nematic protein organisation technology: a powerful tool to track drugs toxicity and side effects...
TRANSCRIPT
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
Confidentiel 3
Due to toxicity and lose of efficacy from pre-to clinical studies
opening
or
Confidentiel 4
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
Non-confidential 14
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
Non-confidential 17
PHYSIOLOGY AND PATHOLOGY
Non-confidential 18
Non-confidential 15
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
Non-confidential 20
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
Non-confidential 21
Nematic crystals
Hetero-oligomersation
Specific macromolecularassembly
Drug
NEMATIC PROTEIN ORGANISATION TECHNIQUE
Non-confidential 22
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.
Non-confidential 24
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
Non-confidential 25
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
Non-confidential 26
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
Non-confidential 28
Non-confidential 29
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
-30
-20
-10
0
10
20
30
40
50
-100 0 100 200 300 400 500
CB133 HCQRU
Re
sp
on
se
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) 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
-30
-10
10
30
50
70
-100 0 100 200 300 400 500
CB029RU
Re
sp
on
se
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
-60
-40
-20
0
20
40
60
80
100
-100 0 100 200 300 400 500
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
-10
0
10
20
30
40
50
-100 0 100 200 300 400 500
CB103RU
Re
sp
on
se
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
-20
-10
0
10
20
30
40
-100 0 100 200 300 400 500
CB108RU
Re
sp
on
se
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
-15
-10
-5
0
5
10
15
20
25
-100 0 100 200 300 400 500
CB112RU
Re
sp
on
se
sTime
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
-40
-20
0
20
40
60
-100 0 100 200 300 400 500
CB114RU
Re
sp
on
se
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
-50
50
150
250
350
450
-100 0 100 200 300 400 500
CB133 HCQRU
Re
sp
on
se
sTime
ON Target
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-08 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
-200
0
200
400
600
800
1000
-100 0 100 200 300 400 500
CB029RU
Re
sp
on
se
sTime
ka (1/Ms) kd (1/s) Rmax (RU) RI (RU) Conc of analyte KA (1/M) KD (M) Req (RU) kobs (1/s) Chi23,61E+03 3,81E-06 113 9,48E+08 1,05E-09 7,84
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
-20
0
20
40
60
80
100
120
140
160
-100 0 100 200 300 400 500
RU
Re
sp
on
se
sTime
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
-200
0
200
400
600
800
1000
1200
-100 0 100 200 300 400 500
CB103RU
Re
sp
on
se
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) Chi2
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
-50
0
50
100
150
200
250
300
350
400
-100 0 100 200 300 400 500
CB108
RU
Re
sp
on
se
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) Chi2
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
-20
30
80
130
180
-100 0 100 200 300 400 500
CB112RU
Re
sp
on
se
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) Chi2
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
-100
0
100
200
300
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600
-100 0 100 200 300 400 500
CB114RU
Re
sp
on
se
sTime
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
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INOVIEM SCIENTIFIC SAS8, rue de Verdun
F-67084 StrasbourgTel: 03 68 85 14 38www.inoviem.com
INNOVATION – MORE THAN JUST A WORD
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F-67084 Strasbourg
Inoviem Scientific (ISIS)8 Rue Gaspard Monge
F-67084 Strasbourg
Non-confidential