EP1102580A2 - Formulations renfermant des micro-organismes a effet probiotique - Google Patents

Formulations renfermant des micro-organismes a effet probiotique

Info

Publication number
EP1102580A2
EP1102580A2 EP99938048A EP99938048A EP1102580A2 EP 1102580 A2 EP1102580 A2 EP 1102580A2 EP 99938048 A EP99938048 A EP 99938048A EP 99938048 A EP99938048 A EP 99938048A EP 1102580 A2 EP1102580 A2 EP 1102580A2
Authority
EP
European Patent Office
Prior art keywords
formulation according
microorganisms
lactobacillus
formulation
stabilized
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99938048A
Other languages
German (de)
English (en)
Inventor
Helmut Vierstein
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of EP1102580A2 publication Critical patent/EP1102580A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • 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
    • 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
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A61K9/0007—Effervescent
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • 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

Definitions

  • the invention relates to formulations with probiotic microorganisms.
  • probiotics A General View in: “The Lactic Acid Bacteria, Volume 1", BJB Wood ed. (1992), the probiotic microorganisms (common synonym “probiotics”) are , Elsevier Applied Science, 155/156) describes individual or mixed cultures of living microorganisms which are applied to humans and animals and which have a beneficial effect on the host by supporting the properties of the naturally present intestinal microflora. This means that only products that contain living or viable microorganisms, e.g. as freeze-dried cells, improve the health status of humans and animals and which can develop their effects in the mouth or the gastrointestinal tract.
  • MSB lactic acid bacteria
  • lactic acid bacteria of the genera Lactobacillus, Streptococcus, Lactococcus, Leuconostoc, Enterococcus, Bifidobacterium, Carnobacterium and Sporolactobacillus are in pharmaceutical preparations and food supplements according to the current state of knowledge and current taxonomy. increasingly found.
  • Many subspecies of the genera Lactobacillus, Bifidobacterium and Enterococcus are considered to be necessary natural intestinal inhabitants of humans and animals. They occur - depending on nutritional status, age, state of health and intestinal tract - in dynamically changing amounts.
  • Lactic acid bacteria are assessed according to their abilities in a similar way to other microorganisms. These are: 1. biochemical (eg the utilization of carbohydrate utilization)
  • physiological e.g. stimulation of intestinal peristalsis
  • the microflora of the intestine can be specifically supported by the application of certain MSB via medical indications or the way of supplementing food.
  • MSB medical indications or the way of supplementing food.
  • microorganisms are difficult to stabilize. In preparations, they are usually used dried as viable, lyophilized microorganisms. The display is the responsibility of the culture producers. The continuous production of the microorganisms in the required qualities, as well as their further processing into physiologically effective forms. Definitions and the subsequent storage period determine the biological activity of the material (Brennan et al., J. Food Prot. 46 (1983), 887-892).
  • the object of the present invention is to provide formulations containing probiotic microorganisms which, when properly administered in suspension form, guarantee that physiologically relevant germ contents enter the human or animal intestine and which can therefore be highly effective in the intestinal physiology. Linked to this is the task of maintaining the viability of the probiotics, especially the MSB, in the product itself.
  • formulations themselves can be produced on a large scale and reproducibly as medical formulations for therapeutic indications or as an additional or preventive food supplement for humans and animals.
  • a formulation comprising buffer substances and microorganisms stabilized by lyophilization and. optionally technologically necessary auxiliaries, wherein the microorganisms stabilized by lyophilization are dry incorporated into the matrix-forming buffer substances and these formulations can be dry, for example either in powder form or as tablets.
  • the preparations according to the invention develop their effect, which are predominantly in powder form or as a tablet, after appropriate dissolution and ingestion by the patient or consumer.
  • the phase of the disintegration of the formulation can be carried out, for example, in water without a substantial loss of the number of bacteria, even if acidic reactions occur when dissolved.
  • the (spatial) close contact between microorganisms and buffer substances is sufficient to survive even the acidic processes during the dissolution process if the formulation according to the invention is formulated as effervescent powder and substances such as organic acids (e.g. tartaric acid, citric acid), acidifying agents, various (acidic) ) Flavors, etc.
  • the buffer substances are also able to exert this protective effect when passing through the stomach.
  • the formulation according to the invention guarantees protection of the microorganisms stabilized by lyophilization from the natural acid barrier of the stomach, the microorganisms being protected by the action of the buffer substances of the formulation after their reconstitution.
  • the microorganisms therefore enter the intestine in a viable form, where they can develop their effectiveness.
  • formulations according to the present invention are outstandingly suitable for the production of preparations for a wide variety of medical indications and as an additional or preventive food supplement for humans and animals.
  • the microorganisms stabilized by lyophilization preferably contain auxiliaries for lyophilization, as well as residues from the nutrient medium. These lyophilisate compositions determine the hygroscopic properties of the lyophilisate.
  • Preferred lyophilisates have a w values in the range from 0.2 or below, in particular from 0.1 to 0.2, after the freeze-drying process. As a result, they are long-lasting, but they are also able to absorb large quantities of water quickly from the ambient air, which of course must be avoided during storage got to.
  • This problem can be technologically solved by further processing - tableting, filling, packaging - immediately after the introduction of lyophilized microorganisms into the effervescent matrix, so that contact with the ambient air is minimized.
  • the packaging must have an a w value of 0.1 to 0.2 in the long term so that free water is not available for active life processes in cells.
  • Packaging in composite films, sachets or for tablets in aluminum sleeves with desiccants are particularly suitable for this.
  • the storage temperature should be chosen so that 20 ° C is not exceeded.
  • the formulation according to the invention advantageously has an a w value of 0.1 to 0.2, in particular 0.1 to 0.15. Therefore, buffer matrices which are composed of completely anhydrous mineral constituents are preferably also used according to the invention. In principle, these are easy to prepare, but have so far not been used in the production of probiotic formulations.
  • Preferred buffer systems in the context of the present invention are phosphate buffers, citrate buffers, carbonate buffers, malate buffers and tartrate buffers with alkalis and alkaline earths (pH 4-7.5).
  • Strains from the families Lactobacillus, Enterococcus, Bifidobacterium and Enterobacteriaceae, preferably from the genera Enterobacter and Escherichia, are used in the formulations according to the invention as microorganisms stabilized or preserved by lyophilization, in particular strains from the genera Lactobacillus delbschreibii subs. bulgaricus, Lactobacillus acidophilus, Lactobacillus casei GG, Lactobacillus casei subsp. casei, Lactobacillus helveticus, Lactobacillus lactis, Lactobacillus salivarius, Lactobacillus plantarum, Streptococcus salivarius subsp.
  • thermophilus Enterococcus faecium, Bifidobacterium bifidum, Bifidobacterium infantis, Bififobacterium longum (as well as other Bifidobacterium species) and physiologically important subspecies of Escherichia coli and mixtures of two or more species of these microorganisms are particularly preferred.
  • yeast strains in particular Saccharomyces bulardii for the human field, or other probiotic yeasts, for example Kluyveromyces marxianus for animals, are suitable for the production of the formulations according to the invention.
  • microorganisms are those whose effects can be used for oral active immunization.
  • the formulations according to the invention advantageously comprise further formulation auxiliaries or technically necessary combinations of adjuvants.
  • antioxidants and amino acids may be necessary.
  • the substance classes mentioned are typically part of all natural systems of organic origin and are therefore also obtained on an industrial scale and used as additives. They are present in the concentrations used for the formulation according to the invention below the limit values permitted in medications and nutritional supplements and are completely harmless.
  • Auxiliaries used with preference are L-ascorbic acid and L-cysteine.
  • the decisive technological step for the formulations according to the invention lies in the use of buffer substances which, after being dissolved in a suitable solvent, such as water, fruit juices, mineral waters, etc., offer the microorganisms, which were also dry until then, optimal conditions for cell wall reconstitution and these then allow passage through the upper digestive tract, in particular the stomach, with the hydrochloric acid present in the stomach being buffered for a short time.
  • a suitable solvent such as water, fruit juices, mineral waters, etc.
  • composition When dispersing the pharmaceutical form, a composition should preferably be produced which corresponds to the currently valid WHO recipe for rehydration during or after diarrhea.
  • the formulation according to the invention is made available as a shower, the buffer substances being part of the dry shower bases.
  • the formulation according to the invention disintegrates, CO 2 is formed , which displaces the oxygen present in the solution, and thus leads to more favorable survival conditions for anaerobic and microaerophilic germs.
  • the effervescent basics are preferably divided as effervescent granules (cf., for example, European Pharmacopoeia, 3rd edition (1997), 1847/1848 and Hagers Handbook of Pharmaceutical Practice, 5th edition, page 723).
  • the buffer granules are manufactured using the technologically customary granulation methods, e.g. by thermal granulation in heatable mixers, by granulation with reactive or non-reactive liquids in suitable commercial devices, whereby granulation can also be carried out in a vacuum (e.g. evacuable mixer with chopper with subsequent drying and screening, the TOPO® process or fluidized bed granulation) . Spiked with the intended microorganisms, these granules can then be tableted or filled into sachets.
  • a vacuum e.g. evacuable mixer with chopper with subsequent drying and screening, the TOPO® process or fluidized bed granulation
  • the formulation according to the invention preferably contains one or more active pharmaceutical ingredients, in particular an antidarrheal active ingredient, as a result of which a superadditive, anti-diarrhetic effect results from the combination of these substances while at the same time restoring a healthy intestinal flora.
  • active ingredients for the formulation according to the invention can be, for example, loperamide, domperidone and ofloxacin, which themselves have no germ-damaging effect on lactic acid bacteria.
  • the active formulation according to the invention preferably preferably comprises
  • 0.1 - 30% by weight of microorganisms stabilized by lyophilization in particular 0.1 to 5% by weight, and - 10 - 99.9% by weight, in particular 70 to 99% by weight of buffer substances (shower base) and, if necessary, 0 - 80% by weight, in particular 0 to 2% by weight, of auxiliaries.
  • the buffer substances, but also auxiliary substances and lyophilisates, are preferably in granular form or in powder form.
  • a particularly preferred formulation contains microorganisms of the genus Lactobacillus casei GG stabilized by lyophilization.
  • the present invention relates to the use of the formulations of the intestinal physiologically active according to the invention for the preparation of preparations for medical indications, in particular for gastrointestinal disorders, or as a food supplement and animal food supplement.
  • formulations according to the invention are particularly suitable in all cases in which the natural intestinal flora has been impaired or destroyed, for example in the effective accompanying and aftertreatment during or after administration of antibiotics.
  • formulations according to the invention can also include glucose and / or oligosaccharides and / or other saccharides which are favorable for the growth of germs in the intestinal lumen, as well as polysaccharides and related substances (inulins, dietary fibers).
  • Example 1 Stable formulations of effervescent granules (tablets) with lactic acid bacteria as an effective principle
  • Lactobacillus casei GG (LGG, Valio, Finland) approx. 10 8 CfU (colony forming units) / dose
  • Enterococcus faecium (Ef) (strain M74, Medipharm, Sweden) approx.
  • Bifidobacterium lactis (strain Bb 12, Hansen 's Biosystems / Denmark) approx. 10 10 CfU / dose
  • Example 2 Incubation of the Formulations According to the Invention in Artificial Gastric Juice (aMS)
  • the buffer substances are mixed with bacterial lyophilisates in a weight ratio of 249: 1.
  • One part of lyophilisate is sufficient to achieve a germ density of approximately 10 8 -10 9 CfU / g of formulation mass.
  • germ densities in ranges above 10 12 CfU / g are achieved, which are of course preferred for commercial products according to the invention.
  • the incubation is carried out at a body temperature of 37 ° C to simulate as physiological conditions as possible.
  • SWB shaking water bath
  • REF reference sample
  • the course of the reduction in the number of bacteria is shown in Table 1.
  • Column 2 of Table 1 shows the course of the bacterial count as it results from the use of the buffer substance and LGG according to the invention in the case of incubation in aMS.
  • Column 3 contains the REF data.
  • Column 4 shows how aMS affects unprotected lactic acid bacteria in the form of a lyophilisate (Lyo).
  • composition of the effervescent mixture (according to Bauer et al. "Pharmaceutical Technology", 5th edition, page 316):
  • Table 4 below lists the survival germ counts of LGG after incubation in artificial gastric juice and in parallel after incubation in water (REF).
  • Example 4 Effect of domperidone on the survivability of lactic acid bacteria
  • Table 6 shows that the active ingredient domperidone has very little influence on the survivability of the lactic acid bacteria during the incubation. -The combination of domperidone and artificial gastric juice causes a slightly greater decrease in the number of bacteria than water. E.g. Effect of ofloxacin on the survivability of lactic acid bacteria

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  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Mycology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
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  • Communicable Diseases (AREA)
  • Immunology (AREA)
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  • General Engineering & Computer Science (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicinal Preparation (AREA)

Abstract

L'invention concerne une formulation renfermant des micro-organismes stabilisés et des substances tampon. Cette formulation facilite le passage des micro-organismes, après leur dispersion et leur absorption dans le tube digestif, à travers l'estomac, tout en maintenant leur vitalité.
EP99938048A 1998-08-06 1999-07-29 Formulations renfermant des micro-organismes a effet probiotique Withdrawn EP1102580A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
AT136098 1998-08-06
AT0136098A AT407008B (de) 1998-08-06 1998-08-06 Formulierungen mit probiotisch wirksamen mikroorganismen
PCT/AT1999/000192 WO2000007571A2 (fr) 1998-08-06 1999-07-29 Formulations renfermant des micro-organismes a effet probiotique

Publications (1)

Publication Number Publication Date
EP1102580A2 true EP1102580A2 (fr) 2001-05-30

Family

ID=3512210

Family Applications (1)

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EP99938048A Withdrawn EP1102580A2 (fr) 1998-08-06 1999-07-29 Formulations renfermant des micro-organismes a effet probiotique

Country Status (4)

Country Link
EP (1) EP1102580A2 (fr)
AT (1) AT407008B (fr)
AU (1) AU5270499A (fr)
WO (1) WO2000007571A2 (fr)

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ES2636439T3 (es) 2011-03-09 2017-10-05 Regents Of The University Of Minnesota Composiciones y métodos para el trasplante de microbiota de colon
WO2013176774A1 (fr) 2012-05-25 2013-11-28 Arizona Board Of Regents Marqueurs de microbiome et thérapies pour troubles du spectre autistique
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AU2015245942A1 (en) * 2014-04-09 2016-10-20 Kambouris Shares Pty Ltd Methods and compositions for delivering a probiotic
CN114159476A (zh) 2015-05-14 2022-03-11 克雷斯顿沃控股公司 用于粪便菌群移植的组合物以及用于制备和使用它们的方法和用于递送它们的装置
WO2016191356A1 (fr) 2015-05-22 2016-12-01 Arizona Board Of Regents On Behalf Of Arizona State University Procédés pour le traitement d'un trouble du spectre de l'autisme et de symptômes associés
US20170360848A1 (en) 2016-06-15 2017-12-21 Arizona Board Of Regents On Behalf Of Arizona State University Methods for treating autism spectrum disorder and associated symptoms
US10849936B2 (en) 2016-07-01 2020-12-01 Regents Of The University Of Minnesota Compositions and methods for C. difficile treatment
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US11213549B2 (en) 2016-10-11 2022-01-04 Finch Therapeutics Holdings Llc Compositions and method for treating primary sclerosing cholangitis and related disorders
US10092601B2 (en) 2016-10-11 2018-10-09 Crestovo Holdings Llc Compositions and methods for treating multiple sclerosis and related disorders
WO2018071537A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes pour traiter la sclérose en plaques et des troubles associés
US11040073B2 (en) 2017-04-05 2021-06-22 Finch Therapeutics Holdings Llc Compositions and methods for treating diverticulitis and related disorders
US11433102B2 (en) 2017-04-05 2022-09-06 Finch Therapeutics Holdings Llc Compositions and methods for treating Parkinson's disease (PD) and related disorders
JP2020521760A (ja) 2017-05-26 2020-07-27 クレストヴォ・ホールディングス・エルエルシー 糞便微生物ベースの治療剤を含む凍結乾燥組成物ならびにそれを製造および使用する方法
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WO2020069280A1 (fr) 2018-09-27 2020-04-02 Crestovo Holdings Llc Compositions et méthodes de traitement de l'épilepsie et de troubles apparentés

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Also Published As

Publication number Publication date
AU5270499A (en) 2000-02-28
WO2000007571A2 (fr) 2000-02-17
ATA136098A (de) 2000-04-15
AT407008B (de) 2000-11-27
WO2000007571A3 (fr) 2000-05-11

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