Nematic protein organisation technology:

Nematic protein organisation technology: A powerful tool to track drugs toxicity and side effects

already at early preclinical drug development

3rd International summit on Toxicology & applied Pharmacology

INOVIEM SCIENTIFIC – AN OBSERVATION

Opening Light at the end of the tunnel

Confidentiel 2

Today’s drug development

No issue

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Due to toxicity and lose of efficacy from pre-to clinical studies

opening

or

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Drug candidat or a ligand

INOVIEM SCIENTIFIC – UNE FORCE

Inefficiency

cure inefficiency

Human Being/ PatientAnimalHuman Tissue (1µg)

Healthy or from patient

Molecule

Ino

viem

Risque

100 0

Human Being/ Patient

Clinical PhasePreclinical PhaseR&D

Cla

ssic

al

Market

Animal

Classical in vitro techniques

Molecule

INOVIEM’S APPROACH

Preclinical Phase

R&D MarketPreclinical Phase

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PIMS : Physiological Intermolecular Modulation Spectroscopy

NPOT : Nematic Protein Organisation Technology

MUXI : Multiplex ‘Cross-Sandwich’ Immunoassay

In Practice :From as low as 1 µg of a tissue, we pinpoint the target of your compound in each organ

OUR TECHNOLOGIES

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PIMS:Visualising

Discriminating

NPOT:Isolating

Identifying

MUXI:Reconstructing in 3D

Representing

1 4 0 01 3 7 2

1 3 4 41 3 1 6

1 2 8 81 2 6 0

1 2 3 21 2 0 4

1 1 7 61 1 4 8

1 1 2 01 0 9 2

1 0 6 41 0 3 6

1 0 0 89 8 0

9 5 29 2 4

-0 .2

-0 .1 5

-0 .1

-0 .0 5

0

0 .0 5

0 .1

0 .1 5

0 .2

3 7 -1 0 1 5

3 7 -3 7 -2 5 -1 5

-1 0 -5 0 1 0

1 5 2 5 3 7

HumantissueExtract

Lead or drug

FROM CRUDE HUMAN TISSUE TO THERAPEUTIC TARGET

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OUR SERVICES

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Pre

clin

ical

Isolation and identification of therapeutic target

« hit » to « lead » discrimination

Target identification (ON Target )

Toxic target identification (OFF Target)

Lead optimisation

•

•

•

•

•

Biomarker identification•

Inoviem Scientific brings its expertise in the process of drug development and offer

a full range of tailored services :

OUR SERVICES

Inoviem Scientific brings its expertise in the process of drug development and offer

a full range of tailored services :

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Individualized medicine: Post market patient stratification to responders and non-responders

•Individualized medicine: Biomarker identification, validation and selection &

associated companion test development

Cli

nic

al

•

•

•

•

Pre-phase II patients stratification & optimisation for phase II clinical trial

Drug rescue

Drug-up: optimisation of ON/OFF target interaction ratio

DYNAMIC AND STATIC

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A * B = B* AO2 * H2 = H2 * O2

* = *

≠* *

THEORY AND BIOLOGY

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http://www.world-builders.org/lessons/less/les4/les4gifs/quartzcrystal.gif


http://www.hydrogenboyz.com/pictures/hydrogen1.jpg






Equilibriumt=0

Equilibriummissorder

t=1Disorder

ΔGD > ΔGE0 = ΔGE1

FROM ORDER TO DISORDER & MISS-ORDER

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Equilibriumt=1

1st Disorder

ΔGD1 < ΔGD2 > ΔGE1

Distorted order

FROM MISS-ORDER TO DISTORTED ORDER

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THE VERY SAME

MOLECULAR

COMPOSITION

physiologicMeta-Stable state

22°C

solidMeta-Stable state

-20°C

SolidStable state

>45°C

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PHYSIOLOGY AND PATHOLOGY

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LIFE’S PHILHARMONIC

1. Dynamic

2. Environment specific

3. Take into account order of molecular interaction

4. Take into account the impact of a disruption

5. Take into account molecular energy state

REQUIREMENTS FOR MAIINTENANCE OF PHYSIOLOGICAL CONDITIONS

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PIMS : Physiological Intermolecular Modulation Spectroscopy

NPOT : Nematic Protein Organisation Technology

In Practice :From as low as 1 µg of a tissue, we pinpoint the target of your compound in each organ

OUR TECHNOLOGIES

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Nematic crystals

Hetero-oligomersation

Specific macromolecularassembly

Drug

NEMATIC PROTEIN ORGANISATION TECHNIQUE

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Putamen

Atropine

Serotonine

Prefrontal cortex Cerebellum

Solvent

EXAMPLES

NPOT Identifies a drug or a xenobiotic target & its partners in a physiological condition.

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R4

R3

R1R2

Compound 1 Compound 2 Compound 3 Lead Solvent

R

O

NH

R'

Small molecule

Peptide

EXAMPLES

NPOT Identifies a drug or a xenobiotic target & its partners in a physiological condition.

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MACROMOLECULAR AUTO -ASSEMBLY OF TOTAL HUMAN PLACENTA MEMBRANE WITH RECOMBINANT TGFβ (rTGFβ)

In absence of rTGFβ In presence of rTGFβ

Macromolecular assemblies due to the presence of 1µl of rTGFβ is shown withblue arrow. In absence of rTGFβ no macromolecular assemblies are formed

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Destrin

G-actin

Spectrin beta non- erythrocytic

Proton pump

rTGF-β family

Receptor partner 3

Receptor partner 2

Receptor partner 1

SCHEMATIC REPRESENTATION OF FAMILY rTGFβ MOLECULAR INTERACTION

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A

Isolation ProteomicsExtracted

Specific ON/OFFTargets

-Prot. 1-Prot. 2-Prot. 3-…

Identification

NPOT: a highly reproducible, rapid and safe technique for isolating & identifying drugs’ ON/OFF targets

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(Leponex)Clozapine

(Plaquenil)

Hydroxychloroquine LabEx

Identified

On Target

Identified

OFF Target

Molecular Design

Originalmolecules

Partner

Charité Berlin

Start 09/2012

Patent application 03/2013

DEMONSTRATION OF CONCEPT

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DRUG UP

PATHOLOGY: LUPUS

Treatment: Hydroxychlorquine (HCQ)

Clinical efficacy of HCQ:

• About 30% responder with no vasculitis, skin & joint inflammation

• Retinopathy with chronic treatment

Clinical study PBMCs from 20 patient all on HCQ and subjected to NPOT

Patients Sex Treatment Pathology Isolated protein with solvent Isolated protein with HCQ

HEH F MTT LUPUS HSP70 Annexin VI, Myosin 9 cytoplasmic

LB F HCQ LUPUS HSP70 ON, Myosin 9 cytoplasmic, histone H3

SX F HCQ LUPUS Actin beta, HSP70 Annexin VI, ON,Myosin 9 cytoplasmic, ATP synthetase

ECR F HCQ LUPUSAnnexin VI, Myosin 9 non

sytoplasmic,HSP70 Annexin VI, ON,Myosin 9 cytoplasmic, ATP synthetase

DMN F HCQ LUPUSActin,pyryvat kinase, platelet

membrane glp IibAnnexin VIII, ON,Myosin 9 cytoplasmic, ATP

synthetase,transgelin 2

CTS F HCQ LUPUS Actin,pyryvat kinase, HSP60Annexin VIII, ON,Myosin 9 cytoplasmic, ATP

synthetase,L-plastin

PXM M HCQ LUPUS Actin, HSP70 ON, Myosin 9 cytoplasmic

SLG F HCQ LUPUSAktin, Pyruvat kinase, HSP70,

coagulation factor XIII HSP70, ON, Myosin 9,SP40,40

JGB M MTT LUPUS HSP70, Actin Myosin 9 cytoplasmic, FLJ00279

JWW F HCQ LUPUS HSP70, myosin 9 heavy chain Myosin 9 cytoplasmicSFR F HCQ LUPUS Actin beta, Actinin alpha ON, Myosin 9 cytoplasmic,SWI/SNF

MHW F HCQ LUPUS Actin,myosin 9 heavy chainON,Myosin 9 cytoplasmic,platelet membrane

glycoprotein IIb, lactoferrin

GLL F MTT LUPUS Actin,myosin 9 heavy chainMyosin 9 cytoplasmic,platelet membrane glycoprotein

IIb

ABL F MTT LUPUS Actin,myosin 9 heavy chainMyosin 9 cytoplasmic,platelet membrane glycoprotein

IIIa

CGG F HCQ LUPUS Actin, vinculinON,platelet membrane glycoprotein Iib, protein

disulfide isomerase

F=13 ON=10

M=2

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CB133 HCQRU

Re

sp

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ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi24,52E+03 9,90E-06 12,1 4,56E+08 2,19E-09 0,371

20,2 1,67E-02 1,50E-07 11,9 6,87E-0412,7 -4,24E-04 3,00E-07 12 1,37E-038,29 -1,25E-02 6,00E-07 12 2,72E-03

2,93 -0,027 1,25E-06 12 5,66E-03

OFF Target

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-10

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30

50

70

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CB029RU

Re

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sTime

ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi21,89E+03 6,60E-05 14 2,87E+07 3,49E-08 0,162

5,43 -5,07E-03 1,50E-07 11,4 3,50E-0413,3 2,70E-03 3,00E-07 12,6 6,34E-0421,4 -5,27E-03 6,00E-07 13,3 1,20E-03

35,1 -0,0109 1,25E-06 13,7 2,43E-03

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Tim e s

Response

RU

ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi23,80E+03 5,98E-04 46,7 6,36E+06 1,57E-07 2,57

3,53 1,50E-07 22,8 1,17E-0310,3 3,00E-07 30,7 1,74E-0318,3 6,00E-07 37 2,88E-03

30,3 1,25E-06 41,5 5,35E-03

CB072

-20

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0

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CB103RU

Re

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sTimeka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi21,75E+04 0,0117 13,4 1,50E+06 6,68E-07 0,188

5,03 1,54E-03 1,25E-07 2,12 0,01399,36 5,27E-03 3,00E-07 4,16 0,01715,7 8,38E-03 6,00E-07 6,36 0,022224,5 0,0169 1,25E-06 8,75 0,0336

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0

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CB108RU

Re

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sTimeka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi21,15E+04 9,37E-03 8,49 1,23E+06 8,16E-07 0,16

2,67 -2,14E-03 1,50E-07 1,32 0,01115,48 2,24E-03 3,00E-07 2,28 0,01289,28 5,61E-03 6,00E-07 3,6 0,0163

16 9,73E-03 1,25E-06 5,14 0,0237

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CB112RU

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ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi22,32 1,14E-05 28,2 2,04E+05 4,90E-06 0,359

-0,0134 1,50E-05 21,3 4,62E-051,38 3,00E-05 24,2 8,10E-052,76 6,00E-05 26,1 1,51E-04

5,01 1,25E-04 27,1 3,01E-04

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CB114RU

Re

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sTime

ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi2520 0,0115 111 4,52E+04 2,21E-05 0,276

1,96 -2,12E-03 1,50E-07 0,749 0,01165,43 -4,33E-04 3,00E-07 1,49 0,011710,1 -2,04E-03 6,00E-07 2,94 0,0118

18,6 -2,61E-03 1,25E-06 5,95 0,0122

ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi24,40E+03 3,13E-03 186 1,41E+06 7,11E-07 8,09

104 1,50E-07 32,4 3,79E-03155 3,00E-07 55,1 4,45E-03225 6,00E-07 85,1 5,77E-03

302 1,25E-06 118 8,64E-03

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250

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CB133 HCQRU

Re

sp

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sTime

ON Target


104 1,50E-07 32,4 3,79E-03155 3,00E-07 55,1 4,45E-03225 6,00E-07 85,1 5,77E-03

302 1,25E-06 118 8,64E-03

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0

200

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CB029RU

Re

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36,1 1,50E-07 112 5,46E-0437,1 3,00E-07 112 1,09E-0350,9 6,00E-07 113 2,17E-03

70,4 1,25E-06 113 4,52E-03

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RU

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CB072

ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Drift (RU/s)Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi2

3,31E+04 0,012 369 2,76E+06 3,62E-08 22184 0,12 1,50E-07 108 0,0169318 0,235 3,00E-07 167 0,0219481 0,416 6,00E-07 230 0,0318

683 0,578 1,25E-06 286 0,0533

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1200

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CB103RU

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4,77E+03 6,58E-03 225 7,24E+06 1,38E-08 4,5672,5 -0,0459 1,50E-07 22 7,30E-03108 -7,75E-03 3,00E-07 40,1 8,01E-03168 0,0177 6,00E-07 68,1 9,44E-03

254 0,0441 1,25E-06 107 0,0125

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CB108

RU

Re

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3,49E+04 0,0111 77,3 3,15E+06 3,17E-07 2,6932,9 0,0167 1,50E-07 24,8 0,016338,2 0,0301 3,00E-07 37,6 0,021556,4 0,0811 6,00E-07 50,6 0,032

70,2 0,119 1,25E-06 61,7 0,0547

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CB112RU

Re

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250 1,50E-05 1,59E+03 1,67E+07 5,99E-08 14,861,1 -0,0626 1,50E-07 1,13E+03 5,25E-05125 -0,0897 3,00E-07 1,32E+03 8,99E-05229 -0,144 6,00E-07 1,44E+03 1,65E-04

410 -0,285 1,25E-06 1,51E+03 3,27E-04

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CB114RU

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Molecule Kd (M)ON target

Kd (M)OFF target

Molecule Kd (M)ON target

Kd (M)OFF target

HCQ

3.0e -7 2.9e -8 CB108 3.6e-8 5.5e-7

CB0293.5e-8 1.8e-8

CB1122.0e-7 4.9e-6

CB0723.7e-9 1.7e-6

CB1145.99e-8 3.5e-5

CB1031.1e -8 3.4e-7

FROM PRECLINICAL TO CLINICAL:LEADING EXPLORER OF DRUG-TARGET INTERACTION

Siège social:

INOVIEM SCIENTIFIC SAS8, rue de Verdun

F-67084 StrasbourgTel: 03 68 85 14 38www.inoviem.com

INNOVATION – MORE THAN JUST A WORD

Inoviem Scientific (IPCB)15 Rue René Descartes

F-67084 Strasbourg

Inoviem Scientific (ISIS)8 Rue Gaspard Monge

F-67084 Strasbourg

Non-confidential

Documents

Nematic protein organisation technology: