Predictive in vivo Pharmacology - TransCure … · Predictive in vivo Pharmacology ... • Both HAART treatments induced comparable decreases of viral load ... current trend

Predictive in vivo Pharmacology

FullFunc'onalHumanImmuneSystemMouseModels

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Patrick Nef, PhD, CEO Sebastien Tabryun, PhD, CSO Tel: +33 4 56 44 81 21 [email protected] www.tcbioservices.com TCSOverviewHIV-BIOEUROPEFALL2016

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WhoWeAreandWhatWeDo

Exper'se•  Fullrecons'tu'onofthehumanimmunesystem/func'oninmousemodels•  Protocoldesigns,tailor-madeApplica'on•  PreclinicalCandidateSelec'on•  EfficacymodelsforONCO,HIV,IBD,Lupus,fullAg/ImmuneResponses•  Immunosafety•  Combina'onstrategy

Aboutus•  EuropeanCROlocatedinArchamps,FrancenearGeneva(Switzerland)•  Teamof8FTE/experts(5PhD)•  BioSafetyLevelBSL-2andBSL-3animalandlab.cer'fiedfacili'es•  Highqualityandflexibility•  FeeforServices•  Interna'onalrecurringcustomers(Pharma,Biotech)

TCSOverviewHIV-BIOEUROPEFALL2016

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2013-2016

•  25AccountsinUSA,EUROPE&JAPAN•  45projectsexecuted•  >1’600huMiceusedforR&Dprojects•  huMouseModels(numberofmice):

§ HIV>250§ InflammatoryBowelDisease>300§ Cancer>550§ Lupus>40§ Vaccina'onandfullImmuneResponses:new>30

§  AccreditedR&DCréditImpôtResearch(CIR)


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StudyDesign

ProtocolExecu'on

DataAnalysis

FinalRepor'ng

§  Study:Pilot,Main§  Tailor-madeprotocols§  SOPdriven§  ClearRoleandResponsability§  Quotes§  Timelines

§  FinalReport§  Datatransfer§  Feedback

collec'on

§  BSL2&3animalfacility

§  Stateoftheartcytometryandcellbiology

§  Rawdatacollec'on

§  DataAnalysis§  DrajReport

FullFee-for-Services


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TheTeam•  Patrick Nef, PhD, Founder and CEO, has over 25 years of experience in

R&D, and Business Development. HewasGlobal BusinessDirector,Vice-President&DiseaseAreaHeadforCNSatF.Hoffmann-LaRocheLtd.,CEO,CSOorCBOinseveralbiotech.companies,andearlierPatrickwasAssistantProfessor in theDepartmentofBiochemistryat theUniversityofGeneva.HeholdsaPhDinneurosciencesfromtheUniversityofGeneva,Switzerland

•  Sébastien Tabruyn, PhD, Chief Scientific Officer, has 12 years of

international experience in Oncology, Molecular Biology and in vivo models. Project leader at UCSF (California, USA), the Center for CancerBiology (Adelaide, Australia) and theUniversity of Liege (Belgium), he hasmore than 30 publica'ons in the field of Tumor, Vascular biology andInflamma'on. Sébas'en holds a PhD in molecular biology from theUniversityofLiege,Belgium


•  Stéphane Legastelois, PhD, President, has over 20 years experience with

the management of Service and Product companies in Life Sciences. Hebeganhis carrier as aR&DManager ina start-up company,before joiningMerck KGaA as Sales & Marke'ng Manager. Stéphane is also Founder &President of the CMO Indicia Produc'on, and President at Mabdesign, anon-profit organiza'on dedicated to the therapeu'c an'body industrialsector.StéphaneholdsaPh-D in Immunology fromtheUniversityofLyon,France.

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+/-growthfactorexpression

ForbestClinicalCandidateSelec'on

HumanImmuneSystemRecons'tu'on

+/-growthfactorexpression

Predic6veDrugTes6ng


NOG/NSG/BRG

CD34+StemCellOriginforHumaniza'on

CordBlood

-  FromEFS(EtablissementFrançaisduSang)-  Informedconsentsignedbytheparents-  Anonymiza'on-  Maximum48hajerdelivery-  Volume:+/-100ml-  TotalNumberofcells(around2X109)-  %ofCD34:around0.5%

AffinitySepara'on

between5x105and3x106Purity:>90%

HaplotypeSelec'on

CD34+


TypicalHematopoie'cRecons'tu'on

0 5 10 15 20 250

20

40

60

80

100

Hematopoietic Reconstitution

Time (Weeks)

% a

mon

g hC

D45

T cellsB CellsMyeloid CellsNKDendritic Cells


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-  Highrateofhumaniza'on(20-90%)-  Lifelongstability-  Recons'tu'onkine'csfullyestablished@14weeks-  Func'onalB&T-cells

Op'mizedProtocolforHumaniza'on


Humaniza'onProcessandQualityControl

-  Chemoabla'on(DX)

CD34injec'on(DX)

Time(Week) O 14

4weekoldNOG

Mou

seCD4

5

HumanCD45

-  QualityControl-  (FlowCytometry)

-  OnlymicewithhRate>25%entertheStudy

CustomerStudy


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FlowCytometryCharacteriza'onatW15

Bcells39%

CD19

SSC

CD3

SSC

Tcells37%

CD4

CD8

humanCD45 TcellsCD3+

CD8+71%

CD4+20%

CD14

SSC

Monocytes10%

CD56

SSC

NKcells1,5%


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HumanCelltype ExpressedinHIS Marker

Leucocytes ✔ CD45

T-Lymphocytes ✔ CD3,CD4,CD8,CD25

B-Lymphocytes ✔ CD19

NKcells ✔ CD16,CD56

Dendri'ccells ✔ CD1c,CD303,CD141,CD11c

Monocytes ✔ CD14

Macrophages ✔ CD11b

Ac'vatedTand/orNKcells ✔ HLA-DR,CD69

HumanCellsinPeripheralBlood

DetectedbyFlowcytometry


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HumanCytokines ExpressedinHIS MethodofDetecPon

huTNF-α ✔ IHC

IFN-γ ✔ FC,RT-PCR

IL-2 ✔ FC

IL-4 ✔ FC

IL-17 ✔ FC,RT-PCR

IL-23 ✔ RT-PCR

IHC(Immunohistochemistry),FC(FlowCytometry),qRT-PCR(RealTimeTaqMan)

HumanCytokinesExpressedinhu-Mice


Inflamma&on/AutoimmuneDiseasesEx:IBD,RA,Lupus,SLE,…

Infec&ousDiseasesEx:HIV,Dengue,…

Cancer:ImmunotherapyEx:Solidtumors,leukemia,PDX.

=

Mul'pleIndica'ons


Vaccines&ImmunoSafety100%humanIgG,prophylaxis,

15TCSOverviewHIV-BIOEUROPEFALL2016

HIVMouseModels

HumanImmunodeficiencyVirus/AIDS

•  Retrovirusfamily,Len'virusgenus•  HIVinfec'onleadstoAIDS&developmentofopportunis'c

co-infec'ons(TB,HBV,HCV,Plasmodiumfalc.)•  >35millionspeopleinfectedworldwide•  >1milliondeathannually•  UnmetNeeds:Nocureoreffec'vevaccine•  StandardTreatment:HAART(HighlyAc'veAn'Retroviraltri-

Therapy)lifelongmanagement•  Sideeffect:adverseevents,resistance,latency•  MainstrainHIV-1ismorevirulentandprevalentthanHIV-2


Rebound

HIVMouseInfec'onModelasinHuman

An'-HIVTreatment

STOPAn'-HIVTreatment


Infec'onwithR5-HIVvirus(Yu2)

Protec'veAn'-HIV•  Efficacy•  Combotherapy

Rebound/Latency

Resistance

HIVMouseModels

HIV-1strain(Yu2,NL4-3)§  Tropism:R5,X4,orcombined§  CytopathicorCytoly'cstrains

TCSOverviewHIV-BIOEUROPEFALL2016 18

RoutesofInfecPon§  IP§  IV§  Vaginal

EndpointsorDiseaseProgression

§  HIVViralloadinbloodbyqPCR§  Leukocytepopula'onbycytometry

§  Survivalrate,weightloss

RoutesofTreatment§  IP§  IV§  Gavage§  FoodPellet/Drinkingwater

ProfilingofanP-HIVdrugcandidates§  Efficacy,Rebound,Resistance,Latency§  Vaccina'on


§  Sampling:Bloodcollec'onfromTailvein/Retro-orbitalvenoussinusUpto100µl/weekAnesthesianotrequired

§  RNAextracPon:frombloodorplasmaAutomatedprocess(Arrow)§  HIVviralload:InHouse–SOPprotocolGenericHIVChargeViral(BiocentricHIVPCRKit)Sensibility:1’000copiesper50µlofplasmasampleStandardcurveestablishedfromspikedplasma(extractedinparallelwiththesamples)Valida'oncriteria:

Standardcurveslope(between-3.6and-2.9)Linearity>0.95Nega'veandposi'vecontrols

HIVViralLoadDetec'onbyqRT-PCR

VaccinPrime

VaccinBoost HIVInfec'on

BloodSamplingHIVLoad(QRT-PCR)

-42 -21 1 7142128

(Resistancelevel:HIVviralload<500copies/ml)

0 10 20 30 40 500

100000

200000

300000

400000

Time (days post infection)

HIV

load

(cop

ies/m

l)

HIV Load

Group 1Group 2


Prophylac'cTreatmentInfec'onwithR5-HIVvirus(Yu2)


HAARTTreatment

•  HAART:Raltegravir,Lamivudin,Tenofovir(foodpelletatlibidum)•  95%decreaseofviralloadinducedbyHAART•  Observablerebound(+/-10daysajerHAARTstop)•  ViralloadajerHAARTstopcomparabletoini'alvalues

0 10 20 30 40 50 60 70 80 90 100 110 120103

104

105

106

107

Time (Days)

Vira

l Loa

d (c

opie

s/m

l)HAART (from day 53 to 78)



DoubleHAARTTreatment+An'-ViralCompound

•  BothHAARTtreatmentsinducedcomparabledecreasesofviralload•  Firstreboundobservablelessthan1weekajerHAARTstop•  Secondrebounddelayedandwithdecreasedviralloadduetoaddi'onal

an'-viraltreatment

0 10 20 30 40 50 60 70 80 90 100 110 120103

104

105

106

107

Time (Days)

Vir

al L

oad

(cop

ies/

ml)

HAART Anti-Viral HAART



ComboHAART+An'-ViralCompound

•  Delayedreboundwhenmicearetreatedwithan'-viralandHAART.

0 10 20 30 40 50 60 70 80 90 100 110 120103

104

105

106

107

Time (Days)

Vira

l Loa

d (c

opie

s/m

l)HAART

Anti-Viral


ComparableHIVloadforIPandvaginalinfec'on


HIV Vaginal Infection

Week post 1st infection

HIV

infe

ctio

n

0 5 100

50

100

Kine'cofvaginalinfec'onrate(Depo-Proveratreatment)

Percen

tageofInfectedmice

Infec'onwithR5-HIVvirus(YU2)VaginalInfec'oninhu-Mice


0 20 40 60 80 10010 2

10 3

10 4

10 5

10 6

10 7 R 5 /Y u2

X 4 /N L4 -3

HIV Load (copies/ml)

Day Post Infection

X4-HIV(NL4-3)Infec'oninhu-Mice

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Day Post Infection

% of hCD4 (among hCD3+)

0 20 40 600

20

40

60

80

100

X 4 /N L4 -3

R 5 /Y u2

X4-HIV(NL4-3)Cytolysisinhu-Mice


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Infec'onwithX4-HIVvirus(NL4-3)

An'-ViralTreatmentinX4-HIVMice

0 10 20 30 40 50102

103

104

105

106

107

Day of Treatment

Vehicle

Tested Compound

HIV Load (copies/ml)


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Restora'onofCD4+CellPopula'oninX4-HIVMiceInfec'onwithX4-HIVvirus(NL4-3)

% of hCD4 (among hCD3+)

0 20 400

20

40

60

Day of Treatment

% in

CD

3+ c

ells

Vehicle

Tested Compound


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Publica'onsSelec'on•  HumanizedmouseasanappropriatemodelforacceleratedglobalHIV

researchandvaccinedevelopment:currenttrend.Ibehetal.,

Immunopharmacol2016,Sept7:1-32.

•  HIV-1cellularand'ssuereplica'onpazernsininfectedhumanized

mice.Araingaetal.,Scien'ficReport2016;6:23513.

•  Topicalgelformula'onofbroadlyneutralizingan'-HIV-1monoclonal

an'bodyVRC01confersprotec'onagainstHIV-1vaginalchallengeina

humanizedmousemodel.Veselinovicetal.,Virology2012:432(2).

•  HumanizedmicerecapitulatekeyfeaturesofHIV-1infec'on:anovel

conceptusinglong-ac'ngan'-retroviraldrugsfortrea'ngHIV-1.

Nischangetal.,PlosOne2012;7(6).


StrengthsofHIVMouseModels

•  FullblownHIVinfec'on(106-107HIVload/ml)within4-5weeks

•  95%decreaseofHIVloaduponHAARTtreatmentin<7days

•  IP,IVorvaginalroutesofinfec'on•  R5andX4tropismrecons'tu'on

•  Cytoly'cac'vityofX4-HIVstrainasinhuman

•  Combotherapystrategyforselec'onofbestclinicalcandidates/

schedule/regimen

•  Latencyandreboundprotocolsfordiseasemanagementwith

mAb&smallNCE’sTCSOverviewHIV-BIOEUROPEFALL2016 30

R&DPartnershipsforGrants


Grant Subject Partner

Eurostars2013 AML Advancedbiodesign(FR),CNRS(FR)

Eurostars2014 DCBasedVaccin:Glioblastoma

AmalTherapeu'cs(CH)

Eurostars2015 DCBasedVaccin:BreastCancer

DCPrime(NL)

•  SaveTimeandBudget

•  Focusonthemostpromisingclinicalcandidates

•  HuDatausefullforINDfilling

•  Onestopshopforinvivopharmacologyexper'se

•  DeriskingforHeadofTransla'onalProjects

•  FlexibilityandAdaptability

Customer’sAdvantages


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Q&A

Happytoansweryourques'ons/concerns

PatrickandSebas'en

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Patrick Nef, PhD, CEO Sebastien Tabruyn, PhD, CSO Tel: +33 4 56 44 81 21 [email protected] [email protected] www.tcbioservices.com


Documents

Predictive in vivo Pharmacology - TransCure … · Predictive in vivo Pharmacology ... • Both HAART treatments induced comparable decreases of viral load ... current trend