KR0141648B1 - Pharmaceutical composition for the treatment of b-type hepatis - Google Patents

Abstract

The present invention is a pharmaceutical for the prevention and treatment of hepatitis B virus (HBV) hepatitis B containing a thioketene derivative represented by the following general formula (1) or a nontoxic salt thereof as an active ingredient and a pharmaceutically acceptable additive To a red formulation.

(Ⅰ)

Wherein Y is "CH2" or "CH = CH", A is NH or O, and R1 is C1C4 straight or branched alkyl. When A is O, R 2 is C 1 C 4 straight or branched alkyl, and when A is NH, R 2 is benzyl, 2pyridyl, 2thiazolyl, 4methylthiazol 2yl, 4carboxythiazol 2yl, Or 1, 3, 4 thiadiazol 2 day.

Description

Pharmaceutical preparations for the prevention and treatment of hepatitis B virus (HBV)

The present invention relates to a pharmaceutical preparation for the prevention and treatment of hepatitis B virus (HBV), more specifically containing a thioketene derivative represented by the following general formula (1) or a non-toxic salt thereof as an active ingredient And a pharmaceutical preparation for preventing and treating hepatitis B virus (HBV) hepatitis, which contains a pharmaceutically acceptable additive.

(I)

Wherein Y is "CH2" or "CH = CH", A is NH or O, and R1 is C1C4 straight or branched alkyl. When A is O, R 2 is C 1 C 4 straight or branched alkyl, and when A is NH, R 2 is benzyl, 2pyridyl, 2thiazolyl, 4methylthiazol 2yl, 4carboxythiazol 2yl, Or 1, 3, 4 thiadiazol 2 day.

Viral hepatitis, one of the representative diseases of liver disease, is known according to the type of the causative virus. Hepatitis caused by hepatitis B virus (hereinafter referred to as HBV) is the highest incidence of viral hepatitis, and more than 300 million are infected worldwide. The infection of HBV is known not only to induce hepatic diseases such as acute and chronic hepatitis B, but also to induce fatal diseases such as cirrhosis and hepatocellular carcinoma (HCC).

On the other hand, the present inventors have developed thioketene derivatives or their non-toxic salts as a compound having an excellent hepatic disease treatment and prophylactic effect, patent application No. 45984 (1991), patent 55163 (1992) I have registered in year). In addition, US Pat. No. 4,035,387 discloses malotilate and derivatives thereof, and effects on hepatic diseases have been known. (Effect on CCI4 induced induced injury in rats: Y. Imaizumi et al., Japan. J.) Pharmacol. 31, 15 (1981); Enhancement of rat liver protein synthesis: eidem, ibid. 32, 369 (1982); Clinical evaluation in liver cirrhosis: Gastroenterol. Japan 23, 639 (1988).

The thioketene derivatives or non-toxic salts thereof disclosed in the above-described patents are not only excellent in treating hepatic diseases in the hepatic experimental animal model induced by carbon tetrachloride (CCI4), but also rarely toxic at high doses.

The present inventors have studied the pharmacological effects of the thioketene derivatives or their non-toxic salts from various angles, and these derivatives exhibit excellent antiviral effects against HBV, which is the causative agent of hepatitis B viral infection, and also have high toxicity. The present invention was completed by discovering that it can be effectively applied to the treatment and prevention of hepatic diseases caused by HBV.

Unlike the thioketene derivatives or non-toxic salts thereof described above, the thioketene derivatives or their non-toxic salts are HBV, unlike the detoxification effect of the patents 45984, 55163 or US Pat. No. 4,035,387. It also has an effective effect on the viral disease caused.

Accordingly, an object of the present invention is to provide a pharmaceutical preparation for the prevention and treatment of hepatitis B virus (HBV) containing thioketene derivative as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention is a pharmaceutical for the prevention and treatment of hepatitis B virus (HBV) hepatitis B containing a thioketene derivative represented by the following general formula (1) or a non-toxic salt thereof as an active ingredient and a pharmaceutically acceptable additive To a red formulation.

(I)

Wherein Y is "CH2" or "CH = CH", A is NH or O, and R1 is C1C4 straight or branched alkyl. When A is O, R 2 is C 1 C 4 straight or branched alkyl, and when A is NH, R 2 is benzyl, 2pyridyl, 2thiazolyl, 4methylthiazol 2yl, 4carboxythiazol 2yl, Or 1, 3, 4 thiadiazol 2 day.

The inhibitory effect on HBV is particularly excellent among the derivatives, and more preferred derivatives are listed as follows.

1. Diisopropyl 2 '(1,3'dithiol­2­ylidene) malonate or a non-toxic salt thereof

2. Isopropyl 2 '(1,3'dithiol­2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate or a non-toxic salt thereof

3. Isopropyl 2 '(1,3'dithiol­2'ylidene)' 2 '[N' (1,3,4'thiadiazol­2­yl) carbamoyl] acetate or a non-toxic salt thereof

4. Isopropyl 2 '(1,3'dithiol'2'ylidene)' 2 '[N' (2'pyridyl) carbamoyl] acetate or a non-toxic salt thereof

5. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate or a non-toxic salt thereof

6. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(2'pyridyl) carbamoyl] acetate or a non-toxic salt thereof

7. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(4'carboxythiazol­2­yl) carbamoyl] acetate or a non-toxic salt thereof

8. Ethyl 2 '(1,3'dithiethane­2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate or a non-toxic salt thereof

9. Isopropyl 2 '(1,3'dithiethane­2'ylidene) ­2' [N '(2'thiazolyl) carbamoyl] acetate or a non-toxic salt thereof

10. Isopropyl 2 '(1,3'diethaneethane­2­ylidene) ­2' (N­benzylcarbamoyl) acetate or a non-toxic salt thereof

The present inventors have described the antiviral effect of the thioketene derivative represented by the general formula (1) or its non-toxic salt against HBV in a known 2.2.15 cell line (Sells, Ma et. Al. Proc. Natl. Acad. Sci., USA). , 84, 10051009, 1987).

2.2.15 Cell line inoculated HBV DNA into Hep G2 cells, which is an approved liver cancer cell. That is, the 2.2.15 cell line has many advantages, such as high virus replication rate, stable DNA morphology, presence of virus-specific RNA transcripts and proteins in appropriate size and shape, and high release of penetrated HBV virus. It is a well-suited cell line for drug efficacy evaluation on HBV (Korba and Milman, Antiviral Research, 15, 217228 (1991)).

Thioketene derivatives represented by the general formula (1) or nontoxic salts thereof can effectively inhibit the replication of HBV DNA in 2.2.15 cell lines, as can be seen in the following examples, and their toxicity is very low. Safty zone is very wide.

Thioketene derivatives represented by the general formula (1) or non-toxic salts thereof can be easily prepared by the preparation methods disclosed in the inventors of Patent Nos. 45984 and 55163, and the preparation methods disclosed in US Pat. No. 4,327,223. Can be.

In addition, thioketene derivatives or non-toxic salts thereof represented by General Time (1) may be prepared into pharmaceutical formulations of various formulations by containing pharmaceutically acceptable additives.

Pharmaceutically acceptable additives depending on the formulation include all forms of diluents or solvents, fillers, scattering agents, binders, dispersants, disintegrants, surfactants, lubricants, excipients or wetting agents. In addition, it may contain a solubilizer, buffer, present agent, colorant, flavoring agent, sweetener and the like which are commonly used if necessary.

The dosage unit form of the pharmaceutical preparation according to the present invention is not particularly limited and may be widely selected according to various therapeutic purposes. For example, formulations of tablets, capsules, granules, emulsions, suspensions and the like can be selected.

The various formulations described above can be prepared by conventional methods. For example, in preparing oral preparations such as tablets, capsules, granules, etc., excipients such as white sugar, lactose, glucose, starch, mannitol; Binders such as syrup, gum arabic, sorbitol, trigacanth rubber, hydroxypropyl cellulose, polyvinylpyrrolidone and the like; Disintegrants such as starch, carboxymethyl cellulose and its calcium salt, microcrystalline cellulose, crospovidone, sodium starch glycolate and the like; Lubricants such as talc, magnesium stearate, calcium stearate, silica and the like; It may be prepared by conventional methods using a humectant such as sodium lauryl sulfate, glycerol and the like.

The dosage of the pharmaceutical preparation according to the present invention using the thioketen derivative of Formula (1) or a non-toxic salt thereof as an active ingredient may be determined by the method of administration, the type of preparation, the age of the patient, the weight of the patient, the sensitivity of the patient, and the condition of the disease. It may be appropriately selected according to the, but is not particularly limited, the amount of the active ingredient may be administered once or several times 20mg to 1g per day in oral administration and the unit dosage form may be 20 to 200mg. Of course, the dose of active ingredient may be in an amount outside the above range.

Hereinafter, the present invention will be described in more detail through test examples and examples. However, the present invention is not limited thereto.

Test Example 1. HBV DNA replication inhibitory effect

HBV DNA replication inhibitory effect of the pharmaceutical preparations according to the present invention is known using a 2.2.15 cell line (Sells, MA et. Al. Proc. Natl. Acad. Sci., USA, 84, 10051009, 1987). (Korba and Gerin, Antiviral Research, 19, 5570, 1992).

In other words, the test compounds were divided into four concentrations (n = 6) to measure the antiviral effect, and the test compounds were as follows.

1.Diisopropyl 2 '(1,3'dithiol­2­ylidene) malonate

2. Isopropyl 2 '(1,3'dithiol­2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate

3. Isopropyl

4. Isopropyl 2 '(1,3'dithiol­2'ylidene) ­2' [N '(2'pyridyl) carbamoyl] acetate

5. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate

6. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(2'pyridyl) carbamoyl] acetate

7. Isopropyl

8. Ethyl 2 '(1,3'dithiethane­2'ylidene) ­2' [N '(4'methylthiazol­2­yl) carbamoyl] acetate

9. Isopropyl 2 '(1,3'dithiethane­2'ylidene) ­2' [N '(2'thiazolyl) carbamoyl] acetate

10. Isopropyl 2 '(1,3'dithiethane­2'ylidene) ­2' (N'benzylcarbamoyl) acetate

The test compound was dissolved in DMSO, and prepared on the day of the test at concentrations of 7, 20, 67, and 200 μM, respectively, and then treated in a culture solution prepared in a 24-well flat1bottomed tissue culture plate at room temperature and immediately 2.2.15 Cell line was added. After 9 days of continuous treatment of the test compounds by concentration, the cells were analyzed by Southern blot hybridization assay (Korba and Gerin, Antiviral Research, 19, 5570, 1992). Extracellular HBV DNA level was measured and the results are shown in Table 1 below. Here, the HBV DNA level of the control (no control) was 81 ± 9 pg / ml, 1 pg extracellular HBV DNA level of 1 ml corresponds to 3 × 105 viral particles.

Table 1

In general, it is considered that statistical significance (p0.05) is observed when the measured extracellular HBV DNA level is 3 times higher than the extracellular HBV DNA level of the untreated group. do. Therefore, as can be seen in the results of Table 1, it can be seen that thioketene derivatives represented by the general formula (1) have a very good antiviral effect.

Test Example 2 Cytotoxicity Test

Toxicity test of the pharmaceutical preparations according to the present invention was carried out using the absorbency test method (Neutral No. 941886) of the neutral red dye which is widely used in various viral host systems including HSV and HIV. Was carried out.

In other words, the test compounds were divided into four concentrations (n = 6) to measure the antiviral effect, and the test compounds were as follows.

1.Diisopropyl 2 '(1,3'dithiol­2­ylidene) malonate

2. Isopropyl

3. Isopropyl 2 '(1,3'diethane ethane 2'ylidene)' 2 '[N' (4'thiadiazol-2-2) carbamoyl] acetate

4. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' [N '(2'pyridyl) carbamoyl] acetate

5. Isopropyl 2 '(1,3'diethane ethane 2'ylidene) ­2' (N'benzenecarbamoyl) acetate

The amount of neutral red dye captured by the host (2.2.15 cell line) was measured at 510 nm using a spectrophotometer (A510), and the average A510 of the untreated control group was 100. The result showing the percentage (mean ± standard deviation) is shown in Table 2 below.

TABLE 2

As can be seen from the results of Table 2, the thioketene derivative of the general formula (1), which is an active ingredient of the pharmaceutical preparation according to the present invention, has almost no cytotoxicity due to the increase in treatment concentration. It can be seen that almost none, in particular, very safe in the effective concentration range.

Example 1 Preparation of Tablets

As an active ingredient, 100 mg of isopropyl 2 (1,3diethaneethane2ylidene) 2 [N (4methylthiazol2yl) carbamoyl] acetate was weighed into the ingredients in Table 3, according to a known production method. Tablets were prepared. That is, after isopropyl lactose, gelling starch, sodium lauryl sulfate, crospovidone, and colloidal silicon dioxide are mixed, the granules are formed into an aqueous polyvinylpyrrolidone solution, the dried granules are passed through a No. 20 sieve, and then magnesium stearate It was mixed with and tableted.

TABLE 3

Example 2. Preparation of Tablets

In Example 1, by using the microcrystalline cellulose instead of gelling starch and weighed by the component content of Table 4 to prepare a tablet in the same manner as in the production method of Example 1.

Table 4

Example 3. Preparation of Capsule

As an active ingredient, 100 mg of isopropyl 2 (1,3diethaneethane2ylidene) 2 [N (4methylthiazol2yl) carbamoyl] acetate were weighed together with the ingredients in Table 5 below, according to a known production method. Hard gelatin capsules were prepared.

That is, isopropyl lactose, microcrystalline cellulose, sodium lauryl sulfate, crospovidone, and colloidal silicon dioxide are passed through a sieve (20), mixed with magnesium stearate, and filled with gelatin capsules (1,000 capsules) for oral administration. The agent was prepared.

Table 5

Example 4 Preparation of Capsule

Oral administration gelatin capsules were prepared in the same manner as in Example 3 using 8 g of sodium starch glycolate instead of crospovidone.

Example 5. Preparation of Granules

As an active ingredient, 100 mg of isopropyl 2 (1,3diethaneethane2ylidene) 2 [N (4methylthiazol2yl) carbamoyl] acetate was weighed into the ingredients in Table 6 and granulated according to a known production method. Was prepared.

That is, isopropyl lactose, microcrystalline cellulose, crospovidone and colloidal silicon oxide were mixed, and then kneaded by adding polyvinylpyrrolidone aqueous solution, granules were prepared using an extrusion granulator, and then dried to prepare granules. .

Table 6

Claims (3)

A pharmaceutical preparation for the prevention and treatment of hepatitis B virus (HBV) hepatitis B, which contains a thioketene derivative represented by the following general time (1) or a nontoxic salt thereof as an active ingredient and a pharmaceutically acceptable additive. (I) Wherein Y is "CH2" or "CH = CH", A is NH or O, and R1 is C1C4 straight or branched alkyl. When A is O, R 2 is C 1 C 4 straight or branched alkyl, and when A is NH, R 2 is benzyl, 2pyridyl, 2thiazolyl, 4methylthiazol 2yl, 4carboxythiazol 2yl, Or 1, 3, 4 thiadiazol 2 day. The compound of claim 1, wherein Y is CH 2, A is NH, R 1 is ethyl or isopropyl, R 2 is benzyl, 2pyridyl, 2thiazolyl, 4methylthiazol 2yl, 4carboxythiazol 2yl, Or 1, 3, 4 thiadiazol 2 days, characterized in that the pharmaceutical preparations for the prevention and treatment of hepatitis B virus (HBV) hepatitis. The compound of claim 1, wherein Y is CH = CH, A is NH, R 1 isopropyl, R 2 is benzyl, 2pyridyl, 4methylthiazol 2yl, or 1, 3, 4thiadiazol 2yl. A pharmaceutical preparation for the prevention and treatment of hepatitis B virus (HBV).