JPH0959151A - Nf-kappa b activation suppressing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an NF-κ B activation suppressing agent useful as an antiviral agent and an immunosuppressive agent. SOLUTION: This suppressing agent contains one or more kinds of agents selected from (a) a gallic acid ester of formula I (R is a hydrolyl-substitutable 1-24C alkyl or alkenyl) (e.g. methyl gallate), (b) tannins having galloyl group (e.g. 1,2,3,6-tetragalloylglucose), (c) tannins having hexahydroxydiphenoyl group expressed by formula II (e.g. stenophysnin A) as active ingredients. The gallic acid ester and tannins are directly administered or orally or parentally administered together with a conventional preparation carrier. The daily dose is 0.01-500mg/kg.weight as gallic acid ester or tannins.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an NF-κB activation inhibitor useful as an antiviral agent and an immunosuppressant.

[0002]

2. Description of the Related Art Higher-order life phenomena such as development, differentiation, proliferation, and maintenance of homeostasis are precisely performed according to a specific genetic program, and regulate the expression of specific genes in individual cells. It is regulated by activation, differentiation and proliferation at the cell level. These changes are initiated by the stimulation of cytokines and hormones from outside to cells where the expression of genetic information should occur, binding to receptors on the cell membrane, and ultimately through various biochemical reactions. To cause a change in gene expression. It is known that such control of gene expression is mainly performed at the transcription level of the gene.

[0003] Genes whose expression is induced by external stimuli are in a state where they can be quickly reactivated by the stimuli. Which gene is selectively activated is determined by a special nucleotide sequence existing in the expression control region of the gene and whether or not there is a transcriptional regulatory factor that specifically binds to these cis elements. That is, gene expression occurs when the transcription regulatory factor is quantitatively or qualitatively activated by an external stimulus.

[0004] Among the transcriptional regulators, NF-κB is p50
And a protein consisting of two subunits, p65 and p65, and binds to the inhibitory protein I-κB during non-stimulation and is present in the cytoplasm. When a stimulus such as IL-1 or TNF-α is applied to the cytoplasm, I-κB is inactivated by the kinase activated by the stimulus, and the released NF-κB translocates to the nucleus and is transcriptionally transcribed. Activation is believed to occur.

Genes which are activated by NF-κB, that is, which have a cis element to which NF-κB binds to its expression control sequence, are IL-1, IL-6, IL-8 and IFN.
It is known that many of those involved in immune reaction, such as -β, ELAM-1, and ICAM-1, are involved. By suppressing the activation of NF-κB, it is possible to suppress the expression induction of these cytokines and adhesion molecule genes. Therefore, the NF-κB activation inhibitor is a disease accompanied by the activation of NF-κB. , Especially as a therapeutic agent for various autoimmune diseases,
Alternatively, it is considered to be effective as an immunosuppressant. In addition, human immunodeficiency virus (HIV), human T-cell leukemia virus (HTLV-I), cytomegalovirus (CMV), adenovirus and the like are NF-κ of host cells.
It is believed that B activates its transcription, and promotes the growth of virus and spread of infection. Therefore, the NF-κB activation inhibitor is expected to be effective in the treatment of viral infections such as HIV, HTLV-I, CMV, and adenovirus (Experimental Medicine: Vol. 9, No. 16, p68-, 1
991, Annu. Rev. Immunol ,: 12
Vol., P141-, 1994, Advances in
Immunology: 58, 1).

N-acetyl cysteine and pyrrolidine dithiocarbamate (The Journal of Immunol) are substances that suppress the activation of NF-κB.
, 1994, 153: 2681), aspirin and sodium salicylate (Science, 1994, 2).
65 (12): p956 ~), but it is not sufficient in terms of efficacy, and no effective substance has been found yet.

[0007]

Therefore, an object of the present invention is to provide an NF-κB activation inhibitor having high potency, and an excellent antiviral agent or immunosuppressant based on this action.

[0008]

[Means for Solving the Problems] In view of such a situation,
The present inventor has conducted diligent research, and as a result, gallic acid ester,
NF-κB excellent in tannins having a galloyl group or tannins having a hexahydroxydiphenoyl group
The present invention has been completed by finding that it has an activation suppressing action.

That is, the present invention provides (a) the following general formula (1):

[0010]

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A gallic acid ester represented by the formula [wherein R represents a linear or branched alkyl or alkenyl group having 1 to 24 carbon atoms which may be substituted by a hydroxy group]; (b) galloyl Group-containing tannins; and (c) Formula (2)

[0012]

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There is provided an NF-κB activation inhibitor, an antiviral agent and an immunosuppressant containing, as an active ingredient, one or more compounds selected from tannins having a hexahydroxydiphenoyl group represented by: To do.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Here, it is known that (a) gallic acid ester has an antioxidant action, a platelet aggregation inhibiting action, a whitening action, a fibrosis inhibiting action, a collagenase inhibiting action, a peroxidase inhibiting action and the like. Although it is
Nothing is known about the effect of suppressing NF-κB activation. In addition, (b) a tannin having a galloyl group or (c) a tannin having a hexahydroxydiphenoyl group (2) has a protein aggregation action, a hemostatic action, an antioxidant action, a Staphylococcus aureus, a Pseudomonas aeruginosa and the like. It is known that some have bactericidal action against bacteria, but N
Nothing is known about the inhibitory effect on F-κB activation.

In the gallic acid ester (a) represented by the general formula (1), R represents a linear or branched alkyl group or alkenyl group having 1 to 24 carbon atoms which may be substituted with a hydroxy group. However, an alkyl group which may have a hydroxy group having 1 to 10 carbon atoms is particularly preferable. Specific examples of R include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, t-butyl group, n-pentyl group, isopentyl group, neopentyl group, t -Pentyl group, heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, nonadecyl group,
Isohexyl group, 2-ethylhexyl group, propenyl group, butenyl group, pentenyl group, oleyl group, linoleinyl group, 3,5,5-trimethylhexyl group, 3,7-
Dimethyloctyl group, 3,7,11-trimethyldodecyl group, 3-methyl-2-butenyl group, 3,7-dimethyl-2,6-octadienyl group, cyclohexylmethyl group, cyclohexylethyl group, 2-hydroxypropyl group, Examples thereof include 8-hydroxyoctyl group, 10-hydroxydecyl group, 11-hydroxyundecyl group, 12-hydroxydecyl group, and 12-hydroxyoctadecyl group. In the present invention, a linear or branched alkyl group is preferable among them. That is, as gallic acid ester (a), methyl gallate, ethyl gallate,
N-propyl gallate, isopropyl gallate, butyl gallate, octyl gallate and the like are preferable. These compounds can be synthesized by a known method.

The tannins (b) having a galloyl group include gallotannin (galloyl tannin), that is, tannins that produce gallic acid as a constituent acid by hydrolysis, and a galloyl group and a hexahydroxydiphenoyl group. Tannins can be mentioned, and more specifically, glucose, hammamellose, xylose, methyl glucoside, salidrociode, 1,5-anhydroglutitol, protoquercitol, quinic acid, and shikimic acid have different hydroxyl groups to various extents. And the like, and various mixtures thereof called tannic acid. Specific examples of the gallotannin include 1,2,3,6-tetragalloylglucose, 1,2,3,4,6-pentagaloylglucose, 3,4-digaloylshikimic acid, and hammamelitannin. . Other examples of tannin having a galloyl group include assamicine A and assamicine B.

The tannins (c) having a hexahydroxydiphenoyl group (2) include erroditannin,
That is, a tannin having a hexahydroxydiphenoyl group obtained by oxidatively condensing two galloyl groups can be mentioned.
Specific examples include stenofinin A, punicalagin, and sangiin H-11. Examples of tannins having a galloyl group and a hexahydroxydiphenoyl group include eugenin, punicacortein A, punigluconin, stenophylanine A and the like.

Since all of these various tannins are contained in plants such as gallic and quintile, they can be extracted and isolated according to a conventional method, and in addition, hydroxyl groups such as glucose, xylose and shikimic acid are normally extracted. It can be obtained by appropriately gallyating by the method.

Although the gallic acid ester and the tannins having a galloyl group or hexahydroxydiphenoyl group in the molecule as described above are effective as NF-κB activation inhibitors, the straight chain alkyl ester of gallic acid is used. Is particularly effective. Further, (a) gallic acid ester, (b) tannins having a galloyl group, and (c) tannins having a hexahydroxydiphenoyl group are useful as antiviral agents and immunosuppressants based on the suppression of NF-κB activation. Is.

The gallic acid esters and tannins described above can be administered to animals and humans either by oral route or parenteral route for the treatment of the above-mentioned diseases as they are or together with conventional pharmaceutical carriers. This dose may be appropriately determined according to the age of the patient or animal, the degree of disease, etc.
Usually 0.01 to 500 mg / kg body weight as gallic acid ester or tannin per day, preferably 0.5 to 50
It can be in the range of mg / kg body weight.

The formulation of the present invention is usually used as a composition containing an excipient and other additives usually used in the formulation. As examples of these, solid ones include lactose, kaolin, sucrose, crystalline cellulose, corn starch, talc, agar, pectin, stearic acid, magnesium stearate, lecithin, sodium chloride, and the like, and liquid ones. Is glycerin, peanut oil, polyvinylpyrrolidone, olive oil, ethanol,
Benzyl alcohol, propylene glycol, water and the like can be mentioned.

The dosage form may be any form, for example, solid preparations such as tablets, powders, granules, capsules, suppositories, troches; syrups, emulsions, soft gelatin capsules, creams, lotions, gels. , Liquid preparations such as paste, spray, ointment and injection.

[0023]

【The invention's effect】

The (a) gallic acid ester, (b) tannins having a galloyl group, and (c) tannins having a hexahydroxydiphenoyl group have an excellent NF-κB activation inhibitory action and are also highly safe. Therefore, it is useful as an antiviral agent and an immunosuppressive agent.

[0024]

Next, the present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.

Example 1 NF-κB activation inhibition test: Confluent vascular endothelial cells were treated with the test substances (10 μM) shown in Table 1 for 15 hours, and then IL-1α (2.5 ng / ml: final concentration) )
I was stimulated by. After 1 hour, the cells were collected and nuclear proteins were extracted according to a conventional method. The obtained nuclear protein is Poly (dI
-DC) in the presence of a DNA probe (NF-κB Consensus Oligonucl) labeled with ³² P in advance.
After the reaction was stopped, the mixture was subjected to polyacrylamide gel electrophoresis, and after completion of autoradiography, the effect of suppressing NF-κB activation was evaluated based on the change in the mobility of the DNA probe. Evaluation is based on the bioimaging analyzer BAS2000.
(Manufactured by Fujifilm Corporation), the radioactivity of each band was measured, and each test was carried out from the radioactivity value of NF-κB when IL-1 was not stimulated and the radioactivity value when only IL-1 was stimulated. It was performed by calculating the degree of NF-κB activation when treated with the substance as the inhibition rate. Table 1 shows the results.

[0026]

[Table 1]

As a result, it was found that gallic acid ester, tannins having a galloyl group and tannins having a hexahydroxydiphenoyl group have an excellent NF-κB activation inhibitory action.

Example 2 1000 g of ethyl gallate, 800 g of hydroxypropyl cellulose, 200 g of light anhydrous silicic acid, 500 lactose
g, crystalline cellulose 500 g, and talc 500 g were formed into tablets having a diameter of 9 mm and a weight of 200 mg by a conventional method.

Example 3 Propyl gallate 1000 g, crystalline cellulose 1000
g, lactose 1500 g, and light anhydrous silicic acid 200 g were made into capsules by a conventional method.

Example 4 200 g of propyl gallate, 200 g of lactose, 300 g of hydroxypropyl cellulose and 15 g of talc were made into granules by a conventional method.

Example 5 0.2 g of 1,2,3,6-tetragalloyl glucose,
Cholesterol 0.5 g, cholesteryl isostearate 1 g, polyether modified silicone 1.5 g, cyclic silicone 20 g, methylphenyl polysiloxane 2 g,
Methyl polysiloxane 2g, magnesium sulfate 0.5
g, vitamin C 0.2 g, 55% ethanol 5 g, carboxymethyl chitin 0.5 g, and purified water (remaining amount) were mixed to give a cream.

Example 6 0.5 g of 1,2,3,6-tetragalloyl glucose,
Cholesteryl isostearate 3g, liquid paraffin 1
An ointment was prepared by mixing 0 g, α-tocopherol 0.1 g, glyceryl ether 1 g, glycerin 10 g, and white petrolatum (residual amount).

Example 7 15 mg of octyl gallate and 100 mg of glucose were dissolved in 5 ml of distilled water for injection and sterilized by heating to obtain an injection.

Example 8 1,2,3,4,6-pentagaroyl glucose 15 m
g, 15 mg of octyl gallate and 100 mg of glucose were dissolved in 5 ml of distilled water for injection and sterilized by heating to obtain an injection.

Example 9 1 g of 1,2,3,6-tetragalloyl glucose, 1 g of hamamelitannin, 1 g of glycerin monostearate,
Ethanol 15 g, propylene glycol 4 g, isopropyl palmitate 3 g, lanolin 1 g, ceramide 0.
5 g, methyl paraoxybenzoate 0.1 g, vitamin C
A lotion was obtained by mixing 0.5 g, a fragrance, a small amount of dye and 72 g of purified water.

Claims (3)

[Claims] 1. (a) The following general formula (1): [In the formula, R represents a linear or branched alkyl or alkenyl group having 1 to 24 carbon atoms which may be substituted with a hydroxy group], a gallic acid ester; (b) having a galloyl group Tannins; and (c) formula (2) An NF-κB activation inhibitor containing, as an active ingredient, one or more compounds selected from tannins having a hexahydroxydiphenoyl group represented by: 2. An antiviral agent comprising the compound according to claim 1 as an active ingredient. 3. An immunosuppressive agent comprising the tannins having a galloyl group (b) according to claim 1 and / or the tannins having a (c) hexahydroxydiphenoyl group as an active ingredient.