry of described saponifera anthraquinone;

Fig. 2 is the high-resolution mass spectrum of the anthraquinone of the saponifera;

Fig. 3 is the 400MHz ¹ H-NMR spectrum of the anthraquinone of the saponifera;

Fig. 4 is the 600MHz ¹ H-NMR spectrum of the anthraquinone of the saponifera;

Fig. 5 is the 100MHz ¹³ C-NMR and DEPT spectra of the flavana anthraquinone;

Fig. 6 is the 150MHz ¹³ C-NMR spectrum of the anthraquinone of the saponifera;

Fig. 7 is the HSQC spectrum of described saponifera anthraquinone;

Fig. 8 is the ¹ H- ¹ H COZY spectrum of the anthraquinone of the saponifera;

Fig. 9 is the HMBC spectrum of the anthraquinone of the saponifera;

Figure 10 shows the structural formulas and key correlations of the compounds.

detailed description

The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, but this does not constitute a limitation to the protection scope of the present invention.

Example 1

(1) The samples of Aurantium saponifera used were collected in Jinxiu Yao Autonomous County, Guangxi. Get 30kg of saponifera samples, pulverize them respectively with ethanol and water with volume concentrations of 95%, 80%, 60%, and 40% as solvents to extract saphophyllium by percolation, saphophylla and each solvent The mass ratios of both are 1:8, the combined extracts are concentrated under reduced pressure, the solvent is recovered until there is no alcohol smell, and the total extract is obtained.

(2) Add water and suspend the total extract to a concentration of 0.02-1g/ml, then sequentially extract with petroleum ether, chloroform, ethyl acetate, and n-butanol at 60-90°C, concentrate and recover the solvent under reduced pressure, respectively, and obtain petroleum Ether part dry extract, chloroform part dry extract, ethyl acetate part dry extract and n-butanol part dry extract.

(3) Use 100-200 mesh silica gel to separate the dry extract from the chloroform part by column chromatography, and use the chloroform/acetone mixed solution for gradient elution. The volume ratio of chloroform/acetone in the chloroform/acetone mixed solution is: 100:1→80 :1→50:1→20:1→10:1→5:1→2:1→1:1; the collected elution fraction is eluted from the mixture of chloroform/acetone with a volume ratio of 50:1 The color of the solution was detected by thin-layer chromatography, and the same fractions were combined, concentrated under reduced pressure, and dried to obtain dark green feathery crystals.

(4) The dark green feathery crystals were separated by 200-300 mesh silica gel column chromatography, and the silica gel column chromatography was eluted with a gradient of chloroform/methanol with a volume ratio of 5:1-1:2 to obtain yellow-green feathery crystals , and then repeatedly purified by Sephadex LH-20 column chromatography, and then eluted with a chloroform/methanol mixture with a volume ratio of 1:1 to obtain light green feathery crystals, which are the saphophyllium anthraquinone compounds.

Example 2:

(1) Pulverize the whole herb of C. saponifera and place it in a solvent for extraction. The weight of the solvent is 4 to 12 times the weight of C. fragrans, and the solvent is methanol; Alcoholic taste, to obtain the extract, take a small amount of extract and dry it completely to obtain a dry paste of the extract, and weigh it, and calculate the net weight of the total extract according to the quality of the obtained dry paste of the extract;

(2) Dilute the extract obtained in step (1) with water to a concentration of 0.02 to 1 g/ml according to the actual net weight, and then use the solvent petroleum ether, chloroform, ethyl acetate, and n-butanol in sequence from small to large polarity The corresponding extracts were obtained by extraction, the extracts were respectively concentrated under reduced pressure, and the dry extracts were collected from the chloroform parts;

(3) Separating and purifying the dry extract from the chloroform part obtained in step (2) by normal phase silica gel column chromatography, the particle size of the silica gel is 200 to 300 meshes, and obtaining the anthraquinone compound of the saponifera; the normal phase The eluent of silica gel column chromatography is a chloroform/acetone mixture with a volume ratio of 100:1 to 5:1 or a chloroform/methanol mixture with a volume ratio of 200:1 to 6:1.

Structural identification of anthraquinone compounds from P.

The compound is light green feathery crystal, easily soluble in chloroform. TLC detection, development in three different development systems, iodine vapor color development, showing a yellow single spot, the results are as follows: petroleum ether-ethyl acetate with a volume ratio of 1:1, R _f =0.28; chloroform with a volume ratio of 5:1 - acetone, R _f =0.53; chloroform-methanol in volume ratio 10:1, R _f =0.64.

The mass spectrum is shown in Figure 1, and the high-resolution mass spectrum is shown in Figure 2, EI-MS m/z(%): 314(M ⁺ ,100), 299,281,253,239,225; HR-EI-MS m/z: 314.0791[M] ⁺ , the molecular formula calculated by high-resolution mass spectrometry is C ₁₇ H ₁₄ O ₆ . At the same time, from the ¹³ C-NMR and DEPT spectrum data, it can also be speculated that the molecular formula is C ₁₇ H ₁₄ O ₆ and the degree of unsaturation is 8.

TMS was used as internal standard for ¹ H-NMR and ¹³ C-NMR spectra, and the solvent was CDCl ₃ . ¹ H-NMR (400MHz) of the compound is shown in Figure 3, ¹ H-NMR (600MHz) is shown in Figure 4, ¹³ C-NMR and DEPT (100MHz) are shown in Figure 5, ¹³ C-NMR (150MHz) As shown in Figure 6, HSQC is shown in Figure 7, ¹ H- ¹ H COZY is shown in Figure 8, and HMBC is shown in Figure 9.

Through the analysis of Figures 1-9, the structural formulas and key correlations of the compounds shown in Figure 10 are obtained.

Its ¹ H-NMR, ¹³ C-NMR data assignment and two-dimensional spectral correlation are shown in Table 1.

Table 1 The one-dimensional and two-dimensional NMR spectra of anthraquinone compounds in Auranthus fragrans (solvent CDCl ₃ , internal standard TMS, 600/150MHz and 400/100MHz)

Note: ^* The serial number is shown in Figure 10; ^** Only DEPT is 100MHz, and others are 600/150MHz; ***Confirm the corresponding relationship between ¹ H signal and ¹³ C signal through HSQC spectrum; ^**** Brackets indicate two key relationship.

Claims (8)

1. a preparation method for Herba Hedyotis Lanceae anthraquinone analog compound, described Herba Hedyotis Lanceae anthraquinone analog compound structural formula is:

It is characterized in that, preparation method comprises the following steps:

(1) being pulverized by Herba Hedyotis Lanceae, be placed in solvent extraction, weight of solvent is 4 ~ 12 times of Herba Hedyotis Lanceae weight, and solvent is for having Machine solvent or organic solvent and the mixture of water, organic solvent is methyl alcohol or ethanol；By gained leaching liquor reduced pressure concentration, reclaim molten Agent, to without alcohol taste, obtains extract；

(2) be diluted with water as the aqueous solution that concentration is 0.02 ~ 1 g/ml by extract obtained for step (1), then with polarity by little to Big solvent petroleum ether, chloroform, ethyl acetate, n-butanol extract successively and respectively obtain corresponding extract, respectively by extract Reduced pressure concentration, collects chloroform extract powdered extract；

(3) step (2) gained chloroform extract powdered extract employing column chromatography is carried out isolated and purified, it is thus achieved that described sword-like leave ear Grass anthraquinone analog compound；The carrier of described column chromatography is silica gel, aluminum oxide, polyamide or octadecylsilane, and eluent is Volume ratio is the chloroform/acetone mixed liquor of 100:1 ~ 5:1 or chloroform/methanol mixed liquor that volume ratio is 200:1 ~ 6:1.

2. the preparation method of Herba Hedyotis Lanceae anthraquinone analog compound as claimed in claim 1, it is characterised in that described step (1) Extraction be: respectively with ethanol that volumetric concentration is 95%, volumetric concentration be 80% ethanol, volumetric concentration be 60% ethanol, body Volume concentrations be 40% second alcohol and water as solvent, Herba Hedyotis Lanceae carried out seepage pressure effects, Herba Hedyotis Lanceae and the quality of each solvent Than being 1:8, merging extract, by gained extract reduced pressure concentration, recycling design, to without alcohol taste, obtains extract.

3. the preparation method of Herba Hedyotis Lanceae anthraquinone analog compound as claimed in claim 1, it is characterised in that described step (3) Column chromatography use purification on normal-phase silica gel column chromatography.

4. the preparation method of the Herba Hedyotis Lanceae anthraquinone analog compound as described in claim 3, it is characterised in that also include step Suddenly target stream part that (3) obtain through purification on normal-phase silica gel column chromatography utilizes Sephadex LH-20, reversed-phase silica gel column chromatography or high-efficient liquid Phase chromatogram is purified, it is thus achieved that described Herba Hedyotis Lanceae anthraquinone analog compound.

5. according to the preparation method of the Herba Hedyotis Lanceae anthraquinone analog compound one of claim 1 to 4 Suo Shu, it is characterised in that institute The granularity stating silica gel is 200 ~ 300 mesh.

6. the preparation method of Herba Hedyotis Lanceae anthraquinone analog compound as claimed in claim 1, it is characterised in that described step (3) Also include: step (2) gained chloroform extract powdered extract is carried out pillar layer separation, then uses chloroform/acetone mixed liquor ladder Degree wash-out, in chloroform/acetone mixed liquor, the volume ratio of chloroform/acetone is followed successively by: 100:1 → 80:1 → 50:1 → 20:1 → 10:1 →5:1→2:1→1:1；The wash-out stream part collected be chloroform/acetone volume ratio be the solution that the mixed liquor of 50:1 elutes, Again through thin-layer chromatography detection colour developing, merging same stream part, reduced pressure concentration, with dry, obtains dirty-green feathery crystalline；Again through silicon Plastic column chromatography separates, the chloroform/methanol gradient elution using volume ratio to be 5:1 ~ 1:2 in silica gel column chromatography, obtains yellow green plumage Hairy crystallizes, more repeatedly purifies through Sephadex LH-20 column chromatography, then with the chloroform/methanol mixed liquor that volume ratio is 1:1 Wash-out, obtains light green feathery crystalline, is described Herba Hedyotis Lanceae anthraquinone analog compound.

The preparation method of Herba Hedyotis Lanceae anthraquinone analog compound the most according to claim 6, it is characterised in that described column chromatography Adsorbent with 100 ~ 200 mesh silica gel.

8. the preparation method of the Herba Hedyotis Lanceae anthraquinone analog compound as described in claim 1, it is characterised in that described sword-like leave ear Grass uses the herb of Herba Hedyotis Lanceae plant.