CN109718200B

CN109718200B - A silymarin compound and its water dispersion system

Info

Publication number: CN109718200B
Application number: CN201910141865.2A
Authority: CN
Inventors: 孙锐杰
Original assignee: Shanghai Pharmaceutical School
Current assignee: Shanghai Pharmaceutical School
Priority date: 2019-02-26
Filing date: 2019-02-26
Publication date: 2021-04-23
Anticipated expiration: 2039-02-26
Also published as: CN109718200A

Abstract

The invention provides a silymarin compound and a silymarin water dispersion system prepared by adopting the silymarin compound; the water dispersion of silymarin is stable in thermodynamics and kinetics, and can be stored for a long time in a liquid form; more importantly, the pharmacokinetic experiment proves that the water dispersion system of the silymarin is more completely released in vivo, the bioavailability is obviously improved, the drug effect is obviously improved, and the water dispersion system of the silymarin can be used for preparing liquid oral agents or injections of the silymarin, and the like.

Description

A silymarin compound and its water dispersion system

Technical Field

The invention relates to a silymarin compound and a water dispersion system adopting the compound, belonging to the technical field.

Background

Silymarin (silymarin) is an antioxidant extracted from Milk Thistle (Milk Thistle) plant, and has effects of stabilizing liver cell membrane, protecting liver cell from free radical damage, maintaining integrity of liver cell, preventing toxin from penetrating and damaging liver, accelerating synthesis of liver cell DNA, accelerating production of new liver cell, and self-repairing damaged liver cell.

The most prominent contribution of silymarin is the stabilization of the hepatic cell membrane, protection of the liver cells from toxic substances, e.g., liver damage caused by carbon tetrachloride, galactosamine, alcohols and other hepatotoxins, protection of the liver from the invasion of alcohol and environmental pollutants (pesticides, heavy metals, etc.), prevention or avoidance of the loss of soluble cellular components (e.g., transaminase), and limitation of the penetration of certain hepatotoxic substances (e.g., α -amanitin) into the interior of the cell. Thus, silymarin is called "natural liver-protecting drug" and is clinically used in acute and chronic hepatitis, early cirrhosis, toxic liver injury, and the like.

The silymarin formulations on the market today are tablets or capsules, i.e. all used in solid form, such as: silymarin tablets, liver benefiting tablets, silybin lecithin compound hard capsules and the like, and the bioavailability of solid medicines entering a human body is low; however, in the liquid state, silymarin is difficult to exist in a stable state, and the injection thereof needs to be made into a silymarin derivative such as: the production cost of the derivative is greatly increased and the cost performance is low.

Therefore, it is desired in the art to develop a silymarin aqueous dispersion which can be applied to liquid dosage forms such as oral liquid and injection.

Disclosure of Invention

In order to solve the technical problems, the invention provides a silymarin compound on one hand, wherein the silymarin compound is a compound of silymarin and poloxamer 188, and is prepared by the following method: adding 0.5-1.5 g of silymarin and 2-8 g of poloxamer 188 into 100ml of dispersion medium, performing azeotropic distillation for 0.5-1.5 hours, and adding 1-4 ml of softener to prepare the silymarin compound; the dispersion medium adopts polyethylene glycol, and the average molecular weight of the polyethylene glycol is 400-700; the softening agent is polyethylene glycol, and the average molecular weight of the polyethylene glycol is 400-700.

Preferably, polyethylene glycol 400 is used as both the dispersion medium and the softener.

Preferably, the silymarin compound is prepared by adding 1g of silymarin and 6g of poloxamer 188 into 100ml of dispersion medium, and adding 1.5ml of softener after azeotropic distillation for 1 hour to obtain the silymarin compound.

In another aspect of the present invention, there is provided a water dispersion of silymarin, wherein the water dispersion of silymarin is: the silymarin complex is dissolved in purified water to form milky water dispersion.

Preferably, in the water dispersion system of the silymarin, the volume concentration of the silymarin compound is 0.03-0.09 ml/ml.

The invention further provides a silymarin liquid medicament, wherein the liquid medicament is prepared by adopting the silymarin water dispersion system.

Preferably, the liquid medicament is an oral liquid.

Preferably, the liquid medicament is an injection.

Detailed Description

The present invention will be further described with reference to the following examples, but the present invention is not limited to these specific embodiments.

Example 1

Compound for preparing silymarin

The silymarin complex of example 1 is a silymarin-poloxamer 188 complex, which is prepared by the following method:

taking 100ml of dispersion medium (constant volume to 100ml, liquid polyethylene glycol PEG400 as dispersion medium, purchased from F990406 product of Shanghai chemical reagent company of China medicine (group), the same below), adding 1g of silymarin (provided by Huidong pharmaceutical Life engineering research institute of Shanghai city, lot number: 980826S, the same below) and 6g of poloxamer 188 (purchased from F991709 product of Shanghai chemical reagent company of China medicine (group), the same below), and keeping azeotropic state for 1 hour;

then, 1.5ml of a softener (liquid polyethylene glycol PEG400 as the softener, a product F990406 available from Shanghai chemical agents of China medicine, Ltd., the same applies hereinafter) was added thereto to prepare the silymarin complex of example 1.

Example 2

The silymarin complex of example 2 is a silymarin-poloxamer 188 complex, which is prepared by the following method:

100ml of dispersion medium (constant volume is 100ml, the dispersion medium adopts liquid polyethylene glycol PEG700, and is prepared by mixing PEG400 and PEG6000 according to a proportion, wherein the proportion of PEG400 is 94.64 percent, the proportion of PEG6000 is 5.36 percent, the PEG400 and the PEG6000 are respectively purchased from Shanghai chemical reagent company of Chinese medicine (group), the type of PEG400 is F990406, the type of PEG6000 is F990103), 1g of silymarin and 2g of poloxamer 188 are added into the dispersion medium, and the azeotropic state is kept for 1.5 hours.

Then 1.5ml of softener (the softener is liquid polyethylene glycol PEG700 and is prepared by mixing PEG400 and PEG6000 according to a proportion, wherein the proportion of PEG400 is 94.64 percent, the proportion of PEG6000 is 5.36 percent, PEG400 and PEG6000 are respectively purchased from Shanghai chemical reagent company of Chinese medicine (group), the type of PEG400 is F990406, and the type of PEG6000 is F990103) is added into the water plant thistle compound of the embodiment 2.

Example 3

taking 100ml of dispersion medium (constant volume to 100ml, liquid polyethylene glycol PEG600 as dispersion medium, product F990809 from Shanghai chemical reagent company of Chinese medicine (group)), adding 1g of silymarin and 8g of poloxamer 188, and keeping azeotropic state for 0.5 hr;

then, 1.5ml of a softener (liquid polyethylene glycol PEG600, product F990809 from Shanghai chemical reagent company, China medicine) was added to the mixture to obtain the silymarin compound of example 3.

Example 4

Aqueous dispersion for the preparation of silymarin

Taking 7mL of the silymarin compound prepared in example 1, adding purified water to a volume of 100mL, and stirring at room temperature for 20 minutes to obtain milky white water dispersion, which is the water dispersion of silymarin in example 4.

Example 5

Aqueous dispersion for the preparation of silymarin

Taking 3mL of the silymarin compound prepared in example 2, adding purified water to a volume of 100mL, and stirring at room temperature for 20 minutes to obtain milky white water dispersion, which is the water dispersion of silymarin in example 5.

Example 6

9mL of the silymarin complex prepared in example 3 was taken, purified water was added to a volume of 100mL, and stirred at room temperature for 20 minutes to obtain a milky white aqueous dispersion, which was the aqueous dispersion of silymarin of example 6.

Thermodynamic and kinetic stability testing of aqueous dispersions of silymarin examples 4-6

30ml of the milky white dispersions obtained in examples 4 to 6 were subjected to heat and pressure sterilization in an autoclave, and no phase change occurred; then, the materials are respectively placed for 7 days, and the phase change phenomenon still does not occur; this indicates that the milky white dispersions of examples 4 to 6 are stable in both thermodynamic and kinetic aspects, can be stored in liquid form for a long period of time, and can be used for the preparation of oral preparations or injections in liquid form.

For comparison, the inventor adds 1g of silymarin into 30mL of boiling water to obtain a transparent and clear aqueous solution, and carries out heating and pressurizing sterilization treatment in an autoclave, so that the phase change phenomenon appears; adding boiling water to obtain clear solution, standing for 7 days to obtain phase transition phenomenon; this indicates that silymarin is directly dissolved in water, is thermodynamically and kinetically unstable, and cannot be made into oral liquid or injection.

Bioavailability test

1. Test medicine

The tested drugs are: aqueous dispersions of silymarin of examples 4, 5, 6.

Comparison of drugs: silybin tablets (available from Shanghai Koghui pharmaceutical factory under the trademark "YIGANNING", production lot 970814) and Silybin powder (available from Dian jin pharmaceutical second factory).

2. Test method

1) Healthy rabbits (half of each male and female) are adopted, about 2.5 kilograms of the rabbits are fed conventionally, water and food are freely drunk, and the rabbits are fed with a complete ration without antibacterial drugs, so that the clinical manifestations are healthy. The healthy rabbits were randomly grouped into groups of 6 rabbits each (each half of the male and female).

Rabbit group one: the aqueous dispersion of silymarin of example 4 was administered at a dose of 36.2mg/kg (the administration equivalent of "silymarin" per kg of rabbits was 36.2mg) by oral gavage;

a second group of rabbits: the aqueous dispersion of silymarin of example 5 was administered at a dose of 36.2mg/kg (the administration equivalent of "silymarin" per kg of rabbits was 36.2mg) by oral gavage;

rabbit group three: the aqueous dispersion of silymarin of example 6 was administered at a dose of 36.2mg/kg by intragastric oral administration (the administration equivalent of "silymarin" per kg of rabbits was 36.2 mg);

fourth group of rabbits: administering commercially available silymarin tablets at a dose of 36.2mg/kg ("silymarin" administration equivalent of 36.2mg per kg rabbit);

rabbit of the fifth group: a commercially available silymarin powder was administered at a dose of 36.2mg/kg (the administration equivalent of "silymarin" per kg of rabbits was 36.2 mg).

2) Fixing the rabbit on an operation plate in a supine position, and taking blood from the anterior vena cava; collecting blank blood before administration; blood samples were taken at 5, 10, 15, 20, 30, 45 minutes and 1, 1.5, 2, 2.5, 3, 4, 6 hours post-dose, respectively, plasma drug concentrations were measured and pharmacokinetic parameters were calculated, and Mean ± standard deviation (Mean ± SD) was calculated for 6 data per group of rabbits.

See table 1 below for results.

TABLE 1

Note: t is_maxAnd represents the time of peak of blood concentration in "hour".

Cmax denotes the highest blood concentration, i.e. the blood concentration value (ng/ml) when the blood concentration reaches a peak.

AUC0- ∞ represents the area under the plasma drug concentration-time curve, which represents the total amount of absorption (h. ng/ml) after one dose, and reflects the extent of drug absorption.

As can be seen from Table 1, the pharmacokinetic test data of the silymarin tablets of the fourth group and the silymarin powder of the fifth group in healthy rabbits prove that the existing silymarin tablets and silymarin powder have low blood concentration, poor absorption, rapid elimination and low bioavailability in vivo. Compared with the existing silymarin tablets and silymarin powder, the silymarin water dispersion systems of the embodiments 4, 5 and 6 of the application have the advantages that the blood concentration in the body is remarkably improved, and the relative bioavailability is respectively as high as 270.1%, 140.3% and 185.4%; this shows that compared with the existing silymarin tablets and silymarin powder, the aqueous dispersion of silymarin of the present application is more completely released in vivo, and the peak reaching concentration is significantly improved, and the bioavailability is significantly increased.

Pharmacodynamic test

Purpose of the experiment: the test is used for detecting the liver protection effect of the water dispersion of silymarin on the carbon tetrachloride liver injury model in the application example 4.

The tested drugs are: aqueous dispersion of silymarin of example 4 of the present application

Comparative drug 1: yiganning (silymarin tablet), produced by Shanghai morning glory pharmaceutical factory, batch number: 970814, respectively;

comparative drug 2: bifendate, supplied by Shanghai balance pharmaceutical manufacturers, 15 mg/pill, Shanghai Wei Standard (1995) No. 011049.

The test animals were:

SD rats, provided by Shanghai family planning institute, were randomly divided into 7 groups of 20 animals (animal quality control number: third quarter-49-2-024/2 in the medical word) after weighing.

Of 7 groups, one group was normally bred; in the other 6 groups, liver injury models were respectively given carbon tetrachloride toxicants, wherein,

normal control group: normal SD rats, normal rearing.

Negative control group: model SD rats, gavage with saline.

Positive control group 1: model SD rat, Yiganning gavage, the administration dosage is 600 mg/kg;

positive control group 2: model SD rat, the bifendate is perfused, the administration dose is 600 mg/kg;

experimental group 4-low: model SD rats, the aqueous dispersion of silymarin of example 4 was gavaged at a dose of 150mg/kg (the administration equivalent of "silymarin" per kg of rats was 150 mg);

experimental group 4-in: model SD rats, the aqueous dispersion of silymarin of example 4 was gavaged at a dose of 300mg/kg (300mg of the administration equivalent of "silymarin" per kg of rats);

experimental group 4-high: model SD rats, the aqueous dispersion of silymarin of example 4 was gavaged at a dose of 600mg/kg (the administration equivalent of "silymarin" per kg of rats was 600 mg).

The test comprises the following main steps:

for each model, the administration (gavage) was performed as described above. The medicine is taken once a day for 20 days continuously, and after the medicine is stopped for 10 days, the animals are sacrificed, the liver is weighed, and pathological examination and radiography are carried out; and observing the pathological section conditions of different groups.

The experimental results are as follows:

the pathological examination result shows that: the positive pathological changes are mainly diffuse focal water sample degeneration (balloon-like cell formation, necrosis of individual liver cells, infiltration of small amount of inflammatory cells, nepheloid degeneration of liver cells, and the like). The treatment groups still had slight focal water-like degeneration, measured by the severity of the lesions, with the physiological saline group being the heaviest, the bifendate and YIGANNING groups being slightly lighter, and the Silybum marianum dispersions (300mg/kg and 600mg/kg) being the lightest.

TABLE 2

Note: ++ - +++: diffuse focal hepatocellular edema degeneration; +++: diffuse focal hepatocellular watery degeneration; ++: the swelling and degeneration of the liver cells are obvious, but no degeneration exists; + - ++: the liver cell swelling is obvious, and the liver sinus disappears but does not denature; -: no pathological changes; +: indicating but not significant hepatocyte swelling.

The experimental results show that: compared with the silymarin tablet (Yiganning) and the bifendate with clinical equivalent dose, the severity of the liver injury of the experimental hepatotoxicity model of the toxicant (carbon tetrachloride) can be obviously reduced after the rat model is subjected to intragastric administration by adopting the silymarin water dispersion system of the embodiment 4 of the application. The aqueous dispersion of silymarin of example 4 of the present application shows better hepatoprotective effect than silymarin tablet (Yiganning) and bifendate. Especially, the liver damage caused by the silymarin dispersion system is the lowest degree and the liver protection effect is the best when the silymarin dispersion system is used at medium dose (300mg/kg) and high dose (600 mg/kg).

The above-listed detailed description is only a specific description of a possible embodiment of the present invention, and they are not intended to limit the scope of the present invention, and equivalent embodiments or modifications made without departing from the technical spirit of the present invention should be included in the scope of the present invention.

Claims

1. A silymarin complex, characterized in that:

the silymarin compound is a compound of silymarin and poloxamer 188, and is prepared by the following steps:

adding 0.5-1.5 g of silymarin and 2-8 g of poloxamer 188 into 100ml of dispersion medium, performing azeotropic distillation for 0.5-1.5 hours, and adding 1-4 ml of softener to prepare the silymarin compound;

the dispersion medium adopts polyethylene glycol, and the average molecular weight of the polyethylene glycol is 400-700;

the softening agent is polyethylene glycol, and the average molecular weight of the polyethylene glycol is 400-700.

2. A silymarin complex according to claim 1, wherein:

polyethylene glycol 400 is adopted as the dispersion medium and the softener.

3. A silymarin complex according to claim 2, wherein:

the preparation method of the silymarin compound comprises the steps of adding 1g of silymarin and 6g of poloxamer 188 into 100ml of dispersion medium, performing azeotropic distillation for 1 hour, and adding 1.5ml of softener to prepare the silymarin compound.

4. An aqueous dispersion of silymarin characterized in that:

the water dispersion system of the silymarin is as follows: a milk-white aqueous dispersion of a silymarin complex according to any of claims 1 to 3 dissolved in purified water.

5. An aqueous dispersion of silymarin according to claim 4, wherein:

in the water dispersion system of the silymarin, the volume concentration of the silymarin compound is 0.03-0.09 ml/ml.

6. A silymarin liquid medicament is characterized in that: the liquid agent is prepared by using the water dispersion of silymarin described in claim 4 or 5.

7. The liquid pharmaceutical agent of silymarin according to claim 6, which comprises: the liquid medicament is oral liquid.

8. The liquid pharmaceutical agent of silymarin according to claim 6, which comprises: the liquid medicament is an injection.