JP2003026587A - Bacterial removing agent for helicobacter pylori - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a bacterial removing agent for Helicobacter pylori. SOLUTION: This bacterial removing agent for the Helicobacter pylori comprises a Chinese medicine extract or a fermentation treated substance of a fruit mixture.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to H.264. pylori
The present invention relates to an oral preparation which has both a bactericidal action and an immunomodulatory action against A.

[0002]

[Prior Art] Helicobacter pylori
pylori) is a spiral gram-negative bacterium having a size of about 2-5 μm. Helicobacter pylori (hereinafter referred to as "H. pylori") is flagella, urease, catalase, adhesin (adhesion factor), vacuolating toxin (VacA), heat shock protein (HSP), toxin-related protein (Cag).
It produces pathogenic factors such as A). Currently, H. pylori
Gastric mucosal damage due to infection is considered to be caused by a multifactorial mechanism. And H. py
Oral administration of lori causes gastritis on human gastric mucosa,
H. It has been proved that eradication of pylori cures gastritis. In addition, H. Regarding the relationship between pylori and gastric cancer, in 1994, IARC (Int
based on an epidemiological study conducted by the H.E. pylori is a group 1 (definite carcinogen) that is clearly associated with carcinogenesis
Is specified in. Thus, H. The relationship between pylori infection and gastritis / gastric cancer is close. Moreover, Helicobacter pylori is said to be a bacterium that lives in the stomach for the rest of its life unless it is sterilized. Therefore, H. py
It is highly expected that the eradication of Lori can prevent or treat the development of gastritis and gastric cancer.

Antibacterial agents mainly composed of antibiotics are used for Helicobacter pylori infection. Currently, a combination therapy using a proton pump inhibitor (omeprazole, lansoprazole) that suppresses the secretion of gastric acid and two antibacterial agents (amoxicillin, clarithromycin, etc.) is showing good eradication results.

[0004] So far, since antibiotics against the bacterial species spectrum of both Gram-positive bacteria and Gram-negative bacteria are effective against the bacteria, it is said that there is no urgent need for eradication measures against the bacteria at this time. I have an opinion. However, there is a fear that resistant bacteria will be induced if therapeutically and prophylactically continue to use antibiotics. In addition, side effects (diarrhea, liver damage, renal damage, etc.) may occur,
It should not be unnecessarily increased in frequency of use. Therefore,
H. It is desired to develop a substance that has a bactericidal effect against pylori and that can replace conventional antibiotics.

[0005]

SUMMARY OF THE INVENTION The present invention is based on the H.264 standard. pyl
It is an object of the present invention to provide an oral preparation having both a bactericidal action against ori and an immunomodulatory action.

[0006]

Means for Solving the Problems As a result of intensive research to solve the above-mentioned problems, the present inventors have found that oral treatments such as Juzen-taiho-to, Toki-rokuo-to, Hochuekki-to and fermented foods Herbal medicines intended for use are H. It has been found that it exhibits an antibacterial action against pylori and at the same time has an immunomodulatory action
The present invention has been completed.

That is, the present invention is a disinfectant for Helicobacter pylori, which contains a Chinese herb extract or a fermented product. Furthermore, the present invention includes an immunostimulant (eg, human peripheral leukocyte granulocyte /
Lymphocyte ratio regulator). Examples of Kampo extract include Juzentaihoto, Hochuekkito or Toki Rokuoto. Hereinafter, the present invention will be described in detail.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, a Chinese herb extract is H. py
Completed the discovery that it has antibacterial activity against Lori, especially bactericidal activity, and also has immunostimulatory activity mainly for the regulation of human peripheral leukocytes (particularly regulation for optimizing the granulocyte / lymphocyte ratio). It is a thing. Of course, H. No report has been made on the effect of eradicating pylori. In the present invention, "disinfection"
Means bactericidal action or growth inhibition. In particular,
The number of bacteria when the disinfectant of the present invention is used is 20% or less of the number of bacteria before use, preferably 1% or less, more preferably 0.
It means decreased to 001% or less (that is, 80% or more of the number of bacteria before use is eradicated).

By the way, as a countermeasure against bacterial infection, it is ideal that both the parasite and the host are treated simultaneously.
That is, in the treatment of bacterial infections, it is preferable to use a drug effective on the one hand for eradication of bacteria and on the other hand for increasing the defense power of the host. The inventor of the present invention has so far searched for a method of controlling the immune capacity of a host by using an oral immunomodulator. As a result, it was found that the herbal medicine and the like have a bactericidal effect against intestinal infectious microorganisms and can quantitatively and qualitatively regulate the host's immunocompetence.

Examples of the medicinal component in the disinfectant and / or immunostimulant of the present invention include natural product-derived components such as Chinese herb extract. In addition, this natural product-derived component,
It may be one kind or two or more kinds. In addition, fermented foods are It can be used as a disinfectant and / or an immunostimulant for P. pylori.

1. Kampo extract As the above-mentioned Kampo extract (herbal medicine), for example, Juzentaihoto,
Toki Rokuoto, Hochuekkito, Kakkonto, Kakkonto, Kagawa Kyukan, Otsujiyu, Anchusan, Jumi-Seito, Hachimijiogan, Daisaikoto, Shosaikoto, Saikokeishito. , Saiko Karyukotsu Oyakuto, Hangeshashinto, Orengedokuto, Hangekobokuto, Goreisan, Shoseiryuto, Hokikoito, Seifusan, Tokishakuyakusan, Kamishoharusan, Katsura Eshibukuryomaru, Keishikaryukotsu Oyakuto, Maoyu, Bakumondoyu, Shigyakusan, Keikeijutsu Amanoyu, Keishiyu, Shichimonokoyu, Geigakubetsuyu, Kiyokamifufu, Hofutsushosei. San, Megami-san, Shakuyaku-kanzo-to, Higasi-san, Daio-kanzo-to, Toki-inko, Rokumi-maru, Jiu-boku, Kokenchu-to,
Daikenchutou, Shinreiseikanto, Goshigurenkimaru, Saireito, Keireiingohangatsubokuto, Hourento, Tokikenchutou, Keishibukuryomaruka, well saijin, asagojinmaru, maotsuki Mention may be made of Kosaishinto, Keishikashakuyakudaioto, Seikatsukikito, Kikyoto, etc., but Juzentaihoto, Toki-rokuoto and Hochuekkito are preferred. However, it is not limited to the above crude drug.

Examples of the crude drug extract include various aqueous solvent extracts, and hot water extract is preferably used. However, it is not limited to the hot water extract. A specific example of preparation of the extract is a method of extracting the crude drug mixture having the above composition with hot water and filtering the obtained extract. This extract can be lyophilized if necessary and used as a dry powder.

[0013] For example, Juzen-taiho-to is a yellow radish (Ougi), a cinnamon (keishi), a ground yellow (dio), a peony (peony),
It consists of a mixed crude drug of river cucumber, soy sauce, toki, carrot, carrot, bukuryo and licorice. Therefore, when preparing Juzen-taiho-to, add 10 to 100 times the amount of distilled water to the above mixed crude drug, decoct at 80 to 100 ° C for 30 to 60 minutes, filter the obtained extract, and then freeze. Dry to obtain a dried extract. Alternatively, the mixed crude drug may be pulverized and finely powdered, and the obtained preparation may be orally administered as it is. The mixed crude drug is
It may be a plain crude drug that constitutes it, or may be a preparation prepared by fermenting either mixed or plain crude drugs.

2. In the present invention, the fermented food product of the fruit mixture, fermented foods, the raw material apples, mandarin oranges, jujube, peach, apricot, ume, summer mandarin orange, pear, corn, such as fermented processed products mainly 10 And means an oral drink containing polyphenols, carotenes, flavonoids.

For the fermentation treatment of the fruit mixture as the raw material, an existing fermentation method can be adopted. For example, the fruit is thoroughly washed with water and shredded to obtain a pre-fermentation preparation. Add 10-100 volumes of water to the obtained pre-fermentation preparation and heat (60
Boil for 30 to 60 minutes at ℃ to 100 ℃). Repeat this 3 times to perform intermittent sterilization.

After that, aseptic operation is thoroughly performed for fermentation.
For fermentation, commercially available fermenting bacteria such as yeast, koji mold, and lactic acid bacteria are added in an amount of 1-10 g per 1 L of the pre-fermentation standard volume, and 7-10 at 10-20 ° C.
Process for days. After cooling this, it can be concentrated to obtain a fermented product. This is used as a fermented food. Alternatively, commercially available fermented foods can be used in the present invention.

3. When the disinfectant and the immunostimulant galenical drug product or the fermented product is used as a complex or a single agent, it may contain both a pharmaceutically acceptable carrier or additive. Examples of such carriers and additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, xanthan gum, Gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, other surfactants acceptable as pharmaceutical additives, etc. , Artificial cell structures such as liposomes and the like. The additives to be used are appropriately or in combination selected from the above depending on the dosage form of the present invention.

When the preparation of the present invention is used for the purpose of sterilization and immunoregulation, the subject to be used is not particularly limited. For example, it can be used as a therapeutic agent for gastric diseases. Examples of stomach diseases include at least one pathological condition such as gastric ulcer and gastric cancer. These diseases may be a single disease, a concurrent disease, or a concurrent disease other than the above.

The formulation of the present invention can be administered orally. In this case, solid preparations such as tablets, granules, powders, and pills, or liquid preparations such as liquids and syrups applied thereto may be used. Especially granules and powders,
It can be in unit dosage form as a capsule, and in the case of liquid formulations it may be a dry product which is redissolved when used.

Of these dosage forms, oral solid preparations usually contain additives such as binders, excipients, lubricants, disintegrants, wetting agents and the like, which are commonly used in the formulation thereof. .
In addition, liquid preparations for oral use usually include stabilizers, buffers, corrigents, preservatives,
Contains additives such as fragrances and colorants.

In this case, the effective amount of the bactericidal agent and / or immunostimulant of the present invention combined with an appropriate diluent and a pharmacologically usable carrier is an effective amount per body weight per administration. The dose is 20 mg to 2000 mg per kg and is administered at 1 to 4 week intervals. The effective dose is 1 mg to 1000 mg per 1 kg of body weight, and it is administered at 1 to 4 week intervals. The fermented product can be used as a food, and the amount used is 0.1 to 0.5 g / kg body weight / day.

[0022]

EXAMPLE 1 The present invention will be described in more detail below with reference to examples. However, the technical scope of the present invention is not limited to these examples.

[Example 1] H. pylo
Bactericidal action against ri of H. The following experiment was conducted in order to examine the eradication effect on pylori.

(1) H. Strains of pylori isolated cultured gastritis patients (10 persons) were inoculated into HP selective medium and microaerobic (5-10% oxygen concentration lower than normal oxygen concentration in normal air (20%)) 5 The culture was separated for 7 days. The strain that has developed is H. API CAMPY kit (BIM
ERIUX SA, FRANCE).

(2) The test drug, Juzentaihoto, was orally administered at 7.5 gr per day in three divided doses before or during a meal. This formulation mixes ten crude drugs and extracts with hot water. The components are described below. Juzen Daihoyu (JTT
) Is a carrot (3.0 gr), sugi (3.
0 gr), Jutsu ((3.0 gr), Touki (3.0 gr), Bukyou (3.0 gr), Geo (3.0 gr), Senkyu (3.
0 gr), peony (3.0 gr), peony (3.0 gr) and licorice (1.5 gr). Toki Rokuoyu (TRT)
Is touki (0.75 gr), oren (0.75 g) in 5.25 gr
r), Oogon (0.75 gr), Jukujiou (0.75 gr),
It consists of Oataku (0.75 gr), Kangiou (0.75 gr) and Ougi (0.75 gr). Hochuekkito (HET) is 7.5 gr
Carrots (4.0 gr), sandalwood (4.0 gr), sugi (4.0 gr), Japanese smelt (3.0 gr), chimpi (2.0 g)
r), Thyso (2.0 gr), Psycho (2.0 gr), Licorice (1.5 gr), Ginger (0.5 gr) and Ginger (1.0 gr). As the fermented food, a commercially available Otaka enzyme (main component: polyphenol, flavonoid) obtained by fermenting a fruit mixture was used (Otaka Enzyme Co., Ltd.).

(3) Bactericidal action of herbal medicines CO in Colombia medium, ie 5% sheep blood agar (BBL)
₂Pre-incubate with 10% at 35 ℃ for 72 hours, and plate the test bacteria that have grown.
More scraped, 10 with Brucella broth (Difco)⁶ And 10⁸C
What was prepared to be FU / ml was used as the test bacterial solution. each
Includes 2-fold dilution series (200-0.098mg / ml) of crude drug formulations, etc. 5
% Horse defibrinated blood added Brucella agar (Difco)
Inoculate 5 μl of each solution with CO₂Incubate at 10% at 35 ℃ for 72 hours
It was After culturing, the number of colonies on the agar plate was counted. Colony
-The degree of proliferation depends on the number of "+++", "++",
It was separated into "+" and "-". That is, the number of colonies is 10
00+ is "+++", 999-100 is "++", 99-10 is
"+", Less than 10 was "-".

The numbers of colonies on the agar plates containing the respective herbal medicines are shown in Table 1 (Hochuekkito), Table 2 (Toki Rokuoto), Table 3
(Juzen-taiho-to) and Table 4 (fermented food). In this example, the antibacterial agent screening was based on 10 ⁶ and 10 ⁸ CFU / ml commonly used in drug susceptibility tests, but the following more closely resembles the situation of the gastric mucosa in vivo.
The results for 10 ⁶ CFU / ml are mainly described.

[0028]

[Table 1]

[0029]

[Table 2]

[0030]

[Table 3]

[0031]

[Table 4]

H. The maximum dilution factor at which the growth of P. pylori was suppressed was 32 for 2 subjects in Juzen-taiho-to.
Fold, 16 times in 7 subjects, 8 in 1 subject
It was double. In Toki Rokuoto, 512 in 3 subjects
Fold, 256 times in 1 subject, 1 in 6 subjects
It was 28 times. In Hochuekkito, the bacterial growth was observed once at the 16-fold dilution, but at the 32-fold dilution, the bacterial growth was suppressed again in all subjects. On the other hand, the fermented food was doubled in 4 subjects, but the remaining 6 subjects suppressed the growth of bacteria only when it was not diluted.

When the above results are averaged, H.264. piler
The maximum dilution rate at which the growth of i-bacillus was suppressed was 16 times for Juzen-taiho-to and 128-fold for Toki-Rokuo-to. The bactericidal action against P. pylori is strong. In Hochuekkito, the bacterial growth was observed once at the 16-fold dilution, but at the 32-fold dilution, the bacterial growth was suppressed again in all subjects. Therefore, it can be said that the maximum dilution factor in Hochuekkito is 32 times.

From the above, the herbal medicine preparations such as Juzentaihoto, Toki Rokuoto, Hochuekkito are H. An extremely excellent bactericidal effect against P. pylori was recognized, and it is expected to be used as an oral preparation for eradication. On the other hand, when the fermented food was not diluted, that is, when it was 200 mg / ml, the bactericidal effect was obtained. However, it can be said that in the fermented foods rich in sugars, the sterilizing effect was obtained even if the dilution ratio was low, which is remarkable.

Example 2 Leukocyte regulation test 1. Effect of Hochuekkito and Juzentaihoto on the composition ratio of leukocyte subgroups HET for "granulocyte dominant" individuals and JTT for "lymphocyte dominant" individuals 500 mg / 30 kg worth of extract
It was orally ingested daily for a day, and the effect on the composition ratio of leukocyte subgroups of both individuals was examined. Here, the "granulocyte-dominant type" means an individual in which the ratio of granulocytes to the total number of white blood cells accounts for 70% or more, and by administering the preparation of the present invention, granulocytes are 45 to 69%, Preferably, it can be judged that leukocyte regulation was observed when the concentration was changed to 50-60%. Further, the "lymphocyte-dominant type" means an individual in which the proportion of lymphocytes in the total white blood cell count is 40% or more, and the lymphocytes are 15 to 39%, preferably by administering the preparation of the present invention. It can be judged that the white blood cell regulation was recognized when it changed to 20 to 30%.

The "intermediate type" means an individual having less than 69% of granulocytes and less than 39% of lymphocytes, and the ratio of granulocytes and lymphocytes maintains the above values by administering the preparation of the present invention. It can sometimes be judged that leukocyte regulation was observed. Table 5 shows the effects of HET, JTT and TRT on the white blood cell composition ratio.

As a result of administration of HET for 30 days to the "granulocyte-dominant" individual, the total white blood cell count decreased from 6.95 × 10 ³ / μl before administration to 5.88 × 10 ³ / μl after administration, a decrease of about 15%. Admitted.
When this was measured according to the number of fractionated cells, the total number of granulocytes changed from 69.5% to 65.4%, 4.1% compared to before administration.
Diminished. In contrast, the number of lymphocytes is 23.6% to 26.3%
And increased by 2.7% compared to before administration. on the other hand,
As a result of administration of JTT for 30 days to individuals classified as "lymphocyte-dominant type", the total white blood cell count increased from 3.82 × 10 ³ / μl before administration to 5.04 × 10 ³ / μl after administration, an increase of about 32%. Admitted.
When this was measured for each fractionated cell as in the former experiment, the total number of granulocytes was changed from 50.8% to 55.7%, and increased by 4.9% compared with before administration. On the other hand, the lymphocyte count changed from 44.3% to 39.2%, 5.1% compared to before administration.
Diminished.

On the other hand, as a result of administration of TRT for 30 days to an individual classified as “intermediate type”, the total white blood cell count was 4.65 × 10 ³ before administration.
An increase of about 7% was observed from / μl to 4.98 × 10 ³ / μl after administration. When this was measured for each fractionated cell in the same manner as in the former experiment, the total number of granulocytes was 59.2% to 60.3%, which was not different from that before administration. On the other hand, the lymphocyte count is also 3
From 5.6% to 36.9%, there was no change compared to before administration.

[0039]

[Table 5]

2. Effect of Hochuekkito, Juzentaihoto and Toki-rokuto on the detection of CD-positive cells HET was applied to both individuals showing "granulocyte-dominant type" and "lymphocyte-dominant type" as described above. Alternatively, the number of CD-positive cells after administration of JTT for 30 days was compared with the number of CD-positive cells before administration. The results are shown in Table 6.

[0041]

[Table 6]

The variation of B cells (CD19 positive cells) was almost unchanged in all individuals, but T cells (CD2 positive cells) increased in all individuals, and individual A (granulocyte type; HET administration)
13.0%, 17.5% for individual B (lymphocyte type, JTT administration)
I saw an increase. However, there was no change in individual C (intermediate type; TRT administration). Also, helper / inducer T
The cells (CD4 positive cells) also showed the same tendency of increase, but the increase of suppressor inducer T cells (CD8 positive cells) was small in both cases. Therefore, the ratio of CD4 / CD8 was 0.51 to 0.71 for individual A, 1.17 to 1.61 for individual B, and
In C, it rose from 1.11 to 1.2. The phagocytic line centering on neutrophils (CD11b) tended to decrease slightly in both volunteers, but monocyte / macrophage lineage cells (CD14 positive cells) increased in HET and conversely decreased in JTT, The TRT showed that there was no change. CD16 is FcrRIII and N
K cell (FcrRIIIa), monocyte / macrophage system (FcrRIIIa
a) and the sum of neutrophils (FcrRIIIb),
It showed a decreasing tendency by the administration of ET and JTT. In addition, in peripheral blood, CD56, which was mainly expressed in NK cells, was examined, and as a result, administration of HET, JTT, and TRT showed a slight increase in the tendency.

From the above results, when the preparations of the present invention Hochuekkito, Juzentaihoto and Toki Rokuto were administered, T cells, B cells, macrophages and N cells were analyzed through the analysis of CD positive cells.
The number of K cells was increased, and it was found to be effective during "bacterial and viral" infections and "onset of cancer."

3. Effects of Hochuekkito, Juzentaihoto and Tokirokuto on Cytokine Production The quantitative effects of immunocompetent cells were examined by assessing CD-positive cells in the previous section. In this section, we investigated the cytokines produced by CD-positive cells and examined their qualitative effects on immunocompetence.

The traced cytokines are IFN-γ as a cell responsible for cellular immunity and the one that controls NK cell activity, IL-4 as a cytokine for humoral immunity, and IL-1β as that for macrophages. did. As a result, as shown in Table 7, IFN-γ showed a marked increase of 140%, 800% and 584%, respectively, by the administration of HET, JTT or TRT.

On the other hand, IL-4 was significantly increased by administration of HET, JTT or TRT to 133%, 190% and 187%, respectively, but IL-1β was significantly increased in HET, JTT or TRT. The fluctuations before and after administration were 56%, 37%, and 102%,
The result was a decrease or no change.

[0047]

[Table 7]

[0048]

INDUSTRIAL APPLICABILITY The present invention provides a bactericidal agent for Helicobacter pylori and an immunostimulant. The Kampo extract used in the present invention has an antibacterial action against Helicobacter pylori, and has a leukocyte subgroup ratio, that is, granulocytes.
It is useful for treating gastric disorders because it optimizes the lymphocyte ratio.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 35/78 A61K 35/78 Q T W Y 35/84 35/84 A A61P 1/04 A61P 1/04 31/04 31/04 35/00 35/00 37/00 37/00 37/04 37/04 F-term (reference) 4C088 AA04 AB12 AB26 AB32 AB33 AB37 AB38 AB40 AB41 AB51 AB52 AB59 AB60 AB62 AB81 AC01 AC04 AC05 AC11 AC12 BA05 BA07 BA08 BA09 CA05 MA07 MA52 NA14 ZA66 ZA68 ZB26 ZB35 ZC75

Claims (7)

[Claims] 1. A disinfectant for Helicobacter pylori, which comprises Chinese herb extract. 2. The disinfectant according to claim 1, wherein the Kampo extract is Juzentaihoto, Hochuekkito or Toki Rokuoto. 3. A disinfectant for Helicobacter pylori, comprising a fermented product of a fruit mixture. 4. An immunostimulant containing Chinese herb extract. 5. The immunostimulant according to claim 4, wherein the Kampo extract is Juzentaihoto, Hochuekkito or Tokirokuto. 6. An immunostimulant containing a fermented product of a fruit mixture. 7. Immunopotentiation of granulocytes of human peripheral leukocytes /
The immunostimulant according to claim 5 or 6, which is a lymphocyte ratio regulation.