The future of probiotics - drprobiotikum.hu · Different probiotic genera/species/strains have different properties •Multi-strain offers wider protection, and potential for synergy

The future of probiotics:as illustrated by the Protexin company

Peter CartwrightMSc, MA, BSc, BA(Hons)

Structure of lecture

1. Introduction2. Protexin’s history and approach3. Advances in science, and changes in

regulation4. Condition-specific products, and research

programmes

© Protexin • Futrue of Probiotics • Peter Cartwright 2013 Slide 2

1. Introduction: Peter Cartwright

•Microbiologist–MSc Microbiology, BSc Biomedicine

• Human microbiota specialist with Protexin

• Manages human product research programmes

• 17 years’ experience leading patient associations

• Lecturer on probiotics - >50 talks to doctors in 14 countries


Author of books on intestinal health

• Independent author of 4 books:–on probiotics, ulcerative

colitis and diverticulitis

• Including Probiotics for Crohn’s & Colitis


Definition of probiotics

“live microorganisms which when administered in adequate amounts confer a health benefit on the host”

(Joint FAO/WHO, 2002)


2. Protexin’s history and approach

•Exclusively a probiotics company

•Formed in 1992, in Somerset, England

•Farmers recognised anti-pathogen properties of lactic acid bacteria


Probiotics support the gut microbiota

Duodenum10,000/ml

Ileum100,000,000/ml

Stomach1,000/ml

Jejunum100,000/ml

Large intestineUp to 1,000,000,000,000/gm

Human probiotic products developed

•Public readily accept probiotic concept

•Familiarity with yoghurt•Gut protection and accelerated recovery


Principles of products:

• Selecting strains on basis of range of properties• Only ‘natural’ strains used

–species normally found in human gut–no genetic engineering

• Safety of strains checked:–Antibiotic resistance–Toxin production

• High quality production• Associated educational services

–Literature, website and lectures


Lactobacillus spp.

Bifidobacterium spp.

Microbes commonly used as probiotics


Understanding strains further

• Reading University, Berkshire, England

• Research led by Profs. Glenn Gibson & Ian Rowland

• Examining Protexin strains for:–antimicrobial activity–mechanisms of action– inter-strain synergies

• Publications (Chapman et al, 2011; Chapman et al, 2012; Tejero-Sarinena et al, 2012)

• Sample slide of researcher followsProf. Glenn Gibson

Prof. Ian Rowland

Batch Culture fermentationsby Sandra Tejero

In vitro, anaerobic, stirred, pH (6.7-6.9) and temperature (37°C) controlled batch cultures

3 faecal donors

1% (v/v) faecal slurry

Gut model media

againstS. Typhimurium

C. difficile

L. casei PXN 37L. acidophilus PXN 35B. breve PXN 25Mix of 15

The following combinations will be tested in every batch culture run:


High quality production• Pharmaceutical grade

production• Good manufacturing practice

(GMP) – awarded by the UK Govt’s Medicines and Healthcare Products Regulatory Agency


Protexin is member of:

• International Scientific Association for Probiotics and Prebiotics (ISAPP)

• International Probiotic Association (IPA)

• Health Food Manufacturers Association (HFMA)


Different probiotic genera/species/strains have different properties• Multi-strain offers wider protection, and potential for synergy

(Chapman et al, 2012; Timmerman et al, 2004)

• Also, some species found in certain age groupsBifidobacterium infantis more common in babies, due to ability to digest human milk oligosaccharides(Sela et al, 2008)


Evidence of benefit from Protexin products

Day Control(n=27)

Control(n=27)

ProtexinRestore

(n=24)

ProtexinRestore

(n=24)

Significance(p<0.05)

Frequency Mean Frequency Mean

1 131 4.85 118 4.91 0.928

2 125 4.63 63 2.65 0.017

3 74 2.74 18 0.75 0.000

4 40 1.48 6 0.25 0.055

Acute gastroenteritis in infantsFrequency of purging

Yala (2010) PIDSP Journal 11: 86-91.

• Initial dose within six hours of hospital admission• Significant reduction in duration of hospital stay


Helicobacter pylori infection in children(treatment in addition to ‘triple therapy’ of antibiotics and PPI)

H. pylori Protexin Restore

Placebo P. value

Eradicated 30 (90.9%) 23 (69.7%) 0.04

Not eradicated 3 (9.1%) 10 (30.3%)


Protexin Restore reduced side effects of drugs:Nausea/vomiting (p = 0.02)Diarrhoea (p = 0.04)

Khodadad et al (2013) Iranian Journal of Pediatrics 23: 79-84

Benefit from Protexin Balancein constipated adults

Balance(n=31)

Placebo(n=29)

Mean difference

Baseline 2.29 2.17 0.11

Week 2 4.81 3.48 1.32

Week 4 5.45 3.86 1.58


Increased stool frequency (SF) in Protexin Balance group (SF/week) [p=0.02]53% increase after 2 weeks;64% increase after 4 weeks (ex-placebo effect)Study group: constipated males

Fateh et al (2011) Swiss Medical Weekly 141: w13239

3. Changes in science and regulation

• Recent scientific developments:• Improved understanding of the fundamentals• Understanding healthy gut microflora

–Metagenomics (mapping species)–How species inter-relate (eg: by-product of one species used by

another)–Dysbioses in different GI conditions

• Bacterial influences on immune system• Role of bacteria in brain-gut axis (eg: IBS)• Effects on metabolism (eg: obesity)


A change in regulation

• European Commission regulation EC1924/2006

• Health claims on foods• To protect public from false

claims• List of approved claims

came into force July 2012


No probiotic products on first official list

•Classified as food supplement–Idea developed from

yoghurts

•All clinical trials in diseased populations rejected by EFSA

•Food not allowed to make claim about disease


Probiotics as medicines


• Companies and scientists increasingly focus on probiotics as medicine

• Discovering what EMA requires of live biotherapeutic microbes

• Enquiries led by Pharmabiotic Research Institute

4. Condition-specific research• Increased knowledge of gut

microbiota, plus greater interest in probiotics as licensed medicines

• Plans for condition-specific products by Protexin

• Research team based at the Blizard Institute (School of Medicine, Queen Mary College, University of London)

• This research programme started in May 2011


Main personnel(from Blizard Institute)


Prof. Lucinda Hall Microbiologist

Prof. Qasim AzizConsultant Gastroenterologist

Prof. Gareth Sanger Neurophysiologist

Research in four parts• Screening strains for general desirable qualities• Screening strains for condition-specific properties• Human study in healthy volunteers• Two human studies in target patients:

–Pilot trial–Full randomised controlled trial



257 strains for screening process:selection by in vitro assays of mucosal adhesion and acid/bile survival

3 CultureCollections(37 strains)

Protexincurrent

strains (12)

Biopsyisolates

(48 strains)

Czech Uni.(142 strainsscreened)

Shortlist of 10 strains

4 strains forhuman studies

Scottishculture collection

(47 strainsscreened)

Biopsy isolates

•Future strains sourced from healthy human gut mucosal surface

•Routine screening process (colonoscopy)

• Isolation of lactobacilli and bifidobacteria for screening tests


In vitro tests to identify best bacterial strains

•Tests for:–Stomach and duodenum

survival (acid and bile)–Mucosal adhesion (auto-

aggregation, mucin adhesion)

•Anti-pathogen effects relevant to condition


Secondary benefit of research:Protexin ‘Pool’ of superior strains for future research


• Twenty strains superior to ‘gold standard’ strains

• Stored at -80°C in specialist culture collections

Other condition-specific research:prevention and treatment of diarrhoea caused by Clostridium difficile infection• Broad-spectrum antibiotics

disturb gut microflora• Clostridium difficile takes

advantage of damaged microflora to cause diarrhoea

• Eradication of C. difficile is becoming more difficult

– increase of antibiotic-resistant strains

–especially in hospitals


Protexin strain has early evidence of effect against C. difficile• Bacillus subtilis PXN21• Species used widely to treat

diarrhoea• Soil bacterium• Life-cycle includes transit through

mammalian gut (including human)• Forms spores, which protect against

stomach acid


Properties of Protexin strain examined at Royal Hollway College, Univ. of London•Work overseen by Prof. Simon Cutting

•A world leader in bacilli as probiotics

•Examining:–Ability to bind toxins– Immune regulation–Effect against Clostridium

difficile

Further information on probiotics

•Probiotic Allies by Peter Cartwright

•Describes the various microbiota

•Evidence of benefit from probiotics in various gut conditions

•Available from VitaminKosar Kft.


Summary

• Protexin is well established specialist company:–Activities illustrate current and future state of probiotics

• Multi-strain products to GMP standards• Changes in regulation and in scientific knowledge• Moves towards probiotics as medicines• Condition-specific probiotics in the future


References• Chapman, C.M., Gibson, G.R. & Rowland, I. (2011) Health benefits of probiotics: Are mixtures more effective than single

strains? European Journal of Nutrition 50: 1–17.• Chapman, C.M., Gibson, G.R. & Rowland, I. (2012) In vitro evaluation of single- and multi-strain probiotics: Inter-species

inhibition between probiotic strains, and inhibition of pathogens. Anaerobe 18:405-413. • Fateh, R., Iravani, S., et al. (2011) Synbiotic preparation in men suffering from functional constipation: a randomised

controlled trial. Swiss Medical Weekly 141: w13239.• Joint FAO/WHO Working Group on Evaluation of Probiotics (2002) Guidelines for the Evaluation of Probiotics in Food

(London Ontario, Canada). Food and Agriculture Oganization of the UN/World Health Organization. ftp://ftp.fao.org/es/esn/food/wgreport2.pdf [Accessed 13 September 2010]

• Khodadad, A., Farahmand, F., et al. (2013) Probiotics for the treatment of pediatric Helicobacter pylori infection: a randomised double blind clinical trial. Iranian Journal of Pediatrics 23: 79-84.

• Sela, D.A., Chapman, J., et al. (2008) The genome sequence of Bifidobacterium longum subsp. infantis reveals adaptations for milk utilization within the infant microbiome. PNAS 105: 18964-18969.

• Tejero-Sarinena, S., Barlow, J., et al. (2012) In vitro evaluation of the antimicrobial activity of a range of probiotics against pathogens: evidence for the effects of organic acids. Anaerobe 18: 530–538.

• Timmerman, H.M., Konig, C.J., et al. (2004) Monostrain, multistrain and multispecies probiotics – A comparison of functionality and efficacy. International Journal of Food Microbiology 96:219-233

• Yala, E.T. (2010) The clinical efficacy of multi-strain probiotics (Protexin) in the management of acute gastroenteritis in children two months to two years old. PIDSP Journal 11: 86-91.


The future of probiotics - drprobiotikum.hu · Different probiotic genera/species/strains have different properties •Multi-strain offers wider protection, and potential for synergy

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