clelia dispenza - superevent...apr 28, 2017 · dr natascia grimaldi* dr simona todaro* maria di...

Radiation synthesis of nanosized drug delivery devices

CLELIA DISPENZADepartment of Industrial and Digital Innovation

University of Palermo

ITALY

MotivationDevelopment of

“MODULAR” NANOCARRIERS

to address the therapeutic needs of specific pathologic states

Drug ProtectionSite Recognition

Triggered ReleaseTracking

Improved biodistribution and pharmacokineticsReduced administered dosesMinimized side-effectsPersonalized therapies

MULTIPLE FUNCTIONS SEVERAL BENEFITS

WITH A SINGLE NANODEVICE

NanogelsWater-swollen, crosslinked polymer nanoparticles

• Stable aqueous colloids

• Soft and conformable

• Stimuli-responsive

• MultifunctionalDh= 10 - 100 nm

• Controlled particle size and surface electrical charge

• Reactive groups for conjugation of targeting ligands, drugs, etc.

• Simple production schemes

• Biocompatibility

KEY REQUIREMENTS

Radiation synthesis of nanogelsPulsed, e-beam irradiation of semi-diluted aqueous solution of an “inert” polymer.

1. The radiation interacts mainly with water undergoing radiolysis:

2. Water radiolysis products (mainly .OH) react with the polymer, forming polymer radicals:

3. Chemical follow-up reactions lead to the formation of crosslinked polymer nanoparticles

H2O = ∙OH, ∙H, e-aq , HO2∙(O2∙

-)

P+ ∙OH = P∙

Radiation synthesis of nanogels

Institute of Nuclear Chemistry and Technology (INCT), Warsaw, Poland.

NO CATALYSTS OR INITIATORS, ORGANIC SOLVENTS AND SURFACTANTS REQUIRED!

Industrial-type

accelerators

(10 MeV)

Doses ≥ Sterilisation

doses Dose per pulse ≈ 10 Gy

Poly(N-vinyl pyrrolidone)

Key process parametersC

RO

SSLI

KIN

G

....

.. ..

. ..+

CONTROL OF SIZE and DENSITY

INTRAINTER

PVP NG (0.1) NG (0.2) NG (0.3) NG (0.4) NG (0.5)

Dia

me

tri id

rod

ina

mic

i (n

m)

0

20

40

60

80

100

120

140

160

TEM*

0 50 100 150 200

0,05

0,1

0,25

0,5

Dh, nm

Po

lym

er

con

c., w

t% High dose

per pulse

Low doseper pulse

Poly-N-vinyl pyrrolidone nanogels

non irradiated

DLS 40 kGy

*

error bars = width of size distribution

macro

Polymer concentration and dose per pulse

Key process parametersC

RO

SSLI

KIN

G

....

.. ..

. ..+


INTRAINTER


Gel Filtration Chromatography

• Linear polydisperse polymers transform into monodispersenanoparticles;

• Size is insensitive to dose in the 20-80 kGy range.

Can the combination between primary radicals be neglected?

OH +OH = H2O2

If the combination between primary radicals cannot be neglected, then…

OH +OH = H2O2

H2O2 +OH = H2O + HO2

2 HO2 = H2O2 + O2

P + O2 = POO …

Key process parameters

Pulse length of 50 ns and frequency of 25 Hz for N2O saturated water and aqueous solutions containing PVP and terbutanol.

H2O2 formation

0.05%

Dispenza et al. RSC Adv., 2016, 6, 2582

Chemical functionality

0.1 % wt

0.5 % wt

N O HN O

O

OH

HNO

O

OH

NH2

NG-COOH determined by their ability to extract Ni2+ from water NG-NH2 titrated by reaction with fluorescamine

COOH

NH2

NH2

OH

COO-

COOH

COOH

OH

OH

Sabatino et al. Polymer , 2013, vol. 54, 54-64Dispenza et al. RSC Adv., 2016, 6, 2582

Dose-dependant functionalisation

Branching

…

CONTROL OF (SURFACE) CHEMISTRY AND TOPOLOGY

....

...

. +

GR

AFT

ING

functional acrylic monomer (e.g. acrylic acid)

(from water radiolysis).

..... .

.. + H2O2/O2

OX

IDA

TIO

N

…COOH

NH2

NH2

OH

COO-

COOH

COOH

OH

OH

CR

OSS

LIK

ING

....

.. ..

. ..+


INTRAINTER


Polymer concentration, dose per pulse and dose

Key process parameters

Polymer concentration, dose per pulse and dose, reactive solutes

From nanogel to nanodevice

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH

PVP-co-AA 40kGy

z-potential = -25 mVDh = 70 nm PDI = 0.25

ANTIBODIES

Targeting

Ligands

PEPTIDES

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH


ANTIBODIES

NUCLEIC ACIDS

Targeting

Ligands

Therapeutics

-DOXO-S-S-

CHEMOTHERAPEUTICS

PROTEINS

PEPTIDES

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH


ANTIBODIES

NUCLEIC ACIDS

Targeting

Ligands

Therapeutics

-DOXO-S-S-

CHEMOTHERAPEUTICS

PEPTIDES

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH DOTA-[Lys3]BBN

CHELATING AGENTS

Diagnostic

tools

PROTEINS


-DOXO-S-S-

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH DOTA-[Lys3]BBN

conjugation through amide bond(EDC, Sulfo-NHS)

FITC, AminoFluo

TRITC, amino-Atto633

Fluo. probe


-DOXO-S-S-

COOH

COOH

NH2

COO-

NH2

COOHNH2

COO-

COO-

COOH

COOHCOOH

OH

COOH DOTA-[Lys3]BBN

Insulin

1. Biomacromolecules (2012) 13, 1805-1817.2. J. Appl. Polym. Sci (2014) 131(2), 39774-39782.3. Biomaterials (2016) vol. 80, 179-194.4. Molecules (2017) vol 21(11),1594.5. Biological Chemistry (2017) vol. 398(2), pp. 277-287.

[5]

[2, 5] [4]

[4]

[1,3]


Insulin has important effects in the treatment of neurological diseases, such as Alzheimer’s Disease (AD) for the recovery of memory deficit.

Alzheimer’s disease

Insulinresistance

Insulinin CNS

AD is also named Type III diabetes

HealthyAdvancedAlzheimer’s

The ProblemEfficient delivery of insulin to the central nervous system is hampered by the Blood Brain Barrier.

Enhanced nose-to-brain insulin delivery through nanogel carriers


Alzheimer’s disease

Insulinresistance

Insulinin CNS

AD is also named Type III diabetes

HealthyAdvancedAlzheimer’s

The StrategyIntranasal delivery: the delivery occurs mainly by the olfactory and trigeminal nerve pathways.

Insulin has important effects in the treatment of neurological diseases, such as Alzheimer’s Disease (AD) for the recovery of memory deficit.

INTRANASAL DELIVERY LIMITATIONS

• Small sprayed volumes

• Low permeability of nasal mucosa

• Hormone degradation by protolithic enzymes

• Harmful effects on the nasal epithelium


NG-In biological evaluation

NG-Atto633

NG-In

NG-InFITC

NG-Atto633 -InFITC

PVP-co-AA NGe-beam irradiation

(40 kGy)


URINE ANALISYS

BIODISTRIBUTION

NG in blood


No deterioration of NASAL MUCOSA

before after 10 min after 30 min

BRAIN DISTRIBUTION

Free InsulinFITC NG-InFITC

NGs cross NASAL MUCOSA

No brain immunogenic response.

HYPPOCAMPUS CORTEX

• Radiation engineered NANOGELS are versatile, biocompatible NANOCARRIERS.

• NG-In is MUCOADHESIVE and RESISTANT TO PROTEOLYTIC ENZYMES.

• NG-In carries biologically active insulin to the brain with higher efficiency.

• NG-In can be cleared from blood and organs (where do not induce modifications).

• Repeated administration effects and studies of disease models are undergoing.

INSULIN SIGNALINGACTIVATION


DIPARTIMENTO DELL’INNOVAZIONEINDUSTRIALE E DIGITALE (UniPa-Italy) Prof. Giuseppe SpadaroProf. Sabina AlessiDr Maria Antonietta SabatinoDr Natascia Grimaldi*Dr Simona Todaro*Maria Di Filippo, PhD studentLorena Anna Ditta, PhD studentAlessia Ajovalasit, PhD student

ISTITUTO DI BIOLOGIA MOLECOLARE (CNR-Italy)Dr Marta Di CarloDr Pasquale PiconeDr Domenico Nuzzo

ISTITUTO DI BIOFISICA(CNR-Italy)Dr Donatella BuloneDr Daniela GiacomazzaDr Pier Luigi San Biagio

ROYAL INSTITUTE OF TECHNOLOGY(Stockholm, Sweden) Prof. Mats Jonsson

INSTITUTE OF NUCLEAR CHEMISTRY AND TECHNOLOGY (Warsaw, Poland)Dr. Grazyna PrzybytniakDr. Marta WaloProf. Andrzej Chmielewski

€

UNIVERSITY OF MARYLAND(College Park, MD, USA)Prof. Mohamad Al-Sheikhly

*past PhD student

STEBICEF (UniPa-Italy)Prof. Antonella Amato Prof. Flavia Mulè

Acknowledgments

NG-In team

PRIN project “NANOMED”

IAEA CRP 2014- 2018 on “Nanosized Delivery Systems for Radiopharmaceuticals.”

IAEA CRP 2009- 2012 on "Nanoscale Radiation Engineering for Biomedical Applications”.

PON HIPPOCRATES

THANK YOU FOR YOUR ATTENTION!

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organised in collaboration with IAEA

clelia dispenza - superevent...apr 28, 2017 · dr natascia grimaldi* dr simona todaro* maria di...

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