bore-10 vectorisé dans les cellules tumorales pour la ... · ewin godio jacques pliquett amélie...
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BoRE-10VEctorisédanslescellulesTumoralespourlathérapieparcapturedeneutrons
BREVET
G.Kalot1,S.Coninx1-2,B.Busser1,J.L.Coll1,L.Sancey1S.Coninx1-2,V.Cosenza2,R.Auzély-Velty2
A.Godard3,J.Pliquett3,E.Godio3,F.Denat3,C.Goze3U.Köster4
1IABU1209UMR5309UGAGrenoble,2CERMAVUPR5301Grenoble,3ICMUBUMR6302Dijon,4InstitutLaueLangevinGrenoble
Outline
Ø PrincipleofBoronneutroncapturetherapy(BNCT)Ø DescriptionofBREVET’sprojectandinvolvedpartnersØ Mainresultsfromyear-1
Ø Descriptionofthedifferentwork-packagesfor2019
ClinicalRadiationTherapy(6MV)
Toxicforallcrossedtissues
=Normaltissueslimitthe
thedose
SanceyetalBrJRadiol.2014:20140134
IV infusion
Boron accumulation in the tumor
10B[n,α]7Li reaction
Neutron Epithermal 0.4-10 keV
Thermal < 0.4 keV
Alpha particle
7Li recoil nucleus
10B -nucleus
BoronNeutronCaptureTherapy:Principle
4-5µm210keV/µm
9-10µm163keV/µm
GlioblastomaóTumorofprimarytarget
BoronNeutronCaptureTherapy:ExamplesGlioblastoma Locallyrecurrentmalignant
HeadandNecktumor
Innovativeboron-basednanocompounds
• Currentlimitationsofboron-neutroncapturetherapy
Weaktumoraccumulation PoornumberofBoron-atom
Innovativeboron-basednanocompounds
HyaluronicacidbackboneMaltosederivativeLinkerforself-assembling
Prof.R.Auzély-Velty
DrC.GOZE
aza-BODIPY
• Objectives:Developandevaluatetheefficacyofinnovativeboron-containingnanocompoundsforefficientBNCT
LinkerR’
R’
R R
Innovativeboron-basednanocompounds• Phase1:formulationcontainingPhenylboronicacid(PBA)
Hyaluronicacidbackbone=HAMaltosederivativeBoron(PBA)derivative
Prof.R.Auzély-Velty
≈161±7nmPDI0.1
Ø BiocompatibleØ HightumoruptakeØ Highboron-content
PelerinGarciaetalBiomater.Sci.2018,6,1754
HA-PBAnanogel
=>Efficientcellinternalization
=>Nontoxicinabsenceofneutrons
Invitro
0
25
50
75
100
0 1 2 3 4 5 6%Boron
load
edcells
Time(Hour)
A375 Cal33MelanomaHead&Necktumor
A375 Cal33
HA-PBAnanogel
HA-PBAnanogelTumor/Liver=0.2
Tumor/Liver=1
BNCTefficacyunderneutronsexposure
0
10
20
30
40
50
60
Irradiation(IRR)alone
LowBoron(12,5ugNP)+IRR
MediumBoron(50ugNP)+IRR
HighBoron(1mgNP)+IRR
TumorW
eight(mg)
-18% -26%-20%
Tumorweight(mg)
Invivodistributionintumor-bearingmice
FormulationOptimization(reticulation)
BNCT
TumorA375
Productionofnewcompounds
• Phase2:optimizationofthesynthesisof2compounds
1)ProductionofHA-BSHderivatives
2)ProductionofHA-poly(DEGA-co-DAAM)derivatives,forthermosensitiveself-aggregation
HA-BSH:BNCTassayinovo
T24hT0 T30m T2hT1h T5h
0
10000
20000
0,5 5
RLU/pixel/m
s
Ctl T°
Neu
V122 +
Neu
V123 +
Neu
0
20
40
60
80
100
Data 1
Tum
or w
eigh
t (m
g)
BNCT
HA
1)Optimizationofthedesignoftheexperiment:neutronexposureafter1hofincubation
èTumorweigh
reductioninabsenceof
10Benrichment
ILLsession:neutronexposureSeptember
2018
EvaluationofBODIPYloadednanogelsInvitro
30mà6h
0
25
50
75
100
0 2 4 6
%Boron
load
edcells
Time(hrs)
U87-MG A375
Glioma Melanoma
FACSanalysis
A375
U87MG
0 30min 2h1h 4h 6h
EvaluationofBODIPYloadednanogelsInvivo
0
2000
4000
6000
8000
heart
Lungs
Uterus
Ovarie
s
Brain
Gut
Adrenal
Muscle
Kidn
ey
Spleen
Liver
Pancreas
Fat
Skin
Tumor
RLU/pix/m
s
24h
0
2000
4000
6000
8000
heart
Lungs
Uterus
Ovarie
s
Brain
Gut
Adrenal
Muscle
Kidn
ey
Spleen
Liver
Pancreas
Fat
Skin
Tumor
RLU/pix/m
s48h
è Optimizationofthereticulationinprogress
U87MG
Objectivesacquiredin2018Objectives Status
SelectionofoptimizedBODIPYsforopticalimaging
ü SelectionofBODIPYsèPATENTinprogress
10B-BODIPYenrichment èReplacedby10B-BSH
ProductionofBODIPYloadednanogels ü Firstsynthesiswithstrongloadingü Secondbatchunderproduction
InvitroevaluationofBODIPYloadednanogels
ü Efficientcellinternalizationasothernanogels
Inovoevaluationofthetumoruptake ü DesignoftheBNCTexperiment
InvitroandinovoBNCTeffectofenrichedformulations èPlannedin2019inthenexttime-beam
Firstinvivoassays:distribution ü Formulationunderimprovementofoptimizedtumoruptake
• Phase3:Borocaptatederivative(BSH)andBODIPYasboron’sourcesObjectivesfor2019
A
B
WP1-WP4:Synthesisoptimizationandchemicalcharacterizations
SimonCONINXPhDstudentCERMAV-IAB
AmélieGODARDPhDstudentICMUB
• Phase3:Borocaptatederivative(BSH)andBODIPYasboron’sourcesObjectivesfor2019
WP2:Biologicalevaluation
Invitroevaluationofthedistribution,toxicity,internalizationprocess,in2DbutalsoonspheroïdsInovo:- DevelopmentofglioblastomacellsexpressingthefluorescentGFPprotein- Selectionofthebestglioblastomacelllineforreproducibletumordevelopment- Standardizationofthetumorimplantedonegg’smembraneBEFOREneutronexposureInvivodeterminationofthedistribution,pharmacokineticandtoxicityinmice
GhadirKALOTPhDstudentIAB
WP3:BNCTefficacy
Beamtime:11daysinJune/July2019+Submissionofaproposalforearly2020CellsandinovoexperimentwithBODIPY-10B-BSHcomplexesloadedintonanogels
Insummary• Developmentofinnovativeformulations:biocompatible,enriched
inboron-atoms,“theranostic”
• Efficienttumortargeting
• BODIPYsforopticalimaging:fromcellstomice(Patentunderinvestigation)
Ø Incorporationof10B-BSH-BODIPYintothenanogelsforthenextbeamtime
Ø 3PhDstudentsrecruitedfromOctobertoDecember2018
Ø 1commonpublicationunderwriting
AcknowledgmentsIABGrenobleJean-LucCOLLBenoitBUSSERMalikaDAKIRGhadirKALOT
V.JOSSERAND
RachelAUZELY-VELTYVaninaCOSENZASimonCONINX
ChristineGOZEEwinGodioJacquesPliquettAmélieGodard
UlliKÖSTER
GrantsANR-11-INBS-0006(FLI)andInterdisciplinaryCNRS(ISOTOP)programs