TW201124146A - Food and pharmaceutical composition with strains of lactic acid bacteria for treatment of gastric ulcer - Google Patents

Abstract

The invention described relates to novel strains of lactic acid bacteria and to novel compositions containing novel strains or strains not previously known to be useful in treatment of gastric ulcer. The composition may be in the form of foodstuffs or in the form of pharmaceutical compositions.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a food composition and a pharmaceutical composition, and more particularly to a food composition and a pharmaceutical composition of a lactic acid bacteria strain for treating gastric ulcer. [Prior Art] Helicobacter pylori was found in 1982 by Marshall and Warren and is associated with ulcerative inflammation of the digestive organs. Ulcerative inflammation of the digestive organs - it has been thought that due to the pressure and the life style, the stomach age is too strong, which makes the sputum and inflammation anti-Newton. Therefore, the treatment of this disease - directly to inhibit the secretion of gastric juice or to neutralize the acidity of the gastric juice, so that the disease is relieved. It was not until 1982 that the presence of Helicobacter pylori was discovered that the development of antibiotics began to be developed, shortening the time course of treatment for peptic ulcer disease. In 2005, Marshamm was awarded the Nobel Prize in Medicine for "The bacterium Helicobacter pylori and its r〇le in gastfitis and peptic disease". It is directly proved that this contribution is very important for human medicine. Peptic ulcer disease is endless in today's business community. Modern people's stress, lifestyle and diet without mosquitoes are the mainstays in digestive ulcers. Most patients with stomach auger _ silk patients must continue treatment for antibiotics for a period of time, but often because of A busy life has a strategy, which may cause the bacteria to develop resistance. Furthermore, the treatment with antibiotics will __and; the human (10) beneficial bacteria will be eliminated, thus causing adverse symptoms to the human body. Drug treatment of Helicobacter pylori usually involves a combination of drugs, such as antibiotics, such as tetracycline new amoxicillin), macrocyclic antibiotics (bismuth nidaz〇ie) and other salts (bismuth) mmmroton pump inhibitor) > losec , takepron. It is easy to produce resistance in the early use of the species, and it will make these resistant strains sit on the 1 S1 3 201124146 i, and the effect will not be any effect. Therefore, two kinds of anti-supplement will be selected. Or proton::=!·, so _ also use less salt eradication rate, = up to 90 / °. If only one antibiotic is used plus proton pump blocker, then non-right ίί 85~ currently in Taiwan's ship payment is also limited to two weeks, except for 2 special cases requiring extended treatment or re-treatment. In addition to the above drugs, there are also cases where the use of a type B anti-sewn drug replaces the proton-blocking coffee. How to choose the best drug group depends on the patient's condition. But this three-in-one therapy also has its drawbacks,

^ The production of effects, such as nausea, thirst, intestinal qi, dizziness, etc., often affect the obedience of patient service. Generally, products containing milk _ (LAB) have only intestinal health effects. Although tens of thousands of lactic acid bacteria strains exist in nature, only a few lactic acid strains have the property of inhibiting the activity of Helicobacter pylori. A few of these strains have the acid and bile-tolerant ability, and the characteristics of the stomach, the ability of the epidermal cells, and the ability to survive after the stomach is the important basis for screening strains that promote health effects. Up to now, only a few strains of lactic acid bacteria have been confirmed to have an effect of inhibiting gastric pyloric helix (IV) activity. The function of lactic acid bacteria on the health of the body lies in the specificity of the strain rather than the spedes. This (4) has a special function of the human probiotics (Guidelines for the evaIuation of pr〇bi〇tics). In f〇〇d; kiss (10) 〇fj· FAO/WHO working group 〇n drafting guidelines for the evaluation of probiotics in food ; London Ontario, Canada April 30 and May 1, 2002 : 1-7). ’

There are many reports on the reaction between lactic acid bacteria and Helicobacter pylori in the country at home and abroad. In the in vitro study of the ability of lactic acid bacteria to inhibit the activity of Helicobacter pylori, the probiotic linoleic acid produced by probiotics decomposes ΙΚΚ_τ(ΙκΒ kinase; Ικβ kinase) and Hsp90 (Heat shock protein 90; heat shock protein) 90) Complex, initiation/cB (Nudear factor-kappaB; nuclear transcription factor_also) message delivery chain, producing anti-inflammatory activity in gastric mucosal epithelial cells infected with Helicobacter pylori and inflamed (Jung M 201124146

Kim, 2008). Active lactic acid bacteria MLBPLl iacioftaczZ/ws rhamnosus GG, Lactococcus Zactis can reduce the adhesion of Helicobacter pylori to cells by about 50%, and active lactic acid bacteria can inhibit the production of IL-8 by cells infected with Helicobacter pylori (Interleukin-8; Auxin-8), reducing the concentration of il-8 secreted by cells, helps to reduce the inflammatory response of cells and the production of CD74 in the Helicobacter pylori receptor (S. Rokka, 2008). [Summary of the Invention]

The present invention broadly encompasses any of the following biological cultures: Lactobacillus salivarius strain, Food Industry Development Institute, registered waver BCRC 910437 and Lactobacillus johnsonii (toto6ady'〇/2wom7) MH-68 strain 'κκ 〇Industrial Development Institute hosting number BCRC 910438, and a food composition or pharmaceutical composition comprising a physiologically acceptable excipient or diluent. In another specific case, the physiologically achievable physiologicly achievable antibacterial activity of the genus Helicobacter pylori is the following: a retrospective or a blunt biological culture: Lactobacillus saliva AP-32 strain's food industry development The Institute's deposit, number BCRC9i〇437, and WI-68 of the bacterium of the genus B. cerevisiae were recorded as bcrc 9 Li. Further, in another specific embodiment, the present invention can broadly include a lactic acid bacteria strain, a gastric Helicobacter pylori competition, an inhibition analysis, an animal test, and a human clinical trial confirmed to have a stomach stagnation auger-activity- The aforementioned biological culture. The present invention may also broadly include any part of the parts of the specification, the parts and the parts, the constituent elements and the features, and the combination of the parts and features, and if The complete thing has been given the right to know (4), and the equivalent of the scale will be incorporated into this article as a separate matter. [Embodiment] 201124146 The present invention can broadly comprise a biological culture selected from the group consisting of the following: or a combination of any one or more of the following: Lactobacillus saliva - (4) ΑΡ · 32 strains - Food Industry Development Institute No. BCRC 91〇437 and S. cerevisiae (Ζ_Ζ^α7/^·0/^_ϊ)ΜΗ-68 strain, Food Industry Development Institute registration number BCRC 910438. ' ^ Many literatures refer to iij probiotic g can be used to divide the amount of debris f, help the patient to gastric Helicobacter pylori balance the gastric juice pH, destroy the ammonia cloud formed by the Helicobacter pylori invading silver cells, and with the stomach The pyloric helix relies on the space for survival to achieve the effect of inhibiting the activity of Helicobacter pylori.

The cold lactic acid plant of the present invention is cold; the eastern dry culture has been deposited at the Taiwan Food Industry Development Research Institute, and the detailed information of the storage at 33H, Food Road, Hsinchu City, Zhuguan, is shown in Table 1: BCRC910438

------L_^^J The two strains of lactic acid bacteria that have been deposited have been found to have activity against Helicobacter pylori, inhibiting the activity of Helicobacter pylori, and for H. pylori antibiotics such as diarrhea and Stomach discomfort _ such as flatulence, stomach cramps, etc.) have the function of slowing down and treating. Example 1: Anti-gastric micro (4) milk gn morphology and general properties. w _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Rods ^ Round, usually appearing separately, in pairs of short chains. 4 is positive for =_ color, does not produce spores, does not produce gas in both aerobic and anaerobic environments. Quality Dial 1. When the MRS medium is cultured, the ends are square and chained. A Lactobacillus faecalis ΜΗ-68 2· Gram-positive rod _ 'Do not generate lion lion and motility' in both aerobic and anaerobic environments 3: ^ Suitable growth temperature is mi〇c, belongs to both Sexually acidic acid-producing strains do not produce gas during glucose metabolism.

Examples of the effect of lactic acid bacteria against Helicobacter pylori on the activity of inhibiting the production of ammonia and carbon dioxide by Helicobacter pylori in the stomach 'mainly measuring the concentration of ammonia and inhibiting the percentage of Helicobacter pylori in the stomach (the urinary tract classification (4) For the in vitro work platform). The anti-stomach Η 料 _ _ 株 — 魏 魏 魏 魏 魏 魏 魏 32 32 32 32 32 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 对 对 对 对 对 对 对 对 对 对 对 对 对 对 对 对 对g was co-cultured with the aforementioned anti-Helicobacter pylori milk _ separately, and the concentration of the culture medium was measured, thereby verifying the ability of the above strain against H. pylori. The following experimental procedures were used: 1. The bacterial culture of Helicobacter pylori was cultured to reach an appropriate amount of bacteria. S1 2. Take appropriate amount of gastric sputum sputum and add appropriate amount of milk _ culture liquid broth or appropriate 7 201124146 1 month Helicobacter pylori bacteria solution into the appropriate amount of growth of the positive control group). 3. After mixing evenly, imL environment was cultured for 24 hours.

The liquid is added to the oblique fabric base for her culture method, 37. 〇 Anaerobic ring 4. Centrifuge the broth of step 3 at 3 rpm and conduct a sex test. 10 minutes, take the supernatant, carry out urease activity

PBS, pH 6.5. • 6. Take the supernatant from the Chuancui solution and add 3 L urease reaction buffer to the enzyme immunoassay (ELISA) dish and react at 37 ° C for 1 hour. 7. The absorbance value of the measurement example nm is required to use the urease reaction buffer as a blank control group. The absorbance value is the same as the liquid redness test. The better the activity of the Helicobacter pylori, the more urease is secreted. The statistical analysis of the data is shown in Table 3, Table 4, and Figures 1 and 2, and is expressed as the mean standard deviation (Mean ± SD). Fig. 1 and Fig. 2 show that Lactobacillus johnsonii MH-68 strain BCRC 910438 and 1〇8 are respectively taken from 1〇8 to 1〇9 coffee (Colony-Forming Units) Lactobacillus salivarius Αρ·32 strain BCRC91〇43l Up to 109 cfil (c〇k) ny_F_ing ® Units) Helicobacter pylori was co-cultured for 3 hours in an anaerobic environment for 24 hours, and the supernatant was taken for urease activity assay. The positive control group was AmoxiciHin antibiotic and the negative control group was 5.α?/ζ·HB101. As shown in Fig. 1, when the antibiotic concentration is 240 pg/mL, the ability of the Helicobacter pylori to secrete urease to produce ammonia is not inhibited, so the supernatant is alkaline, indicating

It is nearly five times more ammonia than Lactobacillus saliva AP-32 strain BCRC 910437, Lactobacillus johnsonii strain BCRC 910438 and 1200 pg/mL antibiotics, showing L, L. r/zamwioms and 240 pg/mL antibiotics. No Helicobacter pylori 201124146 was lost in biological activity. Fig. 2 uses -·ΗΒ1〇1 as a negative control group to calculate the inhibitory ability of lactic acid bacteria strains and antibiotics. The calculation method is # co-cultured with Helicobacter pylori

The above supernatant liquid sterilized and sterilized with the genus P. aeruginosa, the supernatant absorbs the light value) / five · HB101 and the Helicobacter pylori co-culture after the supernatant absorbance X100%. The calculated value is the percentage of the strain's inhibition of the biological activity of H. pylori. As shown in Fig. 2, the ability of Lactobacillus saliva sp. Ap_32 strain BCRC9l〇437, Lactobacillus johnsonii 68 g strain BCRC 91 coffee and 12 (8) (four) chaotic antibiotics to inhibit gastric pyloric helix (IV) is higher than the other three groups, showing saliva Lactobacillus his 32 strain BCRC all over 37, yolk lactic acid rod g _68 _BCRC 9_8 help to reduce the biological activity of Helicobacter pylori and further inhibit the growth of Helicobacter pylori. Table 3: 44林发败练 and control of the gastrointestinal pyogenes culture, the absorbance of the β liquid and the percentage of inhibition range (means ± SD): Table 3 supernatant absorbance, indicating the supernatant The name of the multi-reported strain containing ammonia concentration is L. gasseri 1.058±0.006 L. rhamnosus " ____________ 1.049±0.023 Lactobacillus saliva AP-32BCRC910437 ----- -—---0.230±0·027 Lactobacillus johnsonii MH-68 BCRC 910438 0.228±0.016 Antibiotic Amoxicillin 240 pg/mL —------ 1.068±0.034 Positive control group Amoxicillin 1200 pg/mL 0.255 ±0.003 - Lily of the biological activity range of H. pylori. Name of the strain - percentage L. gasseri ------- 1.298 ± 0.748 201124146 L. rhamnosus 2.814 ± 3.201 Lactobacillus saliva AP-32 BCRC 910437 79.690 ± 2.621 Lactobacillus MH-68 BCRC 910438 79.1〇7±1.239 Antibiotic Amoxicillin 240 pg/mL 2.325±1.949 Positive control group Amoxicillin 1200 pg/mL 76.183 ± 0.613 --- Example 3; Helicobacter pylori-resistant Helicobacter pylori inh〇use human clinical Test pretest

The urea pylorus secretes urease, decomposes urea, produces silk and carbon dioxide. Therefore, the breath test is to use this special exhibition. The principle of this method is that the gastric pyloric screw g is decomposed to produce isotope-labeled carbon dioxide after the urea sKc or labeled urea is taken down by the subject. Collect exhaled specimens, use liquid scintillation counter or gas nuclear mass spectrometer to mark the carbon dioxide, high sensitivity, quantifiable, patients are painless, the method is simple and fast, there is a small amount of fine for detecting the pyloric Paste 13c urinary gas test method was used to collect 16 patients who had been infected with Helicobacter pylori in the stomach, and a % breath test. Eight patients were identified as infected with Helicobacter pylori: 1. Determined that the patient has no stomach so far. Major symptoms of ulceration or gastric perforation. 2. Stop taking any products related to lactic acid bacteria four weeks before the test. 3. Take S. salivarius ΑΡ·32 g strain BCRC 9 purchase 7 and Lactobacillus johnsonii MH-68 _ strain BCRC 910438 capsules, the capsule content of each capsule is 2χ1〇9 cfli/capsule (C〇l〇ny_F_ing(5)灭 (5). In the morning and evening, each one will continue to take * week 0 4. After four weeks of taking, the uc breath test will be performed after two weeks of inactivity. 5. Observe the change of the 13C breath test value before and after taking the sample. And Figure 3, Figure 4, Figure 3 is the patient 13 (: breath test [S.1 10 201124146

The value distribution is checked; Figure 4 shows the 13C breath test value 'as expressed by Mean Shi SD. Figure 3 shows the distribution and changes of the l3C breath test values before and after administration of Lactobacillus salivarius AP-32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules. As shown in Fig. 4, 8 patients took the mean ± standard deviation of the 13C breath test values before and after taking Lactobacillus salivarius AP-32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules, and the results showed that the patient took After the Lactobacillus saliva AP-32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules, the 13C breath test value was significantly reduced, clearly showing the Lactobacillus saliva AP-32 strain BCRC 910437 and Lactobacillus johnii MH -68 strain BCRC

910438 inhibits the growth and physiological activity of Helicobacter pylori in humans. Table 5 shows the 13C breath test value (means ± SD) before and after the patient took Lactobacillus saliva AP-32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438: Table 5 Human clinical trial pre-test, taking The value of 13c breath test before and after the Lactobacillus saliva sp. Ap_32 strain bcrc 9HM37 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsule. DOB value (13C02 : 12C02 experimental value / 13co2: 12co2 reference 佶 23.71±14.45 18.73±12.59 test status

Taking Lactobacillus saliva AP-32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 before capsule

Taking Lactobacillus saliva AP_32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 After the sac sac, the strain can be found in the human body;; =; in addition to the ability to produce acid, the new person (four); and the reading _ spine The side of the anti-stomach _;; = can: ==: 201124146 The origin of the infection after the medical treatment _ way. The anti-gastric bacterium belonging to the present invention can simultaneously reduce the physiological activity of H. pylori and inhibit its growth. The invention is in the following: the goal is to continue to achieve these hopes or at least provide silk in the stomach pyloric spiral two treatments ^ in addition to antibiotics outside riding selection 'this hair (four) human body no side limbs beneficial ^ anti-stomach scorpion spiral ugly acid g As a new choice for the treatment of stomach pyloric (four) ^. The above-mentioned embodiments are only for explaining the technical idea and characteristics of the present invention, and those who are familiar with the art (4) understand the content of the present invention to implement, when: = limited forest (four) special face g, namely Dafanyi The variations or modifications made by the spirit of the present disclosure are still to be covered by the scope of the present invention. two

[Simple description of the figure] Figure 1 shows that 1 〇 8 to 1 〇 9 cfU (c〇1〇ny F_ing Un(8) saliva milk

Acid bacillus AP-32 strain BCRC 910437 or yolk lactic acid rod mh_68 strain BCRc 2:38 and 1〇8 to 1〇9 cfij(c〇1〇ny_F〇rmingUn(4) Helicobacter pylori culture in anaerobic environment 37 °C '24 small light, take the clear urine urinary surface activity analysis test. Positive control group for Amoxicillin antibiotics. Antibiotic bromine is applied (four) mL, and can not inhibit the stomach _ _ _ _ self-secretion urinary fine lineage Therefore, the supernatant is alkaline, indicating that the supernatant contains a high concentration of ammonia.: wen·, (10) and 24〇Mg/mL antibiotics are all more than Lactobacillus sp. Ap_32 strain BCRC 91〇437, Lactobacillus johnii The MH-68 strain BCRC 910438 and 1200 pg/mL antibiotics were nearly 5 times higher than the ammonia concentration. Figure 2 shows that '1,8 to 1〇9 pools (c〇i〇ny_F〇rmingUnits) Lactobacillus saliva AP- 32 strain BCRC 910437 or Lactobacillus johnsonii MH-68 strain BCRC 910438 and 1〇8 to 1〇9 cfU (Colony-Forming Units) Helicobacter pylori co-cultivation 37 C under anaerobic conditions, 24 hours later, The supernatant was taken for urease activity analysis test. The positive control group was Amoxicillin antibiotic, negative control group. The control method is as follows: (five and stomach secluence 201124146 door spiral (four) co-cultivation after nurturing new milk · milk _ with stomach sec (10) ^ after training Liquid absorbance value) / wu· μ μ i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i · 32 _BCRC91_, yolk lactic acid rod® ΜΗ·68 miscellaneous C91 Gang and deputy kisses have the ability to produce Helicobacter pylori in the stomach more than 76%.

Figure 3 shows the distribution and variation of the % exhalation test values before and after administration of Lactobacillus salivarius from 32 strains of BCRC 91〇437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules. The patient's % breath test value was significantly lower before and after taking Lactobacillus saliva ap_32 strain bcrc 910437 and Lactobacillus johnsonii strain BCRC 91〇438 capsule, clearly showing Lactobacillus saliva sp. Ap_32 strain BCRC 910437 and Lactobacillus johnsonii mh- The 68 strain BCRC 91〇438 inhibits the growth and physiological activity of Helicobacter pylori in humans. Figure 4 shows the mean soil standard deviation of the 13C breath test values before and after administration of the Lactobacillus saliva sp. Ap_32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules. The patient's 13C beer test values were significantly reduced before and after taking Lactobacillus saliva AP_32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 strain BCRC 910438 capsules. 'Clearing expression of Lactobacillus saliva AP_32 strain BCRC 910437 and Lactobacillus johnsonii MH-68 The strain BCRC 910438 inhibits the growth and physiological activity of the pyloric snail in the human body. [Main component symbol description] None [si 13

Claims (1)

201124146 VII. Patent application scope: 1. A food composition comprising: a lactic acid bacteria strain for treating gastric ulcer, the strain being selected from the biological culture of any one or more of the following combinations: Lactobacillus saliva, such as) ^_32 strain, ^ Deposit number BCRC 91〇437 and Yogurt lactic acid pole mail (10) ▲ Qin (four) _·〇 age 68 strain, storage number BCRC91〇438, the above strains are deposited in Taiwan Hsinchu Food Industry Development Institute (FIRDI); and physiologically acceptable Excipient or diluent. 2. The food composition of claim 1, wherein the lactic acid strain is Lactobacillus salivarius AP-32 strain, accession number BCRC91〇437. 3. The food composition according to claim 3, wherein the lactic acid strain is Lactobacillus johnsonii MH-68 strain storage number BCRC91〇438. The food composition according to the above item 1, wherein the excipient or diluent is a food. The food composition of claim 4, wherein the food comprises a lactic acid emulsion milk powder, tea, B plus or a combination of the above. , yoghurt, cheese, 6. The food composition according to claim 1, strain. Wherein the lactic acid bacteria strain is an active bacteria A pharmaceutical composition for treating gastric ulcer, comprising: a lactic acid plant for treating gastric ulcer, the strain is selected from the following __ or redundant: ^mim^mLactobacillussalivanus) AP-32 m > ^ 9 fine and yolk lactic acid Bacillus (z_ is a pharmaceutically acceptable excipient or diluent. 8. The medicinal conjugate as described in claim 7, the side strain, the registered number of the servant 7. The partial strain is Lactobacillus saliva. The pharmaceutical composition according to the above-mentioned item (4), wherein the lactic acid strain is a pharmaceutical composition according to claim 7, wherein the lactic acid bacteria strain is An active strain. 15