JPH0440356B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides novel 1,2
The present invention relates to a dithiol-3-thione compound, a method for producing the same, and a novel immunomodulator containing the same as an active ingredient. In general, immunomodulatory effects have little effect when the immune system is functioning normally, but
It is understood that the function is to enhance immune function when it is low, and conversely to decrease it when immune function is hyperactive, correcting abnormalities in immune function and returning it to a normal state. Immune hyperactivity is also caused by a decline in the suppressive mechanism, but on the other hand, a state of decreased immune function can also be caused by an enhanced suppressive mechanism. Actions that act in the direction of correction can be called immunomodulatory actions. Many side effects have been reported in treatment with conventional immunomodulators (levamisole, thymus hormone, etc.). For example, levamisole causes nausea, rash,
Blood disorders have been reported, and the most serious side effect is granulocytopenia, which disappears when treatment is discontinued, but white blood cell counts should be closely monitored when using this drug over a long period of time. Must be. For this reason, there is a strong need for new immunomodulators that do not have the serious side effects normally observed in known immunomodulators. In recent years, efforts have been made to restore immune function that has been suppressed due to various causes, restore enhanced immune function to normal, and further strengthen and maintain normal immune function to prevent in-vivo infection and proliferation of pathogenic parasites such as viruses and bacteria. to oppose,
In addition, the importance of immunomodulators that strengthen resistance to the growth of foreign substances in the body such as cancer is increasing significantly. Such drugs are also desired to be applied to diseases such as various allergies, rheumatoid arthritis, diabetes, immunodeficiency, multiple sclerosis, and Kiran-Barré syndrome. As a result of searching for various immunomodulators that can be used for this purpose, the present inventors discovered lipoic acid (thioctic acid), which is widely used in the treatment of liver diseases.
He discovered that the DL form (DL form) has excellent immunomodulatory effects, and presented the results (Pharmaceutical Society of Japan, 104th Annual Meeting, Abstracts of Lectures 1984, p. 397, Yamamoto). As a result of further intensive study, we arrived at the present invention, which has the general formula () (In the formula, R represents a hydrogen atom, a halogen atom, a lower alkoxy group, a lower alkyl group, an amino group, an amino group substituted with a lower alkyl group, or a lower alkoxycarbonyl group) The compound of the present invention is surprisingly safe. It was found that it has a high immunomodulatory effect and superior immunomodulatory effect than lipoic acid (thioctic acid DL form). Therefore, the immunomodulatory agent of the present invention is effective against autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, nephritis, and multiple sclerosis, as well as other immediate and delayed allergic diseases, malignant tumors, and severe infections. It can be widely used in the treatment of immunodeficiency, immunosuppression, etc. The immunomodulator of the present invention can be administered in one dose for adults.
The dose ranges from 0.1 to 500 mg, and can be administered once or more per day. For example, it can be administered orally in the form of tablets, capsules, powders, granules, etc., as well as in the form of injections or suppositories. Next, a manufacturing method for obtaining the compound of the present invention will be explained. General formula () (wherein R is the same as in the above general formula ()) using a cinnamaldehyde derivative represented by an organic solvent,
That is, solvents such as aliphatic or aromatic hydrocarbons, halogenated hydrocarbons, alcohols, and ethers,
5-methyl-, preferably in a solvent such as hexane, toluene, or methyl alcohol, in the presence of a base
It can be produced by condensation with 1,2-dithiol-3-thione. Examples of the base used include lower alkoxides of alkali metals and alkaline earth metals such as sodium methoxide, sodium ethoxide, and magnesium methoxide, and organic bases such as triethylamine and DBU. Usually, these bases are used in an amount of 1 to 10 equivalents, preferably 2 to 5 equivalents, relative to 5-methyl-1,2-dithiol-3-thione. The reaction temperature is -10°C to 100°C, preferably 0°C to
The reaction is completed by stirring at 50°C for 1 hour, and the compound of the present invention can be produced in good yield. The method of the present invention, which is a method for producing a compound represented by the general formula () by condensing an aliphatic aldehyde and 5-methyl-1,2-dithiol-3-thione, is a novel production method. More specific examples will be shown below. 5-methyl-1,2-dithiol used as raw material
3-thione is Bull.Soc.Chim.France 1962 2182
(Thurlier, A). (Reference example) Reference example 1 4,4-di(methylmercapto)-3-buten-2-one 60% sodium hydride in 500 ml of anhydrous benzene 40
g (1.0 mol) and add 88.2 g of t-amyl alcohol while making sure that the internal temperature does not exceed 60℃.
(1.0 mol) was added dropwise. After the dropwise addition, the mixture was heated and stirred under reflux for 2 hours and left at room temperature overnight. A mixed solution of 29 g (0.5 mol) of acetone and 38.1 g (0.5 mol) of carbon disulfide was added dropwise while keeping the internal temperature below 10°C. After stirring at room temperature for 5 hours,
141.9 g (1.0 mol) of methyl iodide was added dropwise while stirring under ice cooling. After stirring at room temperature for 3 hours, it was left to stand overnight. Water was added to the reaction mixture, and the organic layer was separated, washed with water, and then dried over magnesium sulfate. The solvent was distilled off under reduced pressure, and hexane was added to the resulting residue for crystallization to obtain 59.7 g of crude crystals. Recrystallized from ethanol to give 4,4-di(methylmercapto)-3
-Buten-2-one was obtained as yellow needle-like crystals. Yield 51.9g (yield 61.4%) Melting point 65-66.5℃ Reference example 2 5-methyl-1,2-dithiol-3-thione 130g (0.58 mol) of phosphorus pentasulfide in 1.3 xylene
51.9 g (0.32 mol) of 4,4-di(methylmercapto)-3-buten-2-one was suspended under reflux.
200 ml of xylene solution was added dropwise. After heating under reflux for 30 min, the reaction mixture was dissolved in diethyl ether.
I poured it into 1.5. Insoluble materials were removed by filtration, and the mother liquor was washed with water and then with a 1% aqueous sodium hydroxide solution. The organic layer was dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. Purified by silica gel column chromatography (hexane: ethyl acetate = 10:1), 5-methyl-1,2-dithiol-3-
Thione was obtained as a red oil. Yield 21.9g (yield 46.0%) NMR ( _CDCl3 , δ _ppn ) 7.0 (1H, S) 2.54 (3H, S) Example 1 5-(4-phenyl-1,3-butadienyl)-
1,2-dithiol-3-thione (compound no.
1) 600mg of metallic magnesium in 40ml of methanol (24
milligram atoms) and heated under reflux for 1 hour to dissolve the magnesium. 1.4 g (10 mmol) of cinnamaldehyde under ice cooling, followed by 5-methyl-
1.56 g (10 mmol) of 1,2-dithiol-3-thione was added, and the mixture was stirred at the same temperature for 1 hour. The precipitated crystals were collected by filtration and added to 300 ml of eluted acetate.
Insoluble matters were removed by filtration, and the mother liquor was washed with water and then dried over magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was recrystallized from benzene-ethanol to obtain 5-(4-phenyl-1,3-butadienyl)-1,2-dithiol-3-thione as dark red crystals. Yield 0.53g (yield 19.2%) Melting point 160-161℃ NMR (DMSO-d ₆ , δ _ppn ) 7.3-7.7 (5H, m) 6.8
-7.28 (5H, m) Similarly to Example 1, the corresponding cinnamaldehyde derivatives (4-chlorocinnamaldehyde, 4-chlorocinnamaldehyde,
4-Methoxycinnamaldehyde, 4-dimethylaminocinnamaldehyde, p-methylcinnamaldehyde,
o-chlorocinnamaldehyde, m-methylcinnamaldehyde) and 5-methyl-1,2-dithiol-3
-thione was reacted to obtain the following compound group. 5-{4-(4-chlorophenyl)-1,3-butadienyl}-1,2-dithiol-3-thione (Compound No. 2) Shape: Red crystals, Yield: 18.9% Melting point: 144-145℃ NMR (DMSO−d ₆ , δ _ppn ) 6.80−7.25 (4H, m)
7.30−7.79 (5H, m) 5-{4-(4-methoxyphenyl)-1,3-
Butadienyl}-1,2-dithiol-3-thione (Compound No. 3) Shape: Black crystals, Yield: 13.6% Melting point: 147-148°C NMR (DMSO-d ₆ , δ _ppn ) 7.28-7.69 (4H, m)
6.78−7.12 (5H, m) 3.79 (3H, s) 5-{4-(4-dimethylaminophenyl)-1,
3-butadienyl}-1,2-dithiol-3
-Thion (Compound No. 4) Shape: Dark green crystal, Yield: 5.3% Melting point: 133-134
°C 5-{4-(p-tolyl)-1,3-butadienyl}-1,2-dithiol-3-thione (Compound No. 5) Shape: Dark green crystals, Yield: 26.7% Melting point: 144-145
°C NMR (CDCL ₃ , δ _ppn ) 2.44 (3H, s) 6.50−7.50
(9H, m) Elemental analysis (C ₁₄ H ₁₂ S ₃ ) Calculated value (%) C; 60.83, H; 4.38, S; 34.79 Actual value (%) C; 60.70, H; 4.11, S; 35.05 5-{ 4-(o-chlorophenyl)-1,3-butadienyl}-1,2dithiol-3-thione (Compound No. 6) Shape: Black-red crystals, Yield: 11.0% Melting point: 161-162
°C NMR (DMSO−d ₆ , δ _ppn ) 7.87 (1H, s) 6.84−
7.70 (8H, m) 5-{4-(m-methylphenyl)-1,3-butadienyl}-1,2 dithiol-3-thione (Compound No. 7) Shape: Black-red crystals, Yield: 23.1% Melting point ;143.5−
144.5℃ NMR (DMSO-d ₆ , δ _ppn ) 2.36 (3H, s), 6.95-
7.51 (9H, m) Specific test examples and formulation examples of the compounds of the present invention are shown below. Test Example A Effect on mouse splenocyte blastogenesis Concanavalin A in mouse splenocytes
The effects of the compounds of the present invention on DNA synthesis induced by Concanavalin A (hereinafter referred to as Con A) were investigated. BALB/C mouse (female 10 weeks old) splenocyte 1
×10 ⁵ conA 2 μg/ml and the compound of the present invention
in RPMI 1640 medium containing 10% fetal bovine serum with 10 ^-6 , 10 ^-5 or 10 ^-4 M, 37°C, 5% CO ₂
After culturing for 48 hours ^under
Cultured for 18 hours. The cultured cells were harvested using a harvester, and the amount of ³ H-thymidine taken into the cells was measured using a liquid scintillation counter. The results are shown in the table. The displayed value is a relative value when the uptake amount of the control well is set as 100. B. Effect of enhancing antibody production The effect of the compounds of the present invention on antibody production against sheep red blood cells (SRBC) in mice was investigated. A group of 4 ICR mice (female, 7 weeks old) was used and 5 x 10 ⁷ SRBCs were injected into the tail vein of the mice for immunization. The compound of the present invention was administered twice on the day of immunization and the next day.
It was suspended in 0.5% methylcellulose and administered orally. Three days after immunization, the number of anti-SRBC antibody producing cells (PFC number) in the mouse splenocytes was measured by the method of Jerne (Science, vol. 140, p. 405, 1963). The results are shown in the table. The displayed value is that of the control group.
This is a relative value when the number of PFC is set to 100. C. Rat experimental allergic encephalomyelitis (EAE)
Chronic relapsing rat EAE, which is a model for autoimmune diseases such as multiple sclerosis, was produced according to the method of Feuer et al., and the effects of the compounds of the present invention were investigated. Lewis rats (female, 7 weeks old, 8 per group)
10 mg of freeze-dried guinea pig spinal cord and 0.2 ml of emulsion from Freund's complete adjuvant (containing 2 mg of Mycobacterium tuberculosis strain H37RA) were inoculated in divided doses into the footpads of both hind legs. The compound of the present invention was suspended in 0.5% methylcellulose and administered subcutaneously a total of 8 times at intervals from 7 days before to 7 days after inoculation. Clinical symptoms were observed until 50 days after inoculation and scored from 0 to 5 according to the method of Feuer et al. The results are shown in the table. As is clear from the average symptom score during the survival period, Compound No. 1 showed an excellent suppressive effect on EAE in rats. D. Acute toxicity test Acute toxicity in mice was investigated using the oral administration method. The compound of the present invention was suspended in 0.5% methylcellulose and orally administered to ddY mice (male, 7 weeks old), each group having 6 mice. The progress was observed for 14 days after administration, and the number of deaths was determined. Compound No. 1 500mg/Kg
When administered, there were no deaths and no effect on body weight fluctuation. On the other hand, lipoic acid (thioctic acid DL form)
When administered at 300 mg/Kg, one out of six animals died.

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【table】

Table: Formulation Example (Tablet) Tablets containing 20 mg of active compound and having the following composition were prepared by conventional techniques. Compound No. 1 20mg Lactose 78mg Cornstarch 50mg Magnesium stearate 2mg Other compounds of the present invention were also made into tablets in the same manner.

Claims (1)

[Claims] 1 General formula () (In the formula, R represents a hydrogen atom, a halogen atom, a lower alkoxy group, a lower alkyl group, an amino group, an amino group substituted with a lower alkyl group, or a lower alkoxycarbonyl group). 2 General formula () (In the formula, R represents a hydrogen atom, a halogen atom, a lower alkoxy group, a lower alkyl group, an amino group, an amino group substituted with a lower alkyl group, or a lower alkoxycarbonyl group). A method for producing a 1,2-dithiol-3-thione derivative represented by the general formula (), which comprises condensing with 5-methyl-1,2-dithiol-3-thione in a solvent in the presence of a base. 3 General formula () (In the formula, R represents a hydrogen atom, a halogen atom, a lower alkoxy group, a lower alkyl group, an amino group, an amino group substituted with a lower alkyl group, or a lower alkoxycarbonyl group) as an active ingredient. .