JPH0137374B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to anti-inflammatory compositions containing novel neutral copper conjugates and methods of use thereof. In recent years, much research has been conducted on the role of copper, especially the role of copper conjugates, in the treatment of inflammatory diseases.
It has been reported. Most recent studies have reported the use of copper conjugates, particularly cupric salicylic acid salts, in the treatment of rheumatic diseases.
(Sorenson &Hangarter;Inflammation; 2
217-238 (1977) describes the production of various copper salicylic acid conjugates by reacting inorganic copper salts with salicylic acid in an aqueous solution. The specific conjugate formed in aqueous solution has been found to depend on the PH of the aqueous solution. Then, it is a pale blue crystalline bis(salicylate)cotpar()tetrahydrate, with the general formula General formula of the compound represented by and olive green salicylate caprate () sodium salt Both anionic conjugates are known, such as the compound represented by The purpose of the present invention is to treat inflammatory diseases by using a composition containing as an active ingredient a novel copper conjugate produced by the reaction of a copper compound and substituted benzoic acid in a non-aqueous solvent. The goal is to provide drugs that do this. According to one aspect of the invention, the general formula Cu[C ₆ H ₄
(X)COO] ₂ ROH (However, ROH represents an alkanol, preferably represents a C ₁ to C ₆ aliphatic alkanol, and X may be located at any position; OH, SH, SeH,
NH ₂ or NHR′, R′ is

【formula】

The present invention provides an anti-inflammatory composition containing a novel neutral conjugate represented by the following formula and a pharmaceutically acceptable non-aqueous carrier. According to the results of biochemical studies, the conjugates of the present invention appear to have the following duplex structure.
Note that the theory is not limited to this theory. Preferred compounds for use as active ingredients in anti-inflammatory compositions according to the invention are of the general formula This is a copper salicylate conjugate thought to be represented by This compound may be prepared by reacting an inorganic copper compound, such as a copper salt or cupric hydroxide, with a suitable substituted benzoic acid in the presence of an alkanol. For example, the reaction of cupric hydroxide with salicylic acid in anhydrous ethanol produces the general formula Cu
A conjugate represented by [C ₆ H ₄ (OH)COO] ₂ ·C ₂ H ₅ OH (hereinafter referred to as general formula Cu[HSal] ₂ ·C ₂ H ₅ OH) is produced. The compound contains excess salicylic acid (approximately
When crystallized from absolute ethanol containing 5M), dark green crystals are formed. This compound has been found to be particularly useful in the treatment of inflammatory diseases in humans and animals. The present invention therefore relates to a method and a medicament thereof, characterized in that an effective amount of the above-mentioned compound is administered to humans or animals. This compound has been found to be particularly effective when used topically in humans and animals in alleviating the symptoms of inflammatory diseases such as rheumatoid arthritis, arthritis, and rheumatoid arthritis. The anti-inflammatory agent of the present invention is preferably used in the form of a composition comprising the above-mentioned compound and a pharmaceutically acceptable carrier. Therefore, the present invention provides an anti-inflammatory composition comprising the above compound and a pharmaceutically acceptable carrier, which composition is suitable for topical application to the human or animal being treated. It is suitable. The composition may be used in the form of a gel, ointment, paste, cream or lotion. The composition of the present invention is particularly preferable because a solution containing the above-mentioned compounds is effective for permeation through the skin. Since the above compounds have low stability in the presence of water, it is preferable to form a composition containing a non-aqueous carrier. A particularly preferred composition is one in which the above compound is dissolved in a non-aqueous liquid auxiliary.
Preferred auxiliaries are, for example, short-chain ( _C1 - _C10 ) and long-chain ( _C12 - _C20 ) alcohols, e.g. monohydric alcohols such as methanol, ethanol, propanol, butaryl, cetyl alcohol;
Dihydric alcohols such as ethylene glycol and
These include monohydric, dihydric and trihydric alkanols such as polyhydric alcohols such as glycerol. A particularly preferred composition of the invention, which has been found to be free of deterioration over very long periods of time, is a solution of the above compound in a liquid auxiliary containing an alkanol, glycerol and salicylic acid. The potency of the above compounds incorporated into the anti-inflammatory compositions of the present invention will vary depending on the inflammatory condition being treated, but will usually range from 0.1 to 15% W/V, preferably 1
A copper binder of 7% W/V is blended. The composition of the present invention, in the form of a pharmaceutical composition as described above, has been found to be effective in alleviating symptoms of inflammatory diseases when applied topically to humans and animals. The composition is believed to be effective in this type of treatment because it penetrates the skin immediately. Furthermore, it is believed that the compositions of the present invention are non-toxic, making them particularly preferred for use in the treatment of inflammatory diseases in humans. this is,
The limited duration of the anti-inflammatory effect of the compounds of the present invention is evidenced by the immediate disappearance of the compounds from treated humans and animals. The anti-inflammatory composition of the present invention may be a mixture of one or more of the above compounds and other active pharmaceutical ingredients such as anti-inflammatory agents and known pharmaceutical excipients. Data regarding the toxicity of the copper salicylate-ethanol conjugate, which is the anti-inflammatory composition of the present invention in Reference Example 1, is as follows. The oral LD50 in female rats is 690 mg/Kg and in male rats it is 800-900 mg/Kg. In addition, the toxicity data regarding ALCUSAL, which is a formulation similar to the formulation prepared in Reference Example 1 except that a gelling agent, surfactant, etc. are added to make the composition into a gel, is as follows. It's like this. Incidentally, Alxal is a preparation that is already commercially available in Australia and other countries. The transdermal LD50 in rabbits is 5 g/Kg or more when the gel is applied topically. Therefore, at these values, this formulation can be said to be non-toxic.
Also, the oral LD50 in rats is 5 g/Kg or more. The present invention will be further explained, but the present invention is not limited to the following examples. Reference example 1 Production method of Cu[HSal] ₂・C ₂ H ₅ OH). 0.97 g of freshly prepared Cu(OH) ₂ was added to a solution of 6.9 g of salicylic acid in 50 ml of absolute ethanol. The suspension was refluxed on a water bath until all the copper hydroxide had reacted. The resulting dark green solution was filtered and green crystals were precipitated by cooling. The precipitate was filtered and green crystals were precipitated by cooling. The precipitate was filtered, washed with cold alcohol and air dried. The yield was 3.2 g, and the results of elemental analysis were as follows. (CuC ₁₆ H ₁₆ O ₇ ) Cu C H Calculated value (%) 16.6 50.1 4.2 Actual value (%) 16.6 50.2 4.2 Reference example 2 Cu[HSal] ₂・CH ₃ OH) and Cu[HSal] ₂・
Method for producing C ₃ H ₇ OH. In the method of Reference Example 1, by replacing anhydrous ethanol with anhydrous methanol or anhydrous n-propanol, Cu analyzed as CuC ₁₅ H ₁₄ O ₇
[HSal] _{2.CH 3} OH and Cu[HSal] _{2.C 3} H ₇ OH analyzed as CuC ₁₇ H ₁₈ O ₇ were produced. Example 1 This example shows a method for producing a composition containing Cu[HSal] _{2.C 2} H ₅ OH. 1.0 g of freshly prepared cupric hydroxide was added to a solution of 7 g of salicylic acid in 80 ml of absolute ethanol. The suspension was heated until the cupric hydroxide reacted. The resulting dark green solution was filtered to separate the insoluble polymerized copper salicylate and added to the 20 ml glycerol filtrate. Dark green 0.1M−Cu obtained by this method
[HSal] ₂ C ₂ H ₅ OH solution was stable for a long period of time.
This was used in the tests described in Examples 2 and 3. Example 2 This example demonstrates the transdermal absorption of the compound Cu[HSal] ₂ C ₂ H ₅ OH. Male Wistar rats (average weight 250 g) were anesthetized with diethyl ether, their spines were shaved over an area of 20 cm ² , and the Cu prepared according to Example 1 was shaved.
[HSal] ₂ C ₂ H ₅ OH composition was applied to each rat.
(20 ml per 3 rats) The rats were returned to their cages, and 2 days later their urine was collected and compared with the urine of control untreated rats using the method described below. 5 ml of urine was acidified with 2M H ₂ SO ₄ and stirred with 2 ml of ethyl acetate. The ethyl acetate layer was centrifuged and analyzed by thin layer chromatography using silica gel. (Solvent: benzene, diethyl ether, glacial acetic acid, methanol 120:60:18:1 volume ratio) Salicylic acid, salicyluric acid, gentisic acid, and ethyl salicylate were used as samples. The results are shown in Figure 1. Figure 1 shows samples 1 to 9.
FIG. 2 is a diagram showing a chromatogram when a solvent is flowed in the direction of the arrow under the above conditions from the original line showing the point where the material is attached. Here, 1 and 9 indicate points in which a chromatogram was performed using a sample of a mixture of salicyl uriquaside a and salicylic acid b for comparison. 2 and 8 indicate the use of urine samples taken from rats treated with the Cu[HSal] ₂ C ₂ H ₅ OH composition. 3 and 7 indicate points using urine samples taken from control untreated rats. 4 and 6 indicate points in which chromatograms were performed using a sample of a mixture of gentisic acid c and ethyl salicylate b for comparison. 5 is Cu[HSal] ₂ C ₂ H ₅ OH used for rat treatment.
The points are shown using samples of the composition. This chromatogram clearly shows the presence of salicylic acid and gentisic acid in the urine of rats treated with Cu[HSal] ₂ C ₂ H ₅ OH, which indicates that the composition does not affect the rat skin. Indicates that it has penetrated. Example 3 In this example, inflammation was artificially induced in rats, and Cu[HSal] ₂ C ₂ H ₅ OH was used to alleviate this symptom.
This figure shows the effects of topical application of the compound. Male Wistar rats (average weight 250 g) were anesthetized with diethyl ether, their spines shaved over an area of 20 cm ² , and the rats were divided into 7 groups. Group 1 is control rats, untreated. The second group of rats was treated with a 4:1 mixture of alcohol and glycerol, the adjuvant used for the compound Cu[HSal] ₂ C ₂ H ₅ OH. Group 3 of rats received 7% salicylic acid dissolved in a 4:1 mixture of alcohol and glycerol.
Treated with W/V solution. Rats in groups 4, 5, 6, and 7 received 154 mg of Cu[HSal] ₂ C 2 H ₅ OH produced by the method of Example 1, and 231 mg of Cu[HSal] 2 C ₂ H 5 OH.
Cu containing mg, 462mg and 770mg respectively
[HSal] ₂ C ₂ H ₅ OH composition was used for treatment. Approximately 6 hours after topical treatment of the rats, 1% carrageenan sodium salt was applied to each rat's paw pad.
Inflammation was induced in the paws of rats in each group by injecting 0.1 ml of physiological saline solution. Inflammation in the paws of rats in each group was observed by measuring paw swelling using a micrometer, screw, and gauge. The results are shown in Figure 2. In Figure 2, the vertical axis is
The increase in rat paw thickness (mm x 10) is shown, and the horizontal axis is the inflammatory agent administration (1% carrageenan sodium
represents the time (h) after 0.1 ml). Each curve shows the increase in rat paw thickness versus time after injection of the inflammatory agent. Curves 1 to 7 show the paw swelling of rats from groups 1 to 7, respectively. Figure 2 clearly shows that the Cu[HSal] ₂ C ₂ H ₅ OH compound has an anti-inflammatory effect when applied topically. Reference example 3 Production method of Cu[C ₆ H ₄ (SH)COO] ₂・C ₂ H ₅ OH. The present compound was produced by the method of Reference Example 1, except that 7.7 g of 2-thiobenzoic acid was used in place of salicylic acid. Yield 2.9g (elemental analysis, actual value Cu15.6; C45.9;
H3.6%, calculated value of CuC ₁₆ H ₁₆ O ₅ S ₂ is Cu15.4;
C46.2; H3.9%) Reference example 4 Production method of Cu[C ₆ H ₄ (SeH)COO] ₂・C ₂ H ₅ OH. The present compound was produced by the method of Reference Example 1, except that 10.0 g of 2-selenibenzoic acid was used in place of salicylic acid. Yield 8.1g (elemental analysis, actual value Cu12.3; C38.1;
H3.5%, calculated value of CuC ₁₆ H ₁₆ O ₅ Se ₂ is Cu12.6;
C37.7; H3.1%) Reference Example 5 Production method of Cu[C ₆ H ₄ (NH ₂ )COO] ₂・C ₂ H ₅ OH. The present compound was produced in the same manner as in Reference Example 1, except that 6.9 g of 2-amino benzoitic acid was used in place of salicylic acid. Yield 5.6g (elemental analysis actual value Cu16.4; C50.6;
H5.0%, calculated value of CuC ₁₆ H ₁₈ O ₅ N ₂ Cu16.8;
C50.3; H4.7%) Reference example 6 Cu [C ₆ H ₄・CO ₂・NH・C ₄ N ₂ H ₂・NH・
Production method of C ₇ H ₄ O ₂ ]・C ₂ H ₅ OH. Except that 16.8 g of 2-[2-carboxyphenyl]amino-4-[2-carboxyphenyl]amino pyrimidine was used instead of salicylic acid.
This compound was produced by the method of Reference Example 1. Yield 13.3g (elemental analysis actual value Cu24.5;
C46.2; H3.2%, the calculated value of Cu ₂ C ₂₀ H ₁₈ O ₅ N ₄ is
Cu24.5; C46.0; H3.5%) Reference example 7 Cu ₂ [C ₆ H ₄・CO ₂・NH・C ₄ N ₂ H ₂・NH・
Production method of C ₈ H ₄ O ₄ ]・C ₂ H ₅ OH. This compound was prepared by the method of Reference Example 1, except that 19.0 g of 2-[2-carboxyphenyl]amino-4-[2,3-dicarboxyphenyl]amino-pyrimidine was used in place of salicylic acid. Manufactured. Yield 14.0g (elemental analysis, actual value Cu22.4;
C44.7; H3.3%, the calculated value of C ₂ Cu ₂₁ H ₁₈ O ₇ N ₄ is
Cu22.6; C44.5; H3.2%) Reference example 8 Cu ₂ [C ₆ H ₄・CO ₂・NH・C ₄ N ₂ H ₂・NNH・
C ₈ H ₄₄ ]・C ₂ H ₅ OH manufacturing method. The present compound was produced by the method of Reference Example 1, except that 19.0 g of 2-[2-carboxyphenyl]amino-4-[2,4-dicarboxyphenyl]amino pyrimidine was used in place of salicylic acid. . Yield 15.3g (elemental analysis, actual value Cu24.7;
C45.8; H3.7%, Cu ₂ C ₂₁ H ₁₈ O ₇ N ₄ , the calculated value is
Cu24.5; C46.0; H3.5%) Reference example 9 Production method of Cu[C ₆ H ₄・CO ₂・C ₄ N ₂ H ₂ Cl] ₂・C ₂ H ₅ OH. The present compound was produced by the method of Reference Example 1, except that 11.7 g of 4-[2-carboxyphenyl]amino-2-chloropyrimidine was used in place of salicylic acid. Yield 9.5g (elemental analysis, actual value Cu12.5; C56.5;
H3.9%, Cu12.7 as CuC ₂₄ H ₁₈ O ₅ N ₄ ; C56.9;
H3.6%) Reference example 10 Production method of Cu [C ₆ H ₄ , CO ₂ , NH, C ₈ H ₉ ] ₂ C ₂ H ₅ OH. The present compound was produced by the method of Reference Example 1, except that 6.8 g of 2-[2,3-dimethylphenyl]aminobenzoic acid was used in place of salicylic acid. Yield 5.5g (elemental analysis, actual value Cu11.2; C64.9;
H5.5%, calculated value of CuC ₃₂ H ₃₄ O ₅ N ₂ Cu10.9;
C65.1; H5.8%)

[Brief explanation of drawings]

FIG. 1 shows thin layer chromatograms of urine from rats treated topically with a compound of the invention and from untreated rats and related samples. FIG. 2 is a graph showing paw swelling curves when carrageenan sodium salt is injected into the paws of rats to which the compound of the present invention has been applied topically and of untreated rats to induce inflammation.

Claims (1)

[Claims] 1 General formula Cu[C ₆ H ₄ (X)COO] ₂ ROH (However, ROH represents an alkanol, and X may be located at any position, including OH, SH, SeH,
NH ₂ or NHR′, R′ is An anti-inflammatory composition for topical use, which contains a neutral copper binder represented by the formula or represented by the formula and a pharmaceutically acceptable non-aqueous carrier. 2. The anti-inflammatory composition of claim 1 for topical application in the form of an anhydrous gel, ointment, paste, cream or lotion. 3 General formula Cu[C ₆ H ₄ (X)COO] ₂ ROH (However, ROH represents an alkanol, and X may be in any position, OH, SH, SeH,
NH ₂ or NHR′, R′ is 2. The anti-inflammatory composition according to claim 1, which is in the form of a solution and comprises a neutral copper conjugate represented by [Formula] or [Formula], an excess molar amount of salicylic acid, and ethanol. 4. The anti-inflammatory composition according to claim 1, wherein the nonaqueous carrier is an alkanol, glycerol, or salicylic acid. 5. Claim 1, wherein the neutral copper binder as an active ingredient is blended at 0.1 to 15% W/V.
The anti-inflammatory composition according to any one of items 1 to 4.