Guterres, 2012

Sílvia Stanisçuaski Guterres Universidade Federal do Rio Grande do Sul

Desafios e perspectivas para a nanobiotecnologia no cenário brasileiro

Guterres, 2012

Comparative scales

Average size = 200 nm

Guterres, 2012

1908 Ehrlich's Magic Bullet 1959

Feynman gives after-dinner talk describing molecular machines

1974

Taniguchi uses term "nano-technology" in paper on ion-sputter machining

1977

Drexler originates molecular nanotechnology concepts at MIT

First technical paper on molecular engineering to build with atomic precision

1986

First book published

First organization formed

1987

First university symposium- MIT

1988

First university course-Stanford

1989

IBM logo spelled in individual atoms

First national conference: Stanford University

1990

First nanotechnology journal

N&N: A brief history

Guterres, 2012

1991

Japan's MITI commits $200 million

Carbon nanotube discovered 1992 Taxol™ (paclitaxel) 1994

Nanosystems textbook used in first university course

1995

First industry analysis of military applications Doxil ™

1996

First European conference: European Nanotechnology Initiative, Copenhagen NASA begins work in computational nanotech

First nanobio conference: California Institute of Technology 1997

First company founded: Zyvex

First design of nanorobotic system

1998

First DNA-based nanomechanical device

1999

First Nanomedicine book published, First safety guidelines

N&N: A brief history

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2000

President Clinton announces U.S. National Nanotechnology Initiative

2001-2017 First 25 Brazilian Networks on Nanotechnology

Brazilian government has invested, through its universities and research centers, approximately R$ 160 million on nanotechnology research 2002

First nanotech industry conference

2003

Congressional hearings on societal implications

2005

Nanoethics meeting 2008 Brazil: INCTs 16 National Institutes of Science and Technology committed to the advancement of knowledge in various nanotechnologies

N&N: A brief history

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2009

An improved walking DNA nanorobot First Brazilian product based on Nanotecnnology

2010

DNA-based 'robotic' assembly begins 2012 130 companies working with research and development 2012 Ministry of Science, Technology and Innovation organized in 2012, the Interministerial Committee for Nanotechnology 2013-2018 Brazil launched an Initiative to make Nanotechnology “the most innovative Brazilian industry” SisNano R$ 440 million (2013 and 2014) 2015 Brazil participates of Nanoreg 2016 New INCTs

N&N: A brief history

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Guterres, 2012

NANOTECNHOLOGY IN HEALTH

Therapeutics

Nanomedicines

Diagnosis Theranostics Personal care

Nanocosmetics

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NANOENCAPSULATION TECHNIQUES products containing coated particles act as drug delivery systems

NAN

O D

ELIV

ERY S

YSTEM

S

To control the drug delivery to the site of

action

To control the kinetics of drug release

To dissolve/deaggregate during or after releasing

the drug

Guterres, 2012

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A comprehensive analysis of the worldwide state of investigational and approved nanomedicine products as of January 2012 has identified 67 commercialized nanodevices and 33

marketed

nanotherapeutics. A total of 25 devices and 122 therapeutics currently in development accounted for 789 ongoing clinical trials.

International Journal of Nanomedicine 2014:9 4357–4373

Ensaios clínicos de nanomedicamentos

Liposomes 23 ago 2016 1865 studies

Ensaios clínicos de nanomedicamentos

Nanoparticles 23 ago 2016 243 studies

Cancer de mama 52

CA Pulmão (células não pequenas) 20 CA Pancreas 19 Tumores sólidos 15 Dental 10 Cancer de cabeça e pescoço 9 Cancer de estomago 5 Melanoma

Glioma 5 CA Fígado 4

CA Próstata 4 Restenose 4

Paclitaxel 120

Nanoparticles 23 ago 2016 243 studies

Nano* and drug 23 fev 2017

Brasil: 14 posição

Nano* and cosmetic 23 fev 2017

Brasil: 6 posição

Nanocarriers in Pharmaceutics

Lipossomas

Fulereno Nanotubo de carbono

SLN NC NS

Dendrímeros

Elevado grau de complexidade

Nanomaterials can be divided into two groups:

Soluble and/or biodegradable nanoparticles which disintegrate into their

molecular components

e.g. liposomes and nanoemulsions

Risk assessment based on mass metrics may be

adequate for the soluble nanoparticles;

Insoluble and/or biopersistent nanoparticles

e.g. TiO2, fullerenes, quantum dots

Risk assessment may require other metrics, such as the number of

particles, and their surface area as well as their

distribution

Organisation for Economic Co-operation and Development (OECD)

N&N

GOVERNMENTAL INITIATIVES

Public policy program consisting of various actions geared towards nanotechnology becoming the most innovative Brazilian industry”.

Nanotechnology can strengthen and “increase the competitiveness of the national economy”.

CENANO

Laboratório de Química de Nanoestruturas de

Carbono

LIN

Let’s begin by showing worldwide data

Market of products that incorporate nanotechnologies was US$135 billion

Expected market: US$3.3 trillion

http://tnsolution.com.br/en/2016/01/26/the-advances-of-nanotechnology-in-brazil/

The big markets

①chemical industry

②semiconductors

③pharmacy

④automotive

⑤defense

⑥antimicrobials

2018

2007 2018

Russian Federation: 500 companies were engaged in manufacturing nanotech (2013, according to the

state corporation Rusnano)

A COMPARISON...

Brazil: 130 companies working with nanotechnoloy (J. Hazard. Toxic Radioact.

Waste, 2016, 20(1): B4014002)

2012 2,500 researchers

3,000 graduate students in the area

Researchers in N&N in Brazil

❶What is the problem to be solved?

❷Are there tools available to solve the problem?

❸Is there expertise to use them?

The rational design a new nanomedicine

LIPID CORE NANOCAPSULES

ROUTES

Parenteral

Skin

Oral Eyes

Brain

Lung Vaginal

Lipid-core nanocapsules (LNC) are formed by an organogel surrounded by poly(epsilon-

caprolactone) and stabilized by polysorbate 80

An original soft nanocarrier

Biodegradability: a critical requirement for nanoparticulate brain delivery systems, i.e.

over a time frame of a few days.

Guterres, 2012

What is the chemical structure of this nanocapsule?

Capric/caprylic triglyceride

Sorbitan monostearate

Poly(e-caprolactone)

Polysorbate 80

-100 -50 0 50 100 150 200

NC-EM

NC-DIC

Heat flow

(exo

)

Temperature (C)

NS-EM

NS-DIC

Cruz, L; et al. Int J Pharm, 2006. Müller, C.R; et al. Pharmazie 2001.

DSC SAXS

TEM

Jager, E. et al; JBN, 2009.

DLS

Venturini, A. et al; COLSUA, 2011

MLS

molecular

macromolecular

supramolecular TEM

Bulcão, R. et al; Toxic Sci, 2013.

200 nm 200 nm

Prof. Adriana Raffin Pohlmann

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ROUTES

Parenteral

Skin

Oral

Eyes

Brain

Lung

Vaginal

Non-toxicity of these nanocapsules per

se is a requirement for their application

ARE THE LIPID CORE NANOCAPSULES SAFE?

Acute cardiotoxicity of lipid core nanocapsules in rats by intravenous administration

Toxicol. Res., 2016, 5, 168–179

Troponin: highly specific marker for myocardial infarction or heart muscle cell death

Aspartate transaminase (AST) Alkaline phosphatase (ALP)

Number of nanocapsules per milliliter per kg.

Rat groups treated with LNC did not demonstrate alteration on electrocardiographical and ecocardiographical analyses

No important differences on biochemical, hematological and histophalogical analysis were found

Electrocardiograms findings Blood pressure evolution

Acute cardiotoxicity of lipid core nanocapsules in rats by intravenous administration

Rats treated with LNC had no toxic

effects and no death occurred

Relative weight of organs

Histopathological analyses

Biochemical parameters in blood and urine

Microalbumin and NAG activity

Serum complement C3 and hs-CRP.

In Vitro Hematological Tests

Toxicological Sciences 132(1), 162–176 2013

IP route

Acute and Subchronic Toxicity Evaluation of Poly(ɛ-Caprolactone) Lipid-Core Nanocapsules in Rats

Do poly(epsilon-caprolactone) lipid-core nanocapsules induce oxidative or inflammatory damages after in vivo subchronic treatments?

Sub-chronic evaluations Oxidative damage? Inflammatory damage?

Protein damage Antioxidant enzymes Lipid peroxidation GSH levels

Cytokines IL-10 1L-6

Tissues Liver Kidney Heart Brain

IP route: no damages were induced

No dose-dependent or systematic alterations observed

Toxicol. Res., 2015, 4, 994

Prof. Solange Garcia

Bender et al., Int J Pharm 426 (2012) 271– 279

control

LNC are hemocompatible

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ROUTES

Parenteral

Skin

Oral

Eyes

Brain

Lung

Vaginal

application

ARE THE LIPID CORE NANOCAPSULES SAFE?

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ROUTES

Parenteral

Skin

Oral

Eyes

Brain

Lung

Vaginal

Some examples of their

effectiveness

ARE THE LIPID CORE NANOCAPSULES EFFICIENT?

BBB: non fenestrated brain

microvessel endothelial cells characterized by tight junctions

which form a continuous almost impermeable cellular barrier

BBB prevents 98% of

neuropharmaceuticals to reach their targets in the CNS.

Nanoparticles as an efficient strategy to deliver drugs to

the tissue?

Our motivation

Rhodamine B-labeled lipid-core nanocapsules (Rho-LNC).

Cerebral microvessels of mice intravenously treated (A and B)

Cerebral microvessels of mice orally treated (C and D)

Intr

avit

al m

icro

sco

py

Lipid-core nanocapsules are efficient shuttles to reach the brain tissue

Prof. Sandra Farsky

Lipid-core nanocapsules can be sterilized and isotonized

Physicochemical

parameters

LNCG LNCGS

Zeta Potential (mV) -10.60 ± 0.61 -13.86 ± 2.92

Particle number density

(particles.ml-1)

1.16 x1013± 5.22 x1011 1.17 x1013± 3.41 x1011

Superficial area (m2.ml-1) 1.41 ± 0.03 1.44 ± 0.03

pH 5.8 ± 0.3 6.3 ± 0.1

Flux index 1.07 ± 0.01 1.05 ± 0.01

Viscosity (cP) 1.18 ± 0.02 1.15 ± 0.06

Frezzing temperature (oC) - 0.520 ± 0.002 - 0.523 ± 0.002

LNC physicochemical characteristics before (LNCG) and after (LNCGS) autoclaving and isotonizing.

AAPS PharmSciTech, Vol. 18, No. 1, February 2017 (# 2016) DOI: 10.1208/s12249-016-0493-3

Our motivation

Prof. Karina Paese

The most common primary tumors of the brain

At least half of all gliomas exhibit aggressive and malignant behavior They are poorly responsive to multimodaly therapeutic interventions, including surgery, radiotherapy and chemotherapy (Deen et al., 1993) Anticancer drugs are toxic to both tumoral and normal cells and the efficacy of chemotherapy is limited by side effects (Brigger et al., 2002) BBB limits the penetration of antineoplasic drugs into brain tumors (Gelperina et al., 2002) LNC

Glioblastoma

Indomethacin-loaded nanocapsules treatment reduces in vivo glioblastoma growth in a rat glioma model

TUMOR SIZE OF IMPLANTED GLIOMAS

Tumor size was measured 20 days after implantation of C6 cells by

three hematoxylin and eosin

Control NC

Glioblastoma

• Increase the intratumoral bioavailability of indomethacin

• Reduce the growth of implanted gliomas

LIPID CORE NANOCAPSULES

Glioblastoma

Prof. Ana Batastini

0

5

10

15

20

25

30

35

40

45

50

IndOH-NC

A1-42

2 M

50 M 100 M 50 M 100 M-

- + + +

-

- -

#

***

***

Control IndOHNC

50 M

IndOHNC

50 M

IndOHNC

100 M

A1-42

2 M

IndOHNC

100 M

% o

f P

I in

co

rpo

rati

on

Neuroprotective effects of indomethacin-loaded nanocapsules in Alzheimer’s Disease models

In vitro model: organotypic hippocampal slice cultures exposed to A1-42 peptide

0

1000

2000

3000

4000

5000

IndOH-NC

A1-42

2 M

50 M 100 M 50 M 100 M-

- + + +

-

- -

*** ***

#

IL-

6 levels

(p

g/m

l)

Neuroprotective effect is mediated by reduction of

neuroinflammation

Positive control Positive control

Negative controls

Negative controls

amyloid beta peptide

INDOH-NC attenuated Aβ-induced cell death

INDOH-NC block the neuroinflammation

Fluorescent image analysisof propidium iodide uptake

Bernardi et al., Int. J. Nanomedicine (2012)

0.00

0.25

0.50

0.75

1.00Object A

Object C

IndOH1 mg/kg

A1-42

2 nmol

- + --

- - - +

IndOH-NC1 mg/kg

+ - --

+

+

+ -

-

+

***

*****

***

Re

co

gn

itio

n in

de

xIndOH-LNC treatment reduces the memory impairment

induced by Aβ1-42 peptide In vivo model: A1-42 peptide was stereotaxically injected bilaterally into

lateral ventricles of Wistar rats

Behavioral analysis

A higher percentage of time spent exploring the novel

object was considered to be an index of enhanced

cognitive performance

Novel object-recognition task

Bernardi et al., Int. J. Nanomedicine (2012)

Guterres, 2012

YES, SAFE AND EFFICIENT ….SO FAR….

LIPID CORE NANOCAPSULES

ROUTES

Parenteral

Skin

Oral Eyes

Brain

Lung Vaginal

From lab to market: report of a successful technology transfer from

University to Industry

Nanocapsules

Buriti oil Octocrilene Avobenzone

Polymeric wall

Concept of Nanophoton Technology

Chronology

2005 2009 2006 2007 2008

Project approved Nanocosméticos

Edital FINEP 2005

Funds avaliable

Agreement Ufrgs--Biolab-Finep

Planning and development of experimental activities

First stage of experimental work

Patent redaction

Round meeting tables

End of the experimental step

Prototypes

Process scaling-up

Patent application

Registration ANVISA

Marketing

Launching event PROTOPROT FPS 10 +++

2010

UFRGS: receipt of royalties

2013

New Products

2015

First ranked in BR market

Photostability

Photoprotection

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•Perspectives in NanoScience

•Brazilian challenges

1 3 4 6 9 7 26 37 98

192

327

457

651

913

1151 1226

0

200

400

600

800

1000

1200

1400

Web of science, nov 2017

Theranostic*

http://academic.hep.com.cn/fcse/CN/10.1007/s11705-014-1425-y

Perspectives Science in N&N

Chalenge: improve innovation target research areas

• Increase innovation in the Brazilian Industry: few of our domestic pharmaceutical companies have internal R&D capabilities

• Improve impact of papers

Desafios Estimular interdisciplinaridade

• Busca sistemática de integração das teorias, dos instrumentos e das fórmulas de ação científica de diferentes disciplinas

• Novas disciplinas surgem devido à integração interdisciplinar

Maior impacto da produção científica

Chalenge: solve regulatory issues

Desafios Burocracia exagerada

Melhorar a comunicação com público leigo

Desafios

Ciência e a mentalidade científica ainda

não estão incorporadas de maneira plena na sociedade.

2017:

Chalenge: improve research funding ensure continuity

Brazil 2017: 44% slash to the federal science budget

Guterres, 2012

Guterres, 2012

silvia.guterres@ufrgs.br