Presentation of our BBB model for hold in France in 2017. Our work has been done in partnership with Servier Industry, France.

1. Contact : nicolas.perriere@brainplotting.com
www.brainplotting.com
Our core business is the study of pharmacokinetics (PK) in the Human brain at
early preclinical stage by using the most accurate predictive models and most
recent tools. To that end, we have developed a proprietary method: the Brain
Exposure Prediction model (BEP) based on primary Human in vitro tools from
fresh Human brain tissues.
The BEP model provides the in vivo time-curves of drug free concentration in
the brain, based on in vitro generated PK brain parameters and PBPK modeling.
Brain Exposure PredictionBrain Exposure Prediction BBB modelBBB model
2. Equilibrium dialysis methods2. Equilibrium dialysis methods
Fresh Human brain slices
Brain tissue is alive and permits to highlight specific
transport mechanisms in neural cells.
We measure (fu) in ECF and ICF to determine drug
concentrations within the brain
Plasma & Brain Homogenates
We use Human plasma and brain tissues to measure
binding to Human lipids and proteins to improve
predictability
Brain extracellular space
Vecf, Cecf, fu,brain ecf
Brain intravascular space
Viv, Civ, fu,plasma
Plasma compartment
Vpl, Cpl, fu,plasma
Dose
Qbr Qbr
CLp
CLinin
CLoutoutoutou
fu,brain icf , fu,brain ecf and fu,plasma : unbound
fraction into the brain intra and
extracellular fluid and plasma
Viv (mL/kg): physiological volume of
the extravascular space
Vicf and Vecf (mL/kg): physiological
volume of the intra and extra-
vascular space
CL (mL/h/kg): clearances
Qbr (mL/h/kg): brain perfusion blood
flow
Vpl (mL/kg): volume of distribution of
the molecule into the plasma
compartment
C (ng/mL): concentration of the
molecule
Brain intracellular space
Vicf, Cicf, fu,brain icf
BBB
We compute in vitro
human data to predict
Human in vivo PK
information.
All results are
obtained from
physiological models
3. PBPK Modelisation (Physiologically Based PK)3. PBPK Modelisation (Physiologically Based PK)
We use brain endothelial cells from gliomas and peritumoral resections.
Tightness of the BBB model is evaluated by fluorescein permeability which
show great tightness results:
Pe AB = 0,12 ± 0,07 & Pe BA = 0,14 ± 0,06 x10-3
cm/min (n=29, n=12)
Prediction of Human PK data based onPrediction of Human PK data based on
innovative Human primaryinnovative Human primary in vitroin vitro modelsmodels
Focus onFocus on BrainPlottingBrainPlotting’s BBB models’s BBB modelsHuman data from preclinical stages
Brain/Plasma
unboundfraction
defined for your product
ConclusionsConclusions
With physiological in vitro BBB models as close as possible to the in vivo
situation, we get optimal permeability coefficients (Pe) and efflux ratio (ER).
1. BBB permeability1. BBB permeability
The level of innovation, relevance and reliability of our BBB proprietary
model makes it a tool of choice for several expert academic partners of
excellence:
- Quantification of protein levels by a LC-MS/MS technique, which
confirms the good expression of ABC transporters and tight junction
proteins (2)
- Lithium transport at the blood-brain barrier in in vivo and in vitro human
models (3)
- Comparative transcriptome analysis of brain and dermal endothelial cells
infected by Neisseria meningitidis : towards the characterization of the
different clinical features observed during meningococcal diseases (4)
- Functional characterization of cationic drug transporters (5)
(1) A. Moreau, C. Denizot, B. Walther ; Technologie Servier, Orléans
(2) D. Gomez, M. Taghi, M.C. Menet ; INSERM UMR-S 1144 Faculté de Pharmacie Paris Descartes
(3) H. Luo, X. Decleves ; INSERM UMR-S 1144 Faculté de Pharmacie Paris Descartes
(4) L. Le Guennec, S. Bourdoulous, PO. Couraud ; Institut Cochin, Paris
(5) S. Cisternino ; INSERM UMR-S 1144 Faculté de Pharmacie Paris Descartes
Fabienne Glacial, Nicolas PerrièreFabienne Glacial, Nicolas Perrière
BEP model predicts Human brain PK from in vitro derived parameters: the Pe
and efflux ratio determined across the BBB and the drug unbound fraction in
plasma (fu,plasma
) and in brain tissue (fu,brain
) determined by the equilibrium
dialysis metho, with brain homogenates and brain slices.
Functional properties are directly
evaluated with the assessment
of permeability.
Drugs are divided in 3 groups:
Efflux, No Efflux and with
combined passages.
The model’s robustness was
validated with 17 out of 20
reference drugs tested which
show good predictability
This evaluation was done in
partnership with Servier Labs(1)
.
We have isolated
endothelial cells from
brains affected with
different pathologies.
Each one has its own
specificities.
On the right, a chart of
TEER vs fluorescein
permeability with each
different model
Isolation of capillaries
from fresh Human
brain resections and
culture of Brain
endothelial cells
Conditioned Medium of
primary glial cells
Vimentin
Brain endothelial
cells
“BLOOD = A”
“BRAIN = B”
To reach its target, the
molecule of interest
has to cross the BBB
and get into the
extracellular (ECF) or
intracellular fluids (ICF)
of neural cells
Endocytosis
or Exocytosis
Facilitated
transport
Efflux
Simple
diffusion
Paracellular
pathwayTight
junctionsBBB
Blood Stream
Brain Parenchyma
fu,plasma
fu,ecf
Brain Extracellular Fluid
(ECF)
fu,icf
Glial & Neuronal cells
Brain Intracellular Fluid (ICF)
We have developed in vitro models of the Human BBB based on Human
fresh resections. In our hands, we obtained relevant reliable tools which
permit the realisation of PK screening studies for one or several compounds,
as well as specific studies which required a totally functional model of the
Human BBB. French academic Laboratories of excellence have already been
conviced to work with us.
• For each compound we are able to offer predictions of the concentration of the free drug in the brain parenchyma and
more particularly in the ICF and ECF.
• To that end we use exclusively physiological in vitro tools, developed at BrainPlotting, based on Human fresh resections
• Depending on yours needs, we can also propose models, from brains affected by different pathologies, whose properties
may differ.
glial cell conditioned
medium (h,i,j,k).
They are then
suitable for further
studies (d,e,f,g).
a b c
j
GFAPSMA (g)/GFAP (r)
k
Isolated Human brain capillaries
are seeded (a). Brain endothelial
cells sprout (b), are positively
selected and shortly amplified (c),
and seeded on insert cultured with
ZO-1
d
Claudin-5
e
Occludin
f
P-gp
g
h i
Imipramin
Quinidin
Sulfasalazin
Ketoprofen
Verapamil
Diazepam
Paracetamol
Ofloxacin
Warfarin
Terbutalin
Carbamazepin
Propanolol
Indomethacin
4 - Digoxin
5 - Indinavir
Prazosin 1- Methotrexate
2 - Cimetidin
3 - Ciprofloxacin
AZT
Efflux
Ratio 1
Efflux
Ratio 2
1
2 3
4
5