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