CN102718647A - Magnesium Danshensu and its preparation method and use - Google Patents

Abstract

The invention discloses magnesium Danshensu and its preparation method and use. The magnesium Danshensu is a drug for preventing and treating cardiovascular and cerebrovascular diseases. The preparation method has the advantages of easily available raw materials, simple processes and high yield. The magnesium Danshensu has a good structure, good curative effects, low toxicity and strong druggability.

Description

The Salvianic acidA magnesium salts

Technical field

The present invention relates to Salvianic acidA magnesium salts and preparation method thereof and the purposes in preparation treatment stenocardia, coronary heart disease, cerebral infarction and cardiac-cerebral ischemia/reperfusion injury medicine, belong to medical technical field.

Background technology

According to World Health Organization's statistics, the cardiovascular and cerebrovascular diseases mortality ratio ranks first, and annual death toll reaches 1,750 ten thousand people, accounts for 30% of all death tolls.Therefore, the medicine of seeking the excellent effect treatment heart, cerebrovascular disease is the difficult problem that current medical circle needs to be resolved hurrily.

For thousands of years; The red sage root is the important component of treatment cardiovascular and cerebrovascular diseases Chinese medicine compound prescription always, and Chinese medicine " Radix Salviae Miltiorrhizae Tabellae " (red sage root), " DANHONG ZHUSHEYE " (red sage root and safflower), " two capsule " (red sage root and Tree Peony Bark), " FUFANG DANSHEN DIWAN " (red sage root, pseudo-ginseng and borneol) all are to be main ingredient with the red sage root.Its activeconstituents comprises Salvianic acidA, poly phenolic acid of Radix Salviae Miltiorrhizae, TANSHINONES, coffic acid etc., and main active ingredient is a Salvianic acidA.Containing two phenolic hydroxyl groups in the Salvianic acidA molecule, is a kind of good oxygen free radical scavenger.

The main pharmacological of Salvianic acidA has: 1, to the effect of cardiac muscle: have the effect of dwindling myocardial infarct size and alleviating the course of disease, simultaneously myocardial ischemia is had provide protection; 2, anticoagulant and anticoagulation: can obviously suppress hematoblastic gathering, and the flowability of platelet increasing film; 3, Azelaic Acid and enhancing body immunization; 4, atherosclerosis and reducing blood lipid: suppress the synthetic of cell endogenous cholesterol, also have the effect of lipotropism matter protein oxidation, reduce blood cholesterol, have protection blood vessel barrier, prevent lipid deposit and atherosclerosis (AS) effect; 5, anti-thrombosis function: have function of promoting blood circulation to disperse blood clots.6, coronary artery dilator effect: ability is coronary artery dilator obviously, and blood flow coronarius is significantly increased; 7, anti-cerebral ischemia damnification effect: obviously dwindle brain infarction area, improve neurologic impairment; 8, Salvianic acidA is to the treatment of pulmonary heart disease: have the effect of expansion artery, and can suppress thrombocyte and discharge the shrinkability material.

In the past, people have ignored mg ion to the active contribution of the red sage root, and the acid-alkali treatment in the red sage compound preparation process thereof especially in the past can destroy the mineral ion effect.In fact, in the clinical heart operation, patient's ubiquity hypomagnesemia has increased the myocardial damage that aorta clamping causes thus.Can cause Ca in the cell during myocardial ischemia ²⁺Increasing of concentration.Research shows Mg ²⁺Can stop Ca ²⁺In cell, flow into, use magnesia mixture and can alleviate ischemia/reperfusion injury.

Summary of the invention

The purpose of this invention is to provide a kind of novel drugs--Salvianic acidA magnesium that cardiovascular and cerebrovascular diseases is had excellent effect prevention and therapeutic action.

Salvianic acidA magnesium salts shown in the formula (I),

。

The mass spectrum M/Z of said Salvianic acidA magnesium salts is 417.50; ¹H-NMR:6.8-7.1 (6H, Ph ^-), 5.5 (4H ,-OH), 4.5 (2H ,-CH), 2.8-3.2 (4H ,-CH ₂), 2.7 (2H ,-OH); X-ray diffraction pattern comprises the characteristic peak of intensity greater than other peak at 2 θ places of 8.15,16.91,19.40,21.94,23.78,24.94,27.90 and 30.35 degree.

Use the EDTA titration, Mg ²⁺Content is 5.6～5.8%.

The preparation method of above-mentioned Salvianic acidA magnesium salts is that Salvianic acidA is dissolved in ethanol, adds alkali and makes Salvianic acidA slough proton, adds the solubility magnesium salts according to Salvianic acidA and mg ion mol ratio 2:1, gets white precipitate Salvianic acidA magnesium salts, through recrystallization or column chromatography purification.

Said Salvianic acidA is that synthetic or from the former medicine of the red sage root, extract obtains.

From the former medicine of the red sage root, extracting Salvianic acidA may further comprise the steps:

(1) the former medicine of the red sage root adds water extraction filtration, and extraction liquid is concentrated into the thick paste shape;

(2) add ethanol to solution and contain the alcohol amount and be 70-80%, cooling, filter, be concentrated into the medicinal extract shape;

(3) Radix Salviae Miltiorrhizae extractum is dissolved in water, and filters and use ethyl acetate extraction, concentrated ETHYLE ACETATE cream;

(4) ETHYLE ACETATE cream carries out silica gel column chromatography, with chloroform-acetone-formic acid wash-out, collects corresponding cut and gets Salvianic acidA.

The invention discloses medicine--the Salvianic acidA magnesium salts of a kind of control and prevention cardiovascular and cerebrovascular diseases, its preparation method raw material is easy to get, technology is simple, productive rate is high.This medicines structure good effect, toxicity is low, becomes the property of medicine strong.

Description of drawings

Fig. 1 is the mass spectrum of Salvianic acidA magnesium salts of the present invention;

Fig. 2 is the X-ray powder diffraction pattern of Salvianic acidA magnesium salts of the present invention;

Fig. 3 is a heart slice map behind the rat myocardial ischemia and reperfusion.

Embodiment

Embodiment 1: the preparation of Salvianic acidA magnesium

(1) preparation method

Get the dry root 2kg of the red sage root, add poach 2 times, filter, be concentrated into the thick paste shape.Adding ethanol to solution contains the alcohol amount and is 70-80%, and cooling, filtration are concentrated into the medicinal extract shape.Get danshen cream and be dissolved in water, filter and use the ethyl acetate extraction aqueous solution, concentrated ETHYLE ACETATE cream.Carry out silica gel column layer,, collect corresponding cut and get Salvianic acidA with chloroform-acetone-formic acid different ratios wash-out.Salvianic acidA is dissolved in ethanol, and adjust pH to 6.8 drips magnesium chloride solution, reclaims ethanol to a small amount of, and room temperature is placed and just separated out white solid Salvianic acidA magnesium salts, recrystallization or column chromatography purification, yield 2-3 ‰.

(2) characterization data

As shown in Figure 1, MS:M/Z 417.50.

¹H-NMR：6.8-7.1（6H，Ph-），5.5（4H，-OH），4.5（2H，-CH），2.8-3.2（4H，-CH ₂），2.7（2H，-OH）。

Mg content, EDTA-2Na titration Mg are surveyed in conventional complexometric titration ²⁺: actual Mg ²⁺%=5.66% (theoretical Mg ²⁺%=5.7%).

X-ray powder diffraction: testing tool: BRUKER EQUZNOX-55, Cu Kα (0.15406 nm), WV: 40 kv, working current: 40 mA, scanning speed: 0.03 s/step, the result is shown in Fig. 2 and table 1.

Embodiment 2: the pharmacodynamic experiment of Salvianic acidA magnesium

Test materials:64 of Sprague-Dawley male rats, weight (230 ± 20) g is provided by The Fourth Military Medical University's Experimental Animal Center; Tn (CK), superoxide-dismutase (SOD), mda reagent (MDA), nitrogen protoxide detect box (NO), Selenoperoxidase testing cassete (GSH-P _X), XOD testing cassete (XOD).Test kit is that biological study institute product is built up in Nanjing.

, the myocardial ischemia-reperfusion injury pharmacodynamic study

The myocardial ischemia-reperfusion Preparation of model

32 rats are divided into 4 groups at random: sham operated rats; Perfusion group again; Salvianic acidA group (30mg/kg); Salvianic acidA magnesium group (30mg/kg).Administration group 10min administration before ischemic (jugular vein injection), dosage is confirmed according to the beneficial effect curve of new drug.

Behind the SD rat anesthesia, adopt ligation ADC blood vessel method to prepare rat MI/R (30min/3h) model: otch promoting the circulation of qi cannula in the positive middle part of neck connects respirator pedestrian worker and breathes.Separate jugular vein and be used for administration.Open chest and expose heart, last 1/3 place in No. 3/0 silk thread ligation ramus descendens anterior arteriae coronariae sinistrae, ECG ST section is raised and is ligation success index.

1. the mensuration of heart function detection and serum CK and SOD of heart tissue, MDA, NO

Ischemic unclamped silk thread after 30 minutes, poured into the J point degree of displacement Δ ST that observes rat electrocardiogram(ECG (EKG) after 3 hours again.Get blood 2mL by carotid artery, as for dry centrifuge tube, after normal temperature left standstill 30min, the centrifugal 15min of 4000r/min got serum, according to the variation of test kit specification sheets operation detection serum CK and the SOD of cardiac muscular tissue, MDA and NO.Experimental result shows, Salvianic acidA magnesium can reduce the displacement of J point ( P<0.01), reduce the serum CK burst size ( P<0.01), the ability SOD activity improving ( P<0.01), minimizing MDA generation ( P<0.01), increase in right amount NO content ( P<0.01).

2. myocardial infarct size is measured

As shown in Figure 3, after pouring into 3h again and finishing, the ligation coronary artery injects 3% azovan coerulen (1～2mL) to left ventricular cavity.Speed is taken out heart, and is frozen in-20 ℃.Be cut into the thick thin slice of 1mm with the heart food slicer perpendicular to long axis of heart, burst places the 12 hole petridish that contain 2mL 1%TTC (pH 7.4), hatches 15min for 37 ℃.Take pictures and import computingmachine with digital camera.Adopt single blind method respectively with azovan coerulen dyeing district (non-ischemic region), TTC dyeing district (red dying, ischemic but still survival tissue) and non-TTC dyeing district (infarcted myocardium), calculate software with the SigmaScan area and carry out computing.Myocardial infarct size is represented with the per-cent that the total infarct size of each heart (INF) accounts for total ischemic region area (AAR).Experimental result shows, Salvianic acidA magnesium can effectively reduce myocardial ischemia zone and infarcted region area ( P<0.01).

, cerebral ischemia re-pouring injured pharmacodynamic study

Cerebral ischemia re-pouring injured model preparation

Make brain I/R model according to reversibility middle cerebral artery occlusion line bolt legal system,, separate left carotid (CCA), external carotid artery (ECA) and internal carotid artery (ICA) with 10% Chloral Hydrate intraperitoneal anesthesia rat.Behind ligation CCA proximal part, the ECA initiating terminal, the of short duration folder of arteria pterygopalatina closes with anti-mis-insertion, inserts the line bolt through CCA, and depth of penetration is remembered 18 mm by furcation.Perfusion is treated to the backguy bolt and makes the end of a thread retreat to external carotid artery again.32 rats are divided into 4 groups at random: sham operated rats; Perfusion group again; Salvianic acidA group (30mg/kg); Salvianic acidA magnesium group (30mg/kg).The administration group is administration again (abdominal injection) behind ischemic 30min, and dosage is confirmed according to the beneficial effect curve of new drug.

1. the TTC staining is measured the cerebral infarction scope

Behind the ischemia-reperfusion, broken end is got brain, rejects rhinencephalon, low brain stem and cerebellum, and-20 ℃ of freezing 15min of refrigerator begin section, every thick 2 mm apart from antinion 3 mm.Rapidly the brain sheet is placed then and contain the 1%TTC aqueous solution, 37 ℃ of temperature of lucifuge are incubated 30 min.After dyed, the ischemic region white colouring, the cerebral tissue that has normal blood to supply is redness.Cerebral tissue put fixing 24 h in 10% the formaldehyde solution.Observe and the turn white size of zone (being the cerebral infarction zone) of cerebral tissue relatively.After taking pictures with digital camera, use pathological image analysis-e/or determining infarct size, according to formula V=(A ₁+ ... + A _n) t/2 calculates Infarction volume, wherein tBe slice thickness, A is an infarct size.Experimental result shows, Salvianic acidA magnesium can effectively reduce rat cerebral tissue Infarction volume ( P<0.01).

2. the anti-oxidant mechanism of brain injury protection

The rat broken end is got the left side pallium behind the ischemia-reperfusion, places-7 ℃ of refrigerators frozen.After process 10% tissue homogenate, press test kit specification sheets operation, measure cerebral tissue t-SOD, GSH-P _X, XOD vigor and MDA content.Experimental result shows, Salvianic acidA magnesium can improve t-SOD in the rat cerebral tissue ( P<0.01), GSH-P _X( P<0.01) and XOD ( P<0.01) activity, reduce MDA ( P<0.01) content.

Claims (10)

1. the Salvianic acidA magnesium salts shown in the formula (I),

。

2. Salvianic acidA magnesium salts according to claim 1 is characterized in that: mass spectrum M/Z is 417.50.

3. Salvianic acidA magnesium salts according to claim 1 is characterized in that ¹H-NMR:6.8-7.1 (6H, Ph ^-), 5.5 (4H ,-OH), 4.5 (2H ,-CH), 2.8-3.2 (4H ,-CH ₂), 2.7 (2H ,-OH).

4. Salvianic acidA magnesium salts according to claim 1 is characterized in that: use the EDTA titration, Mg ²⁺Content is 5.6～5.8%.

5. Salvianic acidA magnesium salts according to claim 1 is characterized in that: its x-ray diffraction pattern comprises the characteristic peak of intensity greater than other peak at 2 θ places of 8.15,16.91,19.40,21.94,23.78,24.94,27.90 and 30.35 degree.

6. the preparation method of the said Salvianic acidA magnesium salts of claim 1; It is characterized in that: Salvianic acidA is dissolved in ethanol, adds alkali and make Salvianic acidA slough proton, add the solubility magnesium salts according to Salvianic acidA and mg ion mol ratio 2:1; Get white precipitate Salvianic acidA magnesium salts, through recrystallization or column chromatography purification.

7. the preparation method of Salvianic acidA magnesium salts according to claim 6 is characterized in that: said Salvianic acidA is that synthetic or from the former medicine of the red sage root, extract obtains.

8. the preparation method of Salvianic acidA magnesium salts according to claim 7 is characterized in that from the former medicine of the red sage root, extracting Salvianic acidA and may further comprise the steps:

(1) the former medicine of the red sage root adds water extraction filtration, and extraction liquid is concentrated into the thick paste shape;

(2) add ethanol to solution and contain the alcohol amount and be 70-80%, cooling, filter, be concentrated into the medicinal extract shape;

(3) Radix Salviae Miltiorrhizae extractum is dissolved in water, and filters and use ethyl acetate extraction, concentrated ETHYLE ACETATE cream;

(4) ETHYLE ACETATE cream carries out silica gel column chromatography, with chloroform-acetone-formic acid wash-out, collects corresponding cut and gets Salvianic acidA.

9. the application of the said Salvianic acidA magnesium salts of claim 1 in preparation prevention and treatment cardiovascular and cerebrovascular diseases medicament.

10. the application of the said Salvianic acidA magnesium salts of claim 1 in preparation prevention and treatment cardiac-cerebral ischemia/reperfusion injury medicine.

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