JPH09301862A - Antiinflammatory agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antiinflammatory agent containing a 2-phenylimidazolo-1- yloxy-3-oxide derivative as an active ingredient, having the action effectively reducing the production of IL-8 which is a prophlogistic cytokine and useful as an excellent antiinflammatory agent. SOLUTION: This antiifnlammatory agent comprises a compound of formula I (R<1> to R<4> are each independently H or a 1-4C alkyl; R<5> is amino, a secondary or tertiary amino having 1-4C alkyls or a quaternary ammonium having 1-4C alkyls) and/or its physiologically acceptable salt. 2-(4-(trimethylamino) phenyl)-4,4,5,5-tetramethylimidazolo-1-yloxy-3-oxide of formula II is exemplified as the compound of formula I. The antiinflammatory agent containing an agent reducing the production of IL-8 is preferably wrapped with a membrane having a small spherical shape and containing a phospholipid.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an IL-8 production inhibitor and an anti-inflammatory agent containing the same as an active ingredient, and more specifically to 2-phenyl-imidazolo-1-yloxy-3.
-An IL-8 production inhibitor containing an oxide derivative as an active ingredient and an anti-inflammatory agent containing the same.

[0002]

2. Description of the Related Art It is said that various substances are involved in inflammation, but the details thereof are not known. Generally, it is said that chemical substances such as histamine and proinflammatory cytokines such as interleukins have a complicated influence on each other. Inflammation caused by chemical substances such as histamine can be alleviated by administration of antihistamines and steroids, but at present, it is not so effective against inflammation caused by proinflammatory cytokines represented by IL-8. No cure has been found.

On the other hand, 2-phenyl-4,4,5,5-tetramethyl-imidazolo-1-yloxy-3-oxide (PTIO) and 2- (4-carboxyphenyl)-
P such as 4,4,5,5-tetramethyl-imidazolo-1-yloxy-3-oxide (carboxy PTIO)
TIO and its derivatives (hereinafter, such 2-phenyl-
Compounds having an imidazolo-1-yloxy-3-oxide skeleton are collectively referred to as “PTIOs”) have excellent nitric oxide scavenging ability, and are effective against cardiovascular diseases and inflammations that are said to be involved. It is speculated that it will work effectively. However, because of their high reactivity, these PTIOs immediately react with blood components and lose their nitric oxide scavenging power, resulting in inactivation. Therefore, their usefulness as pharmaceuticals is low. The fact is that it has not been fully demonstrated, and the relationship between PTIOs and the above-mentioned proinflammatory cytokine, IL-8, has not been confirmed at all.

[0004]

The present invention has been made from the above viewpoints, and an IL-8 production inhibitor having an action of effectively suppressing the production of IL-8 and an anti-inflammatory agent containing the same as an active ingredient. The challenge is to provide.

[0005]

[Means for Solving the Problems] As a result of intensive studies to solve the above problems, the present inventors have found that a specific PTIO
Administration of this compound in vivo in an appropriate dosage form, the production of IL-8 in vivo is significantly suppressed, and further, the production of IL-8 is suppressed in this manner. It was found that an excellent anti-inflammatory effect can be obtained by this, and completed the invention.

That is, the present invention is an IL-8 production inhibitor comprising a compound represented by the general formula (1) and / or a physiologically acceptable salt thereof.

[0007]

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(In the formula, R ¹ , R ² , R ³ , and R ⁴ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, R ⁵ is an amino group, and an alkyl group having 1 to 4 carbon atoms. It represents a secondary or tertiary amino group having an alkyl group or a quaternary ammonium group having an alkyl group having 1 to 4 carbon atoms.) Specific examples of the compound represented by the general formula (1) used in the present invention Specifically, 2- (4- represented by the following formula (2)
(Trimethylamino) phenyl) -4,4,5,5-tetramethyl-imidazolo-1-yloxy-3-oxide (hereinafter abbreviated as “trimethylamino PTIO”), represented by the following formula (3): -(4- (dimethylamino) phenyl) -4,4,5,5-tetramethyl-imidazolo-1-yloxy-3-oxide (hereinafter abbreviated as "dimethylamino PTIO") and the like, In the invention preferably trimethylamino PTI
O is used.

[0009]

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[0010]

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In the present invention, it is also possible to use a physiologically acceptable salt thereof in the same manner as the compound represented by the general formula (1). Examples of such salts include mineral acid salts such as hydrochlorides, nitrates and phosphates, and organic acid salts such as citrates, oxalates and tartrates. Among these salts, a hydrochloride is preferably used in the present invention. Further, among the hydrochlorides, those in which R ^{5 in the} above general formula (1) is a quaternary ammonium chloride are more preferably used.

Further, the present invention provides an anti-inflammatory agent containing the above IL-8 production inhibitor as an active ingredient. In the anti-inflammatory agent of the present invention, the proinflammatory cytokine I
PTIOs having an action of suppressing L-8 production are treated with IL
In order to effectively act in vivo as a -8 production inhibitor, for example, the IL-8 is added to a spherical membrane containing phospholipid.
The anti-inflammatory agent contains an IL-8 production inhibitor in the form of microspheres in which the production inhibitor is encapsulated.

When the anti-inflammatory agent of the present invention contains the IL-8 production inhibitor in the form of the microspheres, the spherical membrane containing the phospholipids of the microspheres is preferably composed of only a saturated compound.

Specific examples of the microspheres include liposomes and niosomes having the characteristics of the present invention. In the present invention, liposomes are preferably used.

Such microspheres preferably used when the anti-inflammatory agent of the present invention contains an IL-8 production inhibitor,
It can be produced by an ordinary method using an encapsulating component containing the compound represented by the general formula (1) and / or a physiologically acceptable salt thereof and a spherical membrane component containing a saturated phospholipid.

Further, the anti-inflammatory agent of the present invention is blended with an optional ingredient for formulation so that the IL-8 production inhibitor can act effectively in vivo like the above-mentioned microspheres. It can be produced in the same manner as a usual formulation method.

The dosage form of the anti-inflammatory agent of the present invention is not particularly limited as long as it is a dosage form taken by an ordinary anti-inflammatory agent, but a skin external preparation is preferably used in the anti-inflammatory agent of the present invention.

The IL-8 production inhibitor of the present invention comprises the compound represented by the general formula (1) and / or a physiologically acceptable salt thereof, and is an inflammatory cytokine IL-
It has the effect of effectively suppressing the production of 8.

The anti-inflammatory agent of the present invention containing the above IL-8 production inhibitor as an active ingredient has an excellent anti-inflammatory effect. Furthermore, when an IL-8 production inhibitor is mixed with an anti-inflammatory agent, it is encapsulated in a spherical membrane containing a phospholipid, thereby allowing the IL-8 production inhibitor to act more effectively in vivo. Is possible, in addition,
If the spherical membrane is composed of only a saturated compound, storage stability as a preparation can be ensured. Here, the spherical membrane is composed only of a saturated compound means that the amount of the unsaturated compound that causes an oxidative reaction with the encapsulating PTIOs to substantially inactivate the IL-8 production inhibitory effect is a spherical membrane. Means not to contain.

[0020]

Embodiments of the present invention will be described below. First, the IL-8 production inhibitor of the present invention will be described. (1) IL-8 production inhibitor of the present invention The IL-8 production inhibitor of the present invention is one of the compound represented by the general formula (1) and / or a physiologically acceptable salt thereof.
It consists of two or more species. P other than the compound represented by the general formula (1) and / or its physiologically acceptable salt
For TIOs, it may be difficult to formulate PTIOs as an IL-8 production inhibitor in order to effectively act in vivo. The compound represented by the general formula (1) and / or a physiologically acceptable salt thereof used in the present invention is as described above, but such PTIOs are already known and a method for producing the same. Is also known.

For example, in order to produce the compound represented by the above general formula (1), a dihydroxylamine derivative is reacted with a para-substituted benzaldehyde as shown in the following reaction formula, and the resulting compound is converted to lead dioxide or the like. Oxidation may be performed using the metal oxide as a catalyst.

[0022]

[Chemical 6]

(However, R ¹ , R ² , R ³ , R ⁴ , and R ⁵ have the same meanings as those in the general formula (1).) The physiology of the compound represented by the above general formula (1) For example, a salt which is acceptable is a compound represented by the general formula (1) and an acid such as a mineral acid such as hydrochloric acid, nitric acid, phosphoric acid and an organic acid such as citric acid, oxalic acid and tartaric acid. It is easily obtained by mixing in a polar or non-polar solvent.

The compound represented by the general formula (1) or a physiologically acceptable salt thereof can be obtained by the above-mentioned method, and these are easily purified by a conventional method such as recrystallization or column chromatography. It is possible.

The PTIOs thus obtained have an effect of effectively suppressing the production of IL-8 which is a proinflammatory cytokine, and are used as the IL-8 production inhibitor of the present invention.

(2) Anti-inflammatory Agent of the Present Invention The anti-inflammatory agent of the present invention contains the above IL-8 production inhibitor as an active ingredient. The anti-inflammatory agent of the present invention includes IL-8 described above.
To incorporate a production inhibitor as an active ingredient, the PTI
The compounding method is not particularly limited as long as the IL-8 production inhibitor composed of O-groups can be compounded so that it can effectively act in the living body.
In the present invention, it is preferable to include the IL-8 production inhibitor in the anti-inflammatory agent in the form of microspheres containing the 8 production inhibitor. In this way, IL-comprising PTIOs
When an 8 production inhibitor is mixed with an anti-inflammatory agent, the IL-8 production inhibitor is given reaction specificity by the spherical membrane and is transferred to the affected area without losing the reactivity. By erasing a specific substance, specifically nitric oxide, through, it becomes possible to exert an IL-8 production inhibitory action.

In the microsphere used for the anti-inflammatory agent of the present invention, when the IL-8 production inhibitor is encapsulated in the globular membrane, the IL-8 production inhibitor may be encapsulated alone. However, it may be included as a suitable solution. Also,
It may be included together with other optional components. IL-8 above
When the production inhibitor is encapsulated in a spherical membrane as a solution, water is preferably used as a solvent, and more preferably physiological saline prepared by adding various components to water, a phosphate buffer solution, a phosphate buffer physiological agent. Saline solution or the like is used.

The content of the IL-8 production inhibitor in the microspheres is preferably 1 to 30% by weight, more preferably 5 to 20% by weight, and even more preferably 7 to 17% by weight.
Further, the phospholipid contained in the spherical membrane of the microspheres,
It is preferably a saturated phospholipid. This is because in the anti-inflammatory agent of the present invention, it is preferable that the spherical membrane of the small spheres is composed only of a saturated compound in terms of storage stability of the IL-8 production inhibitor comprising PTIOs in the preparation. by.

Here, a saturated compound is a compound which does not contain an unsaturated carbon-carbon multiple bond in the compound, and an unsaturated compound is a compound which has an unsaturated carbon-carbon multiple bond in the compound. However, in the following description, a phospholipid classified as a saturated compound will be described as a “saturated phospholipid”, and a phospholipid classified as an unsaturated compound will be described as an “unsaturated phospholipid”.

Examples of such saturated phospholipids used in the present invention include hydrogenated lecithin, dimyristoylphosphatidylcholine, dimyristoylphosphatidylinositol, dimyristoylphosphatidic acid, hydrogenated lysolecithin and the like. Further, among these, hydrogenated lecithin, which is inexpensive and easily available, can be mentioned as a preferred saturated phospholipid in the present invention.
In the present invention, one of these saturated phospholipids may be used alone, or two or more thereof may be used in combination.

The preferable content of the phospholipid in the spherical membrane of the small spheres is 10 to 100% by weight, and 30 to 100% by weight is more preferable, when the preferable saturated phospholipid is used as the phospholipid. Preferably 50 to
100% by weight is more preferred.

When the spherical membrane of the small spheres used in the anti-inflammatory agent of the present invention is composed of only a saturated compound for the purpose of storage stability of the preparation, in addition to the saturated phospholipid, any other saturated phospholipid may be used. It is possible to add a saturated compound, and as such an optional component, for example, 1,3-
Examples thereof include polyhydric alcohols such as butanediol and propylene glycol, POE hydrogenated castor oil, and surfactants such as POE stearic acid ester. Components such as these polyhydric alcohols and surfactants have the advantage that the spherical film can be made flexible, but on the other hand, depending on the content, there is the disadvantage that the PTIOs that are included tend to flow out, so the content is It is necessary to make use of these advantages and set so as not to impair the effects of the invention. Such a content is 1 to 2 in the polyhydric alcohol.
0 wt% is preferable, 3 to 15 wt% is more preferable,
A more preferable example is 5 to 10% by weight. Further, in the surfactant, 0.1 to 10% by weight is preferable,
0.5 to 7% by weight is more preferable, and 1 to 5% by weight is further preferable.

The above-mentioned microspheres contained in the anti-inflammatory agent of the present invention include a raw material component of a spherical membrane containing a phospholipid, an IL-8 production inhibitor and other optional components in a conventional thin film dispersion method or reverse evaporation method. It is possible to produce by treating with the method of. However, in consideration of the storage stability of the medicinal component in the preparation, when the components to be contained in the microspheres are all saturated compounds including the phospholipid, the solvent used in the method of the saturated phospholipid to be blended. Since it has low solubility in water, it is preferable to carry out a dispersion treatment with high-intensity ultrasonic waves or the like before the above treatment. Further, such a spherical film is a small sphere composed of only a saturated compound, it is also possible to be produced by a solution method, a solution-extrusion method, or the like,
These methods are preferable production methods in the case of producing small spheres in which the spherical membrane is composed of only a saturated compound. The method for producing such small spheres will be described more specifically below by taking small spheres whose spherical film is composed of only a saturated compound as an example.

Saturated phospholipids and other optional components for membrane formation and an organic solvent were sonicated with a sonicator, and PTIOs and other optional components contained therein were dissolved in a solvent such as phosphate buffered saline. Add solution and sonicate. The obtained dispersion is subjected to an evaporator to slowly remove the organic solvent under reduced pressure to obtain small spheres. After adding phosphate-buffered saline to the mixture and shaking well, centrifugation is performed to pellet the small spheres. Discard the supernatant and wash the pelleted pellet well. This shaking-centrifugation-washing operation is repeated about 4 to 5 times until the blue-purple color of the PTIO derivative disappears in the supernatant.

In the anti-inflammatory agent of the present invention, the above IL-
As described above, the 8 production inhibitor is preferably mixed in the form of being encapsulated in the microspheres obtained as described above.
The dosage form of the anti-inflammatory agent of the present invention is not particularly limited, and for example, injection agents, powders, granules, tablets, capsules, solutions, external preparations for skin, and the like, which are commonly used, can be used. Although it can be mentioned as a dosage form of the agent, a skin external preparation is preferably used in the anti-inflammatory agent of the present invention.

In order to formulate the anti-inflammatory agent of the present invention as an external preparation for the skin, the above IL-8 production inhibitor, preferably the IL-8 production inhibitor encapsulated in the microspheres, and other usual Optional ingredients used in external preparations for skin, for example, hydrocarbons such as petrolatum and microcrystalline wax, esters such as jojoba oil and gallow, triglycerides such as beef tallow and olive oil, higher alcohols such as cetanol and oleyl alcohol. , Fatty acids such as stearic acid and oleic acid, polyhydric alcohols such as glycerin and 1,3-butanediol, nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, ethanol, carbopol Thickeners, antiseptics, UV absorbers, antioxidants, pigments, powders, etc., in any proportions, and then used for normal skin According to the method of manufacturing a use agent may be a skin external agent.

In addition, when formulating a dosage form other than the external preparation for skin, similarly, in addition to the above IL-8 production inhibitor, an excipient, a binder, a disintegrating agent, a lubricant, a flavoring agent, and an increasing amount. Agents, coating agents, sugar coating agents, emulsifying / solubilizing / dispersing agents, stabilizers, pH adjusting agents, isotonic agents and the like can be used in arbitrary amounts, and any optional ingredients commonly used in pharmaceuticals can be used. The anti-inflammatory agent of the present invention can be obtained by formulating the IL-8 production inhibitor, preferably the IL-8 production inhibitor encapsulated in the microspheres, according to a conventional method.

The dose, administration method, etc. of the anti-inflammatory agent of the present invention are appropriately selected in consideration of the type of disease, symptoms, patient's age, body weight, etc. Then, the amount of the IL-8 production inhibitor (PTIOs) is 1
It is preferable to administer ˜1000 mg once to several times, and for local administration, 1 to 500 mg may be similarly administered once to several times. Examples of the method for administering the injection include intravenous administration, intraarterial administration, intraportal administration, intraperitoneal administration, intramuscular administration, and subcutaneous administration.

[0039]

[Example] 44 mg of hydrogenated lecithin and 3 ml of ether
Was weighed into a test tube, sonicated with a sonicator for 5 minutes, and trimethylamino PTIO was added at a concentration of 2 mM.
10 mM phosphate buffered saline (pH 7.
4) (hereinafter abbreviated as “PBS”) (1 ml) was added, and sonication was further applied for 15 minutes. This was subjected to an evaporator, and ether was slowly distilled off under reduced pressure to obtain small spheres.

0.6 ml of PBS was added to the obtained microspheres, shaken well, and then centrifuged at 14000 G for 15 minutes to pellet the microspheres. The supernatant of the centrifugation was discarded, and the pelleted pellet was washed with PBS. further,
The above-mentioned operations of shaking, centrifugation and washing were repeated 4 to 5 times until the blue-purple PTIO disappeared from the supernatant of the centrifugation.

The electron spin resonance (ESR) of the small spheres obtained in the above example was measured.
The absorption spectrum of the nitric oxide radical peculiar to O was detected. This spectrum has a wide linewidth,
The above-mentioned trimethylamino P is contained in the microspheres obtained in the examples.
It was confirmed that TIO was included. The obtained microspheres were directly used as an anti-inflammatory agent.

<Evaluation of anti-inflammatory agent of the present invention> With respect to the anti-inflammatory agents obtained in the above-mentioned examples, a storage stability test, a safety test and a test for suppressing IL-8 production of PTIOs in the preparation were conducted. went.

(1) Storage stability test The anti-inflammatory agents obtained in the above-mentioned examples were subjected to 1
I left it for a month. When the ESR was measured for one month with time from the start of the test on the microspheres, the ESR measurement result after one month still showed almost the same peak as when the ESR was measured immediately after the production. From these results, in the anti-inflammatory agents obtained in the above-mentioned examples, IL-
8 It was found that PTIOs that act as a production inhibitor can be stably stored for a long period of time.

(2) Safety test The anti-inflammatory agent obtained in the above-mentioned example was used in ICR male mouse 5
100 mg / kg of the amount of trimethylamino PTIO was intraperitoneally administered to a mouse (5-week-old), and life or death was determined 14 days later.

The results showed that all the mice were alive and that the anti-inflammatory agent of the present invention was excellent in safety.

(3) Test for IL-8 production inhibitory action The anti-inflammatory agent obtained in the above-mentioned example was treated with trimethylamino PT.
Human bronchial epithelial cells were cultured in MEM supplemented with 10% fetal bovine serum (FBS) under the condition of presence of 0.1 mM in terms of IO amount or in the absence of the anti-inflammatory agent. . When the amount of IL-8, which is a proinflammatory cytokine, was measured under these conditions, the amount of IL-8 produced was suppressed in the presence of the anti-inflammatory agent of the Example.

[0047]

INDUSTRIAL APPLICABILITY The inhibitor of IL-8 production of the present invention has an action of effectively suppressing the production of IL-8 which is a proinflammatory cytokine. The anti-inflammatory agent of the present invention is the IL-
8 It has an excellent anti-inflammatory effect by containing a production inhibitor.

Claims (7)

[Claims] 1. An IL-8 production inhibitor comprising a compound represented by the general formula (1) and / or a physiologically acceptable salt thereof. Embedded image (Wherein, R ¹ , R ² , R ³ , and R ⁴ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, and R ⁵ represents an amino group or an alkyl group having 1 to 4 carbon atoms. Grade 2 or 3
Represents a quaternary ammonium group having a primary amino group or an alkyl group having 1 to 4 carbon atoms. ) 2. A compound represented by the general formula (1) is a 2- (4- (trimethylamino) phenyl) -4,4,5,5-tetramethyl-imidazolo represented by the following formula (2). The IL-8 production inhibitor according to claim 1, which is -1-yloxy-3-oxide. Embedded image 3. An anti-inflammatory agent containing the IL-8 production inhibitor according to claim 1 or 2 as an active ingredient. 4. The anti-inflammatory agent according to claim 3, which contains the IL-8 production inhibitor in the form of microspheres in which the IL-8 production inhibitor is encapsulated in a spherical membrane containing a phospholipid. 5. The anti-inflammatory agent according to claim 4, wherein the spherical film is composed only of a saturated compound. 6. The anti-inflammatory agent according to claim 4, wherein the microsphere is a liposome. 7. The anti-inflammatory agent according to any one of claims 3 to 6, which is a skin external preparation.