FF

Universidad de Santiago de Compostela

““ NanotecnologNanotecnolog íías y Sistemas de Liberacias y Sistemas de Liberaci óón de n de FFáármacosrmacos ”” (NANOBIOFAR(NANOBIOFAR--USC)USC)

NewNew therapeutictherapeutic strategiesstrategies basedbased onon

nanotechnologiesnanotechnologies: : NanotherapeuticsNanotherapeutics

MarMaríía Josa Joséé AlonsoAlonso

VIII JORNADAS DE LA SOCIEDAD ESPAVIII JORNADAS DE LA SOCIEDAD ESPAÑÑOLA DE QUOLA DE QUÍÍMICA TERAPMICA TERAPÉÉUTICAUTICACarmona (Sevilla), Noviembre 08Carmona (Sevilla), Noviembre 08

HookeHooke observes observes a a cellcell in in anan opticalopticalmicroscopemicroscope

TransmissionTransmissionelectronelectronmicroscopymicroscopy

ScanningScanningelectronelectronmicroscopymicroscopy

MagneticMagneticResonanceResonanceimagingimaging

AtomicAtomic forceforcemicroscopymicroscopy

TwoTwo--photonphotonmicroscopymicroscopy

WatsonWatson and and CrickCrickStructureStructure of DNAof DNA

PolymerPolymerPolymerPolymerPolymerPolymerPolymerPolymer particlesparticlesparticlesparticlesparticlesparticlesparticlesparticlesradiosotopesradiosotopesradiosotopesradiosotopesradiosotopesradiosotopesradiosotopesradiosotopes

IntroductionIntroductionIntroductionIntroductionIntroductionIntroductionIntroductionIntroductionof of of of of of of of liposomesliposomesliposomesliposomesliposomesliposomesliposomesliposomes Drug/Drug/Drug/Drug/Drug/Drug/Drug/Drug/vaccinevaccinevaccinevaccinevaccinevaccinevaccinevaccine

deliverydeliverydeliverydeliverydeliverydeliverydeliverydelivery

withwithwithwithwithwithwithwith polymerpolymerpolymerpolymerpolymerpolymerpolymerpolymer

NanoparticlesNanoparticlesNanoparticlesNanoparticlesNanoparticlesNanoparticlesNanoparticlesNanoparticles

DNA DNA selfself--assemblyassembly

·· OfficialOfficial lunchlunch of of thethe human human GenomeGenomeprojectproject

··FirstFirst FDA FDA approvedapproved attempattemp atat gene gene therapytherapy in in thethe USUS

AdvancedAdvancedAdvancedAdvancedAdvancedAdvancedAdvancedAdvanced DNA/Drug/DNA/Drug/DNA/Drug/DNA/Drug/DNA/Drug/DNA/Drug/DNA/Drug/DNA/Drug/VaccineVaccineVaccineVaccineVaccineVaccineVaccineVaccine

deliverydeliverydeliverydeliverydeliverydeliverydeliverydelivery withwithwithwithwithwithwithwith nanoparticlesnanoparticlesnanoparticlesnanoparticlesnanoparticlesnanoparticlesnanoparticlesnanoparticles

D.AD.A. La Van, . La Van, D.MD.M. . LynnLynn and R. and R. LangerLanger

NatureNature ReviewsReviews, 1, 77, 2002, 1, 77, 2002

16651665 19301930’’s s 19501950’’s s 1953 1953 1965 1965 1969 1969 1976 1976 1981 1981 1986 1986 19891989--90 90

FeymanFeymanFeymanFeymanFeymanFeymanFeymanFeyman’’’’’’’’ssssssss speech onspeech onspeech onspeech onspeech onspeech onspeech onspeech onNanotechnologyNanotechnologyNanotechnologyNanotechnologyNanotechnologyNanotechnologyNanotechnologyNanotechnology

TheThe originorigin ofof drug drug deliverydelivery nanostructuresnanostructures::nanopharmaceuticalsnanopharmaceuticals

NanoparticlesNanoparticles

polymer micellesdendrimers

Liposomes

DNA vectorsNanotubesNanotubes

Courtesy from Ruth DuncanCourtesy from Ruth Duncan

WHY NANOTHERAPEUTICS?WHY NANOTHERAPEUTICS?

-- More More efficaciousefficacious-- LessLess toxictoxic-- EasierEasier toto administeradminister

HOW DO THEY WORK?HOW DO THEY WORK?

-- OvercomingOvercoming barriersbarriers-- TargetedTargeted controlledcontrolled deliverydelivery-- PreservingPreserving stabilitystability-- SolubilizingSolubilizing hydrophobichydrophobic drugsdrugs

TheThe biopharmaceuticalbiopharmaceutical problemsproblems ofofdrugsdrugs andand vaccinesvaccines

SolubilitySolubility

PermeabilityPermeability

DegradabilityDegradability

BiodistribuciBiodistribucióónn

Drug Drug synthesissynthesis::LowLow MwMw compoundscompounds

BiotecnologyBiotecnology::PeptidesPeptides, , proteinsproteinsDNA, DNA, siRNAsiRNA, , ……

TheThe majoritymajority ofof drug drug candidatescandidates do do notnot reachreach PhasePhase I I duedue totosolubilitysolubility //stabilitystability problemsproblems

More More thanthan 40% 40% ofof active active compoundscompounds approvedapproved in in thethe lastlast decadedecadeare are biopharmaceuticalsbiopharmaceuticals : : unstabilityunstability andand difficultiesdifficulties forfor overcomingovercoming biologicalbiological barriersbarriers

THE NEED OF A DELIVERY CARRIERTHE NEED OF A DELIVERY CARRIER

NanotherapeuticsNanotherapeutics

TherapeuticTherapeutic benefitsbenefits

EconomicalEconomical benefitsbenefits

Social Social benefitsbenefits

ImprovedImproved medecinesmedecineseasiereasier toto administeradminister

More More efficaciousefficacious andandlessless toxictoxic nanomedicinesnanomedicines

NewNew medicines, medicines, nucleicnucleic acidacid--basedbased therapiestherapies

SomeSome real real examplesexamples: : cancercancer therapytherapy

DRUGDRUG--POLYMER CONJUGATESPOLYMER CONJUGATES

Fase III

(pulmón, ovario)Polyglutamic acidPaclitaxelXyotax ®

Carcinoma hepatocelular

Polyestiren-maleicacid

Neocarzinostatina

Zinostatin®

(YamanouchiPharm)

Leucemia PEGAsparaginasaOncaspar®

(Enzon)

IndicationPolymerdrugCommercial

name

SomeSome real real examplesexamples: : cancercancer therapytherapy

POLYMER NANOPARTICLES AND MICELLES POLYMER NANOPARTICLES AND MICELLES

Colorectal

Mama

IndicationIndicationDrugDrugCompositionCompositionCommercial Commercial

namename

Cis-platinoPglu-PEG MicellesEloxatin ®

PaclitaxelAlbumin NanoparticlesAbraxane ®

SomeSome real real examplesexamples: : cancercancer therapytherapy

SomeSome real real examplesexamples: : cancercancer therapytherapy

•• MecanismMecanism ofof actionaction: : thethe nanoparticlesnanoparticles leaveleave thethe bloodblood vesselsvesselsby a receptor by a receptor mediatedmediated ((glycoproteinglycoprotein 60) 60) transcitosistranscitosis andandaccumulateaccumulate in in thethe tumoral tumoral tissuetissue duedue toto theirtheir interactioninteraction withwithSPARC (SPARC (proteinprotein overexpressedoverexpressed in tumoral in tumoral cellscells))

•• No No premedicationpremedication isis requiredrequired toto avoidavoid hypersensitivityhypersensitivityreactionsreactions

•• LowerLower infusioninfusion time time

AlbuminAlbumin nanoparticlesnanoparticles

ContainingContaining placlitaxelplaclitaxel

AprovedAproved JanuaryJanuary 2005 FDA2005 FDA

IndicationIndication: :

MetastaticMetastatic breastbreast cancercancer

AdditionalAdditional reasonsreasons forfor nanotherapeuticsnanotherapeutics

Int. Int. Ass.PharmAss.Pharm. . TechTech. . BaselBasel 20072007

Int. Int. Ass.PharmAss.Pharm. . TechTech. . BaselBasel 20072007

Biocompatible Biocompatible andand biodegradable biodegradable nanostructuresnanostructures

100 100 nmnm

OUR WORK AT THE USCOUR WORK AT THE USC

CURRENT WORK AT THE USC:CURRENT WORK AT THE USC:

RationalRational designdesign ofof nanocarriersnanocarriers basedbaseduponupon final final applicationsapplications ((resolvingresolving problemsproblems):):

-- OvercomingOvercoming mucosalmucosal barriersbarriers: : nasal nasal deliverydelivery ofof macromoleculesmacromolecules: : peptidespeptides andand vaccinesvaccines

-- ImprovingImproving efficacyefficacy andand reducingreducing toxocitytoxocity: : anticanceranticancer drug drug deliverydelivery

-- NewNew nucleicnucleic acidacid basedbased therapiestherapies: : pDNApDNA, , siRNAsiRNA

OVERCOMING MUCOSAL BARRIERS:OVERCOMING MUCOSAL BARRIERS:CyclodextrinCyclodextrin//polysaccharidepolysaccharide nanocarriersnanocarriers

PROPERTIES:PROPERTIES:

-- VersatileVersatile

-- ProtectiveProtective effecteffect

-- PermeabilizingPermeabilizing effecteffect

-- AdhesiveAdhesive effecteffect

TECHNOLOGY:TECHNOLOGY:

-- MildMild

-- SimpleSimple

-- FastFast

Cyclodextrin (CD)

+ TPP

Chitosan (CS)

CS/CD Nanoparticle suspension

Preparation of CS/CD Nanoparticles:

Ionotropic gelation

CS/SBE-ββββ-CD nanoparticles

Magneticstirring

FreezeFreeze--dried dried ChitosanChitosan nanoparticles nanoparticles before and after before and after resuspensionresuspension

Nasal Nasal administrationadministration

OVERCOMING MUCOSAL BARRIERS:OVERCOMING MUCOSAL BARRIERS:CyclodextrinCyclodextrin//polysaccharidepolysaccharidenanocarriersnanocarriers

(n = 6)

INSULIN SOLUTION INSULIN SOLUTION <15%<15%

Maximum decrease ofblood glucose levels:

CS/CS/CMCM--ββββββββ--CD CD nanoparticlesnanoparticles:32%:32%

CS/CS/SBESBE--ββββββββ--CD CD nanoparticlesnanoparticles: 36%: 36%

55

65

75

85

95

105

-30 0 30 60 90 120 150 180 210 240 270 300

Time (min)

Pla

sma

gluc

ose

(% o

f ini

tial l

evel

)

*

**

Chitosan-CD nanoparticles: In vivo Studies in Rabbits

Insulin solution CS/CM-ββββ-CD CS/SBE-ββββ-CD

Chitosan coating

Oil core –Immunostimulant

rHBs Ag

Size:100-400 nmAg loading: 90%

OVERCOMING MUCOSAL BARRIERS:OVERCOMING MUCOSAL BARRIERS:Oil/Oil/polysaccharidepolysaccharide nanocarriersnanocarriers: : NanovaccinesNanovaccines

Oil/polysaccharide nanocarriers: intranasal immunization rHBs Ag

1

10

100

1000

1 2 3 4 5 6 7

Time (months)

Ant

i-HB

sAg

IgG

(µg/

ml)

Control NE - 10µg x 2 i.n.NC:Ag 1:12.8 - 10µg x 2 i.n.NC:Ag 1:0.25 - 10µg x 2 i.n.HBsAg-Alum - 5µg x 1 i.m.

*

** **

* NC:Ag 1:0.25 – HBsAg-Alum & NC:Ag 1:12.8

1

10

100

1000

10000

15 43 73 103 133 163 193

Time (days)

Ant

i-HB

sAg

IgG

(µg/

ml)

NC:Ag 1:12.8 - 10µg x 2 i.m.NC:Ag 1:0.25 - 10µg x 2 i.m.HBsAg-Alum - 10µg x 2 i.m.

Oil/polysaccharide nanocarriers: intramuscular immunization rHBs Ag

PaulPaul EhrlichEhrlich

““““““““The maggic bulletThe maggic bulletThe maggic bulletThe maggic bulletThe maggic bulletThe maggic bulletThe maggic bulletThe maggic bullet””””””””

““To go straith to the organisms to To go straith to the organisms to WhichWhich theythey werewere aimedaimed””

The Nobel Prize in Physiology or Medicine 1908

"in recognition of their work on immunity”

IMPROVING EFFICACY/REDUCING TOXICITY:IMPROVING EFFICACY/REDUCING TOXICITY:anticanceranticancer drug drug deliverydelivery

PoliaminoacidPoliaminoacidPoliaminoacidPoliaminoacidPoliaminoacidPoliaminoacidPoliaminoacidPoliaminoacid andandandandandandandand polysaccharidepolysaccharidepolysaccharidepolysaccharidepolysaccharidepolysaccharidepolysaccharidepolysaccharide nanocapsulesnanocapsulesnanocapsulesnanocapsulesnanocapsulesnanocapsulesnanocapsulesnanocapsules

LessLess toxictoxic andandmore more efficaciousefficacious

drugsdrugs

“The week link of gene therapy is paradoxically the vehicle ratherthan the drug itself”

J.P. Behr, Bioconjugate Chemistry (1994)

NEW NUCLEIC ACID BASED THERAPIES: NEW NUCLEIC ACID BASED THERAPIES: pDNApDNA, , siRNAsiRNA

NanobiopharmaceuticalsNanobiopharmaceuticals

ConfocalConfocalfluorescencefluorescencemicroscopymicroscopy

corneacornea

3 d3 díías postas post --instillationinstillation

Green channel = Transfected cells: GFP

HialuronicHialuronic acidacid basedbased nanoparticlesnanoparticles::Ocular gene Ocular gene therapytherapy

flHA

flHA:CSO

2h post-instilación 12h post-instilación

BIODEGRADABILITY OF BIODEGRADABILITY OF HA:CSHA:CS NANOPARTICLES NANOPARTICLES IN CORNEAL TISSUEIN CORNEAL TISSUE

TheThe futurefuture……....

‘‘Inteligent TherapeuticsInteligent Therapeutics’’

‘‘Smart Drug DeliverySmart Drug Delivery’’

-- IdentificationIdentification ofof biomarkersbiomarkers andand targetstargets

-- NewNew biomaterialsbiomaterials: : biomimeticbiomimetic//bioinspiredbioinspired(molecular (molecular recognitionrecognition, , sensitivesensitive toto biologicalbiological changeschanges…….).)

-- NewNew techniquestechniques forfor evaluationevaluation

Gracias!Gracias! MarMaríía Josa Joséé Alonso: ffmjalon@usc.esAlonso: ffmjalon@usc.es

NANOBIOFAR GROUPNANOBIOFAR GROUP

FinnancialFinnancial supportsupport::

NanobiosaccharidesNanobiosaccharides--EUEUVascuplugVascuplug--EUEUGalenosGalenos--EUEUGCGHGCGH-- BillBill andand MelindaMelinda GatesGates FoundationFoundationNanovaccinesNanovaccines-- MICINNMICINNCenit Cenit NanopharmaNanopharma ((FaesFaes, , PharmamarPharmamar))AdvancellAdvancell S.A.S.A.