Yves Henrotin, PT, MT, PhD

University of Liège

Nutraceuticals for joint health and

OA biomarkers

Osteoarthritis an «old disease»Act I: Local mechanical disease

Cartilage degradation

Overloading

Overuse

Joint instability

Malignancy

MMP-3

ADAMTS4

ADAMTS5

AGG

COL2

Catabolism Anabolism

Catabolism

Anabolism

MMP-13

MMP-3

ADAMTS4

ADAMTS5

AGG

COL2

Healthy osteoarthritis

NO

PGE2

IL-6

COX2

iNOS

Act II: a dysregulation of chondrocytemetabolism

IL-1b

Act III: OA affects the whole joint and

surrounding tissue

MuscleAtrophy

↓Strength↓Neuromuscular

control

LigamentLaxity

CapsuleRetraction

Fat pad/Tendons

Inflammation

Synovial membrane

Inflammation

Cartilage/ Meniscus

Degradation

Subchondral

boneSclerosisEdema(BML)

Cartilag

e

Subchondral bone

Hypertrophy/mineralisation

MMP-3

MMP-13

IL-6

VEGF

VEGF

Tidemark

PGE2IL-1, IL-6, TNFaNOMicrocrystalsOsteochondral fragments

Inflammation

MMPsIL-1, TNF-a, IL-6

PGE2, VEGF

Synovial membrane

+

RANKL

IL-6OsteoclastsOsteoblasts

RANKL+

+

ADAMTS-4,5

MMP-1,-3,13

Aggrecan

Collagen II-

IL-1b

NF-kB

+

+

ACT IV: a metabolic and systemic

disease of an organ « the joint »

Matrix peptidesCytokinesProstanoidsOxidized lipids

Chronic low-gradeinflammation

Metabolic syndrome-Hypertension-Type 2 diabetes-Dyslipidemia

Adipokines

Cytokines

Sedentarity

Chronic

Mechanical stress

Aging

Obésité Post-Traumatique

Inflammation Hypertrophique

Bone driven

ACT V: a disease with some

subphenotypes

O•Obesity-

related

•Metabolicsyndrom

I•Inflammatory

driven

•Systemic

•Local

T •Post-traumatic

H •Hypertrophic

•Bone driven

OA treatments and

limitations

Modified Clegg et al. Eur J Orthop Surg Traumatol, 2013

Surgery

Prescription

NSAIDs

Including COXIB

Acetaminophen

OTC NSAIDs

Patient education, physical and

occupational Therapy, weight loss

Limitations

Costly, invasive procedure

Primarily indicated for «end stage » OA

GI bleeding or other complications

CV risks

Renalr complications

GI bleeding or other complications

CV risks

Renal complications

Hepatotoxicity

Poor patient compliance

Drug discovery is protracted, risky and

costly

Nothing new to offer at the

patients and the OA research

community

Clinical trials end-point

Symptoms modification (3 to 6 months)

Pain

Physical function

Patient global assessment

Structure modification (1 to 3 years)

Imaging outcomes

Joint Space Narrowing

The Gold Standard

(Radiography) is inadequate

…We need better methods to predict OA progression and response to therapy

Slide courtesy of Dr A Mobasheri (Nottingham University)

Biomarkers - definition

Genomic/Biochemical

substances

Imaging,

Questionnaire,

VAS

RNA,

DNA

Metabolites

Proteins

X-rayMRI Ultrasound

Fragments

Peptides

Soluble or « wet » biomarkers « Dry » biomarkers

A biomarker is a characteristic that is objectively

measured and evaluated as an indicator of

normal biologic processes, pathogenic

processes, or pharmacologic responses to a

therapeutic intervention. » Biomarkers Definitions Working Group I. Biomarkers and surrogate endpoints: preferred definitions and conceptual framework. Clin Pharmacol Ther 2001; 69: 89-95.

Performed tasks

THE CURRENT SITUATION

28 identified biochemical makers1

1 WE van Spil et al. Osteoarthritis cart, 2010; 18: 605-12

Bone

metabolism

PYD, DPD,

NTX-I, CTX-I,

osteocalcine,

BSP

Cartilage

metabolism

CTX-II, Coll2-1,C2C,

C1,2C, ARGS

epitope, KS, COMP,

D-COMP

Synovium

metabolim

Glu-Gal-PYD,

HA, PIIINP,

COMP

14

Coll2-1 and Coll2-1NO2:

two cartilage specific biomarkers

Coll2-1NO2

Coll2-1 HRGYPGLDG

NH2

Coll2-1NO2HRGY(NO2)PGLDG

NO + O2. ONOO-

+

Specific of degradated cartilage

Multiple pathological processes

(inflammation + degradation)

Not confounded

Coll2-1

Hartley Guinea Pig

65 male HA guinea pigs

2/3 w old + 1w acclimatation

– 5 « Baseline », sacrified at T0 (group E)

– 15 group A Oleuropein

– 15 group B (control)

– 15 group C Rutin

– 15 group D Rutin + Curcumin

Arr

iva

l To (w4) W10 W16 W22 W28(1month)

(8 months)

Blood

EuthanasiaUrine

Each week : weighing

W35

W7/8 W19/20 W30 W35

Control Control A C C+D0

100

200

300

400

500

600

700

800

Coll2-1

W4 W35

**

*****

nM

Biomarkers in serum : Coll2-1

0

100

200

300

400

500

600

700

800

2 4 6 8 10121416182022242628303234

Co

ll2

-1 (

nM

)

Age (Weeks)

A

B

C

D

* * *** ** **

N=53

W35

A= Oleuropein

B = Control

C= Rutin

D = Rutin + Curcumin

Control Control A C C+D0

10

20

30

40

50

Coll2-1 NO2

W4 W35

**

****

*

nM

0

5

10

15

20

25

30

35

40

2 4 6 8 10121416182022242628303234

Co

ll2

-1N

O2

(n

M)

Age (Weeks)

A

B

C

D

Biomarkers in serum : Coll2-1NO2

* * *

* ** * ** *

* ** ** ** **

A= Oleuropein

B = Control

C= Rutin

D = Rutin + Curcumin

TIFLEXY Study

Bio-optimized curcuminoids (BOC)

0

20

40

60

80

100

120

140

160

180

200

0 1 2 3 4 5 6 7 8 9 10 11

12 h

eure

s

Flexofytol 2 caps

Curcumine 12 gr.

Curcuminoids /Low avaibility

POLYSORBATE

CURCUMINE

MICELLE

Bio-optimized curcuminoids BOC

« Proof-of-concept study »

-22 knee OA patients

-2x3 caps (42 mg BOC)/days

-3 months treatment

Henrotin Y, Priem F, Mobasheri A, Springerplus, 2013

Criteria of efficacy

Primary end-point : Biomarkers

variation– Coll-2-1 (nmol/L)

– Coll-2-1NO2 (nmpm/L)

– Fib3-1 (pmol/L)

– Fib3-2 (pmol/L)

– CRP (mg/L)

– CTX-II (ng/L)

Secondary end-point: – Variation of pain on movement during the last 24H

– Variation of Global Patient Assessment on disease activity

TIFLEXY Study

A proof-of-concept studyHenrotin et al., BMC Complem Altern Med,2014

Baseline 84 days of treatment p-Value

sColl2-1 (nmol/L) 302.21+/-53 257.84 +/-52.78 0.002*

sColl2-1NO2 (nmol/L) 0.71 +/- 0.78 0.80 +/- 0.24 NS

sCTX-II (ng/L) 11.81 +/-7.98 13.17+/-4.96 NS

sFib3-1 (pmol/L) 707.05 +/- 178.79 765.20 +/- 261.90 NS

sFib3-2 (pmol/L) 580.58 +/- 103.59 636.74 +/- 119.73 NS

sCRP (mg/L) 10.42 +/- 30.27 3.10 +/- 2.40 NS

sMPO (ng/ml) 27.20 +/- 29.05 21.96 +/- 14.65 NS

Time (days)

0 7 14 28 84

26

02

80

30

03

20

Co

ll-2

-1 (

nm

ol/L

)

p = 0.02

Coll-2-1 (nmol/L) between day 0 and day 84

%

-120 -100 -80 -60 -40 -20 0 20 40

51

01

52

0

92%

FDA/OARSI Recommendation to advance the

science of biomarkers

The co-development (Drug + biomarker)

pathway should be determined early in

development.

Used biomarkers in combination rather than

individually

Used in combination with imaging (MRI)

Biomarkers may serves as titration tools;

falicitating dose setting in early clinical study

Appropriate analytical validation of

immunoassay

Thank you for your attention !

Team

International collaborations:F Blanco (La coruna, Spain)

T Conrozier (CHU Lyon, France)

V Kraus (Duke University, USA)

L Punzi (University of Padova, Italy)

A Mobasheri (University of Notttingham, UK)

J Monfort (Hospital del mare (Spain)

P Richette (Lariboisiere, France)

J Runhaar (Erasmus MC, Rotterdam)

Design

J0

Pain assessmentBiomarker

assays

J7

Pain assessment

J14

Pain assessmentBiomarker

assays

J28

Pain assessment

J84

Pain assessment

Blood sampling

Flexofytol 2 X 3 caps/days during 3 months