JP2002241292A - Interleukin 15 production promoter, digestive disorder prevention / treatment agent, food and drink - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enhance the production of interleukin-15, to be effective in the prevention and treatment of digestive disorders such as diarrhea due to the side effects of anticancer drugs, and to have a safe and usable daily routine with very few side effects. Get the material. SOLUTION: The genus Biffidobacterium
containing at least one cell selected from lactic acid bacteria containing ium ) as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to interleukin-1
The present invention relates to a 5 (IL-15) production promoter, a prophylactic / therapeutic agent for preventing / treating digestive disorders caused by side effects of an anticancer drug, and a food or drink. More specifically, the present invention relates to a drug having an effect of promoting the production of IL-15 by using a combination of an anticancer drug and a microbial cell such as a lactic acid bacterium to prevent or ameliorate diarrhea caused by side effects of the anticancer drug.

[0002]

2. Description of the Related Art In the treatment with an anticancer drug, there is a concern about the problem of side effects of the anticancer drug used constantly. The side effects of these anticancer agents are diverse, and for example, anticancer agents such as glucarone, mitobronitol, tamoxifen citrate, sobuzoxan, flutamide, or antimetabolites such as 5-FU, furtulone, methotrexate, etc., induce diarrhea as a side effect and increase the strength of the patient. Stealing is a problem. ("Therapeutic Remedies of the Day" (2000 edition), published on May 25, 2000, the third edition of the 22nd edition, edited by Yu Mizushima, Nankodo Co., Ltd.).

[0003] In order to reduce diarrhea caused by the side effects of these anticancer drugs, antibiotics, herbal medicines and the like have been studied so far (Jpn. J. Pharmacol. 75, 407-413, 1997., Jpn.
J. Cancer Res., 86, 978-974, 1995.). As an example of such a treatment method, irinotecan hydrochloride (CPT-11)
It has been reported that the combination of the antibiotics streptomycin and penicillin G or the use of vancomycin is effective in preventing diarrhea due to the side effects of cancer.
s., 56, 3752, 1996.).

[0004] However, in recent years, antibiotics have been heavily used due to overuse of bacteria, particularly methicillin-resistant Staphylococcus aureus (Meth).
icillin-resistant Staphylococcus aureus; MRSA
) Or vancomycin-resistant enterococci (Vancomycin re
sistant enterococci (VRE) has emerged, limiting the use of antibiotics.

[0005] Therefore, treatment or prevention methods other than antibiotics are also being studied. For example, in the case of diarrhea caused by the side effect of CPT-11, treatment with an antidiarrheal drug such as loperamide or acetorphan is considered to be effective. ing. However, this treatment was not effective in improving the symptoms of all patients (J. Natl. Cancer. Inst., 86, 446, 199).
4., J. Clin. Oncol., 13, 2144, 1995., J. Clin. Onc.
ol., 16, 2745, 1998.).

On the other hand, interleukin 15 (IL-1
5) is a novel 15-K cloned in 1994.
Da cytokine, interleukin 2 receptor (I
L-2R) is known to exert an IL-2-like physiological action. In addition, IL-15 acts on peripheral blood mononuclear cells (PBMC), NK cells, γδ T cells, promotion of differentiation of B cells, and production of cytotoxic T cells and LAK.
For example, by enhancing the ability of intestinal epithelial cells (IEC) to produce IL-15, the ability to induce differentiation of IEL is promoted, the resistance to invasion of bacteria and viruses from the digestive tract is increased, and the biological defense ability is increased. (Immunologic Re
search, 20 (3), 219-235, 1999.).

[0007] Further, IL-15 is required for promoting differentiation of NK cells.
Is known to be essential, and by promoting the differentiation of NK cells by IL-15, an effect of inhibiting the growth of cancer cells or inhibiting cancer metastasis by enhancing the damage activity to host cancer cells can be obtained. Furthermore, N-15 activated by IL-15
IFN-γ produced by K cells promotes differentiation of Th1 cells, and IL-12 produced by the
Activate the cells. It is known that induction of differentiation into Th1 cells by such a pathway results in an antiallergic effect. On the other hand, in animal experiments, a gene therapy method using IL-15 is known.

[0008]

It is known that IL-15 has a therapeutic effect on diarrhea caused by irinotecan hydrochloride (CPT-11) (CANC
ER RESEARCH, 58, 3270-3274, 1998).
This required 15 large doses, which was not practical in terms of cost. Therefore, attempts have been made to promote the production of IL-15 in vivo and obtain the same effect, but no effective drug has been found.

On the other hand, it has not been known that lactic acid bacteria containing Bifidobacterium sp. Enhance the production of IL-15, and it is also known that they are effective in preventing and treating diarrhea due to side effects of anticancer drugs. I didn't.

Under the circumstances described above, the present invention provides an IL-
An object of the present invention is to obtain a safe material which is effective for preventing and treating digestive disorders such as diarrhea caused by side effects of an anticancer agent, has very few side effects, and can be used on a daily basis.

[0011]

According to the first aspect of the present invention, there is provided an interleukin-15 production promoter, which comprises one or more bacterial cells selected from lactic acid bacteria containing the genus Biffidobacterium. As an active ingredient.

The interleukin-15 production promoter according to the invention described in claim 2 is characterized in that the lactic acid bacterium described in claim 1 is selected from the group consisting of Lactobacillus , Lactococcus , and Pediococcus. Pediococc
us), Leuconostoc (Leuconostoc), Bifidobacterium spp. (Biffidobacterium), Enterococcus
( Enterococcus ), Streptococcus
), Which is one or more selected from the group consisting of:

[0013] A prophylactic / therapeutic agent for gastrointestinal disorders according to the invention described in claim 3 comprises the interleukin 15 production promoter according to claim 1 or 2 as an active ingredient and is administered to a patient with an anticancer drug-induced gastrointestinal disorder. It is characterized by being performed.

[0014] The preventive / therapeutic agent for gastrointestinal disorders according to the invention described in claim 4 is characterized in that the anticancer agent described in claim 3 is irinotecan hydrochloride.

According to a fifth aspect of the present invention, there is provided a food or drink comprising the interleukin-15 production promoter according to the first or second aspect.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION The present inventors stimulated intestinal epithelial cells by specific microbial cells, so that
The present inventors have found that the production of -15 is promoted, and reached the present invention. That is, in the present invention, the genus Bifidobacterium
(Biffidobacterium), one or more selected from lactic acid bacteria including lactic acid bacteria as an active ingredient, IL
It enhances the production of -15 and is effective for the prevention and treatment of digestive disorders such as diarrhea due to side effects of anticancer drugs.

Specifically, the lactic acid bacterium containing the genus Bifidobacterium used in the present invention is not particularly limited as long as it is a strain having an effect of promoting IL-15 production. Lactobacillus , lactobacillus, Coccus ( L
actococcus), the genus Pediococcus (Pediococcus), Leuconostoc (Leuconostoc), Bifidobacterium spp. (Biffidobacterium), Enterococcus (Enteroc
occus ) and one or more bacterial cells selected from strains such as Streptococcus .

More specifically, examples of such strains include, for example, Lactobacillus casei YIT 9019 (Biotechnology and Industrial Technology Research Institute Article No. BP-665, contracted on December 18, 1978 (Micro Engineering) Lactobacillus casei YIT 9029 (Biotechnology and Industrial Technology Research Institute, Jyoro BP-1366, contracted on January 12, 1981 (Microtechnical Laboratories No. 5852) Transfer)), Lactobacillus reuteri YIT
0197, Bifidobacterium breve YIT 4064
(Biotechnology and Industrial Technology Research Institute Article No.BP-2824, 1996.2
Contracted on March 29 (transferred from Microtechnical Laboratory P. No. P-15488), Bifidobacterium breve YIT 4065
(Biotechnology and Industrial Technology Research Institute Article No.BP-6223, 1989
Contracted on April 19 (transferred from Microtechnological Laboratory P. No. P-10672), Lactococcus lactis subspecies species Holdnier ATCC29071, Pediococcus acidilactici ATCC33314, Leuconostoc lactis ATCC19256, Enterococcus faecalis ATCC1
9433, Streptococcus thermophilus ATCC19258
And the like.

These lactic acid bacteria containing the genus Bifidobacterium may be used alone or in combination of two or more. Further, at the time of use, any of live cells, dead cells, or cell constituents thereof can be suitably used.

[0020] In the present invention, the interleukin-15 production promoter is used as an active ingredient and is administered to a patient with an anticancer drug-induced gastrointestinal disorder.
Provide a therapeutic agent and food and drink. This makes it possible to obtain a gastrointestinal disorder prevention / treatment agent effective for prevention and treatment of digestive disorders such as diarrhea, and safe foods and drinks that can be used on a daily basis.

That is, the preventive / therapeutic agent for gastrointestinal disorders of the present invention has an effect on gastrointestinal disorders caused by anticancer drugs.
Anticancer drugs that cause such side effects include irinotecan, cisplatin, adreamycin, ketotifen, cisplatin, glucarone, mitobronitol,
Tamoxifen citrate, sobuzoxane, flutamide,
Or antimetabolites such as 5-FU, furtulon, and methotrexate.

The timing of administration of the lactic acid bacterium containing Bifidobacterium is not particularly limited, but it is preferable to start the administration of the lactic acid bacterium containing Bifidobacterium before administering the anticancer agent. It is preferred that administration of lactic acid bacteria containing Bifidobacterium be started one day before. Further, these lactic acid bacteria may be used alone,
A combination of more than one species may be used.

The dose of the lactic acid bacterium containing Bifidobacterium may be any commonly used amount.
The dose may be from 03 mg / kg to 1 g / kg, preferably 1.
5 mg / kg to 200 mg / kg, more preferably 2 mg / kg to 1
00 mg / kg, particularly preferably 5 mg / kg to 50 mg / kg may be administered.

The lactic acid bacterium disclosed in the present invention has been conventionally used in foods and the like, and has high safety. Therefore, the dosage form and dosage can be arbitrarily selected, and the lactic acid bacterium is added to and blended with foods and pharmaceuticals. be able to. The material of the present invention is a single product, or compounded with a liquid or solid carrier, and if necessary, a solvent, a dispersant, an emulsifier, a stabilizer, an excipient, a binder, a disintegrant, a lubricant, etc., For example, a desired dosage form such as a tablet, a granule, a powder, a powder, a capsule and the like can be obtained.

[0025]

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

Example 1 (IL-15 production promotion test) Lactobacillus casei YIT 9029 (hereinafter referred to as YIT9029), Lactobacillus reuteri
YIT 0197 (hereinafter referred to as YIT0197) was used. Each cell was cultured overnight in an MRS medium (manufactured by Difco), collected by centrifugation, washed, and heated at 100 ° C. for 30 minutes. The obtained cells were freeze-dried to obtain sample cells.

The obtained sample cells (50 μg / ml) were added to DMEM.
The cells were suspended in a medium (manufactured by Sigma) and the rat intestinal epithelial cell line IE
C6 (ATCC CRL-1592; hereinafter referred to as IEC6) 1x
The cells were added to 10 ⁶ cells / ml and cultured. As a control group, those to which no strain was administered were used. 24 hours later, IEC
6 was recovered and Sigma was extracted from mRNA extracted according to a standard method.
RT-PCR was performed according to the method described above, and the PCR product was electrophoresed on an agarose gel.

The gel after electrophoresis was stained with ethidium bromide, and a UV irradiation image was photographed. The photograph of the migration pattern was taken into a computer by an image analyzer, and the band intensity of the target PCR product and the band intensity of β-actin were measured by analysis software. From the obtained measurement results, the amount of IL-15 relative to the amount of β-actin was calculated and used as the expression level.
The results are shown in FIG.

FIG. 1 is a diagram showing the results of an IL-15 production promotion test. As shown in FIG. 1, YIT9029 and YIT01
97, the induction of IL-15 expression was confirmed.
T9029 showed a value about 4.4 times higher than YIT0197. On the other hand, other strains hardly expressed IL-15.

Example 2 (Comparison between strains for improvement of diarrhea caused by CPT-11) Lactobacillus casei YIT 9018 (hereinafter referred to as YIT9018) and Bifidobacterium breve YIT 4065 Heat-killed cells were prepared using Bifidobacterium breve YIT 4064 (hereinafter referred to as YIT 4064) in the same manner as in Example 1, and used as sample cells. As antibiotics, C
PT-11 (Kampt injection, manufactured by Yakult Honsha) was used.

Each group of 5 to 6 week old Slc: SD rats (male, manufactured by SLC Japan) was treated as YIT901.
8 and YIT4065 0.1% and 1.0%, YIT4064
Was mixed with a feed (F-2, manufactured by Funabashi Farm) at a concentration of 0.05%, and CPT was administered 2 weeks before CPT-11 administration.
They were fed on a diet for 3 weeks until the third day after -11 administration. The control group received only F-2 diet. CPT-11 was administered to all rats at a dose of 60 mg / kg once daily for 4 days in the tail vein.

The fecal properties were determined as diarrhea in loose or watery stool that had broken down and contained a large amount of water. Also, CPT-
Acute diarrhea that was observed within 3 hours after administration during the 11 administration period (Day 1 to Day 4 in FIG. 2) and 3 days (Day 5 to Day 7 in FIG. 2) during the drug holiday after administration What was done was called delayed diarrhea. The results are shown in FIG.

FIG. 2 is a graph showing the results of a comparative test between strains on the improvement of diarrhea caused by CPT-11. As shown in FIG. 2, there was no difference between the groups in the occurrence of acute diarrhea during the CPT-11 administration period. On the other hand, in the onset of delayed diarrhea, the frequency of occurrence and the degree of decrease in the frequency of occurrence of the diarrhea were observed in all of the YIT9018, YIT4065 and YIT4064 administration groups as compared with the control group. In particular, the effect of the YIT9018 diet group was remarkable, and in the 1.0% diet group, only one watery stool was observed on the second day of drug holiday.

Example 3 (Comparative test of administration concentration for improvement of diarrhea caused by CPT-11) YIT9018 was used as a strain. Four groups of 5 to 6-week-old Slc: SD rats (male, manufactured by Japan SLC) were provided, each group consisting of 5 rats, from 0, 75, 150, 3 days before CPT-11 administration for 10 days. YIT9018 at 300mg / kg / day
Was administered by gavage three times a day. CPT-11 was administered at a dose of 60 mg / kg once a day for 4 days in the tail vein as in Example 2, and the fecal properties were determined as in Example 2. The results are shown in FIG.

FIG. 3 is a graph showing the results of a comparative test of the administered concentration for improvement of diarrhea caused by CPT-11.
As shown in FIG.
The frequency of acute diarrhea on Day 4 decreased. Also, Day 5
From day 7 to day 7, delayed onset of diarrhea was clearly reduced in frequency and degree. In particular, no onset of delayed diarrhea was observed in the 150 mg / kg / day administration group.

Example 4 (Comparative test of administration time for improvement of diarrhea caused by CPT-11) YIT9018 was used as a strain. 5-6 week old Slc: SD
Rats (male, manufactured by SLC Japan) were divided into four groups each consisting of 10 rats, three of which were 150 mg / kg / da.
Y was administered by gavage YIT9018 three times a day by gavage, and depending on the administration timing, 3 days before CPT-11 administration for 9 days (pre-treatment group) and 6 days from CPT-11 administration start date (simultaneous treatment group) , And 3 days from the last day of CPT-11 administration (post-treatment group). Another group was a control group not administered with YIT9018.

CPT-11 was used in the same manner as in Example 2 at 60 mg /
Example 2 was administered in a tail vein at a dose of kg once a day for 4 days.
The fecal properties were determined in the same manner as in. FIG. 4 shows the results. FIG.
Fig. 3 is a graph showing the results of a comparative test of administration timing on improvement of diarrhea caused by CPT-11.

As shown in FIG. 4, there was no difference in the frequency and extent of acute diarrhea between Day 1 and Day 4 between the YIT9018 administration group and the control group. On the other hand, YIT9018 administration reduced the frequency and extent of delayed diarrhea.

Example 5 Preparation of Food and Beverage Formulation Example 1 10% (W / V) glucose liquid sugar was added to a 20% (W / V) solution of skim milk powder, and this was used in Example 3 as a substrate. A culture solution of lactic acid bacteria was added at 0.5% (V / V) and cultured at 37 ° C. for 18 hours to prepare fermented milk. In addition, the obtained fermented milk had a good taste, and was comparable to ordinary fermented milk.

Formulation Example 2 Using the lactobacillus cells used in Example 1, bread was prepared with the following composition.

The resulting bread had a characteristic sour odor flavor, good taste, and was comparable to ordinary bread.

As described in detail above, the prevention and treatment of diarrhea caused by an anticancer drug such as CPT-11 by administering one or more selected from lactic acid bacteria including Bifidobacterium. Is possible. In addition, administration of these cells enhances not only antigen-presenting cells but also the intestinal epithelial cells (IEC) IL-15 producing ability, and as a result, promotes the ability to induce differentiation of intestinal interepithelial lymphocytes (IEL). However, it can be expected to increase resistance to invasion of bacteria and viruses from the digestive tract.

It is also expected to promote the differentiation of NK cells and to inhibit the growth of cancer cells or to inhibit metastasis by enhancing the damaging activity on host cancer cells. Further, activated NK
IFN-γ produced by the cells is expected to promote the differentiation of Th1 cells and exhibit an antiallergic effect.

[0044]

As described above, the present invention relates to IL-15.
Is effective in preventing and treating diarrhea due to side effects of anticancer agents, and has the effect of obtaining a safe material that can be used on a daily basis with very few side effects.

[Brief description of the drawings]

FIG. 1 is a diagram showing the results of an IL-15 production promotion test.

FIG. 2 is a graph showing the results of a comparative test between strains on improvement of diarrhea caused by CPT-11.

FIG. 3 is a graph showing the results of a comparative test of administration concentrations for improvement of diarrhea caused by CPT-11.

FIG. 4 is a graph showing the results of a comparative test of administration timing for improvement of diarrhea caused by CPT-11.

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Shoji Onoe 1-1-19 Higashi-Shimbashi, Minato-ku, Tokyo Yakult Honsha Co., Ltd. (72) Inventor Yasunobu Yoshikai 1-9-9 Sonoyamacho, Chikusa-ku, Nagoya-shi, Aichi Prefecture -2 (72) Inventor Tetsuya Matsuguchi 2-66-116 Yada-cho, Higashi-ku, Nagoya-shi, Aichi F-term (reference) 4B001 AC31 BC14 EC99 4B018 MD86 MD87 ME11 4B065 AA30X AA39X AA49X CA42 CA44 4C087 AA01 AA02 AA04 BC56 BC59 BC61 NA14 ZA66A

Claims (5)

[Claims] 1. The genus Biffidobacterium
One or interleukin 15 production promoting agent comprising as an active ingredient two or more cells selected from lactic acid bacteria including ium). Wherein said lactic acid bacteria, Lactobacillus (Lacto
bacillus ), Lactococcus ( Lactococcus ), Pediococcus ( Pediococcus ), Leuconostoc ( Leu
conostoc ), the genus Biffidobacteriu
m ), one or more bacterial cells selected from the genus Enterococcus and the genus Streptococcus, wherein the interleukin 15 production promoter according to claim 1, characterized in that: 3. A prophylactic / therapeutic agent for a gastrointestinal disorder, which comprises the interleukin 15 production promoter according to claim 1 or 2 as an active ingredient and is administered to a patient with a gastrointestinal disorder caused by an anticancer agent. 4. The preventive / therapeutic agent for gastrointestinal disorders according to claim 3, wherein the anticancer agent is irinotecan hydrochloride. 5. A food or drink comprising the interleukin-15 production promoter according to claim 1 or 2.