FAPESP WEEK

October 24th – 26th, 2011

WASHINGTON, DC

PROF. JORGE KALIL

Instituto Butantan | 1

AGENDA

BUTANTAN - SCIENCE IS THE BASIS

BUTANTAN – SOME PROJECTS

Instituto Butantan |

In 1901 Butantan was established to produce serum against the bubonic plague

Vital Brazil, the first director, investigated antivenoms against snake bites

Currently, Butantan is the main public producer of vaccines, antivenoms,

antitoxins in Latin America

Fully dedicated to develop scientific research and production of immunebiological

products for public health

2

BUTANTAN – A PUBLIC INSTITUTION OF THE STATE GOVERNMENT OF SÃO

PAULO

Instituto Butantan | 3

Strategy

Brazil decided in the mid 80's to become self-sufficient

in vaccines and immunization programs

Decision

This was a State decision rather than a government decision

Too important to depend on availability and pricing

Why

BRAZIL HAS BECOME AN INTERNATIONAL REFERENCE IN IMMUNIZATION

Instituto Butantan | 4

Smallpox

Poliomyelitis

Measles (autocne transmission)

Neonatal tetanus

Accidental tetanus

Tuberculosis

Diphtheria

Pertussis

Hepatite B

Influenza

Pneumococcus

Eradication

Under

Control

PNI – Founded in 1973

NATIONAL IMMUNIZATION PROGRAM (PNI)

193 MM inhabitants

43 types of immunobiological

26 Vaccines

13 sera from animal

4 sera from human

77% produced in Brazil

~ 300 MM doses of vaccines per year

30 K vaccination rooms

Expansion of national self-sufficiency

PNI – General Information of Brazil (2010)

Source: SVS/Ministry of Health, National Immunizatation Program, Brazil, 2010.

Instituto Butantan |

Butantan

51%

‘Market Share’ per Suppliers(1)

(2010)

NATIONAL SUPPLIERS OF VACCINES FOR THE MINISTRY OF HEALTH

DTP DT dT

Anti Rabies Hepatitis B

Influenza (Flu)

5

Biomanguinhos

42%

FAP

4%

FUNED

3%

1Source: Ministry of Health, 2010

Products

Note: Part of Butantan’s production was sent to other Institutes,

such as Biomanguinhos. Not computed in the analysis

Instituto Butantan | 1Source: Ministry of Health, 2010

Butantan

56% FUNED

19%

‘Market Share’ per Suppliers(1)

(2010)

Snakes AV

Scorpion AV Spider AV Caterpillar AV

Tetanus AT Diphitheria AT Anti Rabies

Botulism AB & E

Bee AV

NATIONAL SUPPLIERS OF ANTIVENOMS AND ANTITOXINS FOR THE MINISTRY OF

HEALTH

6

IVB

25%

Products

Note: Part of Butantan’s production was sent to other

Institutes, such as FUNED and CPPI. Not computed

in the analysis

Instituto Butantan |

EXPORTING PRODUCTS

7

Instituto Butantan | 8

Cooperation Projects

NIH-PATH Rotavirus (pentavalent)

NIH-DVI Dengue (tetravalent)

Sabin Vaccine Institute - George Washington University Necator - Schistosoma

Children’s Hospital Harvard - PATH Pneumococcus (cellular)

Infectious Diseases Research Institute Leishmaniosis (for dogs)

Ludwig Institute for Cancer Research Adjuvant for ovarian cancer

BR Foods Lung Surfactant

Universidade de São Paulo – Medical School Recombinant OncoBCG for bladder

cancer

Institut Pasteur – Paris / Novartis - Siena /

Albert Einstein College of Medicine Recombinant BCG-Pertussis

VACCINES WITH EXTERNAL COOPERATION

Instituto Butantan |

0

50

100

150

200

250

0

500

1000

1500

2000

2500

1980 1985 1990 1995 2000 2005 2010

sum of times cited (TOTAL)

Papers Published

Poly. (sum of times cited (TOTAL))

Expon. (Papers Published)

INDEXED SCIENTIFIC PUBLICATIONS

PAPERS PUBLISHED AND CITED

9

Instituto Butantan |

ANALGESICS

ANTI- INFLAMMATORY

ANTICOAGULANTS

ANTITUMOR

ACTION IN THE NERVOUS

SYSTEM

ANTIVENINS

ANTIHIPERTENSIVE

Venom Composition (Transcriptomics and Proteomics)

Pharmacological activities

SCREENING OF BIOACTIVE COMPONENTS OF POISONOUS ANIMALS

10

Species

Instituto Butantan |

Derived from animal secretions (32)

LOPAP (4 PATENTS APPLICATIONS) • Prothrombin activator,

• defibrinogenating/ antithrombotic agent

• kit for diagnosis of dysprothrombinemias

• anti-apoptotic activity

TICKS’ SALIVARY GLANDS • A new anticoagulant and anti-tumoral agent

• Microarray analysis

Lonomia obliqua

Amblyomma cajennense

11

PATENTS

Instituto Butantan |

New developments in Pertussis Vaccines with Appropriate technologies

Whole Cell Pertussis Vaccine

Low LPS content- Whole Cell Pertussis less reactogenic – low cost

B. pertussis fermentation Tangential

filtration

WCP Organic extraction

WCPLow

Patent

MPLA Adjuvant

Patent

Clinical Trials 2007

Infants

Acellular Pertussis Vaccine

Patent

Adolescents

Recombinant BCG-Pertussis

Immunization at birth

Under GMP production

Clinical Trials planned 2009

Neonates

Patent

Biotechnology (16)

IMMUNOGENIC COMPLEX FORMED BY VACCINAL

ANTIGENS ENCAPSULATED BY NANOSTRUCTURED

MESOPOROUS SILICA

C R I S T A L O G R A P H Y L A B O R A T O R Y

PATENTS

12

Instituto Butantan |

RESEARCH & DEVELOPMENT LABORATORIES

13

~21 scientific labs

~180 Researchers

85% are PhD

1 Biotechnology Center

Multiple laboratories

1 Hospital (10 hospital beds)

1 Central Animal Facility

Training programs (PAP)

Graduate studies in Toxicology

Masters and PhDs

Instituto Butantan |

INDUSTRIAL COMPLEX

14

7 Main Industrial Plants (Buildings)

Anaerobic vaccines (tetanus and botulinic )

and Anatoxin Purification

Biological control

Aerobic Vaccine (Diphtheria and Pertussis)

Hepatitis

Influenza

Rabies

Blood Products (under construction)

Control, Serums, Formulation and Filling

6 Pilot Plants Dengue / Rotavírus (Under Construction)

Recombinant (BCG)

Monoclonal Antibodies

Influenza

Blood Products

Instituto Butantan | 15

AGENDA

BUTANTAN - SCIENCE IS THE BASE

BUTANTAN – SOME PROJECTS

Instituto Butantan | 16

Area Projects 1

Vaccines for immunepreventable

diseases

Rotavirus (pentavalent)

Dengue (tetravalent)

Research and improvement Adjuvant BpMPLA

Lung Surfactant

Silica nanostructure mesoporous – vaccine

antigens encapsulated

Biopharmaceuticals Monoclonal antibodies (anti-human CD3)

Clotalfina (analgesic product)

Amblyomin-X (anti cancer)

Lopap (antithrombotic agent)

Presentation and discussion of some Projects

1 Not exhaustive

Instituto Butantan | 17

Composition

– Attenutated virus

– Sorotypes: G1, G2, G3, G4 e G9

Technology of Production

– Cell substrate: Vero cells

– Reassortment – Human/bovine

– Nº lots produced: 09 (6.324 doses)

Phase of Development

– Phase I: 2010

Results: safe and immunogenic

– Phase II: 2012

• Partnership

– NIH / PATH / BNDES

Product – Pentavelent Rotavirus Vaccine

Challenges:

– To perform Phase II and III - non-inferiority

study

– To find funding for:

Clinical Trial and laboratory assay – Phase II / III

Objective:

– Pentavalent low cost vaccine

Butantan –

Rotavirus Vaccine - Overview

Instituto Butantan | 18

Composition

– Attenuated virus

– Sorotypes: DEN1, DEN2, DEN3, DEN4

Technology of Production

– Cell substrate: Vero cells

– Recombinant DNA technology

– Chimeric

– Nº lots produced: 06 (12.640 doses)

Phase of Development

– Phase I: 2011/2012

• Partnership

– NIH / DVI / BNDES / FAPESP

Product – Tetravalent Dengue Vaccine Butantan –

Dengue Vaccine - Overview

Challenges:

– To speed up Phase I, II, and III – (to avoid

non-inferiority study)

– To find funding for:

Clinical Trial and Laboratory assay - Phase II

and III

Equipment

Plant

Maintenance of “The Global Solutions for

Infectious Disease” support

– To define target population for immunization

– Production capacity x national and

international demand

Objective:

– Tetravalent low cost vaccine

Instituto Butantan | 19

Dengue Vaccine - technical and scientific aspects Attenuation strategies for Dengue virus and Pre-Clinical studies

30 nucleotide deletion in DENV 3’ UTR = 30

mutation

rDEN1 30 and rDEN4 30

The D30 mutation partially attenuated DEN2 and

did not attenuated DEN3 for rhesus monkeys

Antigenic chimeras - rDEN4 30 background

rDEN2/4 30 – infectious in monkeys and

humans

rDEN3/4 30 – infectious in monkeys and

poorly infectious in humans

DEN3/4 30

Additional 3’-UTR deletion mutations

rDEN3 30/31 – infectious in monkeys and

humans

Attenuation strategies for Dengue virus

vaccine candidates

SCID-HuH-7 mice

The candidate vaccine viruses were attenuated,

replicating to a peak titer from 100.9 up to 102.4

PFU/mL

Rhesus macaques

Low/undetectable levels of viremia in (101.0 to 101.3

PFU/mL)

Immunogenic

Mosquitoes

Vaccine viruses restricted ability to infect the midgut

and to cause a disseminated infection (Ae. Aegypti -

Toxorhynchites splendens)

Ae. albopictus fed on viremic subjects - vaccine

virus was not recovered from any mosquitoes

Pre-Clinical Studies

Instituto Butantan |

Composition

– BpMPLA derived from LPS of B.pertussis

Production Technology

– Purification of B.pertussis followed by LPS

hydrolises

Phase of Development

– Scale-up

Product - Monophosphoryl lipid A (BpMPLA) Butantan –

Challenges:

– Scale-up

Objectives:

– To optimize immunolgical response of pre-

exisiting and new vaccines

– To increase production capacity

Adjuvant – Monophosphoryl lipid A (BpMPLA)

BpMPLA

– Clinical trial

Pandemic H1N1 + BpMPLA

– Pre-clinical

Human rabies

– Animal Study

Dog Leishimania

– In development

Hepatites B + BpMPLA

Seazonal Influenza + BpMPLA

21

Instituto Butantan | 22

Adjuvant – Monophosphoryl lipid A (BpMPLA)

Monophosphoril Lipid A (MPLA)

derived from Bordetella pertussis

as a new vaccine adjuvant

Pathogen-Associated Molecular

Patterns (PAMPs) and Danger-

Associated Molecular Patterns are

potential new adjuvants

Instituto Butantan | 23

Composition

– Origin porcine

– Phospholipids + Proteins SP-B and SP-C

Production Technology

– Porcine pulmonary extract adsorbed on a

cellulose derivative followed by organic

extraction, solvent evaporation, lyophilize

drying and final formulation

Phase of Development

– Licensed in Brazil in 2009

Product – Surfactant Butantan –

Challenges:

– To test a new process of production

(ultracentrifugation) and formulation – 2nd

generation of surfactant

Objectives:

– To make available a low cost surfactant to

Brazilian Public Maternities

Lung Surfactant

Instituto Butantan | 24

Lung Surfactant - technical and scientific aspects R

esp

irat

ory

Dis

tres

s S

ind

rom

e N

orm

al

Su

rfac

tan

t

Alveolus

Lung Parenchyma

Instituto Butantan | 25

The SBA-15 possesses hexagonal porous uniformity (3.1 – 6.5 nm)

Thermal and hydrothermal stability

Exhibits potential applications for selective adsorption and catalysis

Features

Silica (SBA-15) Immunogenic complex formed by vaccinal antigens encapsulated by

nanostructured mesoporous silica

Instituto Butantan | 26

Adjuvant effect of SBA-15

Instituto Butantan | 27

IM VO

Source: Carvalho et al. 2010.

Modulation of antibody response - Silica 10 μg/animal de BSA:250 μg/animal de SBA-15 ou Al(OH)3

Instituto Butantan | 28

Monoclonal Antibodies Anti-human CD3

AcMo Murine AcMo Quimeric AcMo Humanized

Crohn's disease

Encephalomyocarditis autoimmune

Psoriatic arthritis

Diabetes Type I

Lupus erythematosus

Control of acute rejection in kidney transplant

Prevention of rejection in transplanted pancreatic islet

Steroid-refractory acute GVHD

POTENTIAL

CLINICAL USES

anti-human CD3

Instituto Butantan |

Clotalphine Potent analgesic drugs from animal venoms and toxins

29

Venom of

Potent analgesic

long-lasting analgesic action

(2-5 days)

Crotalphine

Molecular mechanisms involved in

the analgesic action

release of

endogenous

opioids

Activation of peripheral and

opioid receptors

intracellular

signaling

pathways

activation

Activation of NO-cGMP-PKG

and opening of potassium

channels

Recent studies show involvement of PKC

which activates MAPKs (Mechanism that may contribute to its long-lasting

analgesic action)

Activation of CB2 cannabinoid

receptors

Crotalus

Instituto Butantan | 30

Amblyomin-X ®

An anti-cancer from Amblyomma cajannense

® = Patents (INPI, PCT)

Kunitz type Inhibitor

Amblyomma cajennense

Scale-up Production

Experiments were performed

in C57BL/6J mice

Tumor implants - Suspensions of B16F10 (2.5 x 104)

cells subcutaneously injected

A

Experimental model: Macroscopic aspect of nude dorsal Mia-PaCa-2 and SKMel-28 tumors

Partnership

– União Química / BNDES / FAPESP

Phase of Development

– Pre-clinical and clinical studies

Source: Chudzinski-Tavassi et al. Toxicon 2010

Amblyomin-X ®

Instituto Butantan | 31

4- USPTO Peptide, compositions, and uses thereof. Chudzinski-Tavassi AM,

Carrijo-Carvalho LC, Ventura JS, inventors. US patent application. 2009.

Lopap

Lopap ® – Patents Application A prothombin activator from Lonomia obliqua

Prothombin

Activator

1- INPI (PI0200269-8, Brazil, 2002)

Chudzinski-Tavassi AM, Reis CV. Native prothrombin activator.

WO03/070746 / AU2003208190 / CAN2,471,410 / EP1482969 / JP2003-

569653 / MX04007344 / US10/501,238

2- INPI (PI0403882-7, Brazil, 2004).

Chudzinski-Tavassi AM, Reis CV, Ramos CR, Ho PL. Recombinant molecule

and its use as defibrinogenating/ antithrombotic agent and a kit for diagnosis

of dysprothrombinemias PCT/BR2005/000171

Antithrombotic

Agent

3- INPI (PI0504199-6, Brazil, 2005).

Reis CV, Maria DA, Falsi M, Chudzinski-Tavassi AM.

Anti-apoptotic activity of CNF 021.01 and pharmaceutical formulations

Anti apoptotic

activity

USPTO

L. obliqua

Instituto Butantan | 32

Av. Vital Brasil, 1500 - Butantã

São Paulo – SP

Zip Code - 05503-900

(+ 55 11) 3726.7222

www.butantan.gov.br