JP7361752B2 - Complex probiotic composition for relief of overactive bladder related symptoms - Google Patents

Description

NPMD NITE BP-03316 CGMCC CGMCC 15203 FIRDI BCRC 910919 NPMD NITE BP-03311 FIRDI BCRC 910876

The present invention relates to a probiotic composition, and particularly to a composite probiotic composition for alleviating symptoms associated with overactive bladder, which can delay urination frequency, reduce bladder pressure, increase bladder capacity, and the like.

Overactive bladder syndrome (OAB) is the sudden non-autonomous contraction of the bladder that occurs during the urine storage process, resulting in an urgent need to urinate immediately, and patients generally do not reach the toilet in time to urinate. Refers to the phenomenon of leakage. Common noticeable symptoms include urinary urgency, increased frequency of urination (also called frequency), nocturia, and urge urinary incontinence. According to the International Continence Society (ICS), frequent urination refers to urinating eight or more times a day and urinating more than once during bedwetting. Nocturia refers to interrupting sleep and getting up to urinate in the middle of the night due to a sudden urge to urinate. Urinary urgency refers to the sudden and strong desire to urinate. Urinary incontinence refers to the involuntary passing of urine, or the inability to reach the toilet in time.

Overactive bladder can have a serious impact on quality of life, including decreased sexual intercourse, inability to travel, exercise, work, feelings of guilt or decreased self-esteem, and has a negative impact on body, mind, and spirit. . In general, the incidence of overactive bladder attacks increases with age, and age-related decreases in bladder elasticity (detrusor relaxation), decrease in bladder capacity, weakening of pelvic floor muscles due to loss of estrogen, and urinary resistance due to prostate enlargement. , related to neurological disorders, etc. According to Taiwanese statistics, about 18% of the adult population over the age of 20 suffers from these problems, given that older people in Chinese society are more conservative in seeking treatment for urinary tract-related diseases. However, the actual proportion of patients with overactive bladder is likely to be much higher.

Treatment for overactive bladder can be divided into behavioral therapy and drug therapy. Behavioral therapy includes improving your physical health (improving other complications), controlling your eating habits, controlling your fluid intake, regular urination, and improving your bladder to avoid the overall causes of overactive bladder. These include training, pelvic floor exercises, weight loss, and smoking cessation. Medication is used to treat patients with mild symptoms. Common drugs include muscarinic antagonists, anticholinergics, calcium antagonists, muscle relaxants, and the like. Most of them achieve their modulatory effect by blocking some of the transmissions of the nervous system. However, these therapeutic agents inevitably have side effects such as dry mouth, constipation, dizziness, blurred vision, and impaired cognitive function, are difficult to use for a long period of time, and are not suitable for everyone.
Many functions of probiotics are gradually being discovered and reported, and the functions and characteristics vary depending on the strain. Studies have shown that in an obese mouse model, Lactobacillus plantarum helps reduce mouse body weight and metabolic indicators such as blood lipids (Hus et al., 2019), and in an asthmatic animal disease model, Lactobacillus plantarum helps reduce allergy. It has the effect of improving immunity and improving immunity (Shih et al., 2019). A study also found that Lactobacillus plantarum (L. plantarum) has the ability to slow muscle loss in a mouse model of sarcopenia (Li et al., 2020). Research has also found that Lactobacillus paracasei can delay aging indicators in aging mice and has an anti-aging effect (Lin et al., 2021). In recent years, many scholars have further linked the brain-gut axis to explain the mechanism of probiotics for neuromodulation. For example, studies have indicated that feeding Lactobacillus brevis or Lactobacillus fermentum can improve abnormal behavior in mice with depressive disorder caused by confinement. (Chen et al., 2021; Lin et al., 2021).

Hus et al. ,2019

However, there has been little research on probiotics for overactive bladder, and there is an urgent need to develop a composite probiotic composition and its uses for alleviating symptoms associated with overactive bladder.

Accordingly, one aspect of the present invention provides Lactobacillus brevis strain GKJOY, L. fermentum strain GKF3, L. plantarum strain GKK1, and Pediococcus lactis acidilactic ) A composite probiotic composition for alleviating symptoms associated with overactive bladder comprising strain GKA4 is provided.

Another aspect of the present invention provides a composite probiotic for use in alleviating symptoms associated with overactive bladder, and continuously administers an oral composition containing an effective dose of the composite probiotic containing the above-mentioned strain. It involves administering it to a subject over a period of time.

According to the above aspects of the invention, a combined probiotic composition for alleviation of overactive bladder related symptoms is provided. In one embodiment, the composite probiotic composition for alleviation of overactive bladder related symptoms comprises Lactobacillus brevis strain GKJOY, L. fermentum strain GKF3, Lactobacillus plantarum (L. Lactobacillus brevis strain GKJOY, including L. plantarum strain GKK1 and Pediococcus acidilactici strain GKA4, was approved by the National Institute of Technology and Evaluation on November 6, 2020. Chnology and Evaluation ; NITE) National Patent Microorganisms Depositary, Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818, and the accession number is NITE BP-0331 6 . Lactobacillus fermentum strain GKF3 was deposited at the China Common Microorganisms Storage Management Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, China on January 12, 2018, and the accession number is CGMCC 15203. Bacillus plantarum strain GKK1 was deposited with the Food Industry Development Institute Biological Resource Conservation Research Center (BCRC, No. 331 Food Road Hsinchu, Taiwan) on July 18, 2019, and the accession number is BCRC 910919. Diococcus lactis strain GKA4 was deposited at NITE on November 6, 2020, and the accession number is NITE BP-03311.

In the above embodiments, the composite probiotic composition may be an oral composition, such as a pharmaceutical composition or a food composition. In the above example, the composite probiotic composition further includes pharmaceutically and food-acceptable carriers, excipients, and/or additives.

In the above embodiments, the dosage form of the composite probiotic composition may include, for example, a powder, tablet, granule, suppository, microcapsule, ampoule, liquid spray, or suppository.

In the above embodiments, the overactive bladder-related symptoms include increased urination frequency, increased bladder pressure, and/or decreased bladder capacity.

Another aspect of the invention includes an oral composition containing an effective dose of a complex probiotic, wherein the oral composition is continuously administered to a subject for at least 30 days, and the complex probiotic is , Lactobacillus brevis strain GKJOY whose accession number is NITE BP-0331 6 , Lactobacillus fermentum strain GKF3 whose accession number is CGMCC 15203, Lactobacillus plantarum strain GKK1 whose accession number is BCRC 910919, and Lactobacillus plantarum strain GKK1 whose accession number is BCRC 910876. A composite probiotic composition for alleviating symptoms associated with overactive bladder is provided, comprising Pediococcus lactis strain GKA4.

In the above example, the subject may be, for example, an adult, and the effective dose of the complex probiotic for this subject is, for example, 150 mg/60 kg body weight/day to 600 mg/60 kg body weight/day. good.

In the above embodiment, the subject may be, for example, a rat, and the effective dose of the complex probiotic to this subject is, for example, from 5 mg/kg body weight/day to 20 mg/kg body weight/day. good.

The composite probiotic composition of the present invention for alleviating the above-mentioned overactive bladder-related symptoms, comprising Lactobacillus brevis strain GKJOY, Lactobacillus fermentum strain GKF3, Lactobacillus plantarum strain GKK1, and Pediococcus lactis strain GKA4. And if its application is applied, it can be continuously administered to subjects for a certain period of time to effectively delay urination frequency, reduce bladder pressure and increase bladder capacity, and further prepare various compositions as active ingredients. It can be used for

It should be understood that the foregoing general description and the following detailed description are exemplary only and are intended to provide further explanation of the claimed invention.
In order to make the above and other objects, features, advantages and embodiments of the present invention more clearly understood, the accompanying drawings are described below.

FIG. 1A is a curve diagram showing the bladder pressure of animals in a control group (FIG. 1A) and an experimental group (FIG. 1B) before and after administration of physiological saline according to an embodiment of the present invention, and the figure number ▲ indicates the time point of administration of physiological saline. shows. FIG. 1A is a curve diagram showing the bladder pressure of animals in a control group (FIG. 1A) and an experimental group (FIG. 1B) before and after administration of physiological saline according to an embodiment of the present invention, and the figure number ▲ indicates the time point of administration of physiological saline. shows. FIG. 2 is a curve diagram showing the bladder pressure of animals in the control group (FIG. 2A) and the experimental group (FIG. 2B) after administration of physiological saline and after a stabilization time, respectively, according to another embodiment of the present invention, and the figure number is ▲ indicates the time point of saline administration. FIG. 2 is a curve diagram showing the bladder pressure of animals in the control group (FIG. 2A) and the experimental group (FIG. 2B) after administration of physiological saline and after a stabilization time, respectively, according to another embodiment of the present invention, and the figure number is ▲ indicates the time point of saline administration. 1 is a bar graph showing the bladder capacity of animals in a control group and an experimental group according to an embodiment of the present invention. 1 is a bar graph showing the urination frequency of animals in a control group and an experimental group according to an embodiment of the present invention.

If a definition or usage of a term in a cited document is inconsistent with or contrary to the definition of the term herein, the definition for the term herein shall apply in place of the definition of the term in the cited document. In turn, singular terms may include plural terms and plural terms may include singular terms, unless the context clearly dictates otherwise.

As mentioned above, the present invention, after being continuously administered to a subject for a certain period of time, can effectively delay urinary frequency, reduce bladder pressure and increase bladder capacity in the treatment of overactive bladder related symptoms. A complex probiotic composition for palliation is provided.

In one example, "complex probiotics" as referred to herein may refer to probiotics that include different species or different genera, such as Lactobacillus brevis strain GKJOY, Lactobacillus fermentum ( L.fermentum strain GKF3, Lactobacillus plantarum strain GKK1, and Pediococcus acidilactici strain GKA4. Lactobacillus brevis strain GKJOY was deposited at the Food Industry Development Institute Biological Resource Conservation Research Center (BCRC, No. 331 Food Road Hsinchu, Taiwan) on July 18, 2019, and the accession number is BCRC 910920. The survival of the strain was confirmed on July 26th. Lactobacillus brevis strain GKJOY has been separately deposited at the National Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (NITE), an independent administrative agency. ositary, address: 292-0818 Kazusakama, Kisarazu City, Chiba Prefecture, Japan 2-5-8 Room 122), the deposit date is November 6, 2020, and the accession number is NITE BP-03316, and the survival of the strain was confirmed on November 11, 2020. Ta. Lactobacillus fermentum strain GKF3 was deposited with BCRC on February 12, 2018, and the accession number is BCRC 910824, and the survival of the strain was confirmed on March 1, 2018. Lactobacillus fermentum GKF3 was separately deposited at the China Common Microorganism Species Storage Management Center, No. 3, Hospital No. 1, Beichen West Road, Chaoyang District, Beijing, China, with the deposit date being January 12, 2018, and the accession number being CGMCC. It is 15203. Lactobacillus plantarum strain GKK1 was deposited with BCRC on July 18, 2019, and the accession number is BCRC 910919, and the survival of the strain was confirmed on July 26, 2019. Pediococcus lactis strain GKA4 was deposited at NITE on November 6, 2020, and the accession number is NITE BP-03311, and the survival of the strain was confirmed on November 11, 2020. It should be noted that in the present invention, it is necessary to select a composite probiotic of a specific strain, and only by this, the resulting composite probiotic composition can exert effects such as alleviating symptoms related to overactive bladder. can do. Complex probiotics cannot be expected to be effective in alleviating symptoms of overactive bladder if they are not selected from the specific strains listed above, if the strains are increased or decreased, or if some or all of them are replaced with other strains of the same species. .

The term "overactive bladder-related symptoms" as referred to herein refers to the main symptoms of overactive bladder and symptoms caused by it. The primary symptom is, for example, urine urgency, and associated symptoms may include, but are not limited to, increased urinary frequency, increased bladder pressure, and/or decreased bladder capacity.

In one embodiment, the content of each strain of the composite probiotic is not particularly limited. In one example, the weight of Lactobacillus brevis strain GKJOY (BCRC 910920), Lactobacillus fermentum strain GKF3 (BCRC 910824), Lactobacillus plantarum strain GKK1 (BCRC 910919), and Pediococcus lactis strain GKA4 (NITE BP-03311) ( mg) ratio or bacterial count (CFU) ratio may be, for example, 1:1:1:1, but in other examples, the four strains described above may have a ratio other than 1:1:1:1. May be adopted.

When applied, the complex probiotic composition is an oral composition, such as a pharmaceutical composition or a food composition. The usage modes of the composite probiotics may include, but are not limited to, whole fermentation broth, bacterial slurry, supernatant liquid, and lyophilized powder. In the above examples, the entire fermentation liquid refers to a product containing bacterial cells and a culture liquid. The bacterial slurry refers to the product after removing the supernatant from the entire fermentation liquor. The supernatant liquid refers to the product after removing the bacterial slurry from the entire fermentation liquid. The freeze-dried powder refers to a freeze-dried powder prepared from the entire fermentation liquid, bacterial slurry and/or supernatant liquid.

Examples of applications for the pharmaceutical composition may include, but are not limited to, pharmaceuticals. Examples where complex probiotics are applied to food compositions may include, but are not limited to, general foods, health foods, beverages, nutritional supplements, dairy products, or feeds. In the example of the oral composition, the composite probiotic composition may optionally further include pharmaceutically and food-acceptable carriers, excipients and/or additives. In other examples, dosage forms of the composite probiotic composition may include, but are not limited to, powders, tablets, granules, suppositories, microcapsules, ampoules, liquid sprays, or suppositories.

When using the composite probiotics described above for alleviating symptoms associated with overactive bladder, an oral composition containing an effective dose of the composite probiotics can be continuously administered to a subject over a certain period of time. The effective dosage varies depending on the subject and is not particularly limited. For example, when the subject is an adult, the effective dose of the composite probiotic for adults is, for example, 150 mg/60 kg body weight/day to 600 mg/60 kg body weight/day, but 225 mg/60 kg body weight/day to 450 mg. /60 kg body weight/day, and more preferably about 300 mg/60 kg body weight/day. In another example, if the subject is a rat, an effective dose of the complex probiotic to this subject is, for example, 5 mg/kg body weight/day to 20 mg/kg body weight/day, but 7.5 mg/kg body weight/day. Preferably, the amount is between 15 mg/kg body weight/day, and more preferably about 10 mg/kg body weight/day. In other examples, the period of administration can be, for example, at least 30 consecutive days.

It should be understood that the following specific strains, specific formulations, specific dosages used, specific detection methods, aspects, illustrations and examples are illustrative only and are not subject to the limitations of the present invention. do not have. Without departing from the spirit and scope of the present invention, the technical characteristics necessary for the present application can be easily determined, and the present invention can be made with various changes and modifications and applied to different uses and conditions. Ru.
Example 1
1.1 Bacterial species source

The bacterial strains used in this example include a specific combination of probiotics, including Lactobacillus brevis strains GKJOY, Lactobacillus fermentum of the genus Lactobacillus spp. These include strain GKF3, Lactobacillus plantarum strain GKK1, and Pediococcus acidilactici strain GKA4 of the Pediococcus spp. Lactobacillus brevis strain GKJOY was deposited at the Food Industry Development Institute Biological Resource Conservation Research Center (BCRC, No. 331 Food Road Hsinchu, Taiwan) on July 18, 2019, and the accession number is BCRC 910920. The survival of the strain was confirmed on July 26th. Lactobacillus brevis strain GKJOY has been separately deposited at the National Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (NITE), an independent administrative agency. ositary, address: 292-0818 Kazusakama, Kisarazu City, Chiba Prefecture, Japan 2-5-8 Room 122), the deposit date is November 6, 2020, and the accession number is NITE BP-03316, and the survival of the strain was confirmed on November 11, 2020. Ta. Lactobacillus fermentum strain GKF3 was deposited with BCRC on February 12, 2018, and the accession number is BCRC 910824, and the survival of the strain was confirmed on March 1, 2018. Lactobacillus fermentum GKF3 was separately deposited at the China Common Microorganism Species Storage Management Center, No. 3, Hospital No. 1, Beichen West Road, Chaoyang District, Beijing, China, with the deposit date being January 12, 2018, and the accession number being CGMCC. It is 15203. Lactobacillus plantarum strain GKK1 was deposited with BCRC on July 18, 2019, and the accession number is BCRC 910919, and the survival of the strain was confirmed on July 26, 2019. Pediococcus lactis strain GKA4 was deposited at NITE on November 6, 2020, and the accession number is NITE BP-03311, and the survival of the strain was confirmed on November 11, 2020.

The above bacterial species are all from pickles from different regions. As an example, strain GKJOY was isolated from sauerkraut in the Taoyuan region. Strain GKF3 was isolated from Taiwanese kimchi in the Hsinchu region. Strain GKK1 and strain GKA4 were isolated from sour pepper in the Hualien region.

In some examples, the strain GKF 3 may be collected from traditional pickled fermented kimchi commercially available in Beipu, Hsinchu, Taiwan. Various fruit and vegetable kimchi products that had been naturally fermented for more than 30 days were sampled, collected, packaged, labeled, stored in a refrigerator at low temperatures, and transported to the laboratory on the same day to isolate bacterial strains. Then, the filtrate or leaves of vegetables and fruits in the sample bag were extracted aseptically and homogenized with sterilized water equivalent to 10 times the volume of the sample. Take 1 mL of the diluted solution, place it in MRS medium containing 0.1% bromocresol green, and perform enrichment culture at 37°C for 16 hours to increase the number of Lactobacillus contained in the sample. , which enhanced the probability that low-number lactic acid bacteria strains in the screening samples would be isolated. Next, a sample was taken after the enrichment culture and a sequence dilution operation was performed. After diluting the sequence to 100,000 times and 1 million times, take 0.1 mL of each and place them in a sterile petri dish, and place in each dish Rogosa medium agar, BIM-25 agar, potato starch + yeast extract agar, bromocresol green MRS. Agar and bromocresol purple agar were added. After 40 hours of anaerobic cultivation at 37°C, single colonies of 15 to 20 bacterial strains were selected from each type of culture medium. The single strain culture after isolation was incubated in MRS broth at a specific temperature for 16 hours before adding 50% strain cryopreservation solution (glycerin).

The Lactobacillus or Pediococcus bacteria were each inoculated onto a solid medium to activate the bacterial species. In a preferred embodiment, the solid medium is MRS agar. After colonies were generated, a single colony was taken and placed in a liquid medium for liquid culture. In a preferred embodiment, the culture temperature of the strain is 25°C to 40°C, and the liquid culture is carried out for 16 to 24 hours under conditions such as nitrogen or carbon dioxide with an aeration rate of 0 to 1 vvm, and a shaking rate of 250 rpm to 1000 rpm. The number of viable bacteria reached the maximum after 18 hours. The liquid medium is MRS (deMan, Rogosa, and Sharpe) liquid medium. After the liquid culture was completed, fermentation culture was carried out, and the composition of the fermentation medium was shown in Table 1.

１．２凍結乾燥粉末の調製

1.2 Preparation of freeze-dried powder

After the growth of the above-mentioned Lactobacillus or Pediococcus is completed by fermentation culture, the whole fermentation liquid including the bacterial cells and the culture liquid is collected and centrifuged at a speed of 1000 rpm to 15000 rpm to obtain a cell pellet. was gotten. The obtained bacterial cell pellets and a protective agent (ie, 6% to 50% by weight skim milk powder) were mixed and then freeze-dried. The freeze-drying was performed by pre-freezing in a gradient setting mode, sequentially storing at 20°C to 0°C for 1 to 4 hours, and then storing at 0°C to -20°C for 4 to 8 hours. and then stored at -196°C to -30°C for more than 8 hours. In a preferred embodiment, the freeze-drying temperature and time are -40°C for 2 hours, -20°C for 2 hours, 0°C for 2 hours, and finally 20 hours. It was stored at °C for about 10 hours or more. The lyophilized powder obtained above was then stored at low temperature. In another preferred embodiment, the cryopreservation temperature is between -30°C and 4°C. The stored lyophilized powder was used as the raw material for a composite probiotic composition administered to experimental animals in the following animal experiments.
1.3 Experimental animals

For the following examples, 0 week old Wistar male rats, each weighing approximately 300 grams, purchased from BioLASCO (LASCO Biotechnology Company) were selected. Rats are housed in open cages with 2-3 rats per cage. The rat rooms had an average temperature of approximately 22±2° C., an average humidity of 35±5%, normal 12 hours of daylight and 12 hours of darkness, and were provided with sufficient drinking water and standard chow. All animal experimental procedures were reviewed and approved by the Institute of Agricultural Science and Technology Laboratory Animal Care and Use Committee (IACUC), and approval number Jidooji No. 109099 was obtained. After one week of acclimatization, the test animals were randomly divided into control and experimental groups, with 7 animals in each group.
1.4 Experimental design

After holding the experimental animals with bare hands, 0.9% physiological saline (control group) or the composite probiotic composition (experimental group) was orally administered daily for 30 consecutive days. The dose of the composite probiotic composition administered to the experimental animals in the experimental group was 10 mg/kg body weight (B.W.), which corresponds to an oral dose of 300 mg per day for an adult weighing 60 kg. After one month of continuous administration, experimental animals were anesthetized with 3-4% isoflurane. A small incision was made in the lower abdomen of the experimental animal to expose the bladder and a small opening was cut in the upper end of the bladder with ophthalmic scissors. Insert the PE50 catheter (BB31695-PE/3) into the upper opening of the bladder and connect the opening to the PE catheter with a surgical tie wire (SW SERALON) until the pressure value of the MP150 monitor system (MP150 Systems, Biopac) is stable. , saline was injected into the bladder along the PE50 catheter at a rate of 0.04 ml/min using an infusion pump (Syringe Pump, KD Scientific® Syringe Pump, KDS230) to control the bladder pressure ( bladder pressure, bladder capacity, urination frequency, and urination delay index.
1.5 Data statistics

The numerical values described in the text are expressed as mean ± standard deviation (mean ± SD) and were analyzed using one-way ANOVA and commercially available software, and the statistical results were determined by p < 0.05. This indicates that there is a significant difference in terms of
Example 2
2.1 Detection of bladder pressure

This example uses the MP150 detection system to detect pressure values in the bladder of experimental animals without injecting physiological saline, pressure values in the bladder after injecting physiological saline, and pressure values in the bladder after injecting physiological saline and after stabilization. Bladder volume pressure (cystomertrogram) including bladder pressure values over time was detected. Bladder pressure was expressed in millimeters of mercury (mmHg). The stabilization time refers to the time required for the bladder pressure to stabilize, and the stabilization time for experimental animals in the control group was 3 minutes, while the stabilization time for experimental animals in the experimental group was 1 minute.

Please refer to FIGS. 1A and 1B, which are curve diagrams showing bladder pressures of animals in a control group (FIG. 1A) and an experimental group (FIG. 1B) before and after administration of physiological saline according to an embodiment of the present invention, respectively; The figure number ▲ indicates the time point at which saline is injected.

As can be seen from the results in Figure 1A, before the saline was administered to the experimental animals in the control group, their bladder pressure values were relatively low (<5 mmHg), and after the saline was administered, their bladder pressure values were relatively low (<5 mmHg). The pressure value increased immediately, and the disturbance in the pressure value continued for about 3 minutes, and the maximum bladder pressure value detected exceeded 20 mmHg, and after about 3 minutes it gradually stabilized, and the bladder pressure value was less than 10 mmHg. Became.

As can be seen from the results in Figure 1B, the experimental animals in the experimental group had their bladder pressure values (7.5 mmHg) higher than the control group, but within the standard range, before saline was administered. . After saline was administered, the pressure value gradually increased up to 17 mmHg, which was significantly lower than the control group and stabilized within 1 minute (approximately 8 mmHg).

Refer to FIGS. 2A and 2B, the bladders of animals in the control group (FIG. 2A) and experimental group (FIG. 2B) after administration of saline and after a stabilization period, respectively, according to another embodiment of the present invention. It is a curve diagram showing pressure, and the figure number ▲ indicates the time point at which physiological saline is injected.

As can be seen from the results in Figure 2A, the experimental animals in the control group had a bladder pressure value of about 9 mmHg after being continuously administered with physiological saline, and the pressure was still stable, but the pressure curve Bandwidth is disrupted.

As can be seen from the results in Figure 2B, after the experimental animals in the experimental group were continuously administered physiological saline, the bladder pressure value was also about 9 mmHg, but the band of the pressure curve was stable and regular. It is.

According to the above results, when test animals were given a complex probiotic composition for a certain period of time and then given physiological saline, the bladder pressure value was less affected, and urination was influenced by the bladder pressure value. This shows that behavior can be moderated.
2.2 Detection of bladder capacity

In this example, the first micturition time of each experimental animal was recorded and its bladder capacity was obtained after calculating at a flow rate of 0.04 mL/min. Bladder capacity was expressed in milliliters (mL) and the results are shown in Figure 3.

Please refer to FIG. 3, which is a bar graph showing the bladder capacity of animals in a control group and an experimental group according to one embodiment of the present invention. As can be seen from the results in Figure 3, although a statistically significant difference in bladder capacity between the experimental group and the control group was not reached, the experimental group of experimental animals to which the composite probiotic composition was administered The capacity was greater than that of the control group and there was a tendency to increase bladder capacity.
2.3 Detection of urination frequency

In this example, the time and frequency of urination of each experimental animal was recorded, and the statistics were analyzed using the number of times of urination per minute (times/min). Concerning the observation of animal urination, continuous discharge from the urethral meatus is defined as a micturition movement, and if only the urethral meatus is moist and there is no continuous discharge, it is not considered urination, and the results are shown in Figure 4. shown.

Please refer to FIG. 4, which is a bar graph showing the urination frequency of animals in a control group and an experimental group according to an embodiment of the present invention. As can be seen from the results in Figure 4, although a statistically significant difference in urination frequency between the experimental group and the control group was not reached, the experimental group's experimental animals to which the composite probiotic composition was administered showed a higher urinary frequency. compared with the control group, and there was a tendency to reduce the frequency of urination.
2.4 Detection of micturition delay index

This example compares the bladder capacity of experimental animals in the experimental group and the control group to obtain a urinary delay index, which is expressed as a fold. As can be seen from the results of the micturition delay index, the experimental set of experimental animals administered the composite probiotic composition had a bladder capacity approximately 1.2 times that of the control group, which increased micturition delay (i.e. tended to delay urinary frequency).

In summary, in the present invention, a specific strain, a specific formulation, a specific dosage used, a specific detection method, or a specific evaluation method are used to describe the complex probiotics for alleviation of overactive bladder related symptoms, by way of example only. It describes the composition and its uses. However, those skilled in the art will recognize that other strains, other formulations, other dosages, other detection methods, or other methods of evaluation of overactive bladder-related symptoms may be used without departing from the spirit and scope of the present invention. Applicable and not limited to the palliative complex probiotic composition and its uses. By way of example, the complex probiotic composition may be, for example, a pharmaceutical composition or a food composition and may optionally include pharmaceutically and food-acceptable carriers, excipients and/or additives; It may be manufactured into dosage forms such as powders, tablets, granules, suppositories, microcapsules, ampoules, liquid sprays, or suppositories.

According to the above examples, the composite probiotic composition for alleviating overactive bladder-related symptoms of the present invention has the following advantages, including Lactobacillus brevis strain GKJOY, Lactobacillus fermentum strain GKF3, and Lactobacillus plantarum. Containing strain GKK1 and Pediococcus lactis strain GKA4, it is continuously administered to subjects for a certain period of time, effectively delaying urination frequency, reducing bladder pressure and increasing bladder capacity, and in the future, it can be used as an active ingredient in various It can be used in the preparation of compositions.

Although the invention has been disclosed above in several specific embodiments, other embodiments may also be implemented. Therefore, the spirit and scope of the following claims should not be limited to the description of the embodiments contained herein.

[Deposit of biological materials]
Domestic deposit information (please note in order of depository organization, date, and number)
Lactobacillus brevis strain GKJOY was deposited at the Bioresource Conservation Research Center of the Food Industry Development Institute, China Foundation, No. 331 Hsinchu Food Road, Taiwan, and the deposit date was July 18, 2019, and the accession number was BCRC 910920, and the survival of the strain was confirmed on July 26, 2019.
Lactobacillus fermentum (L.fermentum) strain GKF3 was deposited at the Biological Resources Conservation Research Center, Institute of Food Industry Development, China Foundation, No. 331 Hsinchu Food Road, Taiwan, and the deposit date was February 12, 2018. The strain is numbered BCRC 910824, and the survival of the strain was confirmed on March 1, 2018.
Lactobacillus plantarum (L. plantarum) strain GKK1 was deposited at the Biological Resource Conservation Research Center, Institute of Food Industry Development, China Foundation, No. 331 Hsinchu Food Road, Taiwan, and the deposit date was July 18, 2019. The number is BCRC 910919, and the survival of the strain was confirmed on July 26, 2019.
Pediococcus acidilactici strain GKA4 was deposited at NITE on November 6, 2020, with the accession number NITE BP-03311, and the survival of the strain was confirmed on November 11, 2020.
Foreign deposit information (please note in the following order: country of deposit, organization, date, number)
Lactobacillus brevis strain GKJOY has been deposited at the National Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (NITE), an independent administrative agency in Japan. t Microorganisms Depositary, Address: 292-0818 Kisarazu, Chiba Prefecture, Japan Ichikazusa Kamatari 2-5-8 Room 122), the deposit date is November 6, 2020, the accession number is NITE BP-03316, and the survival of the strain was confirmed on November 11, 2020. has been confirmed.
Lactobacillus fermentum GKF3 has been deposited at the China Common Microorganism Species Storage Management Center, No. 3, Hospital No. 1, Beichen West Road, Chaoyang District, Beijing, China, and the deposit date is January 12, 2018, and the accession number is CGMCC 15203. It is.

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Claims (9)

Lactobacillus brevis strain GKJOY, L. fermentum strain GKF3, L. plantarum strain GKK1, and Pediococcus acidilact ici) containing strain GKA4, the Lactobacillus Brevis strain GKJOY was submitted to the National Institute of Technology and Evaluation (NITE) National Patent Microorganisms Depository on November 6, 2020. Depositary, Address: 292-0818 Chiba Prefecture, Japan The Lactobacillus fermentum strain GKF3 was deposited at Kazusa Kamatari 2-5-8 Room 122, Kisarazu City, China, and the accession number is NITE BP-03316. The Lactobacillus plantarum strain GKK1 was deposited at the China Common Microorganism Species Storage Management Center, No. 3, Hospital No. 1, Road 1, and the accession number is CGMCC 15203. The Pediococcus lactis strain GKA4 was deposited with the Biological Resource Conservation Research Center (BCRC, No. 331 Food Road, Hsinchu, Taiwan) with the accession number BCRC 910919, and the Pediococcus lactis strain GKA4 was deposited with NITE on November 6, 2020. A composite probiotic composition for alleviation of overactive bladder-related symptoms, numbered NITE BP-03311, wherein the composite probiotic composition is an oral composition, and the overactive bladder-related symptoms include: , a complex probiotic composition for the relief of overactive bladder-related symptoms, including increased bladder pressure. The composite probiotic composition for alleviating symptoms associated with overactive bladder according to claim 1, wherein the composite probiotic composition is a pharmaceutical composition or a food composition. The composite probiotic composition for alleviating overactive bladder-related symptoms according to claim 1, further comprising pharmaceutically and food-acceptable carriers, excipients and/or additives. The combined probiotic composition for alleviation of overactive bladder related symptoms according to claim 1, wherein the dosage form of the combined probiotic composition comprises a powder, a tablet, a granule, a suppository, a microcapsule, an ampoule, or a liquid spray. biotic composition. The composite probiotic composition for alleviating overactive bladder-related symptoms according to claim 1, wherein the overactive bladder-related symptoms further include increased urination frequency and/or decreased bladder capacity. an oral composition containing an effective dose of a complex probiotic, said oral composition being continuously administered to a subject for at least 30 days, and said complex probiotic having accession number NITE BP-03316. Lactobacillus brevis strain GKJOY, Lactobacillus fermentum strain GKF3 with accession number CGMCC 15203, Lactobacillus plantarum strain GKK1 with accession number BCRC 910919, and Pediococcus lactis strain GKA4 with accession number BCRC 910876. A composite probiotic composition for the relief of overactive bladder-related symptoms, wherein the overactive bladder-related symptoms include increased bladder pressure. . 7. The composite probiotic composition for alleviating overactive bladder-related symptoms according to claim 6, wherein the overactive bladder-related symptoms further include increased urination frequency and/or decreased bladder capacity. Overactive bladder related symptoms according to claim 6, wherein the subject is an adult, and the effective dose of the composite probiotic to the subject is 150 mg/60 kg body weight/day to 600 mg/60 kg body weight/day. A complex probiotic composition for the relief of. Overactive bladder related symptoms according to claim 6, wherein the subject is a rat, and the effective dose of the composite probiotic to the subject is 5 mg/kg body weight/day to 20 mg/kg body weight/day. A complex probiotic composition for the relief of.

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