KR100359176B1 - The composition and preparation method of 2-allythiopyrazine·ascorbic acid salt which has a hepatoprotective effect - Google Patents

Abstract

The present invention relates to a novel hepatoprotective agent and a method for preparing the same, and more particularly, pharmaceutical preparation through change in physical properties induced by binding to ascorbic acid to 2-allylpipyrazine to a salt structure. The present invention relates to a derivative and a method for preparing the same, which aim to increase the hepatoprotective action, which is easy and pharmacological effect. Incorporation of ascorbic acid into water-soluble 2-allylthiopyrazine, which was converted to a water-soluble substance, could increase the pharmaceutical application and pharmacological efficacy. As in the embodiment of the present invention, it can be seen that 2-allylthiopyrazine-ascorbate has an increased hepatoprotective effect in mice compared to 2-allylthiopyrazine. In other words, 2-allylthiopyrazine-ascorbate reduced the levels of aspartate-aminotransferase (GOT) and alanine-aminotransferase (GPT), which are indicators of hepatotoxicity, in the carbon tetrachloride experimental model. The tendency to However, in the acute toxicity test of mice, the mortality of 2-allylpipyrazine-ascorbic acid tended to increase slightly compared to 2-allylchipyrazine. This may be due to increased body absorption or increased rate of absorption compared to pyrazine.

(I)

Description

The composition and preparation method of 2-allylthiopyrazine ascorbate having the effect of preventing and treating hepatic diseases and the method of preparing the same {The composition and preparation method of 2-allythiopyrazine · ascorbic acid salt which has a hepatoprotective effect}

The present invention relates to the development of a substance having a prophylactic and therapeutic effect on liver disease. The main active ingredient of the present invention is 2-allylthiopyrazine-ascorbate, which synthesizes ascorbic acid in the existing 2-allylthiopyrazine and has the structure of Formula (I).

(I)

The method for producing 2-allylthiopyrazine-ascorbate represented by the formula (I) is known from 2-chloropyrazine according to a known domestic patent (Domestic Patent Registration No. 0138005, registered date Feb. 14, 1998). It is synthesized as a salt by reacting ascorbic acid (vitamin C), which is an organic acid, with 2-allylthiopyrazine, a conventional substance synthesized through mercaptopyrazine.

Since 2-allylchipyrazine, the main active ingredient of known domestic patents (Domestic Patent Registration No. 0138005, date of registration February 14, 1998), is a water-insoluble substance that is insoluble in water, Restricted Therefore, if this problem is solved by changing the physical properties of 2-allylpipyrazine, it is possible to increase the commercial value of the material, and also to synthesize the present material to induce the effect of increasing the liver protective effect.

The synthesis method of 2-allylthiopyrazine salt according to the present invention is as follows.

Example 1

Preparation of 2-allylthiopyrazine

15.0 g of 2-chloropyrazine was dissolved in 120 ml of dimethylformamide, and 15.2 g of sodium hydrosulfide was added to the solution, followed by heating to 60 ° C. for 3 hours. The solution was cooled at room temperature and then cooled in ice water. The solid produced while cooling was filtered in vacuo (preparation of 2-mercaptopyrazine). 20.0 g of 2-mercaptopyrazine was dissolved in 245 ml of dimethylformamide, and 27 ml of triethylamine was added thereto. 20.8 ml of allyl bromide was added to the solution and stirred at 0 ° C. for 2 hours. The reaction solution was poured into iced water, extracted with ethyl ether, and concentrated. The residue was distilled under vacuum to give a pale yellow and oily liquid (preparation of 2-allylthiopyrazine). (23g, 82%)

IR (paraffin oil) 1500, 1466, 1379 (cm ^-1 )

Example 2

Production of 2-allyl pyroazine-ascorbate

15.2 g of 2-allylthiopyrazine prepared in Example 1 was dissolved in methanol, and 17.6 g of ascorbic acid was added thereto, followed by stirring and dissolution. The solution was distilled under reduced pressure and dried to prepare a yellow 2-allylpipyrazine-ascorbate salt, which was recrystallized from ethyl acetate. (31.2g, 95.1%)

IR (paraffin oil) 3525, 1754, 1500, 1466, 1379 (cm ^-1 )

Example 3

Production of 2-allylpipyrazine Ascorbate Injection

Dissolve 30 g of 2-allylpipyrazine-ascorbate in 200 ml of water, filter it with a bacterial filter (Millipoa), discard the supernatant, and place the filtrate directly into a sterile 30 ml vial and lyophilize for injection. Prepared as a formulation.

Example 4

Preparation of 2-allylthiopyrazine. Ascorbate Tablets

2-allylthiopyrazine. Ascorbate 100mg lactose 20mg, corn starch 6mg, hydroxypropyl cellulose 12mg, sodium starch glycolate 2mg, gelatin 2mg, potato starch 2mg, magnesium stearate 1mg as a method of manufacturing a conventional tablet. Tablets were prepared by mixing and tableting.

Example 5

Preparation of 2-allyl thiopazine. Ascorbate capsules

2-allylthiopyrazine. Ascorbate 100 mg of lactose, 113 mg of corn starch, 4 mg of magnesium stearate were mixed in a conventional method for preparing capsules, and then filled into gelatin capsules to prepare capsules.

Example 6

Preparation of 2-allyl chiopyrazine. Ascorbate soft capsule

2-allyl chiopyrazine.Ascorbate 100mg, white lead 5.0mg, lecithin 6mg, soybean oil 100mg, gelatin 140mg, concentrated glycerin 55.0mg, sorbitol liquid 12.0mg, paraoxybenzoic acid profile 0.03mg, ethyl vanillin, suitable amount of pigment The soft capsule agent was prepared according to the manufacturing method of the normal soft capsule agent.

Example 7

Preparation of 2-allylthiopyrazine. Ascorbate Liquid

2-Allylchiopyazine. Ascorbate 100mg 6g per 100g, citric acid 9mg, paraoxybenzoic acid propyl 6mg, a suitable amount of purified water, mixed with a conventional method of preparing a solution, sterilized and filled into a 100ml bottle and sealed Was prepared.

Example 8

Preparation of 2-allylthiopyrazine. Ascorbate Granules

2-Allylchiopyrazine.Ascorbate 100mg, 27mg per bag, talc 19mg, gum arabic, proper sage oil, proper amount of peppermint oil, suitable amount of hard fluid paraffin, proper amount of paraffin, suitable amount of coloring agent, and granules according to the general method of preparing granules. Was prepared.

Test Example 1

Acute Toxicity Test of 2-allylpipyrazine Ascorbate

To evaluate the acute toxicity of 2-allylpipyrazine-ascorbate, we investigated the change of mortality and mortality (LD ₅₀ ) over time after drug administration.

To observe changes in mortality over time, 25 ± 2 g of ICR male mice were fasted for 24 hours, followed by 2-allylchipyrazine.ascorbate 2,700mg / kg (1,250mg / kg as 2-allylchipyrazine). Was orally administered and observed for 48 hours. After 48 hours, the mortality rate was almost the same as that of the administration group of 1,250mg / kg of 2-allylpipyrazine.

In addition, 25 ± 2 g of ICR male mice were examined in groups 3 (group 1; control, group 2; 2-allylthiopyrazine, group 3; 2-allylthiopyrazine-as) to investigate the lethal dose (LD ₅₀ ) of each drug. Corbate), fasting for 24 hours, 0.25 ml of drinking water in the first group, 2-750 allylchiopyrazine in the second group, 750 mg / kg, 1,000 mg / kg, 1,250 mg / kg, 1,500 mg / kg In group 3, 1,620 mg / kg, 2-160 mg / kg, 2,700 mg / kg, and 3,240 mg / kg 2-allylpipyrazine-ascorbate (750 mg / kg, 2-1000 mg / kg as 2-allylpipyrazine) , 1,250 mg / kg and 1,500 mg / kg were orally administered and the lethal dose was measured, and as a result of the measurement, the lethal dose (LD ₅₀ ) of 2-allylpipyrazine was about 1,250 mg / kg and 2-allylchipyrazine-as The lethal amount (LD ₅₀ ) of corbate salt was about 2376 mg / kg (1,100 mg / kg as 2-allylthiopyrazine).

Test Example 2

Changes in Activity of Aspartate-Aminotransferase (GOT) by Administration of 2-allylchipyrazine Ascorbate

25 ± 2 g of ICR male mice were divided into five groups (group 1: control group; group 2: carbon chloride alone group; group 3: carbon tetrachloride administration + ascorbic acid; group 4: carbon tetrachloride administration + 2-allylthiopyrazine; group 5: carbon tetrachloride) Administration + 2-allylpipyrazine, ascorbate) divided into 0.25 ml of drinking water in groups 1 and 2, 58 mg / kg of ascorbic acid in group 3, and 50 mg of 2-allylchiopyrazine in group 4. / Kg, and group 5 was administered orally with 2-allylthiopyrazine-ascorbate 108 mg / kg once daily for 3 days. Groups 2, 3 and 4 after 1 hour after the last 3 doses And 0.15 ml / kg of carbon tetrachloride were administered to soybean oil. After 24 hours, the mice were anaesthetized, blood was taken, and centrifuged to take a sample for changing the activity of aspartate-aminotransferase. The activity was measured through photometer 7010 and compared with the control group. Comparison of Aspartate-Aminotransferase Levels Ascorbic acid, 2-allylthiopyrazine, and 2-allylthiopyrazine-ascorbate group were 38.2% and 45.2%, respectively, compared with carbon tetrachloride alone group. 50.1% was inhibited (Table 1).

Table 1. Changes in Activity of Aspartate-Aminotransferase (GOT)

Group 1 Group 2 Group 3 4th group 5 groups Activity (U / L) 68 3350 2070 1835 1670

Group 1: control group,

Group 2: carbon tetrachloride alone group,

Group 3: carbon tetrachloride + ascorbic acid,

Group 4: carbon tetrachloride + 2-allylthiopyrazine,

Group 5: Carbon Tetrachloride + 2-allylchipyrazine Ascorbate

Test Example 3

Changes in Activity of Alanine-Aminotransferase (GPT) by Administration of 2-allylpipyrazine Ascorbate

25 ± 2 g of ICR male mice were treated with 5 groups (group 1: control group; group 2: carbon tetrachloride alone group; group 3: carbon tetrachloride administration + ascorbic acid; group 4: carbon tetrachloride administration + 2-allylchipyrazine; group 5: carbon tetrachloride administration) + 2-allylpipyrazine and ascorbate) divided into 0.25 ml of drinking water in groups 1 and 2, 58 mg / kg of ascorbic acid in group 3, and 50 mg / of 2-allylthiopyrazine in group 4. Kg and Group 5 were administered orally with 2-allylthiopyrazine-ascorbate 108mg / kg once a day for 3 days, and 1 hour after the last 3rd administration, Group 2, Group 3, 4 and In the experimental group of 5, 0.15ml / kg of carbon tetrachloride was diluted and administered to soybean oil. After 24 hours, the mice were anaesthetized, blood was taken, and centrifuged to take a sample for changing the activity of alanine-aminotransferase, and the activity was measured through a photometer 7010 and compared with the control. Compared with alanine-aminotransferase (GPT) levels, ascorbic acid, 2-allylthiopyrazine, 2-allylthiopyrazine and ascorbate group were 38.9% and 42.1, respectively. %, 49.7% inhibition (Table 2).

Table 2. Activity change of alanine-aminotransferase (GPT)

Group 1 Group 2 Group 3 4th group 5 groups Activity (U / L) 48 2825 1725 1635 1420

Group 1: control group,

Group 2: carbon tetrachloride alone group,

Group 3: carbon tetrachloride + ascorbic acid

Group 4: carbon tetrachloride + 2-allylthiopyrazine,

Group 5: Carbon Tetrachloride + 2-allylchipyrazine Ascorbate

The present invention has increased the range and ease of use in laboratory and industrial conditions by increasing the water solubility by preparing a salt from the compound of 2-allylchiopyrazine as in the Examples, and the pharmaceutical application such as injection, homemade It can be seen that the range can be broadened, and the pharmacological effect is also increased. In the acute toxicity test, lethal dose tends to increase slightly compared to 2-allylchipyrazine, a substance described in Patent Publication No. 0138005, (Date 14, 1998), which is 2-allylchio than 2-allylchipyrazine. It is expected that the absorption rate of pyrazine and ascorbate in the body increases or the rate of absorption increases.

Claims (3)

Ascorbic acid (vitamin C) salt of 2-allylthiopyrazine of formula (1) A process for preparing 2-allylthiopyrazine of formula (I) by reacting 2-allylthiopyrazine of formula II with ascorbic acid of formula III. 2-allylthiopyrazine of formula (I). Ascorbate is added to excipients, adjuvants, preservatives, diluents, and flavoring agents commonly used in the pharmaceutical industry, and tablets, capsules, injections, solutions, dusts A pharmaceutical preparation having a hepatoprotective action prepared by preparing a pharmaceutical preparation selected from capsules and granules.