DE202020104290U1 - Composition of Streptococcus thermophilus from strain ST4 as an adjuvant of a chemotherapeutic agent for amelioration of diarrhea, appetite disorders and for weight maintenance - Google Patents

Abstract

A composition for use as an adjuvant composition for reducing side effects of a chemotherapeutic agent, which comprises an effective amount of a Streptococcus thermophilus from the strain ST4, which has been deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH with the depot number DSM33165 on May 28, 2019.

Description

BACKGROUND OF THE INVENTION

Technical area

The present invention relates to a composition from a Streptococcus thermophilus strain ST4 and in particular for use as an adjuvant composition for ameliorating chemotherapy side effects.

State of the art

Although cancer drugs are constantly improving, traditional chemotherapy drugs like 5-fluorouracil (5-FU) and irinotecan are still the primary drugs used in cancer treatment, and their mechanism of action is to kill the rapidly dividing cells. Therefore, in addition to poisoning and killing cancer cells, conventional chemotherapy drugs will also rapidly kill dividing normal cells of the patient, such as intestinal epithelial cells and mucosal cells, causing mucositis, decreased absorption of water and nutrients, and associated clinical symptoms such as abdominal pain, diarrhea, Causes loss of appetite and weight loss.

Chemotherapy drugs reduce nutrient absorption capacity and cause loss of appetite, which leads to weight loss in patients. When the weight is greatly reduced, chemotherapy must be stopped, which will lower the rate of cancer cure. Statistics show that 20% of cancer patients die from malnutrition, not from the cancer itself. The study also shows that during chemotherapy, maintaining the chemotherapy patient's appetite and weight are the most important keys to determining the success or failure of chemotherapy. Patients with better appetite and weight maintenance not only show better treatment results, but also recover faster after treatment.

Streptococcus thermophilus is a common probiotic species and is widely used to make sour milk, yogurt, or as a dietary supplement. However, previous studies have examined the effect of Streptococcus thermophilus on the side effects of chemotherapy drugs, and found that Streptococcus thermophilus has no beneficial effect on weight maintenance (Cancer Biology & Therapy, 2009, 8.6: 505-511; Cancer Biology & Therapy, 2006, 5.6: 593-600; Scandinavian journal of gastroenterology, 2013, 48.8: 959-968; Cancer Biology & Therapy, 2011, 12.2: 131-138). Rather, Streptococcus thermophilus can cause decreased appetite (Cancer Biology & Therapy, 2011, 12.2: 131-138), which shows that Streptococcus thermophilus could not improve the appetite and weight of chemotherapy patients, and even have a negative effect on chemotherapy patients. Patient could have. However, the characteristics and abilities of different tribes are variable. Therefore, through screening, it is possible to develop strains of Streptococcus thermophilus that can promote appetite, maintain body weight, and improve intestinal inflammation and diarrhea in chemotherapy patients.

SUMMARY OF THE INVENTION

It was unexpectedly found in the present disclosure that a new Streptococcus thermophilus strain ST4 isolated from the microbial flora of raw milk can be used as an adjuvant composition for reducing chemotherapy drug side effects and chemotherapy symptoms such as diarrhea Can effectively relieve weight loss, loss of appetite, and bowel inflammation.

Accordingly, it is the object of the present disclosure to provide a composition for use as an adjuvant composition for reducing the side effects of a chemotherapeutic agent in a cancer patient, which comprises an effective amount of a Streptococcus thermophilus of the strain ST4.

According to the present disclosure, the Streptococcus thermophilus strain ST4 was deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany, with the depot number DSM33165 on May 28, 2019.

In one embodiment of the present disclosure, the Streptococcus thermophilus strain is contained vital or dead in the composition. In a preferred embodiment of the present disclosure, the strain is dead.

In one embodiment, the composition can be made in the form of a pharmaceutical composition or a food composition. In one example of the present disclosure, the Streptococcus thermophilus strain ST4 is ingested with the chemotherapeutic agent.

In one embodiment, the chemotherapeutic agent is selected from the group consisting of 5-fluorouracil (5-FU), irinotecan, oxaliplatin, doxorubicin, methotrexate, bleomycin and capecitabine. In a preferred embodiment, the chemotherapeutic agent is 5-FU.

In one embodiment, the effect of the composition is to reduce the side effects of the chemotherapeutic agent, the side effects of the chemotherapeutic agent including weight loss, loss of appetite, intestinal inflammation and / or diarrhea.

Figure list

• 1 zeigt die Zellform von Streptococcus thermophilus Stamm ST4.
• 2 zeigt die Analyseergebnisse des VITEK-Identifikationssystems von Streptococcus thermophilus Stamm ST4.
• 3 zeigt, dass die Gewichtszunahmewirkungen von Streptococcus thermophilus vom Stamm ST4 hilfen, konzentrierte Nahrungsmittel von speziell hochkalorischer Rezeptur aufzunehmen. Normal: Füttern von normalem Futter für adulte Ratten LabDiet 5001, das 5 Gew.-% Fett enthält, um 3,3 kcal/g zu liefern; HF: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern; ST2: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5 × 10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST2; ST3: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5 × 10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST3; ST4: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5 × 10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST4. *p<0,05.
• 4 zeigt, die Organ-Gesamtfettbildungswirkungen der verschiedenen Streptococcus thermophilus Stämme bei der Aufnahme von konzentrierten Nahrungsmitteln speziell hochkalorischer Rezeptur. Normal: Füttern von normalem Futter für adulte Ratten LabDiet 5001, das 5 Gew.-% Fett enthält, um 3,3 kcal/g zu liefern; HF: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern; ST2: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5 × 10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST2; ST3: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5 × 10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST3; ST4: Füttern von konzentriertem hochkalorischem Futter, um 22,5 Gew.-% Fett und 4,4 kcal/g zu liefern, und tägliche Sondenernährung von 5×10⁸ CFU/Tag/Ratte von Streptococcus thermophilus ST4. *p<0,05.
• 5 zeigt das Flussdiagramm des experimentellen Designs für den Streptococcus thermophilus vom Stamm ST4, der die Nebenwirkungen von Chemotherapeutika reduziert.
• 6 zeigt, dass der Streptococcus thermophilus Stamm ST4 durch 5-FU induzierten Gewichtsverlust verringert. Verglichen mit der 5-FU-Gruppe, $p<0,05 für die normale Gruppe, #p<0,05 für die Glutamin-Gruppe, *p<0,05 für die ST4-Gruppe.
• 7 zeigt, dass der Streptococcus thermophilus Stamm ST4 durch 5-FU induzierte Appetitlosigkeit mildert.
• 8 zeigt, dass der Streptococcus thermophilus Stamm ST4 durch 5-FU induzierte Diarrhoe mildert.
• 9 zeigt, dass der Streptococcus thermophilus Stamm ST4 durch 5-FU induzierte Darmentzündungen mildert.

The foregoing description and the following detailed description of the disclosure will be better understood when read in conjunction with the accompanying drawings. For the purpose of explaining the present disclosure, presently preferred embodiments are shown in the drawings.

• 1 shows the cell shape of Streptococcus thermophilus strain ST4.
• 2 shows the analysis results of the VITEK identification system of Streptococcus thermophilus strain ST4.
• 3 shows that the weight gain effects of Streptococcus thermophilus strain ST4 help to consume concentrated foods of specially high calorie formulations. Normal: feeding normal diet for adult rats LabDiet 5001 containing 5 wt% fat to provide 3.3 kcal / g; HF: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g; ST2: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 x 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST2; ST3: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 × 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST3; ST4: Feed concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 x 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST4. * p <0.05.
• 4th shows the total organ lipid formation effects of the various Streptococcus thermophilus strains when consuming concentrated foods, especially high-calorie recipes. Normal: feeding normal diet for adult rats LabDiet 5001 containing 5 wt% fat to provide 3.3 kcal / g; HF: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g; ST2: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 x 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST2; ST3: feeding concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 × 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST3; ST4: Feed concentrated high calorie feed to provide 22.5 wt% fat and 4.4 kcal / g and daily tube feeding of 5 x 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST4. * p <0.05.
• 5 Figure 11 shows the experimental design flow diagram for Streptococcus thermophilus strain ST4, which reduces the side effects of chemotherapeutic agents.
• 6th shows that Streptococcus thermophilus strain ST4 decreases weight loss induced by 5-FU. Compared to the 5-FU group, $ p <0.05 for the normal group, #p <0.05 for the glutamine group, * p <0.05 for the ST4 group.
• 7th shows that Streptococcus thermophilus strain ST4 ameliorates 5-FU induced loss of appetite.
• 8th shows that Streptococcus thermophilus strain ST4 ameliorates 5-FU induced diarrhea.
• 9 shows that Streptococcus thermophilus strain ST4 ameliorates intestinal inflammation induced by 5-FU.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as conventionally understood by those skilled in the art to which the present disclosure is directed. It should be understood that the terminology used herein is for the purpose of describing specific embodiments only, and is not intended to be limiting.

As used herein, the singular forms “a” and “the” include plural references unless explicitly indicated otherwise. For example, a reference to “a sample” therefore includes several such samples and their equivalents known to those skilled in the art.

As used herein, the term "Colony-Forming Unit" (CFU) is defined as the number of bacterial cells as demonstrated by microbial production on an agar medium.

As used herein, the term "improve" or "improve" refers to the effects of treating, healing, alleviating, mitigating, changing, eliminating, improving, enhancing, or affecting the event or characteristic. An example of the disclosure is reducing the side effects caused by chemotherapy.

As used herein, the term “excipient” refers to a substance that enhances the effects of chemotherapy, including the effects of increasing chemotherapy compliance and reducing the side effects caused by chemotherapy.

As used herein, the term "subject" includes human and non-human animals such as domestic animals (e.g. dogs, cats, etc.), farm animals (e.g. cattle, sheep, pigs, horses, etc.) or experimental animals (e.g. rats, mice , Guinea pigs, etc.).

As used herein, the term "effective amount" refers to an amount of an active ingredient or composition in a subject that is sufficient to produce the above therapeutic effect. The effective amount may vary depending, for example, on the type or dosage of active ingredient or composition and on the weight, age, and state of health of the subject to be treated.

The Streptococcus thermophilus strain ST4 provided by the present disclosure was deposited with the "Taiwan Food Industry Research and Development Institute" with the depot number BCRC910922 and also with the German Collection of Microorganisms and Cell Cultures GmbH with the depot number DSM33165 on May 28, 2019.

As used herein, "Streptococcus thermophilus" refers to a Gram-positive, homofermentative, and facultative anaerobic bacterium, and belongs to Streptococcus viridans. His cytochrome, oxidase, and catalase tests are negative, and α-hemolytic test is positive. It has no mobility and no endospores.

The Streptococcus thermophilus strain ST4 of the present disclosure was isolated from the microbial flora of raw milk. According to the analysis of bacteriological characteristics, the strain is a gram-positive bacterium with a spherical or oval shape, in pairs or long-chain shape, and without sporulation and motility, grows under anaerobic or aerobic conditions, and has no catalase and oxidase activities. Further identification by 16S rDNA sequence analysis and the VITEK identification system shows that the ST4 strain is Streptococcus thermophilus.

The ST4 strain of the present disclosure can be present in the composition in any way, including vital or dead bacteria, and also includes equivalent strains with the same characteristics and thalli or products derived from the strains.

By using the deposited strain provided by the present disclosure as a starting material, those skilled in the art can generally obtain other mutants or derivatives thereof by conventional mutation induction or re-separation techniques, the mutants or derivatives having the relevant characteristics and advantages of the strain forming the composition of the disclosure described here maintain or strengthen.

The present disclosure provides a composition of a Streptococcus thermophilus strain ST4 for use as an adjuvant composition to be taken with a chemotherapeutic agent, wherein the composition contains an effective amount of the Streptococcus thermophilus strain ST4 to support the chemotherapeutic agent.

According to the present disclosure, the adjuvant composition is intended to reduce the side effects of a chemotherapeutic agent.

According to the present disclosure, the side effects of a chemotherapeutic agent include weight loss, loss of appetite, intestinal inflammation, and / or diarrhea.

In accordance with the present disclosure, the chemotherapeutic agent includes, but is not limited to, 5-fluorouracil (5-FU), irinotecan, oxaliplatin, doxorubicin, methotrexate, bleomycin, and capecitabine. In a preferred embodiment, the chemotherapeutic agent is 5-fluorouracil (5-FU).

In some embodiments, the composition of the present disclosure has an effect of reducing side effects caused by chemotherapy. As shown in the examples, it was found that the composition can reduce the side effects of chemotherapy, specifically improve appetite, promote weight loss and prevent intestinal inflammation, when the composition of the present disclosure is administered during cancer chemotherapy,

In another aspect, the present disclosure provides a use of the composition comprising an effective amount of the ST4 strain of the present disclosure for the manufacture of a medical product, medicament, food, edible product, dietary supplement, or medicinal food.

The composition of the present disclosure can be prepared in the solid or liquid form, and can specifically be in the form of gel bars, infusion bags, biscuits, powders, dragees or cheek tablets, lozenges, film formulations, solutions, aerosols, granules, pills, suspensions, Emulsions, capsules, enteric tablets and capsules, syrups, liquids, elixirs, candies, meal replacements, chewable tablets, suppositories, mini-enemas, creams, gels or ointments.

The composition according to the present disclosure can be formulated in a form in which the strain of the present disclosure is the only active substance or is mixed with one or more other active substances and / or with a pharmaceutically acceptable excipient or in the case of a food or edible product is mixed with sufficient additives or ingredients.

The composition according to the present disclosure can be made into various types of food. The products of these foods can be made by any conventional manufacturing technique using conventional excipients or fillers and, if necessary, mixed with additives.

The composition of the present disclosure can be prepared as a dietary supplement that can be readily administered, can be mixed with a suitable drinkable liquid such as water, sour milk, milk or juice, or can be mixed with solid or liquid foods. In some embodiments, the nutritional supplement can be in the form of dragees or cheek tablets, pills, capsules, granules, powders, suspensions, sachets, candies, sticks, syrups and corresponding administration forms, usually in unit dosage form. The dietary supplement comprising the composition of the present disclosure is preferably administered in the form of coated tablets, cheek tablets, capsules or powders, as they are produced in the conventional processes for producing dietary supplements.

The composition of the present disclosure can be prepared in various types of medicaments which can be prepared by a general pharmaceutical technique or a method using a therapeutically effective amount of the composition of the present disclosure and a pharmaceutically acceptable carrier. Administration of the composition of the present disclosure is oral administration, and may further comprise a pharmaceutically acceptable carrier, diluent, and / or excipient. The carrier can be a solvent Dispersion medium, a coating, an antibacterial agent, an antifungal agent, an isotonic agent, an absorption retarding agent, or the like. The diluent can be phosphate buffer, Ringer's solution, glucose solution or the like. The excipient can be calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate, lactose, starch, gelatin, alginic acid, stearic acid, magnesium stearate, or the like.

The foregoing description of the present disclosure and the following embodiments explains the content of the present disclosure, but is not intended to limit the scope of the present disclosure.

Example 1 Isolation of Streptococcus thermophilus from strain ST4

The strains were isolated from the microbial flora of raw milk. The raw milk was added to the MRS nutrient medium and cultured at 42 ° C. for 24 hours under anaerobic conditions. The culture was then plated out on MRS agar plates and incubated at 42 ° C for 3 days under anaerobic conditions, and then a single colony appearing on the agar medium was collected and further purified to produce the Gram-positive, catalase-negative, spherical or oval, and to isolate paired or long-chain Streptococcus thermophilus ST4.

Example 2 The bacteriological characteristics of Streptococcus thermophilus from strain ST4

1. (1) Morphologische Charakteristika:
   1. (a) Zellform und Gramfärbung: Nachdem die Zellen in MRS-Nährmedium gegeben und bei 42°C für 24 Stunden unter anaeroben Bedingungen kultiviert wurden, war ihre Erscheinung unter dem Mikroskop, wie in 1 gezeigt, sphärisch oder oval, paarweise oder langkettenförmig ohne Flagellum und Gram-positiv.
   2. (b) Beweglichkeit: keine Beweglichkeit.
   3. (c) Sporenbildung: keine Sporenbildung.
   4. (d) Gramfärbung: positiv.
2. (2) Charakteristika der Kultur:
   1. (a) Medium: MRS-Nährmedium, pH = 6.25.
   2. (b) Kultur-Bedingungen: bei 42°C unter anaeroben oder aeroben Bedingungen.
3. (3) Physiologische Charakteristika:
   1. (a) Katalase: negativ.
   2. (b) Oxidase: negativ.

The bacteriological characteristics of Streptococcus thermophilus from strain ST4 are shown below:

1. (1) Morphological characteristics:
   1. (a) Cell shape and Gram stain: After the cells were placed in MRS nutrient medium and cultured at 42 ° C for 24 hours under anaerobic conditions, their appearance under the microscope was as in FIG 1 Shown spherical or oval, in pairs or long chain-shaped without flagellum and Gram-positive.
   2. (b) Mobility: no mobility.
   3. (c) Sporulation: no sporulation.
   4. (d) Gram stain: positive.
2. (2) Characteristics of the culture:
   1. (a) Medium: MRS nutrient medium, pH = 6.25.
   2. (b) Culture conditions: at 42 ° C under anaerobic or aerobic conditions.
3. (3) Physiological characteristics:
   1. (a) Catalase: negative.
   2. (b) Oxidase: negative.

Example 3 Strain identification of Streptococcus thermophilus from strain ST4

16S rDNA and VITEK systems were used to aid in strain identification of Streptococcus thermophilus from strain ST4. Two identification methods were used to ensure the accuracy of the identification results.

16S rDNA sequence analysis

The DNA of Streptococcus thermophilus strain ST4 was extracted to amplify the 16S rDNA (ribosomal DNA) fragment, and agarose gel electrophoresis was performed on the resulting PCR product to confirm whether the product was the expected size and a Sequencing was performed. The 16S rDNA sequence of the resulting Streptococcus thermophilus from strain ST4 was shown as SEQ ID NO: 1 and compared with the various sequence alignment data sets (NCBI blast). The sequence alignment results were Streptococcus salivarius subsp. thermophilus ATCC 19258 ^T , in which the similarity was up to 99.67%.

Analysis with the VITEK identification system

According to the analysis result of the semi-automatic VITEK identification system, the Streptococcus thermophilus of the ST4 strain was on the Streptococcus salivarius subsp. thermophiles closest, wherein the similarity, as in 2 shown was up to 99%.

According to the 16S rDNA sequence identification, VITEK alignment results, and the Approved Lists of Bacterial Names 1980 issued by the International Committee on Systematics of Prokaryotes, the Streptococcus thermophilus from the ST4 strain Streptococcus thermophiles (with the synonym Streptococcus salivarius subsp. Thermophilus).

Example 4 Support in weight maintenance by Streptococcus thermophilus of the strain ST4

Experimental methods

The purpose of this test was to simulate the concentrated diet of a special high-calorie formula required by cancer patients after chemotherapy to evaluate the effects of Streptococcus thermophilus strains ST2, ST3 and ST4 on weight gain. Eight week old Sprague-Dawley strain male rats were used, the feeding environment was maintained at 22 ± 2 ° C, every day and night was 12 hours, and the rats were fed ad libitum. The normal control group was fed LabDiet 5001, a normal diet for adult rats containing 5% (w / w) fat to provide 3.3 kcal / g during the experiment; and the control group and the experimental groups (ST2, ST3 and ST4 groups) were fed concentrated high calorie feed to provide 22.5% (w / w) fat and 4.4 kcal / g. The experimental groups were accordingly fed daily with 5 × 10 ⁸ CFU / day / rat of Streptococcus thermophilus ST2, ST3 and ST4 by gavage. During the animal experiment, feed consumption was measured and recorded daily, the rats were weighed precisely once a day before feeding, the experiments ran for 8 weeks.

Data processing and statistical methods

The statistical analysis, variation analysis and significance comparison of the experimental data were each carried out with IBM SPSS (Statistical Product and Service Solutions) software, an ANOVA test and a Duncan's multiple range test (* p <0.05). The experimental data are expressed as a mean ± standard deviation (mean ± SD).

Experimental results

There is no significant difference in the average daily feed consumption of each group of animals (n = 5). After the 8-week test period, the average weight gain of the animals in each group at the time of euthanasia is 239.8-315.6 g, with the ST4 group having a significantly higher weight gain compared to other experimental groups ( 3 ), showing that Streptococcus thermophilus strain ST4 has the potential to aid in weight gain. In addition, the results for total organ fat ( 4th ) that the experimental group consumed concentrated high-calorie food significantly higher than the normal control group, and that there was also no significant difference in total organ fat in the experimental groups.

Example 5 Amelioration of chemotherapy-induced weight loss, loss of appetite, or intestinal inflammation by Streptococcus thermophilus strain ST4

Experimental methods

The animals for the experiments were five week old male BALB / cByJNARL mice. The feeding environment was kept at 23 ± 2 ° C, the relative humidity was 40-60%, and the light cycle was adapted to a 12/12 light / dark cycle. Laboratory rodent feed Diet 5001 and drinking water were sufficiently offered to the test animals, with the mice being fed ad libitum.

1. (1) Normale Gruppe, fünf Mäuse insgesamt: Geben von physiologischer Salzlösung + intraperitoneale Injektion (IP Injektion) von physiologischer Salzlösung.
2. (2) 5-FU-Gruppe, fünf Mäuse insgesamt: Geben von physiologischer Salzlösung + intraperitoneale Injektion von 5-FU.
3. (3) Glutamin-Gruppe, fünf Mäuse insgesamt: Geben von L-Glutamin (1 g/kg/Tag) + intraperitoneale Injektion von 5-FU, als die positive Kontrollgruppe.
4. (4) ST4-Gruppe, drei Mäuse insgesamt: Geben von Streptococcus thermophilus vom Stamm ST4 (5 × 10⁸ CFU/Tag) + intraperitoneale Injektion von 5-FU.

The five-week-old mice were randomly divided into four groups after two weeks of domestication:

1. (1) Normal group, five mice in total: giving physiological saline solution + intraperitoneal injection (IP injection) of physiological saline solution.
2. (2) 5-FU group, five mice in total: give physiological saline + intraperitoneal injection of 5-FU.
3. (3) Glutamine group, five mice in total: giving L-glutamine (1 g / kg / day) + intraperitoneal injection of 5-FU, as the positive control group.
4. (4) ST4 group, three mice in total: give Streptococcus thermophilus strain ST4 (5 × 10 ⁸ CFU / day) + intraperitoneal injection of 5-FU.

The experimental period was 18 days in total. During the experiment, the body weight and food consumption of the mice were weighed daily, and the samples were orally fed daily using a gastric tube. PBS or 50 mg 5-FU / kg body weight / day was injected intraperitoneally on days 11, 12, and 13. The mice were euthanized on the 18th day and, as an indicator of intestinal inflammation, the cecum to the colon tissue was harvested to assess the length of the colon. On days 11-17, the diarrhea score γ of each experimental mouse was determined by the appearance of the faeces and anus of the experimental mice according to Bowen et al. (Krebs Biology & Therapy, 2007, 6.9: 1445-1450), with scores ranging from asymptomatic (0 points) to most severe (3 points). The experimental procedure is in 5 shown.

Data processing and statistical methods

The method of calculating weight change uses the weight of the mouse on day 11 as 100% to calculate the amount of weight change of the mouse after intraperitoneal injection. The method of calculating the change in feed intake uses the average feed intake of the mice on days 1-10 as 100% to calculate the change in feed intake after intraperitoneal injection. Data are expressed as mean ± standard deviation (mean ± SD). The difference to the 5-FU group was statistically significant according to the Student's T-test. At p <0.05, the difference to the 5-FU group was considered to be statistically significant.

Experimental results

The weight change is in 6th shown. The weight of the normal group increased slowly during the experiment, while the weight of the 5-FU group continued to decrease and did not recover at the end of the experiment, while the weight of the ST4 group on days 13, 17, and 18 was significantly higher than the 5-FU group and began to increase on days 17 and 18, demonstrating that ST4 can slow down weight loss caused by 5-FU and can promote weight recovery.

The changes in feed intake are in 7th shown. After the intraperitoneal injection, the appetite of the normal group did not change significantly, while the food consumption of the 5-FU group continued to decrease from day 11 to day 15 and did not recover on the 17th day. The food consumption of the ST4 group was higher than or equal to that of the 5-FU group on the 11th to the 16th day, with the appetite recovering significantly on the 17th day, and even higher than the average value of the days 1 to 10, whereby it is shown that Streptococcus thermophilus ST4 can slow down the loss of appetite caused by 5-FU and promote the return of appetite to a normal state.

The 5-FU treatment causes damage to intestinal mucosal tissue and causes diarrhea. The changes in the diarrhea score of the mice is shown in 8th shown. The diarrhea symptoms of the 5-FU-treated mice appeared between days 13-17 and were most severe on days 14, 15, and 17. However, significantly orally administered ST4 can ameliorate the diarrhea caused by 5-FU, and only mild diarrhea occurred on days 13-17, indicating that ST4 can ameliorate the diarrhea caused by chemotherapy drugs.

The bowel shortens when an intestinal inflammatory response occurs. The more severe the inflammation, the shorter the bowel becomes. The length of the intestine is in 9 shown. The intestinal length of the 5-FU group was significantly shorter than that of the normal group, indicating that 5-FU causes intestinal inflammation; while the bowel length of the ST4 group was significantly longer than that of the 5-FU group, indicating that orally administered ST4 can prevent bowel inflammation caused by 5-FU.
Those of ordinary skill in the art will understand that modifications can be made to the specific embodiments described above without departing from the broad scope of the inventive concept leave. Therefore, it should be understood that the present disclosure is not limited to the specific embodiments disclosed, but is intended to embrace the modifications within the scope and scope of the present disclosure as defined by the appended claims.

SUMMARY

The present disclosure provides a composition for use as an adjuvant composition for reducing the side effects of a chemotherapeutic agent in a cancer patient, comprising a Streptococcus thermophilus strain ST4. In particular, the composition alleviates diarrhea and anorexia induced by the chemotherapeutic agent, and maintains the cancer patient's weight.

SEQUENCE LISTING

• <110> Syngen Biotech. Co., Ltd.
• <120>
• <130> SYN0003DE-UM
• <160> 1
• <170> Patent In version 3.5
• <210> 1 <211> 491 <212> DNA <213> Lactobacillus fermentum
• <220> <221> misc_feature <222> (490) .. (490) <223> n is a, c, g, or t
• <400> 1
  

Claims (8)

A composition for use as an adjuvant composition for reducing side effects of a chemotherapeutic agent, which comprises an effective amount of a Streptococcus thermophilus from the strain ST4, which has been deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH with the depot number DSM33165 on May 28, 2019. Composition according to Claim 1 wherein the Streptococcus thermophilus strain ST4 is vital or dead in the composition. Composition according to Claim 2 wherein Streptococcus thermophilus from strain ST4 in the composition is dead. Composition according to Claim 1 wherein the Streptococcus thermophilus strain ST4 is ingested with the chemotherapeutic agent. Composition according to Claim 1 which is in the form of a pharmaceutical composition or a food composition. Composition according to Claim 1 wherein the chemotherapeutic agent is selected from the group consisting of 5-fluorouracil (5-FU), irinotecan, oxaliplatin, doxorubicin, methotrexate, bleomycin and capecitabine. Composition according to Claim 6 wherein the chemotherapeutic agent is 5-FU. Composition according to Claim 6 wherein the side effects of the chemotherapeutic agent include weight loss, loss of appetite, intestinal inflammation and / or diarrhea.

Images