CN101735289A

CN101735289A - Compounds extracted from traditional Chinese medicine of Cassia Seed and applications

Info

Publication number: CN101735289A
Application number: CN200810226864A
Authority: CN
Inventors: 王祝举; 唐力英; 吴宏伟
Original assignee: Institute of Materia Medica of CAMS
Current assignee: Institute of Materia Medica of CAMS
Priority date: 2008-11-19
Filing date: 2008-11-19
Publication date: 2010-06-16
Anticipated expiration: 2028-11-19
Also published as: CN101735289B

Abstract

The invention discloses two new compound structures from traditional Chinese medicine of Cassia Seed, which respectively are 1-hydroxyl-2-acetyl-3, 8-dimethoxy-6-O-[belta-D-apiofuranosyl-(1 to 2) beta-D-glucopyranosyl]-naphthalene (I) and 1-demethylaurantio-obtusin-2-O-beta-D-glucopyranoside (II). Meanwhile, the invention also discloses a method for extracting and separating the two new compounds, and applications of the derivates of the two compounds in preparing oxidation resistant medicines and liver medicines.

Description

Compound that from study of semen cassia, extracts and uses thereof

Technical field

The present invention is a field of phytochemistry, and extraction, separation and the preparation technology field of pharmaceutically active ingredient in specifically belonging to relates in particular to the compounds and methods that from study of semen cassia extraction separation has pharmacologically active.

Background technology

Semen Cassiae (Semen Cassiae) is the dry mature seed of leguminous plants Cassia tora Cassia obtusifolia L. or little Cassia tora Cassia tora L..The chemical ingredients of Semen Cassiae is mainly anthraquinone component, also has other compositions such as Benzofurantone, sterols, fatty acid in addition.Modern pharmacology studies show that Semen Cassiae has hypotensive, and reducing blood-fat protects the liver and antibacterial isoreactivity, is widely used.In addition, Semen Cassiae is again the Chinese medicine that can be used as medicine-food two-purpose, is the good raw material of field of health care food.At present, deep not enough to the chemical constitution study of study of semen cassia, need further to excavate its effective constituent, the relation of clear and definite its pharmacologically active and effective constituent.

Summary of the invention

One of purpose of the present invention is the compound that is different from prior art that extraction separation has pharmacologically active from study of semen cassia, and extraction separation gets 2 new compounds and is:

1-hydroxyl-2-acetyl-3,8-dimethoxy-6-O-[β-D-apiofuranosyl-(1 → 2)-β-D-glucopyranosyl]-naphthalene (new naphthalene compounds, Compound I) and

1-demethylaurantio-obtusin-2-O-β-D-glucopyranoside (new anthraquinone analog compound, Compound I I).

The concrete extraction separation step of study of semen cassia is:

Semen Cassiae dry seed 20kg, pulverize the 95% alcohol diacolation extraction of back with 10 times of amounts of crude drug, be evaporated to and do not have the alcohol flavor, adding suitable quantity of water disperses, use sherwood oil, chloroform, n-butanol extraction successively, reclaim solvent and get petroleum ether extract 100g, chloroform extract 150g, n-BuOH extract 200g and remainder water part.The n-BuOH extract, separate through silica gel (100-200 order) column chromatography, the chloroform-methanol gradient elution, every 500ml collects first-class part, reclaims solvent, wherein 88-90 stream part, filter out the part that is insoluble to methyl alcohol, dried powder is Compound I I, wherein after part merging of 109-113 stream, separates through silica gel (100-200 order) column chromatography, ethyl acetate-methanol-water (10: 2: 1) wash-out, every 100ml collects portion stream part, obtains 74 flow points, wherein 22-33 stream part merging, all separate through polymeric amide (100-200 order) column chromatography, methanol-eluted fractions, the crystallization methanol wash of separating out gets Compound I.The extraction separation process as shown in Figure 1.

The compounds of this invention I is light yellow crystallization (methyl alcohol), UV λ _Max ^MeOHNm:232,275,310 and 392, be the characteristic absorbance of naphthalene compounds.The thin layer plate spray shows red with 5%KOH.The Molish reaction is positive.HRESI-MS (m/z) provides quasi-molecular ion peak: 579.1684, and calculated value (C ₂₅H ₃₂O ₁₄+ Na): 579.1679, so determine that its molecular formula is C ₂₅H ₃₂O ₁₄

¹H-NMR and ¹³The C-NMR data see the following form 1:

Table 1 Compound I NMR data (DMSO-d6, TMS, δ ppm)

By ¹H-NMR, ¹³C-NMR and two-dimensional spectrum determine that compound structure is:

1-hydroxyl-2-acetyl-3，8-dimethoxy-6-O-[β-D-apiofuranosyl-(1→2)-β-D-glucopyranosyl]-naphthalene。

Compound I

The compounds of this invention II is the orange powder, UV λ _Max ^MeOHNm:218,254,278,314 and 438, be the characteristic absorbance of anthraquinone analog compound.The thin layer plate spray shows red with 5%KOH.The Molish reaction is positive.HRESI-MS (m/z) provides quasi-molecular ion peak: 501.1003, and calculated value (C ₂₅H ₃₂O ₁₄+ Na): 501.1024, so determine that its molecular formula is C ₂₅H ₃₂O ₁₄

¹H-NMR and ¹³The C-NMR data see the following form 2.

Table 2 Compound I I NMR data (DMSO-d ₆, TMS, δ ppm)

1-demethylaurantio-obtusin-2-O-β-D-glucopyranoside。

Compound I I

Another purpose of purpose of the present invention, Compound I and Compound I I have purposes aspect the pharmacologically active medicine in preparation.Studies show that Compound I has the effect that the antagonism mouse carbon tetrachloride causes liver injury, prompting has liver-protecting activity, Compound I I has the effect of removing the DPPH free radical, and its activity obviously is better than vitamins C (Vitamin C, VC) (P＜0.01), prompting Compound I I has the activity of anti-oxidant aspect.

Embodiment

The extraction and separation method of embodiment 1 Compound I and II:

Semen Cassiae dry seed 20kg, pulverize the 95% alcohol diacolation extraction of back with 10 times of amounts of crude drug, be evaporated to and do not have the alcohol flavor, adding suitable quantity of water disperses, use sherwood oil, chloroform, n-butanol extraction successively, reclaim solvent and get petroleum ether extract 100g, chloroform extract 150g, n-BuOH extract 200g and remainder water part.The n-BuOH extract, separate through silica gel (100-200 order) column chromatography, the chloroform-methanol gradient elution, every 500ml collects first-class part, reclaims solvent, wherein 88-90 stream part, filter out the part that is insoluble to methyl alcohol, dried powder is Compound I I, wherein after part merging of 109-113 stream, separates through silica gel (100-200 order) column chromatography, ethyl acetate-methanol-water (10: 2: 1) wash-out, every 100ml collects portion stream part, obtains 74 flow points, wherein 22-33 stream part merging, all separate through polymeric amide (100-200 order) column chromatography, methanol-eluted fractions, the crystallization methanol wash of separating out gets Compound I.

The activity of embodiment 2 naphthalene compounds I aspect protecting the liver

1 experiment material

1.1 sample and reagent

Tetracol phenixin is provided by Beijing chemical reagent factory.Alanine aminotransferase (ALT), aspartate transaminase (AST) detection kit is built up biological study by Nanjing is provided.Positive control drug: Biphenylylmethylcarbinol, Dezhou Deyao Pharmaceutical Co., Ltd, lot number 040410.Specimen: the naphthalene compounds I:1-hydroxyl-2-acetyl-3 that above-mentioned separation obtains, 8-dimethoxy-6-O-[β-D-apiofuranosyl-(1 → 2)-β-D-glucopyranosyl]-naphthalene.

1.2 animal

Kunming mouse, male, body weight 18～22g is provided by Chinese department of Chinese medicine institute Experimental Animal Center.

1.3 instrument

752 ultraviolet grating spectrophotometers, Shanghai Precision Scientific Apparatus Co., Ltd; LDZ5-2 type whizzer, Beijing Medical Centrifugal Machine Factory; DKB-8A type electric heating constant temperature tank, the grand experimental installation of last Nereid company limited.

2 experiment contents

Get 90 of Kunming mouses, be divided into 9 groups at random, 10 every group.Grouping and gastric infusion dosage see Table 3, administration every day 1 time, 15d continuously.10h after the last administration, except that the blank group, all the other respectively organize abdominal injection 0.15% tetracol phenixin oil solution 10ml/kg, fasting, freely drink water.Behind the injection tetracol phenixin 16h, the mouse orbit venous plexus is got blood, the centrifugal 10min of 2500r/min, and separation of serum is in the content of 505nm mensuration mice serum ALT, AST.

3 experimental results

Large, medium and small 3 dosage groups all can significantly reduce the content that tetracol phenixin causes acute liver damage mice serum ALT and AST, with model group significant difference are arranged relatively, (P＜0.05 or P＜0.01).The results are shown in Table 3.

Table 3 Compound I to tetracol phenixin cause the chmice acute liver injury provide protection (

, n=5)

Annotate: compare with model group, ^*P＜0.05 or ^*P＜0.01

Above presentation of results Compound I causes the chmice acute liver injury to tetracol phenixin and has significant protective effect under large, medium and small 3 dosage.

The activity of embodiment 3 anthraquinone analog compound II aspect anti-oxidant

1 experiment material

1.1 sample and reagent

DPPH (1, the bitter diazanyl free radical of 1-phenylbenzene-2-), Sigma company product.The positive control drug vitamins C, Sigma company.Specimen: the anthraquinone analog compound II that above-mentioned separation obtains is 1-demethylaurantio-obtusin-2-O-β-D-glucopyranoside

1.2 instrument

UV-T6 new millennium uv-spectrophotometric device, KQ-500E type ultrasonic cleaning machine for medical purpose (Kunshan Ultrasonic Instruments Co., Ltd.), Sartorius ten thousand/electronic balance.

2 experiment contents

2.1 remove the detection method of DPPH free radical

Respectively accurately claim to decide 2.1mgVC, 2.4mg Compound I I is dissolved in respectively and is mixed with concentration with ethanol in the 10mL measuring bottle and is respectively 0.21mg/mL, 0.24mg/mL, and it is standby above liquid to be measured to be diluted to series concentration by 2 times, 6 times, 18 times, 54 times, 162 times.Precision takes by weighing 8.4mgDPPH, is mixed with 0.084mg/mL (2.1 * 10 ^-4Mol/L) standby.

Accurate respectively each liquid 2ml to be measured that draws, the DPPH solution (0.084mg/mL) of adding 2mL shakes up the back and places 30min, is blank with the solvent, measures the Abs value A1 at 517nm place; Measure simultaneously above-mentioned liquid 2mL to be measured mix with the 2mL blank solvent back 517nm the Abs value be designated as A2; Measure again 2mLDPPH solution mix with the 2mL blank solvent back 517nm the Abs value be designated as A3; Be worth according to the clearance rate of following formula calculating three the DPPH free radical.

Clearance rate (%)=[1-(A1-A2)/A3] * 100%.

3 experimental results

Table 4 Compound I I and VC to the clearance rate of DPPH free radical (

N=5)

Annotate: * * represents P＜0.01

Above result shows: Compound I I has the effect of removing the DPPH free radical, and its activity obviously is better than VC (P＜0.01), and prompting Compound I I has the activity of anti-oxidant aspect.

Description of drawings

Fig. 1 is a Semen Cassiae medicinal material extraction separation schema.

Claims

1. new naphthalene compounds is characterized in that having the structure of following Compound I:

Compound I.

2. the purposes of compound as claimed in claim 1 in the anti-visceral organ injury medicine of preparation.

3. purposes as claimed in claim 2 is characterized in that the purposes in the anti-hepar damnification medicine of preparation.

4. the preparation method of compound according to claim 1 is characterized in that:

A. the 50%-95% ethanol percolation that the Semen Cassiae dry seed is doubly measured with 5-10, concentrating under reduced pressure adds suitable quantity of water and disperses;

B. water dispersion is used sherwood oil, chloroform, n-butanol extraction successively, reclaim solvent and get petroleum ether extract, chloroform extract, n-BuOH extract and water section;

C. with the n-BuOH extract,, hang down the polar organic solvent gradient elution through the column chromatography separation, a stream part of every 500ml collection, wherein 88-90 flows part, filters out the part that is insoluble to methyl alcohol, and dried powder is Compound I I;

D. 109-113 flow point merging separates through column chromatography, ethyl acetate-methanol-water wash-out, and every 100ml collects a stream part, obtain 74 flow points, wherein the 22-33 flow point merges respectively, all separates through column chromatography, methanol-eluted fractions, the crystallization methanol wash of separating out gets Compound I.

5. new anthraquinone analog compound is characterized in that having the structure of following Compound I I:

Compound I I.

6. as the purposes of compound as described in the claim 5, it is characterized in that preparing purposes with antioxygenation medicine.