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WO2012146890A1 - Agent prébiotique - Google Patents

Agent prébiotique Download PDF

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Publication number
WO2012146890A1
WO2012146890A1 PCT/GB2012/000380 GB2012000380W WO2012146890A1 WO 2012146890 A1 WO2012146890 A1 WO 2012146890A1 GB 2012000380 W GB2012000380 W GB 2012000380W WO 2012146890 A1 WO2012146890 A1 WO 2012146890A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
improvement
iron
iron chelator
gastrointestinal
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.)
Ceased
Application number
PCT/GB2012/000380
Other languages
English (en)
Inventor
Chris Tselepis
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.)
University of Birmingham
Original Assignee
University of Birmingham
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 University of Birmingham filed Critical University of Birmingham
Publication of WO2012146890A1 publication Critical patent/WO2012146890A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
    • 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
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2013—Organic compounds, e.g. phospholipids, fats
    • A61K9/2018—Sugars, or sugar alcohols, e.g. lactose, mannitol; Derivatives thereof, e.g. polysorbates
    • 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
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2054—Cellulose; Cellulose derivatives, e.g. hydroxypropyl methylcellulose
    • 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
    • A61P35/00—Antineoplastic agents

Definitions

  • This invention relates to methods and medicaments for the enhancement of gastrointestinal health, and/or for the improvement of the gastrointestinal bacterial population.
  • Gl mammalian gastrointestinal
  • Microbes utilised for this purpose include lactic acid bacteria (LAB), bifidobacteria, and certain yeasts and bacilli. It is thought that the main mechanism of action is by adjustment of the microbiota population in favour of non-pathogenic microorganisms and away from pathogenic organisms.
  • LAB lactic acid bacteria
  • yeasts and bacilli certain yeasts and bacilli. It is thought that the main mechanism of action is by adjustment of the microbiota population in favour of non-pathogenic microorganisms and away from pathogenic organisms.
  • a composition for the improvement of gastrointestinal health and/or for the improvement of a gastrointestinal bacterial population comprising an iron chelator, wherein the composition is adapted for the selective targeting of the iron chelator to the colon.
  • a composition for the improvement of gastrointestinal health and/or for the improvement of a gastrointestinal bacterial population comprising an iron chelator, wherein the iron chelator is unable to bind iron until the composition reaches the colon.
  • a chelator binds free metal ions and removes them from solution. Metal ions bound by a chelator molecule are effectively inactivated because they are no longer available to react with other chemical species, or for certain biological actions. Chelators are usually specific for particular metal ions.
  • An iron chelator is a chelator that binds Fe ions. In certain embodiments said chelator may be iron specific.
  • the iron chelator may be naturally occurring, or it may be man-made. In a particular embodiment, the chelator is naturally-occurring.
  • the iron chelator comprises phytic acid, tea flavonoids (catechins), melanoidins, curcumin, capsaicin, polyphenols, alginates, iron-binding proteins (such as for example lactoferrin), bacterial siderophores (such as for example enterobactin) or a combination of any of these.
  • tea flavonoids catechins
  • melanoidins curcumin
  • capsaicin polyphenols
  • polyphenols such as for example lactoferrin
  • alginates iron-binding proteins
  • iron-binding proteins such as for example lactoferrin
  • bacterial siderophores such as for example enterobactin
  • the iron chelator is non-digestible, non-absorbable and non- fermentable in the gastrointestinal tract.
  • a chelator which is not digested or degraded by the body will be more effective at binding excess iron. If the chelator cannot be absorbed, any iron bound by the chelator will also be prevented from being absorbed by the body. The iron will therefore be removed from the body as an iron-chelator complex.
  • compositions for the improvement of gastrointestinal health and/or for the improvement of a gastrointestinal bacterial population comprising a prebiotic, wherein the composition is adapted for selective targeting of the prebiotic to the colon and is substantially free from probiotic.
  • the prebiotic may also be an iron chelator.
  • composition has no iron-chelating effect or other biological (such as prebiotic) activity (as appropriate) until it reaches the colon. This may also be referred to as “colonic delivery”.
  • Colonic delivery of the iron chelator or prebiotic may be achieved by oral or rectal administration of the composition.
  • the composition may be in the form of a suppository incorporating the iron chelator or prebiotic and a carrier such as cocoa butter, or in the form of an enema.
  • the composition may be in the form of discrete units such as capsules, cachets, tablets or lozenges, each containing a predetermined amount of the iron chelator or prebiotic; in the form of a powder or granules; in the form of an emulsion, suspension in an aqueous liquid or non-aqueous liquid.
  • the composition may also be in the form of a bolus, electuary or paste.
  • the composition may be formulated for incorporation into a foodstuff.
  • colonic delivery of the iron chelator or prebiotic may be achieved by ingestion of the iron chelator or prebiotic in an inactivated form, or in a form which otherwise prevents the iron chelator from binding iron, or the prebiotic from having any biological activity, until it reaches the colon.
  • the iron chelator may be ingested in the form of a precursor or prodrug, which is activated or modified in vivo such that the active iron chelator or prebiotic is only released in the colon.
  • the composition comprising the iron chelator or prebiotic is encapsulated by a coating which remains intact while the composition passes through the stomach and small intestine, but which degrades in the colon.
  • colonic delivery is achieved using a coating which is pH sensitive, time- dependent, pressure-dependent or degradable by colonic bacteria.
  • the composition is encapsulated by a biodegradable coating.
  • the coating may be a polymer.
  • the coating comprises colon-targeted microsponges, or a microbially-triggered osmotic pump.
  • the composition may be targeted to the colon by a combination of the above strategies.
  • the coating comprises a biodegradable polysaccharide.
  • the coating may comprise one or more biodegradable polysaccharides selected from the group consisting of albizia gum, alginates, amylose, arabinogalactan, cellulose, chitosan, chondroitin sulphate, curdlan, cyclodextrin, dextran, furcelleran, galactomannan, gellan gum, guar gum, hyaluronic acid, inulin, kara gum, karaya gum, locust bean gum, scleroglucan, starch, pectin, pullulan or xylan.
  • colonic targeting may reduce the incidence of side effects caused by action of the iron chelator or prebiotic in other parts of the Gl tract.
  • the use of colonic targeting may prevent degradation of the iron chelator or prebiotic in other parts of the Gl tract, thereby providing a relative increase in bioavailability of the iron chelator or prebiotic in the colon.
  • a method for the improvement of gastrointestinal health and/or for the improvement of the gastrointestinal bacterial population of a human or animal patient comprising administering to the patient a composition according to any of the first, second or third aspects of the invention.
  • Health promoting bacteria comprise the genera of bifidobacterium (e.g. B. longum, B. reve) and lactobacillus (e.g. L. acidophilus, L. rhamnosus, L. gassen).
  • Pathogenic bacteria comprise the genera Clostridia, bacteriodes, enterobacteriaceae and desulfovibrio.
  • Bacterial levels may be assessed by any suitable method, including for example by high throughput pyrosequencing, such as high throughput Roche-454 pyrosequencing. In brief, this involves extraction of faecal DNA followed by amplification with conserved ID-tagged primers (such as for example those of the V1 -V3 region of bacterial 16S rDNA), followed by sequencing with Roche-454 Titanium amplicon protocols. Typically, thousands of sequence reads per sample can then be obtained routinely to assess the proportions of different bacterial taxa.
  • increasing or reducing the levels of bacteria comprises increasing or reducing the relevant bacterial populations by at least 5%, at least 10%, at least 15%, at least 20% or at least 50%.
  • increasing the levels of bacteria may comprise increasing the bacterial levels above zero, and/or reducing the levels of bacteria may comprise reducing the bacterial levels to zero.
  • the improvement of a gastrointestinal bacterial population is intended to refer to an increase in the ratio of the level of health promoting bacteria to the level of pathogenic bacteria.
  • increasing the ratio of the level of health promoting bacteria to the level of pathogenic bacteria comprises increasing the ratio by at least 5%, at least 10%, at least 15%, at least 20% or at least 50%.
  • the improvement of gastrointestinal health comprises the treatment of irritable bowel syndrome (IBS).
  • IBS irritable bowel syndrome
  • Measurement of treatment of irritable bowel syndrome may be made by any suitable method, such as for example by assessing patient symptom scores based on questionnaires designed to assess quality of life (QoL).
  • QoL assessments may be used in parallel, such as disease specific tools such as the IBS-QOL and generic instruments such as the well-validated 36 item short form general health survey (SF-346) (described in Ware et al., Med Care, 30(6), 1992, 473-83) or the sickness impact profile (SIP) (described in Bergner et al, Med Care, 19(8), 1981 , 787-805).
  • SF-346 well-validated 36 item short form general health survey
  • SIP sickness impact profile
  • the improvement of gastrointestinal health comprises the treatment of inflammatory bowel disease (IBD).
  • IBD inflammatory bowel disease
  • Measurement of treatment of inflammatory bowel disease may be made by any suitable method, such as for example well validated symptom scores for patients with either ulcerative colitis (UC) (such as the simple clinical colitis index - SCCI) or Crohn's disease (CD) (such as the Crohn's disease activity index - CDAI).
  • UC ulcerative colitis
  • CD Crohn's disease
  • QoL tools either specific for IBD (IBDQ) or generic (SF36) allow comparison with studies in other patient groups (such as IBS).
  • the improvement of gastrointestinal health comprises the treatment of colorectal neoplasia.
  • Measurement of treatment of colorectal neoplasia may be made by any suitable method, such as for example by visualisation of adenoma numbers and cancers from colonoscopies and pathological examination of collected biopsy material.
  • the improvement of gastrointestinal health comprises the treatment of antibiotic-associated colitis.
  • Measurement of treatment of antibiotic-associated colitis may be made by any suitable method, such as for example by measurement of production of C. difficile toxin, microbiota populations, and diarrhoea (using the Bristol stool chart).
  • the composition comprises alginate. In some further embodiments, the composition comprises sodium alginate. It will be understood that the alginate (such as sodium alginate) may function as an iron chelator and/or prebiotic. Thus, in an embodiment of the first and second aspects of the invention, the iron chelator comprises alginate. In an embodiment of the third aspect of the invention, the prebiotic comprises alginate. In some further embodiments, the alginate is covalently complexed to a further iron chelator molecule. In particular, the alginate may be covalently complexed to an iron- chelator molecule having a greater affinity for iron than the alginate.
  • the iron chelator molecule may have a greater specificity for iron than the alginate.
  • the composition of the present invention may include one or more additional pharmaceutically acceptable ingredients such as excipients, bulking agents, diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants and preservatives.
  • the amount of alginate administered to the patient is between 0.1g and 4g per day, between 0.2g and 3g per day, between 0.4g and 2g per day or between 0.5g and 1.5g per day, which may be administered as a single dose or multiple doses (e.g. 2, 3 or 4 doses at intervals of e.g. 3, 6 or 8 hours).
  • the composition according to the first and second aspects of the invention comprises a chelator - a chemical which binds iron.
  • Alginates are an example of a naturally-occurring chelator of iron and are used by the UK food industry as gelling, emulsifying and stabilising agents and thus are extensively found in products such as ice creams, jams, sauces and desserts. Alginates are also found in medicines, particularly in anti-reflux medicines. Alginates are therefore known to be safe for consumption by humans and animals. A further advantage of alginates is that they cannot be digested or absorbed by the human body. Both in vitro and in vivo studies have shown that alginates can stably bind iron and may prevent iron absorption. In particular, studies carried out by the inventors have shown that iron- mediated cellular proliferation is inhibited in the presence of alginate (unpublished data).
  • Figure 1 shows the growth kinetics of L. acidophilus in the presence of different concentrations of fragmented alginate
  • Figure 2 shows the growth kinetics of L acidophilus in the presence of 0.1% whole and fragmented alginate
  • Figure 3 shows the growth kinetics of L. acidophilus in the presence of 0.1% whole and fragmented alginate, and 0.1% whole alginate pre-loaded with iron.
  • compositions comprising an iron chelator selected from list A is coated with a colonic- delivery component selected from list B to give compositions 1a to 1 g, 2a to 2g and 3a to 3g.
  • a colonic- delivery component selected from list B to give compositions 1a to 1 g, 2a to 2g and 3a to 3g.
  • a composition comprising microcrystalline cellulose, sugar, propyl paraben and 0.5g sodium alginate is formulated into a 1g tablet and coated with a biodegradable polysaccharide. Two tablets are orally administered to a patient with inflammatory bowel disease three times a day at A hourly intervals.
  • the following strains were used in this study: Staphylococcus aureus, Escherichia coli, Salmonella typhimurium and Lactobacillus acidophilus.
  • the anaerobic bacteria L. Acidophilus
  • the anaerobic bacteria were cultured in anaerobic conditions at 37°C in anaerobic jars (BD anaerobic systems with GasPak) using Wilkins-Chalgren agar (Oxoid) supplemented with 5% defibriginated horse blood or Wilkins-Chalgren broth (Oxoid).
  • Aerobic bacteria (Staphylococcus aureus, Escherichia coli and Salmonella typhimurium) were cultured using Luria Bertani (LB) broth and agar (Sigma). Broth and agar were prepared according to the manufacturers instructions.
  • 'whole' alginate solution 2g of alginate as purchased was dissolved in 100ml of appropriate broth.
  • 2% 'whole' alginate solution was incubated with 10U of alginate lyase (Sigma) in a shaking incubator at 37°C for 12 hours.
  • the 'iron loaded' alginate solutions were prepared by dialysing the 'whole' or 'fragmented' alginate solutions across a 12 KDa membrane (Sigma) against 0.2 mMol iron sulphate dissolved in broth for four hours.
  • the resultant solutions were autoclaved at 121°C for 15 minutes to sterilise.
  • the Minimum Inhibitory Concentration was performed using a microdilution technique. Alginate solutions prepared as described above (initial concentration 2% w/v) were diluted in broth in serial twofold dilutions in a 96 well microtitre plate. An overnight broth of the required bacteria was diluted to 1 :100. 50 ⁇ of diluted bacterial solution was added to 50 ⁇ of the diluted alginate solution, the plate covered and incubated at 37°C in the appropriate atmosphere for 18 hours. The MIC was determined as the lowest concentration of the alginate showing no visible bacterial growth at 18 hours, i.e. the minimum concentration of alginate required to inhibit bacterial growth. The assay was repeated with all bacterial strains and alginate solutions.
  • a 96 well microtitre plate was set up as described for MIC. This was incubated at 37°C on a microplate reader (FLUOstar optima) for 12-18 hours. Absorbance (OD) was measured at 10 minute intervals at 600nm. This was repeated for all species and all alginate solutions. Anaerobes were incubated under a 50 ⁇ layer of sterile mineral oil to replicate anaerobic conditions. This was carried out on 4 independent occasions. Results
  • Sodium alginate was tested against a variety of bacteria ⁇ Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Lactobacillus acidophilus) using a minimum inhibitory concentration (MIC) assay.
  • the whole alginate i.e. whole molecules of alginate which have not been broken down
  • MIC minimum inhibitory concentration
  • the susceptibility of each species to alginate was variable with E. coli demonstrating the most resistance to the effect of alginate, as evidenced by a much higher MIC (1% vs ⁇ 0.5% for other strains).
  • the bacteriostatic effect of alginate was also observed when using fragmented alginate (i.e. alginate molecules which have been cleaved/broken down enzymatically into smaller subunits) in the MIC assay in all species. However, the fragmented alginate was less efficacious, giving a higher MIC.
  • the MIC assay gives a result from a single time point and so in order to investigate the consequences of alginate on bacterial growth curves in more detail, growth kinetics were studied using a microplate absorbance reader. Table 1. Minimum percent concentration of alginate required to inhibit growth

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  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne une composition destinée à améliorer la santé gastro-intestinale et/ou à améliorer une population de bactéries gastro-intestinales. La composition comprend un prébiotique, est conçue pour diriger de manière sélective ledit prébiotique vers le côlon, et est sensiblement exempte de probiotique.
PCT/GB2012/000380 2011-04-26 2012-04-25 Agent prébiotique Ceased WO2012146890A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB1107068.7 2011-04-26
GBGB1107068.7A GB201107068D0 (en) 2011-04-26 2011-04-26 Prebiotic agent

Publications (1)

Publication Number Publication Date
WO2012146890A1 true WO2012146890A1 (fr) 2012-11-01

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PCT/GB2012/000380 Ceased WO2012146890A1 (fr) 2011-04-26 2012-04-25 Agent prébiotique

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WO (1) WO2012146890A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108403970A (zh) * 2018-05-02 2018-08-17 福建省农业科学院农业工程技术研究所 一种益生元组合物及其制备方法和应用
WO2021113542A1 (fr) * 2019-12-03 2021-06-10 Animal Microbiome Analytics, Inc. (D/B/A Animalbiome) Système et méthodes de manipulation du microbiome buccal chez des mammifères
CN115025116A (zh) * 2022-07-13 2022-09-09 纽湃腾（北京）医药科技有限公司 一种显著改善胃食管反流伴肠易激综合征的组合物及其应用

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108403970A (zh) * 2018-05-02 2018-08-17 福建省农业科学院农业工程技术研究所 一种益生元组合物及其制备方法和应用
CN108403970B (zh) * 2018-05-02 2020-10-16 福建省农业科学院农业工程技术研究所 一种益生元组合物及其制备方法和应用
WO2021113542A1 (fr) * 2019-12-03 2021-06-10 Animal Microbiome Analytics, Inc. (D/B/A Animalbiome) Système et méthodes de manipulation du microbiome buccal chez des mammifères
CN115025116A (zh) * 2022-07-13 2022-09-09 纽湃腾（北京）医药科技有限公司 一种显著改善胃食管反流伴肠易激综合征的组合物及其应用
CN115025116B (zh) * 2022-07-13 2023-11-21 纽湃腾（北京）医药科技有限公司 一种显著改善胃食管反流伴肠易激综合征的组合物及其应用

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