CN102250110B - Magnesium cantharidate and preparation method thereof - Google Patents

Abstract

The invention discloses magnesium cantharidate the structural formula (I) of which is shown in the specification. The method comprises the following steps: adding chloroform or isopropanol in pure cantharidin in a weight ratio of 1:(30-50), and heating for dissolving the pure cantharidin; adding magnesium hydroxide in distilled water the volume of which is the same as that of chloroform or isopropanol so as to prepare a turbid liquid, wherein the molar ratio of pure cantharidin to magnesium hydroxide is 3:2; adding the magnesium hydroxide turbid liquid in the chloroform solution or isopropanol solution which is used for dissolving the cantharidin, heating to 60-75 DEG C, and volatilizing chloroform or isopropanol completely so as to obtain white suspension; placing the white suspension ina oven for concentration and drying so as to obtain white crystallized powders; placing chloroform or isopropanol in the white crystallized powders in the weight ratio of (30-50):1, heating to 40 DEGC, and maintaining the temperature so as to dissolve the cantharidin, filtering and drying so as to obtain white crystallized powders. Magnesium cantharidate has the characteristics of high anticanver activity, good safety, stable finished product property and high purity, and the process is simple and is easy to operate.

Description

Magnesium cantharidate and preparation method thereof

Technical field

The invention belongs to technical field of chemistry, particularly a kind of magnesium cantharidate also relates to the preparation method of this magnesium cantharidate simultaneously.

Background technology

China just brings into use Chinese blister beetle to treat tumour in oneth, two century of Christian era.After be proved the Cantharidin of Chinese blister beetle secretion and cantharidin derivative for mainly containing effective substance.Be often used as oral pharmaceutical uses because Cantharidin is water insoluble, limited like this range of application of Cantharidin, rear people have developed cantharidate sodium increases the water-soluble toxicity that reduced simultaneously, utilize this excellent properties, developed the cantharidate sodium injection, can use by direct injection, present cantharidate sodium injection has been successfully applied to clinical, and has obtained significant effect.But cantharidate sodium is falling the hypotoxic while, and antitumour activity is along with great reduction, and this new has more that the cantharidin derivative of high anti-cancer activity solves this problem in the urgent need to developing other.

Summary of the invention

The object of the invention is to overcome above-mentioned shortcoming and a kind ofly having than high anti-cancer activity of providing, technique is simple, easy handling, and security is good, and end properties is stable, the magnesium cantharidate that purity is high.

Another object of the present invention is to provide the preparation method of this magnesium cantharidate.

Magnesium cantharidate of the present invention, its structural formula (I) is:

(I)

The preparation method of a kind of magnesium cantharidate of the present invention comprises the steps:

The weight ratio of (1) pressing 1:30-50 adds trichloromethane or Virahol in the Cantharidin sterling, the heating make its dissolving, get simultaneously the Cantharidin sterling: the magnesium hydroxide mol ratio be the magnesium hydroxide of 3:2 join with trichloromethane or the isopyknic distilled water of Virahol in make suspension liquid;

(2) the magnesium hydroxide suspension liquid is joined in the chloroform soln or aqueous isopropanol of dissolving Cantharidin, be heated to 60-75 ℃ after, trichloromethane or Virahol are volatilized, filter, get white suspension; Put in the baking oven concentratedly, drying gets white crystalline powder;

(3) weight ratio by 30-50:1 joins trichloromethane or Virahol in the white crystalline powder, is heated to 40 ℃ and keeps this temperature to make the Cantharidin dissolving, filters, and drying gets white crystalline powder, and get final product.

Beneficial effect of the present invention, by above technical scheme as can be known, it is simple that the present invention has technique, easy handling, the magnesium cantharidate that obtains is an analog derivative of Cantharidin, has the antitumor mechanism similar to Cantharidin, experimental results show that it has higher antitumour activity.And molecular weight is little, easily enters in the cancer cells, produce cytotoxicity, thereby have stronger antitumor action, and be a kind of comparatively desirable cancer therapy drug.Can be mainly used in clinically the treatment of late primary liver cancer and advanced lung cancer.Its security is good.The finished product stable in properties that present method makes, purity is high.Because it is water-soluble preferably that it has, can be applicable to the antitumor drug production of the multiple formulations such as injection, solution, ointment.

Embodiment

Further specify by the following examples the beneficial effect of the inventive method with experimental example.

Embodiment 1:

A kind of preparation method of magnesium cantharidate comprises the steps:

(1) add Cantharidin sterling 1g in the Erlenmeyer flask of 50ml, add the 30ml chloroform soln, heating makes its dissolving, gets simultaneously magnesium hydroxide 0.2 g and adds 30ml distilled water and make suspension liquid;

(2) the magnesium hydroxide suspension liquid is joined in the chloroform soln of dissolving Cantharidin, be heated to 60 ℃, trichloromethane is volatilized, filter, get white suspension; Put in the baking oven concentratedly, drying gets white crystalline powder;

(3) trichloromethane with 30ml joins in the white crystalline powder, is heated to 40 ℃ of constant temperature 2 hours, makes the Cantharidin dissolving, filters, and drying gets white crystalline powder and is the magnesium cantharidate sterling.

Embodiment 2:

A kind of preparation method of magnesium cantharidate comprises the steps:

(1) add Cantharidin sterling 1g in the Erlenmeyer flask of 100ml, add 50 ml aqueous isopropanols, heating makes its dissolving, gets simultaneously magnesium hydroxide 0.2 g and adds 30ml distilled water and make suspension liquid;

(2) the magnesium hydroxide suspension liquid is joined in the aqueous isopropanol of dissolving Cantharidin, be heated to 75 ℃, Virahol is volatilized, filter, get white suspension; Put in the baking oven concentratedly, drying gets white crystalline powder;

(3) Virahol of 50ml is joined in the white crystalline powder (white crystalline powder is joined in the Virahol of 50ml), be heated to 40 ℃ of constant temperature 2 hours, make the Cantharidin dissolving, filter, drying gets white crystalline powder and is the magnesium cantharidate sterling.

Experimental example 1: product purity inspection

Magnesium cantharidate does not have the quality determination standard of national regulation, so measure at this measuring method with reference to the cantharidate sodium national drug standards [standard No.: WS-10001-(HD-0044)-2002].

Assay

Chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as weighting agent; Methanol-water (51:49) is moving phase; Flow velocity is 1.0ml/min; The detection wavelength is 227nm; Sensitivity is 0.005AUFS.Number of theoretical plate is pressed the Cantharidin peak and is calculated, and should be not less than 2000.

The preparation of reference substance solution: it is an amount of to get the Cantharidin reference substance, accurately weighed, adds methyl alcohol and makes the solution that per 1 ml contains Cantharidin 0.15mg, in contrast product solution.

The preparation of need testing solution: precision takes by weighing this product 10mg, to the 100ml flask, adds 50ml water, adds 1mol/L H again ₂SO ₄Solution 10ml, shake up, put in 65 ℃ of water-baths and to reflux 30 minutes, move in separating funnel after slightly cold, with trichloromethane 25 ml, 15 ml, 10 ml extraction three times, each 10 minutes, combining extraction liquid was put in 70 ℃ of water-baths and is volatilized solvent, residue turns appearance to 50 ml measuring bottles with methyl alcohol, constant volume shakes up, and is need testing solution.

Assay method: precision is drawn reference substance solution and each 20 μ l of need testing solution respectively, the injection liquid chromatography, the record color atlas calculates Cantharidin content with external standard method, multiply by again the molecular weight of the molecular weight/Cantharidin of 1.20(magnesium cantharidate), namely get the cantharidate Mg content.

The magnesium cantharidate purity that detected result embodiment 1,2 obtains is more than 90%.

Experimental example 2: measure magnesium cantharidate to concentration in the inhibition of cancer cells with mtt assay

With 0.02% EDTA solution peptic cell, make cell suspension through piping and druming, collection, after centrifugal, adjusting cell concn is SGC7901 cell or 8 0000/mL of liver cancer QGY-7703 cell, is inoculated in 96 orifice plates with every hole 100 μ L, places 37 ℃, 5%CO ₂Spend the night in the incubator, make cell cover with the bottom surface; Remove nutrient solution, then add respectively the DMEM nutrient solution (the Cantharidin treatment group contains 0.5% acetone) that 100 μ L series contain different pharmaceutical (being respectively Cantharidin, cantharidate sodium, magnesium cantharidate) concentration, each concentration is established 4 holes again; At 37 ℃ of CO ₂After hatching 24 h in the incubator, every hole adds the WST-1 solution 20 μ L that prepared, and lucifuge is in 5% CO ₂In the incubator, cultivate 2h for 37 ℃, microplate reader detects absorbancy (A) value in every hole, wavelength 450nm; Establish respectively without medicine control group (cell is arranged in the nutrient solution, and the Cantharidin treatment group contains 0.5% acetone) and blank group (acellular in the nutrient solution, the Cantharidin treatment group contains 0.5% acetone).

The calculation formula of inhibiting rate is as follows:

Inhibiting rate %=[1-(medicine group OD value-blank OD value)/(contrast OD value-blank OD value)] * 100%

Drug level (IC when asking 50% cell inhibitory rate with regression curve ₅₀).

After testing, magnesium cantharidate is to concentration (IC in the inhibition of liver cancer QGY-7703 ₅₀) be 9.48 μ mol/L, be higher than the activity of Cantharidin and cantharidate sodium sodium.Magnesium cantharidate is to concentration (IC in the inhibition of cancer of the stomach SGC7901 cell ₅₀) be 8.65 μ mol/L, be higher than the activity of Cantharidin and cantharidate sodium.

Claims (1)

1. the preparation method of a magnesium cantharidate comprises the steps:

The weight ratio of (1) pressing 1:30-50 adds trichloromethane or Virahol in the Cantharidin sterling, the heating make its dissolving, get simultaneously the Cantharidin sterling: the magnesium hydroxide mol ratio be the magnesium hydroxide of 3:2 join with trichloromethane or the isopyknic distilled water of Virahol in make suspension liquid;

(2) the magnesium hydroxide suspension liquid is joined in the chloroform soln or aqueous isopropanol of dissolving Cantharidin, be heated to 60-75 ℃ after, trichloromethane or Virahol are volatilized, filter, get white suspension; Put in the baking oven concentratedly, drying gets white crystalline powder;

(3) weight ratio by 30-50:1 joins trichloromethane or Virahol in the white crystalline powder, is heated to 40 ℃ and keeps this temperature to make the Cantharidin dissolving, filters, and drying gets white crystalline powder, and get final product.