Clinically-Documented Probiotics for Human Health

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Clinically-Documented Probiotics for Human Health
Brian Peeters Strategic Sales Manager Chr. Hansen
A/S
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Agenda

• Introduction
• Why Chr. Hansen as Preferred Supplier
• Basics of Probiotics
• Market Trends
• Designing a Probiotic Product for the Military

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Chr. Hansen

• Company founded in 1874
• Fermenting bacteria cultures for more than 100
  years
• 2,500 employees in 30 countries
• Turnover of 500M EUR with double-digit organic
  growth
• 6 of turnover spent on RD
• 225 employees in Research Development
• State-of-the-art production facilities
• GMP pharma food, Kosher, QC, HACCP,ISO 9002
• Production facilities on 5 continents
• Strong partnerships with customers
• Everyday more than 500 million people consume
  products containing Chr. Hansen ingredients

Founder Danish Pharmacist Christian D.A. Hansen
when science matters
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Chr. Hansen Group serving global customers all
over the world with gt25 offices
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Chr. Hansen Organizational Structure
Health Nutrition Division is dedicated solely
to Probiotics
Chr. Hansen
Cultures Enzymes
Natural Colors
Health Nutrition
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Chr. Hansen - Worlds Largest Lactic Acid
Bacteria PlantLocation Avedore, Denmark
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Agenda

• Introduction
• Why Chr. Hansen as Preferred Supplier
• Basics of Probiotics
• Market Trends
• Designing a Probiotic Product for the Military

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Why Partner with Chr. Hansen?

• Documented Efficacy and Safety
• More than 450 research papers supporting Chr.
  Hansen probiotic strains
• More than 80 human clinical studies documenting
  strain efficacy
• Strains selected among the 8,700 strain deposits
  in Chr. Hansens cell bank in Denmark
• Extensive QC programs (cGMP)
• Been producing and selling probiotics for gt20
  years
• Scientific expertise
• 6 of turnover spent on RD
• 225 employees in Research Development
• Patented processes and concepts
• Novel equipment and production technologies
• Strong research team developing new probiotics
  strains

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Why Partner with Chr. Hansen?

• Product Supply
• Market leader with worlds largest lactic acid
  bacteria plant
• Pharmaceutical Production (Pharma cGMP)
• Global quality and safety policy ensuring high
  standard in food production (ISO 9001/HACCP)
• Consumer Friendly
• Well-established safety
• GMO-free products
• Big 8 Allergen-free products available
• Internationally Locally Affiliated
• ISAPP, IPA, Norfood, PROBDEMO, LABIP
• Leading probiotic research groups at universities
  institutes

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Agenda

• Introduction
• Why Chr. Hansen as Preferred Supplier
• Basics of Probiotics
• Market Trends
• Designing a Probiotic Product for the Military

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Definition of Probiotics

• The World Health Organization (WHO) and the Food
  and Agriculture Association (FAO) define
  probiotics as
• Live microorganisms that when consumed in
  adequate amounts confer a health benefit on the
  host.

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Introduction to Probiotics

• Probiotics means for life in Latin
• Probiotics are live bacteria with health benefits

• First scientist to discover health benefits of
  probiotics was immunologist Dr. Eli Metchnikoff
• Received Nobel Prize in 1908

10x more bacterial cells in human body (1014)
than there are human tissue cells (1013)
Gastrointestinal tract is largest immune organ
in human body (80 of Immune System is located
here)

• Probiotics first commercialized via yogurts
• Yakult introduced in Japan in 1935
• Danones Activia introduced in France in 1987,
  U.S. in 2006

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Strain TaxonomyNote Overview not exhaustive
Bacterial genus
Bacterial species
Bacterial sub-species
Bacterial strains
Bacterial group
paracasei subsp. paracasei
paracasei
Lactobacillus
Lactid acid bacteria
CRL-431
acidophilus
Bifidobacterium
LC-01
paracasei subsp. tolerans
F-19
fermentum
Enterococcus
reuteri
Weissella
delbrueckii
Carnobacterium
rhamnosus
Streptococcus
plantarum
Lactococcus
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Probiotics Mechanisms of Action
Probiotic strain
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Probiotics Mechanisms of Action
Probiotic strain
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Agenda

• Introduction
• Why Chr. Hansen as Preferred Supplier
• Basics of Probiotics
• Market Trends
• Designing a Probiotic Product for the Military

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Scientific interest in probiotics has grown
significantly since 2000
PubMed Articles on Probiotics ()
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and so has the number of probiotic trials
Probiotic trials registered on www.clinicaltrials.
gov NIH ()
US trials
Non-US trials
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Increasing interest in probiotics seen in recent
yearsDietary supplements Gone from 16 launches
in 02 to 75 launches in 07
Probiotic product launches globally ()
669
594
Other
491
Cheese
394
Diet. suppl.
242
150
Fermented milks
Source Mintel database Global launches
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Increasing number of dietary supplement probiotic
product launches in 2007 (driven by Europe)
Probiotic product launches globally dietary
supplements ()
75
Rest of World
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Europe
38
31
16
Japan
US
Source Mintel database Global launches
35 average annual growth last 5 years
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Chr. Hansen sees market drivers in probiotic
supplement market continuing to feed a strong
growth
General interest in self-medication
Taboo on intestinal issues declining
Probiotic supplements expected to continue
double-digit growth
Demographic shift
Trend towards natural cure rather than pharma
Consumer awareness below 50 in most countries
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Agenda

• Introduction
• Why Chr. Hansen as Preferred Supplier
• Basics of Probiotics
• Market Trends
• Designing a Probiotic Product for the Military

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Designing a Probiotic Product for the Military
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Indication
2
8
Regulatory / Claim
Other
Strain(s)
Shelf stability
Your product
3
7
Potency
Survival through stomach
Dosage form
6
4
5
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Chr. Hansen offers different indication concepts
1. Everyday Intestinal Balance 2. Antibiotic
Associated Diarrhea 3. Traveler's Diarrhea 4.
Children's Diarrhea 5. Immune Boost 6. Atopic
Eczema 7. Bacterial Vaginosis
Concept supported by fewer clinical studies
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Chr. Hansen offers different indication concepts
1. Everyday Intestinal Balance 3. Traveler's
Diarrhea 5. Immune Boost
Concept supported by fewer clinical studies
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Chr. Hansen offers different indication concepts
1. Everyday Intestinal Balance 2. Antibiotic
Associated Diarrhea 3. Traveler's Diarrhea 4.
Children's Diarrhea 5. Immune Boost 6. Atopic
Eczema 7. Bacterial Vaginosis
Concept supported by fewer clinical studies
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1. Everyday intestinal balance

• Covers a broad range of discomfort such as
  bloating, slow constipation, diarrhea, and slight
  irritable bowel
• Surveys show 15-20 of consumers are concerned
  about their digestive health
• 12 of people worldwide suffer from self-defined
  constipation according to large global omnibus
  study from 2006
• Americas and Asia Pacific suffer nearly twice
  (17) as much as their European counterparts (9)
• 40 of constipation sufferers attempt to treat
  their constipation by changing their nutrition,
  despite research showing that diet and lifestyle
  are not necessarily to blame for the occurrence
  of constipation
• Less than 1/3 of sufferers use laxatives

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Chr. Hansen offers different indication concepts
1. Everyday Intestinal Balance 2. Antibiotic
Associated Diarrhea 3. Traveler's Diarrhea 4.
Children's Diarrhea 5. Immune Boost 6. Atopic
Eczema 7. Bacterial Vaginosis
Concept supported by fewer clinical studies
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3. Traveler's Diarrhea

• Travelers' diarrhea is the most common illness
  affecting travelers. Each year between 20-50 of
  international travelers, an estimated 10 million
  persons, develop diarrhea -Centers for Disease
  Control and Prevention, U.S. Department of
  Health and Human Services
• Duration of travelers' diarrhea usually limited
  to 2-5 days
• In some cases, symptoms may persist for months or
  even years
• Probiotic supplementation reduces the risk of
  diarrhea caused by foreign bacteria from water,
  vegetables, fruit, etc.
• Diarrhea is caused mainly by intake of E. coli,
  Salmonella, or Campylobacter
• Target group Those persons traveling to
  high-risk destinations where the food or water
  supply may contain non-native or pathogenic
  organisms such as E. coli.

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Chr. Hansen offers different indication concepts
1. Everyday Intestinal Balance 2. Antibiotic
Associated Diarrhea 3. Traveler's Diarrhea 4.
Children's Diarrhea 5. Immune Boost 6. Atopic
Eczema 7. Bacterial Vaginosis
Concept supported by fewer clinical studies
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5. Immune Boost

• 80 of the bodys immune cells are located in the
  gastrointestinal tract
• Having the right bacterial balance is important
  for optimizing the immune system
• Increased levels of IgA and phagocytes (immune
  cells and cells which can engulf and destroy
  harmful components), the better the immune system
  fights unwanted components
• Probiotics are proven to be able to strengthen
  natural defenses and stimulate the immune system
• Immune health is the second largest indication
  area within probiotics after intestinal health
• Target group Those who are under consistently
  high levels of stress and/or those who have a
  weak or compromised immune system.

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Chr. Hansens Clinically-Documented Probiotic
Strains

• Bifidobacterium (BB-12)
• Lactobacillus acidophilus (LA-5)
• Lactobacillus paracasei ssp. paracasei (CRL-431)
• Lactobacillus rhamnosus (GR-1)
• Lactobacillus reuteri (RC-14)
• Lactobacillus rhamnosus GG (LGG)
• Streptococcus thermophilus (TH-4)

BB-12
LA-5
CRL-431
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Other Chr. Hansen Strains

• Lactobacillus delbrueckii ssp. bulgaricus LBA-40
• Lactobacillus delbrueckii ssp. bulgaricus LBY-27
• Lactobacillus paracasei ssp. paracasei LC-01
• Lactobacillus rhamnosus LC-69
• Bifidobacterium infantis BB-02
• Bifidobacterium longum BB-46
• Streptococcus thermophilus STY-31

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Chr. Hansen stains have documented efficacygt450
research and clinical evaluations and gt80 human
clinical studies
Strains Intestinal balance Antibiotic Assoc. Diarrhea Traveler's Diarrhea Children's Diarrhea Immune boost Atopic Eczema Bacterial Vaginosis
BB-12 7 2 3 4
BB-12 LA-5 6 4
BB-12 TH-4 4
CRL-431 1
CRL-431 BB-12 1
LGG 2 6 2 15 6 7
BB-12 LGG 1 2
BB-12 LA-5 LBY-27 STY-31 1 2
BB-12 LA-5 LGG 1
GR-1 RC-14 8
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Chr. Hansen stains have documented efficacygt450
research and clinical evaluations and gt80 human
clinical studies
CHR Product Strains Intestinal Balance Antibiotic Assoc. Diarrhea Traveler's Diarrhea Children's Diarrhea Immune Boost Atopic Eczema Bacterial Vaginosis
BB-12 Stick BB-12 7 2 3 4
AB-Cap BB-12 LA-5 6 4
BB-12 TH-4 4
CRL-431 1
CRL-431 BB-12 1
LGG 2 6 2 15 6 7
BB-12 LGG 1 2
Quatro-Cap BB-12 LA-5 LBY-27 STY-31 1 2
BB-12 LA-5 LGG 1
GR-1 RC-14 8
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Chr. Hansen with gt450 scientific publications
Scientific publications registered by strain ()
Source CH internal abstract database
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BB-12 LA-5 for everyday intestinal
balanceClinical study example
BB-12 and LA-5 tested on 15 healthy adults with
tendency to constipation Shioya et al.2000
(days)

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BB-12 for everyday intestinal balanceClinical
study example
BB-12 tested on 209 elderly with constipation
living in residential homes Pitkälä et al. 2007
()


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Quatro-Cap-4 for Lowering Risk of Traveler's
DiarrheaClinical study example
165 tourists going to Egypt were treated with
Quatro caps or placebo 1 cap 3 times daily
starting 2 days prior to departure ending the
last day of travel Black et al. 1989 Black 1995
Share of tourists with diarrhea
Share of tourists with diarrhea


Trial I
Trial II
Trial 2
n 41
n 40
n 45
n 39
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BB-12 for Everyday Immune BoostClinical study
example
Effect of BB-12 on phagocytic activity in 14
healthy adults Schiffrin et al. 1997
Share active phagocytes ()


BB-12
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Dosage Forms

• Standard forms
• Capsules
• Sticks
• Powder blends
• Chewable tablets
• Individual customization
• Capsules
• Sticks
• Chewable tablets
• Sachets
• Tablets
• and more upon request

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Dosage Forms.For the Military
1st Choice

• Standard forms
• Capsules
• Sticks
• Powder blends
• Chewable tablets
• Individual customization
• Capsules
• Sticks
• Chewable tablets
• Sachets
• Tablets
• and more upon request

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Dosage Forms.For the Military
2nd Choice
1st Choice

• Standard forms
• Capsules
• Sticks
• Powder blends
• Chewable tablets
• Individual customization
• Capsules
• Sticks
• Chewable tablets
• Sachets
• Tablets
• and more upon request

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Dosage Form Sticks

• Best dosage form for wide range of ambient
  temperature and humidity fluctuations
• Relatively little loss of viable organisms _at_ 25C
  over 18 months
• Very low sugar content (maximum 0.4 g sugar per
  dose)
• Can be customized to contain higher doses to
  further extend shelf-life over time at various
  temperature ranges
• Consumer Appeal
• Easy to take no tablets to swallow
• Easy to consume only a small amount of powder
  (1g)
• Easy to mix into foods and drinks - cold or
  lukewarm
• Easy to open tear off top
• Easy to carry in your pocket, bag or lunch box
• Easy to store room temperature storage is ok
• Selections
• Taste Vanilla, neutral or per your selection
• Technology Soluble or non-soluble

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Dosage Form Capsules

• Consumer benefits
• Easy to take small capsule size
• Easy to open for powder administration
• Easy to carry Alu-tube or blister card
• Easy to store at room temperature
• Easy to relate to capsules are a traditional
  dosage form for supplements
• Selections for capsule type
• Type Gelatin or Vegetables (Vegetable
  recommended)
• Size 000 5 (size 3 recommended)

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Other Dosage Forms

• Powders
• Delivered in small vial to consumer to take a
  tablespoon and sprinkle onto food or beverage
• Also relevant dosage form when used for infant
  formula
• Chewable Tablets
• Children-friendly
• Sachets
• Sachets are quadrant-shaped bags with powder
• different from sticks being rectangle bags
• Tablets
• Typically 500-1500 mg per tablet

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What is potency?

• What is a CFU?
• CFU Colony Forming Units
• One CFU one live bacteria cell
• Live defined as being able to multiple and
  thereby form a colony
• How to count the cells?
• Small amounts of powder is weighing and diluted
  up to commonly 1,0 E09
• Suitable dilutions with lactic acid bacteria are
  poured onto plates and mixed with medium
• After anaerobic incubation commonly 3 days at
  37oC, the colonies are manually counted
• Different cell count is seen as cells are live
  microorganisms
• Only cells assessed as being colony forming are
  to be counted
• What is potency?
• Potency is minimum number of CFU in one gram
  (powder) or in one dosage (capsules, tablets,
  etc)
• Example Potency of 1.0 E09 per capsule means
  there is minimum 1 billion CFU per capsule
• Which potency to communicate to consumers?
• Best practice towards consumers is to state
  minimum potency at end of shelf life

Chr. Hansen recommends products providing a daily
dose of 1 to 10 Bil CFU
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Chr. Hansen guarantees potency at point in value
chain agreed with customer
Bulk dosage forms
Final dosage form
Powder production
Boxing labeling
Distri-bution
Store shelfing
Consump-tion
End of shelf life
Potency point 1
Potency point 2
Potency point 3
Potency point 4
Illustrative example of potency decline
1B
5B
5.5B
6.5B
Chr. Hansen guarantees CFU count all the way to
potency point 4 when delivering in final
packaging material
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The Shelf-Life Equation



Shelf life
Water Activity
Temperature
Oxygen
Rule-of-thumb recommendation
0.15 aw
24 months

x
x
25OC
5
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What we do to maximize shelf stability
I. Strain

• Strains selected based on superior stability

II. Excipients

• Excipients with extra low aw chosen

III. Handling

• Raw materials and products are handled in such a
  way that we minimize exposure to aw, temperature,
  and oxygen

IV. Production

• We minimize mechanical processing
• We add probiotics as close to final filling as
  possible
• We have for customers preparred Standard
  Operating Procedures

• We discuss choice of product packaging with our
  customers
• Air tight
• Moisture impermeable
• Oxygen free (filled with nitrogen)

V. Packaging
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Types of packaging
Recommended for capsules and tablets
Not recommended
Glass container
Alu-tube
Plastic container w/ desiccant
Plastic Container (PET)
Aclar blister
PVDC-alu blister
Recommended for powder
Not optimal, but best blister type
Stick
Sachet
Alu-alu blister
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Desiccant
Desiccants reduce water activity level
Water activity

• Bentonite
• Activated clay
• Absorbs moisture steadily in given period
• Silicagel
• Chemically produced salt
• Absorbs moisture steadily in given period
• Molecular Sieve
• Crystallized aluminum silicate
• Absorbs moisture quickly within short period (up
  to 20 of its own weight)

Desiccant lid
Desiccant lining
Desiccant bags
(Molecular Sieve)
(Molecular Sieve)
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Moisture Vapor Transmission Rate by type of
packaging material

• Moisture Vapor Transmission Rate (MVTR)
• The rate at which moisture will permeate through
  a barrier over time
• Measures the barrier performance of different
  types of packaging
• Freeze-dried products are very hygroscopic and
  will absorb available moisture quickly
• High-barrier packaging materials (glass,
  aluminum, and plastic vial) are recommended for
  reducing moisture and oxygen migration.

MVTR examples MVTR examples
Glass 0
Aluminum 0
HDPE 3,8
PVDC 4,0
PP 6-10
LDPE 18-23
PET 19-46
Plastic vial with desiccant lining permits 0.5mg
moisture to enter vial, but desiccant absorbs 1g
of moisture
Measured at 37.8C and 100 RH (g/25µ/m2/day)
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Summary

• Supplementation with Probiotics may help soldiers
  prevent pathogen-based GI disturbances such as
  food poisoning, water contamination, etc.
• Probiotics have been shown to increase the
  strength of the immune system of individuals who
  are under chronically-high levels of stress
• In the field, the best dosage form for soldiers
  would be the probiotic stick or sachet
• In the military food service setting, making
  single serve packets of probiotic capsules
  available with meals would be ideal.
• GOAL FOR END RESULT Improved overall health of
  the soldiers and thus possibly significant cost
  savings for the Military (via reduced healthcare
  and sick leave).

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THANK YOU!
For more information, please contact Brian
Peeters usbpe_at_chr-hansen.com Office (415)
503-1834

• Together
• we improve the quality of
• food and health
• for people all over the world