JPH0260672B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to N-(4-phenyl-2-thiazolyl)carbamic acid derivatives and pharmaceutical compositions containing them. More specifically, the present invention has immunomodulatory ability,
Therefore, (4-phenyl-2-thiazolyl)carbamic acid derivatives and pharmaceutical compositions containing the same have therapeutic effects on immune diseases such as rheumatoid arthritis and are also useful for viral diseases or anticancer immunotherapy. relating to things. Traditionally, for autoimmune diseases such as rheumatoid arthritis,
Steroidal and non-steroidal anti-inflammatory drugs
It is also widely used clinically. However, many of these drugs are still not fully satisfactory in terms of their inherent effects, side effects, and toxicity. In contrast, the compounds of the present invention exert specific effects on cells related to immune responses, have the ability to alter the immune response of the host, and have properties that are highly desirable as pharmaceuticals, with little toxic effects. In addition, Japanese Patent Application Laid-open No.
Although the description of No. 54-61172 is known, the compound of the present invention has a substituent different from those and has pharmaceutically important characteristics. N-(4-phenyl-2-thiazolyl) of the present invention
The carbamic acid derivative has the general formula (1) K0423 (wherein R ₁ is a lower alkylthio group, a lower alkylsulfinyl group, a lower alkylsulfonyl group,
(represents a nitro group or an amino group, and R ₂ represents a lower alkyl group), where lower means one having 1 to 4 carbon atoms. Moreover, all compounds can exhibit amidine-type tautomerism, and general formula (1) is intended to include all tautomers. N- represented by general formula (1)
(4-phenyl-2-thiazolyl)carbamic acid derivatives specifically include the following lower alkyl esters of carbamic acid. N-(4-p-methylthiophenyl-2-thiazolyl)carbamic acid, N-(4-o-methylthiophenyl-2-thiazolyl)carbamic acid, N-(4-m-methylthiophenyl-2-thiazolyl) ) Carbamic acid, N-(4-p-methylsulfinyl phenyl-
2-thiazolyl)carbamic acid, N-(4-o-methylsulfinyl phenyl-
2-thiazolyl)carbamic acid, N-(4-m-methylsulfinyl phenyl-
2-thiazolyl)carbamic acid, N-(4-p-methylsulfonylphenyl-2
-thiazolyl)carbamic acid, N-(4-o-methylsulfonylphenyl-2
-thiazolyl)carbamic acid, N-(4-m-methylsulfonylphenyl-2
-thiazolyl)carbamic acid, N-(4-p-nitrophenyl-2-thiazolyl)carbamic acid, N-(4-o-nitrophenyl-2-thiazolyl)carbamic acid, N-(4-m-nitrophenyl-2-thiazolyl) ) Carbamic acid, N-(4-p-aminophenyl-2-thiazolyl)carbamic acid, N-(4-o-aminophenyl-2-thiazolyl)carbamic acid, N-(4-m-aminophenyl-2-thiazolyl)carbamine Acid, These compounds represented by the general formula (1) have the general formula
(2) A 2-amino-4-phenylthiazole represented by K0424 (in the formula, the definition of R ₁ is the same as in general formula (1)) in a dialkyl carbonate ester, preferably 90
Either by heating above ℃ to react, or by using general formula (2)
Compound of general _formula ( ³ )
represents a halogen atom). In the latter method, the reaction is carried out by dissolving or suspending the starting material of general formula (2) (which may be in the form of a suitable acid addition salt) in a solvent to form a compound of general formula (3). It is preferable to add it by a method such as dripping. Examples of solvents include:
Benzene, toluene, xylene, acetone, ethylmethylketone, dioxane, 1,2-dimethoxyethane, tetrahydrofuran, N,N-dimethylformamide and the like are suitable. It is also advantageous to use organic bases such as pyridine and triethylamine or inorganic bases such as sodium carbonate, potassium carbonate, sodium bicarbonate, for the purpose of removing the acid (HX) produced by the reaction. Although this reaction proceeds even at temperatures below room temperature, it is also possible to heat the solvent up to its boiling point in order to accelerate the reaction. 2-, which is a starting material represented by general formula (2)
The method for producing amino-4-phenylthiazoles is described, for example, in the literature (Jacques V. Metzger, ed “The
Cnemistry of Heterocyclic Compounds,
Vol.34: Thiazole And Its Derivatives, Part
Two” John Wiley 8, Sons, Inc. (1979)). In particular, aminothiazoles with methylthiophenyl, methylsulfinylphenyl, and methylsulfonylphenyl groups are described in detail in the literature (Chemia, 27, 99 (1973)).Also, another method for producing the compound represented by the general formula (1) is the general formula (4) K0425 (wherein R ₁ is a lower alkylthio group, a lower alkyl sulfur group). Nyl group, lower alkylsulfonyl group,
α-halogenoacetophenones represented by (representing a nitro group or an amino group, and X represents a halogen atom);
There is a method of reacting with a thiourea represented by the general formula (5) K0426 (in the formula, R ₂ represents a lower alkyl group). The reaction is usually carried out by mixing these two compounds in a solvent such as alcohol, tetrahydrofuran, dioxane, benzene, 1,2-dimethoxyethane, N,N-dimethylformamide. The reaction temperature can be freely selected between usually room temperature and the boiling point of the above solvent. Moreover, the reaction is usually completed in 1 to 6 hours. Among the compounds represented by general formula (1), especially N-
Regarding (4-aminophenyl-2-thiazolyl)carbamate ester, a method of reducing the once synthesized N-(4-nitrophenyl-2-thiazolyl)carbamate ester is more practical and advantageous than the above-mentioned method. The compound represented by the general formula (1) of the present invention is:
Possesses pharmacological activity. It has been particularly surprisingly found by the inventors that the compounds of the invention have immunomodulatory properties. The compounds of the present invention have low toxicity and are extremely useful as medicines. Next, this will be explained using a test example. Many experimental systems are commonly used to test immunomodulatory effects using animals, but the results of the most representative test, the delayed-type hypersensitivity reaction enhancement test, are illustrated below as a test example. . Delayed hypersensitivity induced in mice by applying picryl chloride (2-chloro-1,3,5-trinitrobenzene) to the skin is known as a typical cell-mediated immune phenomenon, and as an experimental system. It is one of the systems widely used worldwide (Asherson. OLand Ptak, W.: Contact and
delayed hypersensitivity in the mouse I.
Active sensitization and passive transfer.
Immunology, 15 , 405-416 (1968)). This experimental system was used in a delayed hypersensitivity enhancement test. Test Example 1 Delayed hypersensitivity reaction enhancement test Test method: ICR male mice weighing approximately 30 g were used in groups of 8 mice. Sensitization was performed by dissolving 3% vinyl chloride in a 4:1 mixture of olive oil and acetone and applying it to the shaved abdomen of the mouse. Simultaneously with sensitization, the compound of the present invention dissolved or suspended in 0.2% carboxymethylcellulose physiological saline was orally administered at a rate of 50 mg per kg of mouse body weight. For the control group, 0.2% carboxymethylcellulose physiological saline was administered in the same manner. To induce delayed-type hypersensitivity (challenge), 7 days after sensitization, a piece of felt soaked in olive oil containing 1% picry chloride was wrapped around forceps and applied to the ear of the mouse. I did it. The thickness of the mouse ears was measured before the challenge and 24 hours after the challenge, and the rate of increase in thickness (average value of both ears of 8 mice) is shown in Table 1. For comparison, the results of a similar test using levamisole hydrochloride are also shown. An F-t test was conducted on the test results, and those with a significant risk ratio of P<0.05 compared to the control group are marked with an asterisk.
Significant values at P<0.01 are marked with **. Results: When the compound of the present invention was administered simultaneously with sensitization, the delayed hypersensitivity response induced by challenge was enhanced. The compound of the present invention was found to have an activity equal to or greater than that of levamisole used for comparison. That is, the compound of the present invention is considered to have an effect of regulating cellular immune response in mice (immunomodulating effect).

【table】

[Table] Next, adjuvant arthritis in rats, which is caused by injection of Mycobacterium tuberculosis, is frequently used as an experimental model for human rheumatoid arthritis. Although the onset mechanism of this disease is not fully understood, it is known that cell-mediated immunity plays an important role. The immunomodulatory potential of the compounds of the invention was investigated using this known adjuband arthritis test. Test Example 2 Adjuvant Arthritis Test (Table 2) Test method: Using 8-week-old SD male rats, 0.4 mg of dried dead Mycobacterium tuberculosis cells was suspended in 0.1 ml of liquid paraffin. It was injected intradermally into the hind paw. The compound of the present invention was subcutaneously administered a total of 9 times before and after injection of the adjuband.
The compound was dissolved or suspended in 0.2% carboxymethyl cellulose saline at a concentration of 5% per kg body weight.
Administered at the rate of mg. The volume of edema in the left hind paw was measured from the day of Aziyuband injection until the end of the test, and the swelling rate was calculated. For comparison, the results of a test using levamisole hydrochloride are also shown. An F-t test was performed on the test results, and those with a significance rate of P<0.05 were marked with an asterisk (*), and P<
Those that are significant at 0.01 are marked with **, and those that are significant at P < 0.001 are marked with ***. Results: The compound of the present invention strongly inhibited secondary inflammation in adjuvant arthritis, and the effect was statistically significant compared to the control group. The compound of the present invention was found to have a higher activity than levamisole used for comparison, and a much higher activity than the compound described in JP-A-54-61172. That is, the compounds of the present invention are considered to have immunomodulatory ability and anti-arthritic activity.

【table】

[Table] As shown in Test Example 1 and Test Example 2, the compound of the present invention has strong activity as an immunomodulator, and is therefore useful for diseases known to be associated with abnormalities in immunoregulatory ability, such as chronic It is effective in treating autoimmune diseases such as rheumatoid arthritis. Next, Test Example 3 shows a toxicity test for the active ingredient of the medicine of the present invention. Test Example 3 Acute toxicity test by oral administration Test method: Using ddY male mice (5 mice per group), the drug was dissolved or suspended in physiological saline and administered orally. The progress was observed for 7 days after administration, and the estimated LD ₅₀ value was determined. I asked for Results: The estimated LD ₅₀ value of the active ingredient of the medicament of the present invention was 1000 mg/Kg or more. This value is much larger than the estimated LD ₅₀ of 200 to 300 mg/Kg for levamisole hydrochloride, and the toxicity of the active ingredient of the present invention is considered to be weak. When using the compound of the present invention as a medicine, it is possible to use it as a pharmaceutical raw material as it is, but it is also possible to use it as a raw material for formulation, taking into consideration stability and ease of formulation. If water solubility is preferred, it can also be used in the drug raw material, for example as a pharmaceutically acceptable salt such as hydrochloride, citrate or phosphate. The medicament of the present invention can be used in the same dosage form and administration method as conventional immunomodulators or anticancer agents. For example, as oral preparations, capsules, granules, pills, fine granules, tablets, syrups, etc. can be used. Suppositories are suitable for intrarectal administration, and subcutaneous, intramuscular, or intravenous injections can be used for injections. Diseases to which the immunomodulator of the present invention can be applied include diseases known to be accompanied by abnormal immune function, such as autoimmune diseases such as rheumatoid arthritis and polymyositis, various infectious diseases, and various cancers. , it can be expected to normalize the immune function of patients with the disease. It is desirable that the administration method and dosage form of the medicament of the present invention be appropriately selected depending on the type of disease, patient's condition, etc. In the case of oral administration, the daily dose per kg of body weight is 0.5 mg to 100 mg, preferably 1 mg to 30 mg, in the case of rectal administration, 1 mg to 100 mg, and in the case of intravenous administration. For subcutaneous or intramuscular administration, 1 mg to 10 mg is appropriate, and 1 mg to 30 mg is appropriate for subcutaneous or intramuscular administration, but these doses depend on the type of disease,
It is desirable to further select an appropriate amount depending on the condition of the disease. Furthermore, depending on the type of disease and the state of the disease, the therapeutic effect of the active ingredient of the present invention can be increased by using other drugs in combination as necessary. For example, cancer chemotherapy drugs such as alkylating agents and antimetabolites have side effects that reduce the patient's immune function, so when administering such drugs, the active ingredient of the present invention may be used in combination. By doing so, it is expected that the side effects of these drugs will be prevented and the therapeutic effects will be synergistically enhanced. Examples of the present invention will be described below. Example 1 4.42 g of 2-amino-4-p-nitrophenylthiazole were suspended in 300 g of tetrahydrofuran,
20 ml of triethylamine was added to this and the mixture was cooled to about 5°C. Add chloroformic acid ethyl ester to this mixture.
10.5 ml was added dropwise. 3.5 then at room temperature
Stir for days. Separating insoluble materials from the reaction mixture;
After concentrating the liquid, the residue was washed successively with water, diluted hydrochloric acid, and water in ethyl acetate. The ethyl acetate solution was then concentrated to about 50 ml, and the resulting crystals were separated and subjected to silica gel column chromatography, eluting with a mixed solvent of benzene and ethyl acetate. The obtained product was further recrystallized from ethyl acetate and N-
2.8 g of (4-p-nitrophenyl-2-thiazolyl)carbamic acid ethyl ester was obtained. Melting point: 224-226℃ Elemental analysis value: C ₁₂ H ₁₁ N ₃ O ₄ S Theoretical value (%) 49.14 3.78 14.33 10.93 Experimental value (%) 49.09 3.79 14.37 10.87 NMR (δ ^DMSO-d6 _TMS , ppm): 1.34 (3H, t: J=7Hz), 4.26 (2H.q: J=7
Hz), 7.88 (1H.s), 8.08 (2H.d: J=8Hz),
8.26 (2H.d: J = 8Hz), 11.88 (1H.s) Example 2 2.22g of 2-amino-4-p-methylthiophenylthiazole was dissolved in 50ml of tetrahydrofuran, 3.6g of triethylamine was added thereto, and the ℃
It was cooled to To this mixture, 3.3 g of chloroformic acid methyl ester was added dropwise. Thereafter, the mixture was stirred at room temperature for 3 hours. Concentrate the reaction mixture to a residue
150 ml of methanol was added and the mixture was heated to 45°C. Insoluble matter was removed, the methanol-soluble portion was concentrated, and the residue was subjected to silica gel column chromatography and eluted with a mixed solvent of chloroform-tetrahydrofuran. The obtained product was further recrystallized from methanol to obtain 1.2 g of N-(4-p-methylthiophenyl-2-thiazolyl)carbamic acid ethyl ester. Melting point: 193.5-195℃ Elemental analysis value: C ₁₂ H ₁₁ N ₂ O ₂ S ₂ Theoretical value (%) 51.41 4.31 9.99 22.87 Experimental value (%) 51.54 4.24 9.91 23.08 NMR (δ ^DMSO-d6 _TMS , ppm): 2.52 (3H, s), 3.78 (3H, s), 7.46 (1H, s),
7.2 to 7.84 (4H, m), 11.8 (1H, s) Compounds of the following examples were synthesized by the method of Example 1 or 2. Example 3 N-(4-p-methylsulfinyl phenyl-
2-thiazolyl)carbamic acid ethyl ester Melting point: 209-211℃ (decomposition) Elemental analysis value: C ₁₃ H ₁₄ N ₂ O ₃ S ₂ as C H N S Theoretical value (%) 50.30 4.55 9.03 20.66 Experimental value (%) 50.11 4.56 8.96 20.61 NMR (δ ^DMSO-d6 _TMS , ppm): 1.28 (3H, t:J=7.5Hz), 2.80 (3H, s),
4.24 (2H, q: J = 7.5Hz), 7.74 (1H, s),
7.66-8.12 (4H, m), 11.88 (1H, s) Example 4 N-(4-p-methylsulfinyl phenyl-
2-thiazolyl)carbamic acid n-butyl ester Melting point: 178-179℃ Elemental analysis value: C ₁₅ H ₁₈ N ₂ O ₃ S ₂ as C H N S Theoretical value (%) 53.23 5.36 8.28 18.95 Experimental value (%) 53.22 5.44 8.19 18.84 NMR (δ ^DMSO-d6 _TMS , ppm): 0.96 (3H, t:J=7.5Hz), 1.22-1.84 (4H,
m), 2.84 (3H, s), 4.24 (2H, t: J = 7.5
Hz), 7.8 (1H, s), 7.72~8.20 (4H, m),
11.92 (1H, s) Example 5 N-(4-p-methylsulfonylphenyl-2
-thiazolyl)carbamic acid ethyl ester Melting point: 248-250℃ Elemental analysis value: as C ₁₃ H ₁₄ N ₂ O _{4 S 2} C H N S Theoretical value (%) 47.84 4.32 8.58 19.65 Experimental value (%) 47.70 4.41 8.40 19.49 NMR (δ ^DMSO-d6 _TMS , ppm): 1.28 (3H, t:J=7.5Hz), 3.24 (3H, s),
4.23 (2H, q: J = 7.5Hz), 7.84 (1H, s),
7.90-8.18 (4H, m), 11.92 (1H, s) Example 6 N-(4-p-methylsulfonylphenyl-2
-thiazolyl)carbamic acid n-butyl ester Melting point: 191-192℃ Elemental analysis value: C ₁₅ H ₁₈ N ₂ O ₄ S ₂ C H N S Theoretical value (%) 50.83 5.12 7.90 18.09 Experimental value (%) 50.62 5.20 7.85 17.98 NMR (δ ^DMSO-d6 _TMS , ppm): 0.95 (3H, t:J=7.5Hz), 1.20-1.84 (4H,
m), 3.18 (3H, s), 4.23 (2H, t: J = 7.5
Hz), 7.90 (1H, s), 7.92-8.26 (4H, m),
12.00 (1H, s) Example 7 N-(4-p-nitrophenyl-2-thiazolyl)carbamic acid methyl ester Melting point: 252-254°C Elemental analysis value: C ₁₂ H ₁₁ N ₃ O ₄ S as C H N S Theoretical value (%) 49.14 3.78 14.33 10.93 Experimental value (%) 49.05 3.67 14.33 11.02 Example 8 N-(4-p-nitrophenyl-2-thiazolyl)carbamic acid n-butyl ester Melting point: 141-144℃ Elemental analysis value: As C ₁₄ H ₁₅ N ₃ O ₄ S C H N S Theoretical value (%) 52.33 4.70 13.08 ． 98 Experimental value (%) 52.43 4.81 13.01 10.00 NMR ( ^CDCl3 _TM , ppm): 0.86 (3H, t), 1.00-1.95 (4H, m), 4.01-
4.18 (2H, m), 7.26 (1H, s), 7.80-8.28
(4H, m), 9.70 (1H, s) Example 9 4 g of N-(4-p-nitrophenyl-2-thiazolyl)carbamic acid ethyl ester was dissolved in 500 ml of ethyl acetate, and 3 g of 10% palladium/carbon catalyst was added. Hydrogen reduction was carried out under normal pressure. The catalyst was removed by filtration, the liquid was concentrated, and the resulting residue was subjected to silica gel column chromatography and eluted with a mixed solvent of chloroform and ethyl acetate. The obtained product was recrystallized from methanol to give N-(4-p
2.6 g of -aminophenyl-2-thiazolyl)carbamic acid ethyl ester was obtained. Melting point: 300℃ or higher Elemental analysis value: C ₁₂ H ₁₃ N ₃ O ₂ S Theoretical value (%) 54.74 4.98 15.96 12.18 Experimental value (%) 54.66 5.13 15.95 12.00 NMR (δ ^DMSO-d6 _TMS , ppm ): 1.29 (3H, t:J=7Hz), 4.22 (2H, q:J=
7Hz), 7.12 (1H, s), 6.58 (2H, d:J=8
Hz), 7.54 (2H.d: J=8Hz) Example 10 N-(4-p-nitrophenyl-2-thiazolyl)
Composition of one tablet containing carbamate ethyl ester as an active ingredient: Active ingredient 100mg Lactose 38mg Corn starch 35mg Crystalline cellulose 20mg Hydroxypropylcellulose 5mg Magnesium stearate 2mg Total 200mg A tablet with the above ingredients was obtained by compression tableting. Example 11 N-(4-p-nitrophenyl-2-thiazolyl)
Composition of one capsule containing carbamate ethyl ester as an active ingredient: Active ingredient 100mg Lactose 95mg Corn starch 60mg Crystalline cellulose 40mg Magnesium stearate 5mg Total 300mg The above ingredients were filled into hard gelatin capsules to obtain capsules. Example 12 N-(4-p-nitrophenyl-2-thiazolyl)
Suppository 1 containing carbamic acid ethyl ester as the active ingredient Composition of the suppository: Active ingredient 0.3g Uitepsol W-35 (Dinamic Nobel Chemicals, West Germany) 1.7g Total 2.0g The above ingredients were once heated and dissolved and mixed thoroughly. rear,
It was cooled and solidified to obtain a suppository.

Claims (1)

[Claims] 1 General formula (1) K0421 (wherein R ₁ is a lower alkylthio group, a lower alkylsulfinyl group, a lower alkylsulfonyl group,
N-(4-phenyl-2-thiazolyl)carbamic acid derivative represented by N-(4-phenyl-2-thiazolyl)carbamic acid derivative, which represents a nitro group or an amino group, and _R2 represents a lower alkyl group. 2 Claims in which R ₁ represents a methylthio group, methylsulfinyl group, methylsulfonyl group, nitro group, or amino group in general formula (1), and R ₂ is a lower alkyl group having 1 to 4 carbon atoms. Derivative according to item 1. 3 General formula (1) K0422 (wherein, R ₁ is a lower alkylthio group, a lower alkylsulfinyl group, a lower alkylsulfonyl group,
A pharmaceutical composition for immunomodulation, characterized in that it contains as an active ingredient an N-(4-phenyl-2-thiazolyl)carbamic acid derivative represented by N-(4-phenyl- ₂ -thiazolyl)carbamic acid (representing a nitro group or an amino group, and R2 represents a lower alkyl group). thing. 4. The immunomodulating pharmaceutical composition according to claim 3, which is combined with a pharmaceutically acceptable diluent or carrier. 5. The pharmaceutical composition for immunomodulation according to claim 3, which is used for the treatment of rheumatoid arthritis. 6. The immunomodulating pharmaceutical composition according to claim 3, which is used in cancer immunotherapy. 7. The immunomodulatory pharmaceutical composition according to claim 3, which is used for the treatment of viral diseases.