JP2003055202A - Prophylactic and therapeutic agent against inflammatory intestinal disease - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an excellent prophylactic/therapeutic agent against inflammatory intestinal diseases. SOLUTION: This prophylactic/therapeutic agent for inflammatory diseases associated with transcription factor NF-κB or inflammatory cytokinin is characterized by comprising curcumin or its derivative as an active ingredient.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a prophylactic / therapeutic agent for inflammatory bowel disease. More specifically, it relates to a prophylactic / therapeutic agent for inflammatory bowel disease involving a transcription factor NF-κB or an inflammatory cytokine, characterized by containing curcumin or a derivative thereof as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION Inflammatory bowel disease is a general term for intestinal diseases accompanied by inflammation, and its main diseases include Crohn's disease and ulcerative colitis.

[0003] Crohn's disease has an unknown cause, and it frequently occurs in young people in their teens and twenties. It consists of a granulomatous inflammatory disease accompanied by fibrosis and ulcer, and can occur in any part of the digestive tract. The clinical symptoms of Crohn's disease include abdominal pain, general malaise, diarrhea, melena, occult blood, fever, weight loss, anemia, ileus, abdominal mass, nausea, vomiting, and peritoneal inflammation. In addition, systemic complications such as nutritional disorders, arthritis, iritis and liver disorders can occur.

Ulcerative colitis is a diffuse nonspecific inflammation of unknown origin, often affecting the mucous membrane, which often forms erosions and ulcers, with lesions occurring in the submucosa. The clinical symptoms of ulcerative colitis include mucous stools, abdominal pain, bloody diarrhea, anemia, fever, loss of appetite, nausea and vomiting.

Glucocorticoids, salazosulfapyridine and 5-aminosalicylic acid have been used as drug treatments for inflammatory bowel disease. However, glucocorticoids have many side effects such as impaired glucose tolerance, decreased adrenal function, and infectious diseases, and are difficult to use in the long term. In addition, there are many patients with hypersensitivity to salazosulfapyridine and 5-aminosalicylic acid.

The cause of inflammatory bowel disease is still unknown, but in recent years, ICAM-1 (Intercellula
It is considered that cell adhesion molecules such as r Adhesion Molecule-1), chemokines such as interleukin-8, and inflammatory cytokines such as tumor necrosis factor (TNF-α) and interleukin-12 are involved. . And
It is the transcription factor NF-κB (Nuclear Factor κB) that triggers the production of these substances.
Therefore, it is considered possible to suppress the disease state of inflammatory bowel disease by suppressing the action of the transcription factor NF-κB.

In fact, the p65 protein of the transcription factor NF-κB is increased in lesional biopsy tissues of patients with inflammatory bowel disease. In addition, although safety has not been established and it has not yet been clinically applied, in experimental animals, NF-κB
The effect of antisense oligonucleotides on is demonstrated.

Regarding the transcription factor NF-κB, Ba
In 1986, ltimore et al.
NF-κB (nuclear factor for) as a transcription factor that binds to enhancers involved in the expression of ght chain) genes.
κ chain gene in B cells) was identified. Transcription factor N
F-κB is a transcription factor distributed in many cells, and it is considered that F-κB translocates to the cell nucleus and promotes expression of various genes with various stimuli. Currently, transcription factor NF-
κB has been shown to be a heterodimer of p50 and p65.

The inventors of the present invention searched for a substance that suppresses the activation of the transcription factor NF-κB and has little toxicity and can be used safely. Among them, the yellow pigment component contained in the plant rhizome of turmeric which is a medicinal plant. I focused on curcumin.

Curcumin has been conventionally known as a substance having an antioxidant effect.

Curcumin is disclosed in JP-A-5-26265.
No. 9 discloses that it is effective for inflammation of intestinal mucosa (ulcerative colitis and Crohn's disease) by simultaneously suppressing the production of leukotrienes and prostaglandins, which are inflammation-promoting mediators generally called eicosanoids. There is. Eicosanoid has 3 unsaturated bonds.
It is constantly synthesized in the cell membrane from fatty acids having 20 or more carbon atoms and is mainly synthesized from arachidonic acid.
Because eicosanoids play important roles in pain, fever, and inflammation, their synthetic pathways are the targets of many therapeutic agents. At the time of filing the patent application pertaining to the same patent publication, leukotriene and other eicosanoids are used for the onset of inflammatory bowel disease because neutrophils infiltrate locally in inflammation and leukotriene is the most potent neutrophil chemoattractant. Is believed to be involved, and many eicosanoid inhibitors have been expected as therapeutic agents. However, in clinical trials, none of the expected therapeutic effects have been observed, and currently, factors other than the aforementioned eicosanoids are being focused on.

The test examples and examples described in JP-A-5-262659 mentioned above describe formulation formulations such as oral administration formulations containing curcumin, but in animal models of inflammatory bowel disease, Data confirming the effect are not described, and there is no description or suggestion that curcumin is effective for inflammatory bowel disease by suppressing activation of the transcription factor NF-κB or production of inflammatory cytokines in the present invention. Absent.

[0013]

Under the background of the prior art described in the preceding paragraph, the purpose of the present invention is to prevent inflammatory bowel disease from being excellent.
To provide a therapeutic agent.

[0014]

The present inventors have surprisingly found that curcumin, as described below, is not the inhibitory effect of eicosanoids described in JP-A-5-262659, but the transcription factor NF. -ΚB activation, tumor necrosis factor (TNF-α) and interleukin-12
It was found that the production of inflammatory cytokines such as the above was suppressed, and the present invention was completed based on such findings.

That is, the present invention relates to a prophylactic / therapeutic agent for inflammatory bowel disease involving the transcription factor NF-κB or inflammatory cytokine, which contains curcumin or a derivative thereof as an active ingredient.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.

Derivatives of curcumin, which is an active ingredient of the preventive / therapeutic agent of the present invention, include tetrahydrocurcumin and demethoxycurcumin.
thoxycurcumin), bis-demethoxycurcumin (bis-dem
ethoxycurcumin), diacetylcurcumin
rcumin), methylcurcumin (metylcurcumin, anisylcurcumin), etc., and any of them can be preferably used in the present invention.Curcumin is orally administered to guinea pigs and rats in an amount of 5 g / kg. It is said that there is almost no toxic effect (the above-mentioned published patent publication) .In addition, 0.5 g, 1 g, 2 g, 4 g or 8 g per day is clinically orally administered to various cancer patients for a long period of time. There was a report that it was safe (Am
erican Society of Clinical Oncology, 1998 Annual M
eeting, Abstract 2140).

The dose of curcumin or its derivative, which is the active ingredient of the prophylactic / therapeutic agent for inflammatory bowel disease of the present invention, is
Patient (body weight 70 kg) usually 0.5 to 8 g per day,
It is preferably 0.5 g to 5 g, more preferably 1 to 4 g. By the way, in the case of the turmeric plant preparation and the drug disclosed in the above-mentioned published patent publication, the maximum daily dose is estimated to be 180 mg, assuming that it is to be administered once in two capsules and three times a day. The therapeutic / prophylactic agent of the present invention is as described above. The reason why such a large difference occurs in the dose is that in the present invention, the clinical dose was estimated based on the therapeutic effect in the animal model and the NF-κB activation inhibitory effect. It is considered that the curcumin content in the drug product was determined based on the in vitro eicosanoid production inhibitory activity without confirming the effect in the animal model.

Administration can be oral or enema. Oral dosage forms include solid or liquid dosage forms such as granules, powders, coated tablets, capsules,
Examples thereof include syrups. Preferably, a core in which curcumin or a derivative thereof and various additives such as a pharmaceutically acceptable excipient is uniformly dispersed is prepared, and the core is a pharmaceutically acceptable film-forming water-insoluble or water-soluble. Enteric delivery formulations coated with one or more polymers selected from active polymers can be prepared. More preferably, since curcumin is water-insoluble, the intestinal delivery preparation can be produced after being subjected to a hydrophilization treatment or the like for improving its gastrointestinal absorption.
The polymeric material can be selected from polyvinylpyrrolidone, methylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, ethylcellulose, cellulose acetate, polyvinylacetate, ethylene-vinylacetate copolymers and copolymers of acrylic acid and methacrylic acid esters.

If desired, curcumin or a derivative thereof may be other pharmaceutically active substance such as anti-adhesion molecule therapeutic agent, anti-free radical therapeutic agent, anti-eicosanoid therapeutic agent, anti-allergic therapeutic agent, immunosuppressive agent. , An immunoglobulin, a sulfated glycosaminoglycan, a precursor of an amino acid such as n-acetylcysteine, a steroid drug, an aminosalicylic acid drug, and the like can also be co-formulated in the production of the dosage form. Preferably glutamine,
At least one selected from salazosulfapyridine, 5-aminosalicylic acid, and glucocorticosteroid
Can be formulated and manufactured with seeds.

Further, curcumin or its derivative is
Proteins isolated from germinated seeds of gramineous plants, insoluble dietary fiber, yeast, vitamins and cysteine, glutamine, amino acids such as glutamic acid, oligopeptide mixture, non-reducing sugars such as trehalose, organic acids such as butyric acid It can be used as a food for patients with inflammatory bowel disease by mixing with one or two or more of them. Furthermore, it can be administered with components that improve the bioavailability of curcumin or its derivatives, such as piperine.

[0022]

EXAMPLES The present invention will be described in more detail with reference to experimental examples.

Experimental Example 1 7- to 8-week-old C57BL / 6 mice (weighing 21 to 24)
g) was fasted overnight and dissolved in 50% ethanol 2.5
mg 2,4,6-trinitrobenzene sulfonic acid (TNB
100 μl of S) was enema to create enteritis. TNBS enteritis persists 2-3 weeks after enema, pathological findings may include ulceration in all layers, and sometimes granulomas are observed, which is considered to be a condition similar to Crohn's disease (Gastroenterology 198).
9; 75: 795). As a control, a group (ET enema group) was prepared in which mice were enema with 50% ethanol instead of TNBS. In the TNBS enema group, mice were fed with a diet containing 0.5% or 2% of curcumin immediately after the enema (treatment group) or a diet containing no curcumin (untreated group). In addition, a group was also set up in which a curcumin-free feed was fed after TNBS enema and administration of 2% curcumin was started from 2 days later. Daily intake of curcumin per body weight is 1g / kg / day in the 0.5% group, and 2%
This was equivalent to 4 g / kg / day in the group.

The mice were sacrificed one week after the enema, and the therapeutic effect was evaluated by determining the weight change of the mice, histological examination of the large intestine, and examination of NF-κB activity by immunohistochemical staining.
We also examined the production of cytokines in the colonic mucosa. The results are as follows.

Weight Change of Mice As shown in FIG. 1, in the control ET enema group, the body weight decreased from overnight fasting to weight recovery after enema, but in the non-treated group, the body weight remained 3 days after enema. The decrease continued. On the other hand, the curcumin-administered group recovered weight faster than the untreated group,
Especially in the 2% treatment group, the body weight after 1 week was ET of the control.
It became almost the same as the enema group.

Further, as shown in FIG. 2, the body weight recovery was earlier in the group to which curcumin was administered 2 days after the TNBS enema than in the non-treatment group.

Histological examination of large intestine The degree of inflammation was classified according to the degree of lymphocyte infiltration and the degree of gland duct destruction by HE staining and classified according to the table shown in Table 1 below.
Five-level evaluation of 0 to 4 was performed (Ann Allergy 1994; 72: 13.
Five).

[0028]

[Table 1]

[Table 1]

As shown in FIG. 3, the higher the curcumin content was 0.5% and 2%, the lower the histological score was.

Examination of NF-κB activity by immunohistochemical staining NF-κB is bound to the inhibitory protein IκB in non-inflammation and is present in the cytoplasm, but when activated, it separates from IκB and enters the nucleus. Migrate and participate in the production of various inflammatory cytokines. When a frozen section of mouse large intestine was prepared and immunohistochemically stained for NF-κB (p65), the nucleus was stained in TNBS enteritis (untreated group), and NF-κB was activated. Was considered to be. However, in the 2% curcumin treatment group, a nuclear staining image was not observed, and it was considered that NF-κB activation was suppressed.

Expression of cytokine-specific mRNA in colonic mucosa The expression of cytokine-specific mRNA in colonic mucosa of mice was examined. See FIG. 4 below. TNBS enteritis (untreated group)
IFN-γ, TNF-α, interleukin 12 (IL12), interleukin 6
Although the expression of inflammatory cytokines such as (IL6) was enhanced, the expression of these cytokines was suppressed in the 2% curcumin treatment group. The expression of muscle protein β-actin not associated with inflammation and interleukin 4 (IL4) involved in humoral immunity did not differ between the three groups.

From the above experimental results, curcumin was found to be TNB.
Suppresses the activation of transcription factor NF-κB in S enteritis,
It was confirmed that suppressing the production of inflammatory cytokines improves enteritis.

[0033]

INDUSTRIAL APPLICABILITY According to the present invention, prevention of inflammatory bowel disease involving transcription factor NF-κB and inflammatory cytokine
Now, an excellent therapeutic agent effective for treatment can be provided.

[Brief description of drawings]

FIG. 1 shows changes in body weight of mice (Experimental Example 1).

FIG. 2 shows changes in body weight of mice (Experimental Example 1).

FIG. 3 shows the results of histological examination of the large intestine (Experimental Example 1).

FIG. 4 shows the results of examination on expression of cytokine-specific mRNA in colonic mucosa (Experimental Example 1).

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 29/00 A61P 29/00 (72) Inventor Hiroyuki Hanai 1-20-3 Izumi, Hamamatsu City, Shizuoka Prefecture (72) ) Inventor Yukio Koide 685-25 F-term, Hamamatsu City, Shizuoka Prefecture F-term (reference) 4C086 AA01 AA02 BA13 BC21 GA02 GA12 NA14 ZA66 ZA68 ZB11 4C206 AA01 AA02 CB18 FA44 NA14 ZA66 ZA68 ZB11

Claims (4)

[Claims] 1. Prevention of inflammatory bowel disease involving the transcription factor NF-κB or inflammatory cytokine characterized by containing curcumin or a derivative thereof as an active ingredient.
Therapeutic agent. 2. A patient 1 in terms of curcumin or its derivative.
The prophylactic / therapeutic agent for inflammatory bowel disease according to claim 1, which is to be administered in an amount of 0.5 to 8 g per day. 3. The prophylactic / therapeutic agent for inflammatory bowel disease according to claim 1, which contains glutamine and / or piperine in addition to curcumin or its derivative as an active ingredient. 4. The preventive / therapeutic agent for inflammatory bowel disease according to claim 1, wherein the inflammatory bowel disease is Crohn's disease.