CN116570629A - Probiotic composition for preventing and treating diarrhea and preparation method thereof - Google Patents

Probiotic composition for preventing and treating diarrhea and preparation method thereof Download PDF

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Publication number
CN116570629A
CN116570629A CN202310325765.1A CN202310325765A CN116570629A CN 116570629 A CN116570629 A CN 116570629A CN 202310325765 A CN202310325765 A CN 202310325765A CN 116570629 A CN116570629 A CN 116570629A
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China
Prior art keywords
probiotic composition
diarrhea
component
preventing
treatment
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CN202310325765.1A
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Chinese (zh)
Inventor
闫述模
钱忠旭
姜子辰
孙圣涛
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Aland Nutrition Taizhou Co ltd
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Aland Nutrition Taizhou Co ltd
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Priority to CN202310325765.1A priority Critical patent/CN116570629A/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/12—Antidiarrhoeals
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a probiotic composition for preventing and treating diarrhea and a preparation method thereof, wherein the probiotic composition comprises a component A and a component B, the component A comprises a probiotic composition consisting of 4 strains, and the component A mainly comprises four efficacy strains of lactobacillus acidophilus CUL34 and bifidobacterium bifidum CUL20, and the component B comprises sugar alcohol, prebiotics, fruit powder, essence, vitamin C and auxiliary materials which are required to be added for preparing common pharmaceutical dosage forms, wherein the common pharmaceutical dosage forms comprise tablets, powder, granules, hard capsules and pills. The probiotic composition for preventing and treating diarrhea and the preparation method thereof provided by the invention can inhibit the growth of potential harmful bacteria during or after antibiotic treatment and reduce the incidence rate of diarrhea caused by clostridium difficile and clostridium difficile toxin positive after antibiotic treatment.

Description

Probiotic composition for preventing and treating diarrhea and preparation method thereof
Technical Field
The invention relates to the technical field of microorganisms, in particular to a probiotic composition for preventing and treating diarrhea and a preparation method thereof.
Background
With the large application of broad-spectrum antibiotics in recent decades, the incidence rate and death rate of diarrhea caused by the antibiotics are continuously improved, and 3% -29% of patients in hospitals can cause diarrhea after taking the antibiotics; clostridium difficile (Clostridium dificile, CD) is the major pathogen responsible for these diarrhea. The colonization rate of clostridium difficile in healthy adults is 3%, but in hospitals is 15% -35%. Clostridium difficile is a gram-positive bacillus, first discovered in 1935 and identified as a conditional pathogen in 1978. Clostridium difficile causes diarrhea symptoms ranging from mild diarrhea to acute diarrhea, toxic colitis, and can even cause death. There are many clinical trials demonstrating that probiotics can aid in the treatment and prevention of clostridium difficile induced diarrhea (Clostridium diffcile-associated diarrhea, CDAD).
Probiotics, according to the description of recent years, can be summarized generally as: living microorganisms, when present in sufficient quantity to reach the human or animal intestine, remain viable and exert beneficial host health effects. In 2001 FAO/WHO has standardized the summary of Probiotics, "Probiotics" are a class of living microorganisms that reach the host intestinal tract in sufficient numbers to colonize and alter the host intestinal colony balance, thereby producing a healthy effect on the host. Probiotics are important microorganisms in the human intestinal tract, and probiotics in the intestinal tract play an important role in regulating immune responses, maintaining mucosal barriers, inhibiting overgrowth of pathogens and anti-inflammatory factor production.
The prior patent CN 111088181B discloses a bifidobacterium breve BK55 which can remarkably inhibit the growth, spore generation, virulence gene expression and toxin yield of clostridium difficile, and can also damage the permeability and integrity of clostridium difficile cell membranes, so that the cell death caused by the leakage of intracellular substances of clostridium difficile cells.
There are still few probiotic compositions that have been found to be useful in the prevention and treatment of diarrhea, and there is a lack of probiotic compositions that can inhibit the growth of potentially harmful bacteria during or after antibiotic treatment and reduce the incidence of diarrhea caused by clostridium difficile and clostridium difficile toxin positivity after antibiotic treatment.
Therefore, we propose a probiotic composition for preventing and treating diarrhea and a preparation method thereof to solve the above problems.
Disclosure of Invention
(one) solving the technical problems
In order to overcome the defects of the prior art, the invention provides a probiotic composition for preventing and treating diarrhea and a preparation method thereof, and solves the problems that few probiotic compositions which can be used for preventing and treating diarrhea are found in the prior art, and the probiotic compositions which can inhibit the growth of potential harmful bacteria during or after antibiotic treatment and reduce the occurrence rate of diarrhea caused by clostridium difficile and clostridium difficile toxin positive after antibiotic treatment are lacking.
(II) technical scheme
The invention adopts the following technical scheme for realizing the purposes:
a probiotic composition for preventing and treating diarrhea and its preparation method comprise component A and component B.
Further, the component A comprises a probiotic composition consisting of 4 strains, and the specific strain comprises lactobacillus acidophilus CUL60 (Lactobacillus acidophilusCUL 60), and the strain deposit number is NCIMB 30157; lactobacillus acidophilus CUL21 (Lactobacillus acidophilusCUL), strain accession number NCIMB 30156; bifidobacterium animalis subspecies CUL34 (Bifidobacterium animalis subsp. Lactis CUL 34), strain deposit No. NCIMB 30172, bifidobacterium bifidum CUL20 (Bifidobacterium bifidumCUL 20), strain deposit No. NCIMB 30153.
Preferably, the composition of 4 strains is designated Lab4 probiotic; wherein lactobacillus acidophilus CUL60 and lactobacillus acidophilus CUL21 are symbiotic fermentation bacteria, and bifidobacterium animalis subspecies lactis CUL34 and bifidobacterium bifidum CUL20 are symbiotic fermentation bacteria; preferably, the bacterial strain performs symbiotic fermentation, so that the stability of the bacterial powder can be improved.
Preferably, the Lab4 probiotic has a total colony count per unit mass of 1.0-4.5X10 11 cfu/g。
Preferably, the daily dosage of Lab4 probiotic is recommended to be 0.2-1.0X10 11 Hundred million cfu total viable bacteria count.
Further, component B comprises sugar alcohol, prebiotics, fruit powder, essence, vitamin C and auxiliary materials which are necessary to be added for preparing common pharmaceutical dosage forms.
Preferably, the prebiotic comprises a resistant dextrin, polydextrose, inulin, xylo-oligosaccharide, fructo-oligosaccharide, isomalto-oligosaccharide, galacto-oligosaccharide, soy oligosaccharide.
Preferably, the sugar alcohol comprises erythritol, xylitol, sorbitol, mannitol, lactitol, maltitol, isomalt.
Preferably, the fruit powder comprises apple juice powder, mango juice powder, passion fruit juice powder, orange juice powder, jackfruit juice powder, litchi juice powder, banana juice powder, mangosteen juice powder, pineapple juice powder, hami melon juice powder, kumquat juice powder, grape juice powder, kiwi fruit juice powder, plum juice powder, watermelon juice powder, juicy peach juice powder, lemon juice powder, grapefruit juice powder, red grape juice powder, sweet orange juice powder, pomegranate juice powder, blackcurrant juice powder, blueberry juice powder, cranberry juice powder, raspberry juice powder, strawberry juice powder, acerola juice powder, brazil berry juice powder, red heart ballon juice powder, green plum juice powder, roselle juice powder, dark plum juice powder, cherry juice powder, snow pear juice powder, mulberry juice powder.
Preferably, the pharmaceutical dosage forms comprise tablets, powders, granules, hard capsules and pills.
Preferably, the auxiliary materials comprise one or more of fillers, binders, wetting agents, diluents, disintegrants, lubricants and flavoring agents which are commonly used in pharmacy
Preferably, the product comprises use in medicine, health products, food.
In order to solve the second technical problem, the preparation method of the probiotic composition preparation for relieving constipation-predominant irritable bowel syndrome mainly comprises the preparation process of powder, tablets and hard capsules.
Preferably, the preparation process of the probiotic composition powder for preventing and treating diarrhea comprises the following steps: 1) Controlling the temperature and the humidity of a workshop; 2) Uniformly mixing the component A and the component B; 3) Then vacuum drying is carried out, and the water activity after drying is controlled; 4) Feeding the materials into a hopper of a racking machine in vacuum, and racking the materials into 3 g/strip by the racking machine.
Preferably, the temperature and humidity requirements of the workshop are as follows: the temperature is 18-26 ℃, and the relative humidity is lower than 30% RH.
Preferably, lab4 probiotics in the component A are more than or equal to 250 hundred million cfu/package.
Preferably, the vitamin C is contained in the above component B in an amount of 1-10mg.
Preferably, the component B comprises 10-50% by weight of prebiotics.
Preferably, the component B comprises 2-20% of fruit powder by weight.
Preferably, in the vacuum drying process, the water activity is required to be 0.05-0.3.
(III) beneficial effects
Compared with the prior art, the invention provides a probiotic composition for preventing and treating diarrhea and a preparation method thereof, and the probiotic composition has the following beneficial effects:
the invention provides a probiotic composition of four main efficacy strains including lactobacillus acidophilus CUL60, lactobacillus acidophilus CUL21, bifidobacterium animalis subspecies lactis CUL34 and bifidobacterium bifidum CUL20, and clinical experiments prove that the probiotic composition can inhibit the growth of potential harmful bacteria during or after antibiotic treatment and reduce the incidence rate of diarrhea caused by clostridium difficile and clostridium difficile toxin positive after antibiotic treatment; the specific expression is as follows: 1) Clostridium difficile positive placebo group patients were 32% different from probiotic group patients in toxin detection; 2) The proportion of positive diarrhea occurring in clostridium difficile-associated toxin in the probiotic group is reduced by 4.35% compared with that in the placebo group; 3) According to the statistical analysis of the flora in the feces after the treatment of the helicobacter pylori patients with antibiotics, it is found that the probiotics can reduce the overgrowth of the potential harmful bacteria; therefore, the use of the extract in the preparation of foods or medicines for preventing and/or treating antibiotic-associated diarrhea or antibiotic-associated diarrhea has a good prospect.
Drawings
FIG. 1 is a schematic diagram of the structure xx of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The invention provides a probiotic composition hard capsule for preventing and treating diarrhea, which is prepared by the following steps: 1) Temperature and humidity of a control workshop: the temperature is 18-26 ℃, and the relative humidity is lower than 30% RH; 2) Uniformly mixing Lab4 probiotics and auxiliary materials; 3) Then vacuum drying is carried out, and the water activity after drying is controlled to be lower than 0.2; 4) Filling and selecting pills, and then sub-packaging into corresponding packaging materials. The hard capsule with the total viable count of the probiotic bacteria of 250 hundred million cfu/granule is prepared.
Test example 1 clinical data
Clostridium difficile assay study: effect of supplemental probiotics on clostridium difficile diarrhea incidence
The study was conducted in the Addenbrooke hospital of Cambridge, recruiting 138 elderly patients in need of antibiotic treatment, and was divided equally into two groups at random. The first group received 250 billions of Lab4 probiotics with their prescribed antibiotics for 20 days per day, and the second group used placebo with their prescribed antibiotics for 20 days. The probiotic or placebo was started within 36 hours after the start of the antibiotic prescription, the probiotic group was started for 26.88 hours and the placebo group was started for 26.4 hours. The efficacy of Lab4 probiotics in preventing CDAD was assessed by recording bowel movement habits and analyzing stool samples.
During the treatment period, 15 diarrhea patients appeared in each of the probiotic group and placebo group, with 5 clostridium difficile toxins positive in the placebo group and 2 clostridium difficile toxins positive in the probiotic group. Statistical analysis of these data showed that the proportion of clostridium difficile-associated toxin positive diarrhea in the probiotic group was reduced by 4.35% compared to the placebo group.
Analysis of the stool sample on day 20 of the analysis found that 20 patients were positive for clostridium difficile, whereas only 4 patients were positive for clostridium difficile when the 138 elderly patients were admitted. Of these 20 clostridium difficile positive patients, 9 out of the placebo group, 7 were detected as clostridium difficile toxin positive and 6 developed diarrhea symptoms; of the 11 remaining probiotics groups, 5 patients were detected positive for clostridium difficile toxin and 2 exhibited diarrhea symptoms. Through data statistical analysis, the toxin detection of clostridium difficile positive placebo group patients and the toxin detection of probiotic group patients are found to be 32 percent different. The data are shown in Table 1 below.
The data in table 1 demonstrate that supplementation with Lab4 probiotics can reduce the incidence of clostridium difficile diarrhea in hospitalized patients.
Test example 2 clinical data
Effect of probiotics on prevention of intestinal flora destruction following antibiotic treatment: double blind, placebo controlled trial study
The study was conducted in Addenbroake Hospital, cambridge, recruiting 30 patients diagnosed with H.pylori infection. Patients are otherwise healthy, with no other gastrointestinal disease other than peptic ulcers, and are considered to be associated with their helicobacter pylori infection. Within 6 weeks prior to the study, they did not receive any antibiotic treatment or any dietary intervention. All participants were given written informed consent to participate in the trial according to the procedure approved by the Cambridge local research ethics committee.
30 volunteers were randomly divided into 3 groups, one for each 10 cases. All participating patients received 7 day eradication triple therapy: amoxicillin 500mg q.i.d, metronidazole 400mg t.i.d. and lansoprazole 30mg b.i.d. for 7 days. Treatment regimen of group I was placebo used on days 1 to 15; group II treatment regimen with placebo on days 1 to 7, with probiotic supplementation on days 8 to 15; treatment regimen of group III was 1 day to 15 days supplemented with probiotics. The probiotics were administered daily in 250 hundred million cfu Lab4 probiotics and the placebo included an inert carrier maltodextrin. During the course of the experiment, since 8 patients were rejected without following the administration requirement, the remaining 22 patients were regrouped: group I had 7 people, including 4 men and 3 women; group II had 9 people, including 1 male, 8 females; group III had 6 individuals, including 2 men and 4 women. Stool samples were taken at 5 time points Day1, day7, day12, day17, day27, etc. for analysis to assess different bacterial populations in the intestinal microbiota. The data are shown in Table 2 below.
Illustration of: data are expressed as mean counts (log cfu/g fecal dry weight) ± standard error, with the amount carried (number of patients carrying a particular bacterial population) shown in brackets. The first group received placebo from day 1-15, the second group received placebo from day 1-7, the first group received probiotics from day 8-15, and the third group received probiotics from day 1-15. Three groups received H.pylori eradication therapy on days 1-7. * Represents P <0.05 compared to the first day, and P <0.01 compared to the first day. y represents P <0.05 compared to day7 and z represents P <0.01 compared to day seven.
Analysis of results: for the total number of facultative anaerobes,
1) The total number of facultative anaerobes in the patients at the end of antibiotic treatment increased significantly (P < 0.05) compared to before antibiotic administration, and the total number of facultative anaerobes remained significantly increased (P < 0.05) up to 27 days after antibiotic treatment (group I).
2) The antibiotic and placebo group patients increased significantly at the end of antibiotic treatment (< 0.05) but the number decreased significantly after the introduction of Lab4 probiotic supplement after antibiotic (P < 0.05) (group II).
3) During the entire study, the Lab4 probiotic was supplemented while the antibiotics were taken, and the total number of facultative anaerobes remained stable (group III).
Conclusion: supplementing Lab4 probiotics reduced overgrowth of potentially harmful bacteria during and after antibiotic treatment.
Example 2
One embodiment of the invention provides probiotic composition powder for preventing and treating diarrhea.
The preparation method comprises the following steps: uniformly mixing 450mg Lab4 probiotics, 4mg vitamin C, 100mg galacto-oligosaccharides, 100mg polydextrose, 100mg fructo-oligosaccharides and 1210mg resistant dextrin, then vacuum drying to reduce the water activity to below 0.2, and finally vacuum feeding and sub-packaging into 2g bags containing 250 hundred million cfu viable bacteria.
Example 3
One embodiment of the invention provides a probiotic composition tablet for preventing and treating diarrhea.
The preparation method comprises the following steps: uniformly mixing 200mg Lab4 probiotics, 50mg vitamin C, 200mg fructo-oligosaccharide, 50mg strawberry, fruit powder, 3mg strawberry essence, 10mg magnesium stearate and 487mg sorbitol, vacuum drying until the water activity is lower than 0.20, pressing into tablets with 1g and 100 hundred million cfu viable bacteria count, and finally packaging into a proper plastic bottle.
Example 4
The invention provides a probiotic composition hard capsule for preventing and treating diarrhea.
The preparation method comprises the following steps: uniformly mixing 200mgLab4 probiotics, 200mg microcrystalline cellulose, 4.5mg magnesium stearate and 45.5mg fructo-oligosaccharide, performing ultra-low water activity vacuum drying process until the water activity is lower than 0.20, then filling capsules to prepare hard capsules with 450mg, 0# and 100 hundred million cfu viable bacteria, and finally filling the hard capsules into a bubble cover plate.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (6)

1. A probiotic composition for the prevention and treatment of diarrhea, characterized in that: the probiotic composition for preventing and treating diarrhea comprises a component A and a component B.
2. A probiotic composition for the prevention and treatment of diarrhea according to claim 1, characterized in that: the component A comprises a probiotic composition consisting of 4 strains, and the specific strains comprise lactobacillus acidophilus CUL60, lactobacillus acidophilus CUL21, bifidobacterium animalis subspecies lactis CUL34 and bifidobacterium bifidum CUL20.
3. A probiotic composition for the prevention and treatment of diarrhea according to claim 1, characterized in that: the component B comprises sugar alcohol, prebiotics, fruit powder, essence, vitamin C and auxiliary materials which are required to be added for preparing common pharmaceutical formulations;
such common pharmaceutical dosage forms include tablets, powders, granules, hard capsules and pills.
4. A probiotic composition according to claim 1 for the prevention and treatment of diarrhea: the probiotic composition for preventing and treating diarrhea is administered in daily dosage recommended by 0.2-1.0X10 11 Hundred million cfu total viable bacteria count.
5. A method for the preparation of a probiotic composition for the prevention and treatment of diarrhoea according to any one of claims 1-4, characterized in that: mainly comprises the preparation process of probiotic composition powder, hard capsules and tablets for preventing and treating diarrhea.
6. A method of preparing a probiotic composition for the prevention and treatment of diarrhea according to claim 5, characterized in that: the preparation process of the probiotic composition powder for preventing and treating diarrhea comprises the following steps:
s1, controlling the temperature and the humidity of a workshop;
s2, uniformly mixing the component A and the component B;
s3, performing vacuum drying, and controlling the water activity after drying;
s4, feeding the materials into a hopper of a racking machine through vacuum, and racking the materials into 3 g/strip by the racking machine.
CN202310325765.1A 2023-03-30 2023-03-30 Probiotic composition for preventing and treating diarrhea and preparation method thereof Pending CN116570629A (en)

Priority Applications (1)

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CN202310325765.1A CN116570629A (en) 2023-03-30 2023-03-30 Probiotic composition for preventing and treating diarrhea and preparation method thereof

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Application Number Priority Date Filing Date Title
CN202310325765.1A CN116570629A (en) 2023-03-30 2023-03-30 Probiotic composition for preventing and treating diarrhea and preparation method thereof

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