JPH02247122A - Anti-inflammatory agent - Google Patents

Abstract

PURPOSE:To obtain an anti-inflammatory agent of reduced toxicity by using 2-methoxy-4-(n-propylthio)aniline or its pharmaceutically permissible acid adduct as an active ingredient. CONSTITUTION:2-Methoxy-4-(n-propylthio)aniline of the formula or its pharmaceutically permissible acid adduct is used as an active ingredient: in case of oral preparations, about 10 to 100mg dose; in injection solution, about 1 to 500mg and in suppository, about 5 to 500mg to give an anti-inflammatory agent of low toxicity, which has excellent anti-active oxygen action to be effective against not only carrageenan edema, adjuvant arthritis to which prostaglandin synthesis inhibitor is effective, but also concanavalin A edema and phorbol ester edema to which prostaglandin synthesis inhibitor is ineffective.

Description

DETAILED DESCRIPTION OF THE INVENTION Technical Field The present invention relates to a novel anti-inflammatory agent. More specifically, the invention provides 2-methoxy-4-(n-propylthio)
The present invention relates to a novel anti-inflammatory agent containing aniline or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.

Prior art> Aminophenylthioethers (French Patent No. 14)
Publication No. 89916 and German Patent No. 2706104
In addition to its use as an intermediate for dyes, the compound known as 1) was known to have acaricidal activity. However, nothing was known about the anti-inflammatory effect of 2methoxy-4-(n-propylthio)aniline, which belongs to aminophenylthioethers.

Since 1960, non-steroidal anti-inflammatory drugs (NSAIDs)
) were developed one after another and came to be used clinically.

Most of these NSA IDs are occupied by acidic anti-inflammatory drugs represented by arylacetic acid derivatives and 2-arylpropionic acid derivatives. A large number of drugs have been created for these acidic anti-inflammatory drugs, but these
All have in common that they exhibit anti-inflammatory effects by inhibiting +ooxy-genase and suppressing prostaglandin (PG) production. Further improvements in terms of action and side effects are almost impossible (Tsurumi: Infection, Inflammation, and Immunology, 16.83 (1986).Also, basic anti-inflammatory drugs have strong analgesic effects but weak anti-inflammatory effects. Therefore, it is not used for chronic inflammation.Therefore, drugs with a different mechanism of action from existing N5AIDs are thought to be useful for clinical applications and basic research on inflammation.

[Summary of the invention]

Summary> The purpose of the present invention is to provide a novel anti-inflammatory agent with low toxicity.

The present inventors focused on the involvement of active oxygen in inflammatory reactions,
As a result of synthesizing compounds with anti-slip oxygen properties and evaluating their anti-inflammatory properties, we found that 2-methoxy-4-
We have discovered that (lopropylthio)aniline has excellent anti-slip oxygen properties and is useful as a basic anti-inflammatory drug with anti-inflammatory properties.

That is, the anti-inflammatory agent according to the present invention contains 2-methoxy-4
-(n-propylthio)aniline or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.

Effect> The anti-inflammatory agent according to the present invention has an anti-inflammatory effect and is less toxic.

In addition, the anti-inflammatory agent according to the present invention has excellent anti-oxygen effects and is effective in an immune complex inflammation model in which indomethacin, a prostaglandin synthesis inhibitor, is ineffective. It can be expected to be used as an inflammatory drug.

[Detailed description of the invention]

2-methoxy-4-(n-propylthio)aniline and its salts> An aminophenylthioether represented by 2-methoxy-4-(n-propylthio)aniline, which has anti-inflammatory effects as described above. be.

This compound can be prepared by any convenient method. An example of this specific production method is to react 5-chloro-2-nitroanisole with n-propylthiol in an aprotic solvent in the presence of a base.
- leading to nitro-5-(n-propylthio)anisole,
This method further reduces the nitro group of this 2-nitro-5-(n-propylthio)anisole to obtain 2-methoxy-4-(n-propylthio)aniline.

Since this compound has a free amino group, an acid addition salt may exist at this position, but the acid addition salt referred to in the present invention should be a pharmaceutically acceptable salt. Acid addition salts which can be used pharmaceutically are, for example, the corresponding mineral salts, such as hydrochlorides, hydrobromides or hydroiodides, sulfates, bisulfates or phosphates, or the corresponding carboxylates. , such as fumarates, maleates, malates, succinates, citrates, gluconates or salicylates, or the corresponding sulfonates, such as methanesulfonates, ethanesulfonates, benzenesulfonic acids salt, p-toluenesulfonate.

2-methoxy- as an active ingredient of the anti-inflammatory agent according to the present invention
4-(n-propylthio)aniline and its pharmaceutically acceptable acid addition salts were used to improve lipid peroxidation in rat brain homogenate (Shingu et al. Japanese Journal of Pharmacology, 87.427 (1)
986)), diphenylpicrylhydrazyl (D
jphenyl picryl-hydrazyl) radical (K, Kubo et al.; Arch, int.

Pharmacodyn, 272.283 (198
4)) elimination and active oxygen production from macrophages (Mibu et al. Japanese Pharmacological Journal, 83.355 (1,984)
) and an increase in plasma deoxidized lipids in mice injected with endotoxin (K, Sugino et al.: Sur
Gery, 101°746 (1987)), it is expected to be useful for inflammatory diseases based on active oxygen.

In addition, PG synthesis inhibitors are not only effective for carrageenan edema and adjuvant arthritis, which are inflammatory models, but also
Immune complex inflammation model in which PG synthesis inhibitors are ineffective (Abe et al.: Inflammation, 1.739 (1981)), Concanavalin A edema (A, J,
Lewis et al.; EurJ, Pharmacol.
40.1 (197B)) and phorbol ester edema (Kura et al., Japan Pharmacological Journal, 93.19p (1989)), and has low toxicity. These compounds can therefore be used as anti-inflammatory agents.

Anti-inflammatory agent> The anti-inflammatory agent according to the present invention contains 2-methoxy-4(n-propylthio)aniline or a pharmaceutically acceptable acid addition salt thereof (hereinafter also referred to as active ingredient compound) as an active ingredient. It is something.

These compounds as anti-inflammatory agents can be applied in various forms and can be used alone or in mixtures with pharmaceutically acceptable diluents or other agents. The dosage form of the active ingredient compound in the present invention as an anti-inflammatory agent is as follows:
Depending on the therapeutic purpose, the drug is appropriately selected from oral preparations such as tablets, capsules, powders, and granules, suppositories, liquid drugs, ointments, and injection preparations such as intravenous injection and intramuscular injection.

Therefore, the anti-inflammatory agent of the present invention can be prepared by any conventionally known production method.

The amounts of 2-methoxy-4-(propylthio)aniline and its pharmaceutically acceptable acid salts, which are the active ingredients to be incorporated into the dosage unit form of the anti-inflammatory agent of the present invention, are fixed depending on the symptoms or dosage form. However, it is generally about 10 to 100 mg for oral use, and about 1 to 500 [1 g,
For suppositories, the amount is preferably about 5 to 500 mg. Also,
Although the dosage of the anti-inflammatory agent according to the present invention per -1 cannot be determined unconditionally depending on the symptoms, it is usually preferable to set the amount of the active ingredient compound in the range of 1 to b.

Experimental Examples The following are production examples, formulation examples, pharmacological test results, and acute toxicity test results of the active ingredient compound of the anti-inflammatory agent of the present invention, but the present invention is not limited by these examples. It's not a thing.

1) Production example (active ingredient compound) 2.6 g of sodium hydride is added to 100 ml of dimethylformamide (DMF), and then 50 ml of a DMF solution containing 5.9 ml of n-propyl mercaptan is added dropwise over 15 minutes or more. (At this time, keep the internal temperature at 0-5°C). After stirring this at 0°C for 40 minutes, the internal temperature was reduced to 0.
While maintaining the temperature at ~5°C, 50ml of a DMF solution containing 10.2g of 5-chloro-2-nitroanisole is added dropwise over 40 minutes. This liquid is stirred at 0° C. for 2 hours. Pour an appropriate amount of cold water into this and adjust the pH to 3-4 with dilute hydrochloric acid.
After adjusting the temperature, extract with ethyl acetate. The extract was washed with water, dried over Glauber's salt, and then the solvent was distilled off under reduced pressure. The resulting amorphous solid is purified by silica gel column chromatography. Ethyl acetate: n-hexane (1:10
), 2-nitro-5-(
8.4 g of n-propylthio)anisole was obtained.

'H-NMR (δ, CDC13): 1.07 (t, 3H
) , 1.75 (m, 2H) , 2.98 (t, 2H
), 3.96 (s, 3H), 6.84 (dd, L
H), 6.88 (d, IH), 7.85 (d, I
H) 2-nitro-5-(n-propylthio)anisole7.
3 g are added to 300 ml of methanol, followed by 150 ml of an aqueous solution of 20 g of sodium dithionite. After stirring at room temperature for 30 minutes, increase the internal temperature to 80-100°C.
Add 10 g of sodium dithionite and stir for 10 minutes. The reaction solution is made acidic (pH 1-2) by adding concentrated hydrochloric acid under water cooling, then made alkaline (pH 11-12) by adding an aqueous potassium hydroxide solution, and extracted with chloroform. After drying the extract with Glauber's salt, the solvent was distilled off under reduced pressure and 2-methoxy-4-(n-propylthio)aniline 3
．． 3g was obtained.

'NMR (δ, CDC13): 0.98 (t, 3H),
1.60 (m, 2H), 2.76 (t.

2H), 3.85 (s, 3H), 6.62 (I
H.

d) , 6.82-6.95 (m, 2H) 2) Formulation example (a) To 50 parts of the tablet active ingredient compound, 100 parts of corn starch, 25 parts of lactose, 2 parts of hydroxypyl cellulose, 2 parts of talc, Add 1 part of magnesium stearate, mix well, and compress the mixture using a tablet machine to form tablets with a diameter of 10 m+n.

(b) 130 parts of cornstarch to 100 parts of the capsule active ingredient compound;
40 parts of lactose, 20 parts of talc, and 10 parts of magnesium stearate are uniformly mixed, and the mixture is made into a powder or fine granules and placed in a capsule container to form a capsule.

(C) 10 parts of the active ingredient compound was added to Wittebusol W35 14
Mix it with 0 parts and fill it into a waste container to use it as a raw material.

3) Pharmacological test The following shows a pharmacological action test of the active ingredient compound of the anti-inflammatory agent according to the present invention (hereinafter also referred to as compound ①).

(1) Inhibitory effect on rat brain homogenate lipid peroxidation Method of Shingu et al. (Japan Pharmacological Journal 93.427 (1986
)) rat brain homogenate (1,7% W/V
) was shaken with the test drug in 1.5 ml of 67 mM phosphate buffer at 37°C for 1 hour, and then 0.5 ml of 20% trichloroacetic acid was added to remove protein. Upper grade 1
．． Add 0.5 ml of 1.2% thiobarbic acid to 5 ml and heat at 100'C.

After heating for 5 minutes, the absorbance at 532 parts m was measured to determine the amount of lipid peroxide produced. The drug concentration (IC50) that suppressed lipid peroxide production by 50% was determined compared to a control in which no test drug was added. As a result, the IC50s for Compound (I) and the control indomethacin were 5.3 (μM) and 100 (μM) or more, respectively.

(2) Diphenylpicrylhydrazyl radical scavenging action diphenylpicrylhydrazyl (D P P H) 1
Add the test drug to 00 μM ethanol solution and incubate for 30 μM at room temperature.
After standing for a minute, the absorbance at 517 parts m was measured (K, K
ubo et al.: Arch, int, Pharmaeo
dyn, 272, 283 (1984) ), D
When we determined the drug concentration (IC0,2) that lowers the absorbance of 517 parts m by PPH by 0.2, we found that the compound (
IC0,2 for indomethacin 1) and the control were 10.0 (μM) and 100 (μM) or more, respectively.

(3) Inhibitory effect on active oxygen production by macrophages 10 ml of liquid paraffin was intraperitoneally administered to rats, and after 4 days, exudate cells were collected in Eagle's MEM medium. 2X
After incubating 106 cells/ml, 63 μg/ml luminol, and the test drug in MEM medium at 37°C for 10 minutes, 160 μz/ml opsonized thymosan was added, and the chemiluminescence generated was measured over time using a luminometer. It was measured. The drug concentration (IC50) that inhibits the integral value of luminescence by 50% for 20 minutes after the addition of thymosan was determined (Chisei et al. = 1-1 This Pharmacological Journal, 83.355 (19
84)). As a result, the IC50 values for Compound (I) and the control indomethacin were 5.5 (μM) and 1500 (μM), respectively.

(4) Effect on endotoxin-induced hyperlipidemia and E. coli (E, colt 055: B5) in mice
30 mg/kg of endotoxin from the plant was administered intraperitoneally,
Blood was collected 16 hours later, and lipid peroxide in the plasma was measured using the fluorescence assay method of Yagi et al.
, Mcd, +15.2]2 (197B)). The test drug was administered intraperitoneally at the time of endotoxin administration and 14 hours later (K, Sugino et al.
: Surgery, 101, 74B (
1987>).

The results are shown in Table 1.

Table 1 Effect control on endotoxin-induced hyperlipidemia (physiological saline) Compound I Normal mouse (mg/kg) (nmol/ml) 29.4 10 21.7 30, 1.9.7 100 14.5 4.5 (5) Effect on carrageenan edema C, A, method of Winter et al. (Proc, Soc
, Exp, Biol.

MQd, III, 544 (1,962))
- 0.1% of 1% Naan on the footprints of La I, who fasted at night.
ml was subcutaneously administered, and 30 minutes later, the test drug was orally administered, and the paw volume was determined at 1 fl11 over time. The results are shown in FIG.

(6) Adjuvan I/Effect on arthritis B, B,
The method of Newbould et al. (J3r, J, Phar
Macol, 21.1.27 (+963)), 0.6 mg of killed Mycobacterium butyricum (M, Butyricum) suspended in liquid paraffin was subcutaneously administered to the rat footprint, and the paw volume was increased over time to 11111. Established. Compound (I) was orally administered at 400 mg/kg/day from day 6 to day 1, day 18.

The results are shown in FIG.

(7) Effect on immune complex inflammation model According to the method of Abe et al. (Inflammation, 1.739 (1981)), 5 x 108 sheep red blood cells (SRBC) were intravenously administered to mice, and 14 days later, the same number was administered again. 5RBC were administered intravenously. After 19 days, 2×1.08 5RBCs were subcutaneously administered into the footprints, and the thickness of the footprints was measured 4 hours later. The test drug was orally administered 30 minutes before administering 5RBC to the footprint. The results are shown in FIG.

(8) Effect on concanavalin A edema and phorbol ester floatation 11m Method of Levi and Cur et al. (A, J, Lewis
et al.: Eur, J. Pharmacol, 40.1
(197B), Kura et al.: Japanese Pharmacological Journal, 93.19P.
(1989)), the test drug was orally administered to rats, and 30 minutes later, 1% concanavalin A or 1 μg
0.1 ml of phorbol-12-myristate-13-acetate was administered subcutaneously to the paw print, and the paw volume was measured over time. The results of the edema-suppressing effect of concanavalin A are shown in FIG. 4, and the results of the edema-suppressing effect of phorbol ester are shown in FIG.

4) Acute toxicity test The following shows an acute toxicity test of the active ingredient compound in the present invention.

Using ICR male mice, the active ingredient compound was dissolved in distilled water and administered orally to a 50% lethal dose L D 5o (
mg/kg) was determined by the Litchfield-Wilcoquilan method. The results are shown in the table below.

[Brief explanation of drawings]

FIG. 1 shows the effect of active ingredient compounds on carrageenan edema. FIG. 2 shows the effect of the active ingredient compound on adjuvant arthritis. FIG. 3 shows the effect of the active ingredient compound on the immune complex inflammation model in comparison with that of other drugs. FIG. 4 shows the effect of the active ingredient compound on concanavalin A edema in comparison with that of other drugs. FIG. 5 shows the effect of the active ingredient compound on phorbol ester edema in comparison with that of other drugs. Applicant's agent Mr. Sato

Claims (1)

[Claims] An anti-inflammatory agent containing 2-methoxy-4-(n-propylthio)aniline or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.