Using probiotics to tackle the

worldwide challenge of obesity

Gregory Lambert

Probiota Asia– October, 23rd, 2019

The challenges of overweight in APAC Regions

Historically, Asian regions had lower rates of obesity and

overweight than US or Europe, however, with high rates of economic

growth…

The rates of overweight & obesity are increasing fast

As technology improves and urbanisation increases, obesity levels rise

Asians tend to have higher amounts of abdominal fat at lower BMIs*

Asians are more likely to develop central obesity, which is associated with

higher risk of developing dyslipidaemia, impaired glucose tolerance, type

2 diabetes mellitus and CVD, as well as more adverse cardiovascular

outcomes.

This increase can also negatively impact the economy

Economic output impacted by loss of productivity of overweight and

obese people

Chronic conditions require expensive lifelong treatment

APAC Statistics

Pacific island countries and areas have a 75% adult prevalence of obesity

In 2014 50% of all overweight children under 5 lived in Asia

75%

50%

8 South-east Asian

countries10% 46%Vietnam Malaysia

Between 2010 and 2016, the adult obesity rate have risen by 33%33%

TargEDys Executive Summary

Based on 15 years of research at Inserm (University of Rouen, France)

Offices in Longjumeau, Paris area & Labs in Rouen, Normandy

Financing

• Raised 9 M€

• Obtained 1.3 M€ in non-dilutive financing

IP: 9 patent families and 6 trademarks

Platform delivers both therapeutic and nutraceutical products

Reduction of food intake

Reduction in body weight

Improvement of body composition

Activation of lipolysis

Decrease of fasting glycemia

(⬈ of glucose tolerance)

Activation of central satiety pathways

Key ProbioSatys® benefits

Sugar

ProbioSatys®

Naturally regulating the appetite via the Microbiome

The concept of molecular mimicry

ClpB / ⍺-MSH Molecular mimicry

Strong sequence homology between an exposed loop on the surface of ClpB and ⍺-MSH1

Confirmation in HRM Mass Spectrometry after immunoprecipitation of enterobacteria proteins with ⍺-MSH antibody2

ClpB is a MCR agonist3

1. Tennoune et al., Transl Psy, 2014

2. Internal report from Biognosys, 2018

3. Ericson et al., Bioorg Med Chem Letters, 2015

534-AEIAEVLA RW TGIPV-548ClpB E.coli

Ac-SYSME HF RW GKPV-NH2a-MSH

DVAEVVS QW TGIPVClpX Lactobacillus casei

ClpC Bifidobacterium animalis IAEV IS QS TGIPV

ClpB Enterococcus fecalis EIAQVVG RL TGIPV

Hsp 104 Saccharomyces cerevisiae ISETAA RL TGIPV

534-VEIAEVLA RW TGIPV-548ClpB H.alvei / ProbioSatys

✓

✗

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✗

✓

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HRM Mass Spectrometry - Volcano plot

ClpB is the only protein with α-MSH aa pattern

ClpB is one of the most abundant proteins amongst candidates

Plasma ClpB levels correlate with

ClpB DNA in gut microbiota in rats

From: Breton et al., Int J Eat Dis 49: 805-808, 2016

ClpB is present in human

plasma

ClpB is endogenously present

and inversely correlates to BMI

ClpB DNA in human feces

inversely correlates with BMI

Submitted Int.J.Obesity

Pr

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Single intraperitoneal Injection (2 pmol): ClpB96 / ClpB25 / control bufferC57Bl/6

2-way Annova, Dunn Post-test, *p<0,05; Mann & Whitney **p<0,01

T e m p s (m in )

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➔ Single injection of ClpB reduces significantly cumulative food intake

In vivo ClpB effect in mice

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Time (min)

Oral gavage with Hafnia alvei

in Genetic (ObOb) and Diet (HFD) mice

obesity models

Submitted Int.J.Obesity

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434445464748495051525354555657

Ctrl H. alvei (2.8x1010CFU/day) Orlistat (80mg/kg/day)

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Orlistat

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Hafnia alvei vs Orlistat

Hybrid Model HF-HSD – ob/ob

Hafnia alvei reduces food intake and glycemia while Orlistat increases it.

Control of appetite is one key to achieve sustainable weight

and metabolic disease management.

0 15 30 45 60 90 1200

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CMC development

Hafnia alvei 4597 - Ex-vivo SHIME model

H. alvei 4597 resists digestion (low sensitivity to pH and enzymes)

H. alvei 4597 adheres to mucus

H. alvei 4597 grows and is metabolically active in colon

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Viable cells pH

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CMC Development

Industrial process - ClpB concentration

5000L1L 30L

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WB band

ClpB profile

CED01 CED02

CMC development

Analytical characterisation

ClpB content reproducibility analysed by WB with

bands extinction.

ClpB profile is similar for both 5000L batches.96kDa = total ClpB protein

CMC development

Formulation - GI resistant capsules

Comparison of 2 GI-resistant formulations

(DRCAPSTM et HPMC) in ex-vivo GI model:

• Study of the release of the bacteria

• Study of the ClpB integrity

0%

10%

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end of gastric

phase

end of

duodenal phase

end of ileal

phase

% cells released in the medium

at each digestion stage

HPMC DRCAPS™

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100%

DRCAPS™ HPMC

ClpB content - Ileal phase

DRCAPS™ provide a delayed bacteria release & protection of the ClpB from degradation.

Finished product

Formulation & control

Stability data on the first commercial

batch at 30°C / 65% HR

Strain

Petri dish cell count (CFU)

Flow cytometry

• Total cell count (bacteria)

• Bacteria in intermediate stage

• Cultivable bacteria

Protein and fragments characterization

ELISA to quantify 𝛼-MSH pharmacophore

Western Blot to characterize degradation

All non-lysed bacteria alive or not

Contain and protects ClpB.

Molecular mimicry is present

even after partial degradation

CMC Development

The Finished Product

Hafnia alvei 4597, THE connected strain

Zinc & Chromium to rebalance your metabolism

The only probiotic with a validated molecular level mechanism of action

Proven efficacy

Gastro-resistant capsules

2 capsules per day (breakfast and lunch)

Feel satiety after only one week

Reduce body weight after one month

Regulate your appetite via your microbiome!

Ingredients Defined Daily Dose

Hafnia alvei 459750 million CFU’s

100 billion cells

Zinc (bisglycinate) 5 mg or 50% of ARs

Chromium (picolinate) 20 μg or 50% of the ARs

Regulatory positionning

ProbioSatys®

PET CARE

Pet Food/ supplement

NUTRA

Food supplement

Ingredient

PHARMA

LBP ClpB

Looking for

partners

On the

market

Nutraceutic based approaches

• Cheese strain = food

• Self affirmed GRAS in the USA

• Dietary supplement

• Functional foods

Therapeutic based approaches

• The ClpB or ClpB fragments as API

• Life Biotherapeutic with Hafnia alvei

Ongoing Clinical Trial

Double-blind, randomised, placebo-controlled study to evaluate benefit of ProbioSatys® on weight

reduction in overweight subjects

Probiotic strain & dosage: Hafnia alvei 4597 – 1.1011 cells/day

12 weeks of treatment, 2 capsules/day – 5 visits

3 centres in Germany

2 arms / 120 subjects par arm - BMI: 25 kg/m2 – 29.9 kg/m2

Main endpoints: • Body weight (kg and %)

• Body fat and fat free mass

• Waist and hip circumference

• Lipid metabolism parameters

A Human POC for ProbioSatys® Nutra and Therapeutic products

• Glucose blood parameters

• Feeling of satiety

• General well-being parameters

• Safety

Confidential

EnteroSatys® consumer study

METHODOLOGYConsumer experience study after 1 and 3 months of EnteroSatys® use

Questionnaire via Google Forms or by phone (10-15 min)

Inclusion criteria: people who bought at least 2 boxes

Based on a voluntary decision to enter the study

Reward: 1 free box per questionnaire

POSSIBLE LIMITATIONSSample – total n=61

People included bought the product on trade-shows or on the website, so sample is slightly different from people who would by in pharmacies)

Sample is probably more motivated, educated and experienced than the common population

Sample size for Q3 after 3 months is limited for the moment (n=27)

Self-assessment

Incentive box

Confidential

Study results

Profile of a typical participant in the

survey

Woman (80%)

Average age: 48 years-old

Overweight or obese (BMI >25 = 70%)

Exercising at least 1x per week

Feels hungry especially in the evening

93% Has some bad eating habitsMostly refill or snacking

Tried restrictive dieting in the past

Primary objective: to loose weight

Study results September 2019; n=61

Confidential

Study results

Eating behaviour

% of people who reported different impacts on

their eating behaviour

55%

58%

56%

65%

0% 20% 40% 60%

SMALLER PORTIONS

LESS SNACKING

LESS SWEET CRAVINGS

DECREASED APPETITE

*reported effect either on eating behaviour or weight loss or both

0%

25%

50%

75%

< 10 days 10 days - 3weeks

After 3 weeks

APPETITE

SNACKING

SWEET CRAVINGS

FOOD SIZE

Lead time to first reported effects

83% are responders*

77% have reported at least 1 effect on their eating behaviour

Majority of people have reported their first effects within first 10 days

Confidential

Study results

Weight loss

Weight loss

After 1 monthAverage loss: - 2.7%

After 3 months

Average loss: - 6.3%

After 1 month

- 2.2kg

After 3 months

- 5.04kg

Average

(kg)

35% have lost more than 3%

65% have lost more than 3%

18% have lost more than 5%

38% have lost more than 5%

Confidential

Study results

Other results and satisfaction

84%WOULD RECOMMEND

THE PRODUCT AFTER

THE FIRST MONTH 100%

96%WOULD RECOMMEND THE

PRODUCT AFTER

3 MONTHS

Waist circumference* (63%)

Digestive comfort (59%)

Feeling better in their body (73%)

Improved relation with food (69%)

Vitality (41%)

Other improvements reported by responders

*Tightened their belt at least by one notch (2 cm)

88% report to be compliant during the 1st month

60% report to be compliant during the 3rd month

Compliance

Contact information

Grégory LambertCEO and VP R&D

Gsm: +33.6.79.39.41.16

Mail: glambert@targedys.com

TargEDysFaculté de Médecine et Pharmacie

22 boulevard Gambetta

76000 Rouen

TargEDysParc Nativelle

1 chemin de Saulxier

91160 Longjumeau

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