FAPESP WEEK WASHINGTON, DC
PROF. JORGE KALIL October 24th – 26th, 2011 AGENDA
BUTANTAN - SCIENCE IS THE BASIS
BUTANTAN – SOME PROJECTS
Instituto Butantan | 1 BUTANTAN – A PUBLIC INSTITUTION OF THE STATE GOVERNMENT OF SÃO PAULO
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
Instituto Butantan | 2 BRAZIL HAS BECOME AN INTERNATIONAL REFERENCE IN IMMUNIZATION
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
Why
Too important to depend on availability and pricing
Instituto Butantan | 3 NATIONAL IMMUNIZATION PROGRAM (PNI)
PNI – Founded in 1973
PNI – General Information of Brazil (2010) . Smallpox Eradication . Poliomyelitis . Measles (autocne transmission) . 193 MM inhabitants
. 43 types of immunobiological
26 Vaccines 13 sera from animal . Neonatal tetanus 4 sera from human Under . Accidental tetanus Control . Tuberculosis . 77% produced in Brazil
. Diphtheria . ~ 300 MM doses of vaccines per year . Pertussis . 30 K vaccination rooms . Hepatite B . Expansion of national self-sufficiency . Influenza . Pneumococcus
Source: SVS/Ministry of Health, National Immunizatation Program, Brazil, 2010. Instituto Butantan | 4 NATIONAL SUPPLIERS OF VACCINES FOR THE MINISTRY OF HEALTH
‘Market Share’ per Suppliers(1) Products (2010)
FUNED 3% FAP 4% DTP DT dT
Butantan Biomanguinhos 51% 42% Anti Rabies Hepatitis B
Note: Part of Butantan’s production was sent to other Institutes, Influenza such as Biomanguinhos. Not computed in the analysis (Flu)
1Source: Ministry of Health, 2010 Instituto Butantan | 5 NATIONAL SUPPLIERS OF ANTIVENOMS AND ANTITOXINS FOR THE MINISTRY OF HEALTH
‘Market Share’ per Suppliers(1) Products (2010)
Snakes AV
IVB 25%
Butantan Scorpion AV Spider AV Caterpillar AV 56% FUNED 19%
Tetanus AT Diphitheria AT Anti Rabies
Note: Part of Butantan’s production was sent to other Bee AV Institutes, such as FUNED and CPPI. Not computed in the analysis Botulism AB & E
1Source: Ministry of Health, 2010 Instituto Butantan | 6 EXPORTING PRODUCTS
Instituto Butantan | 7 VACCINES WITH EXTERNAL COOPERATION
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 Recombinant OncoBCG for bladder Universidade de São Paulo – Medical School cancer Institut Pasteur – Paris / Novartis - Siena / Recombinant BCG-Pertussis Albert Einstein College of Medicine
Instituto Butantan | 8 INDEXED SCIENTIFIC PUBLICATIONS
PAPERS PUBLISHED AND CITED
2500 250 sum of times cited (TOTAL) Papers Published 2000 Poly. (sum of times cited (TOTAL)) 200 Expon. (Papers Published)
1500 150
1000 100
500 50
0 0 1980 1985 1990 1995 2000 2005 2010
Instituto Butantan | 9 SCREENING OF BIOACTIVE COMPONENTS OF POISONOUS ANIMALS
Venom Composition Species Pharmacological activities (Transcriptomics and Proteomics)
ANALGESICS
ANTI- INFLAMMATORY
ANTICOAGULANTS
ANTITUMOR
ACTION IN THE NERVOUS SYSTEM
ANTIVENINS
ANTIHIPERTENSIVE
Instituto Butantan | 10 PATENTS
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
Instituto Butantan | 11 PATENTS
Biotechnology (16)
New developments in Pertussis Vaccines with Appropriate technologies
Whole Cell Pertussis Vaccine
Low LPS content- Whole Cell Pertussis less reactogenic – low cost
Infants B. pertussis WCP WCPLow fermentation Tangential Organic Patent filtration extraction Clinical Trials IMMUNOGENIC COMPLEX FORMED BY VACCINAL 2007 ANTIGENS ENCAPSULATED BY NANOSTRUCTURED Adolescents MPLA Acellular MESOPOROUS SILICA Adjuvant Neonates Pertussis Patent CRISTALOGRAPHY Vaccine Clinical Trials planned 2009 LABORATORY Recombinant BCG-Pertussis Patent Immunization at birth Patent Under GMP production
Instituto Butantan | 12 RESEARCH & DEVELOPMENT LABORATORIES
. ~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 | 13 INDUSTRIAL COMPLEX
. 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 | 14 AGENDA
BUTANTAN - SCIENCE IS THE BASE
BUTANTAN – SOME PROJECTS
Instituto Butantan | 15 Presentation and discussion of some Projects
Area Projects 1 Vaccines for immunepreventable . Rotavirus (pentavalent) diseases . 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) 1 Not exhaustive
Instituto Butantan | 16 Rotavirus Vaccine - Overview
Product – Pentavelent Rotavirus Vaccine Butantan –
. Composition . Challenges:
– Attenutated virus – To perform Phase II and III - non-inferiority study – Sorotypes: G1, G2, G3, G4 e G9 – To find funding for: . Technology of Production Clinical Trial and laboratory assay – Phase II / III – Cell substrate: Vero cells
– Reassortment – Human/bovine . Objective: – Nº lots produced: 09 (6.324 doses) – Pentavalent low cost vaccine . Phase of Development – Phase I: 2010 Results: safe and immunogenic – Phase II: 2012
• Partnership – NIH / PATH / BNDES Instituto Butantan | 17
Dengue Vaccine - Overview
Product – Tetravalent Dengue Vaccine Butantan –
. Composition . Challenges:
– Attenuated virus – To speed up Phase I, II, and III – (to avoid – Sorotypes: DEN1, DEN2, DEN3, DEN4 non-inferiority study)
. Technology of Production – To find funding for: Clinical Trial and Laboratory assay - Phase II – Cell substrate: Vero cells and III – Recombinant DNA technology Equipment – Chimeric Plant – Nº lots produced: 06 (12.640 doses) Maintenance of “The Global Solutions for Infectious Disease” support . Phase of Development – To define target population for immunization – Phase I: 2011/2012 – Production capacity x national and international demand
• Partnership . Objective: – NIH / DVI / BNDES / FAPESP – Tetravalent low cost vaccine
Instituto Butantan | 18
Dengue Vaccine - technical and scientific aspects Attenuation strategies for Dengue virus and Pre-Clinical studies
Attenuation strategies for Dengue virus Pre-Clinical Studies vaccine candidates
30 nucleotide deletion in DENV 3’ UTR = 30 SCID-HuH-7 mice mutation The candidate vaccine viruses were attenuated, rDEN1 30 and rDEN4 30 replicating to a peak titer from 100.9 up to 102.4 PFU/mL The D30 mutation partially attenuated DEN2 and did not attenuated DEN3 for rhesus monkeys Rhesus macaques
1.0 1.3 Antigenic chimeras - rDEN4 30 background Low/undetectable levels of viremia in (10 to 10 PFU/mL) . rDEN2/4 30 – infectious in monkeys and Immunogenic humans
. rDEN3/4 30 – infectious in monkeys and Mosquitoes poorly infectious in humans Vaccine viruses restricted ability to infect the midgut DEN3/4 30 and to cause a disseminated infection (Ae. Aegypti - Toxorhynchites splendens)
Additional 3’-UTR deletion mutations Ae. albopictus fed on viremic subjects - vaccine virus was not recovered from any mosquitoes . rDEN3 30/31 – infectious in monkeys and humans
Instituto Butantan | 19 Adjuvant – Monophosphoryl lipid A (BpMPLA)
Product - Monophosphoryl lipid A (BpMPLA) Butantan –
. Composition . Challenges: – BpMPLA derived from LPS of B.pertussis – Scale-up
. Production Technology . Objectives: – Purification of B.pertussis followed by LPS – To optimize immunolgical response of pre- hydrolises exisiting and new vaccines . Phase of Development – To increase production capacity
– Scale-up BpMPLA – Clinical trial Pandemic H1N1 + BpMPLA – Pre-clinical Human rabies – Animal Study Dog Leishimania – In development Hepatites B + BpMPLA Seazonal Influenza + BpMPLA
Instituto Butantan | 21 Adjuvant – Monophosphoryl lipid A (BpMPLA)
Pathogen-Associated Molecular Patterns (PAMPs) and Danger- Associated Molecular Patterns are potential new adjuvants
Monophosphoril Lipid A (MPLA) derived from Bordetella pertussis as a new vaccine adjuvant
Instituto Butantan | 22 Lung Surfactant
Product – Surfactant Butantan –
. Composition . Challenges:
– Origin porcine – To test a new process of production (ultracentrifugation) and formulation – 2nd – Phospholipids + Proteins SP-B and SP-C generation of surfactant
. Production Technology
– Porcine pulmonary extract adsorbed on a cellulose derivative followed by organic . Objectives: extraction, solvent evaporation, lyophilize drying and final formulation – To make available a low cost surfactant to Brazilian Public Maternities . Phase of Development
– Licensed in Brazil in 2009
Instituto Butantan | 23 Lung Surfactant - technical and scientific aspects
Alveolus
Normal
Sindrome
Respiratory
Surfactant Distress
Lung Parenchyma Instituto Butantan | 24 Silica (SBA-15) Immunogenic complex formed by vaccinal antigens encapsulated by nanostructured mesoporous silica
Features The SBA-15 possesses hexagonal porous uniformity (3.1 – 6.5 nm) Thermal and hydrothermal stability Exhibits potential applications for selective adsorption and catalysis
Instituto Butantan | 25 Adjuvant effect of SBA-15
Instituto Butantan | 26 Modulation of antibody response - Silica
10 μg/animal de BSA:250 μg/animal de SBA-15 ou Al(OH)3
IM VO
Source: Carvalho et al. 2010. Instituto Butantan | 27 Monoclonal Antibodies Anti-human CD3
AcMo Murine AcMo Quimeric AcMo Humanized
. Crohn's disease . Encephalomyocarditis autoimmune POTENTIAL . Psoriatic arthritis CLINICAL USES . Diabetes Type I
. Lupus erythematosus anti-human CD3 . Control of acute rejection in kidney transplant . Prevention of rejection in transplanted pancreatic islet . Steroid-refractory acute GVHD
Instituto Butantan | 28 Clotalphine Potent analgesic drugs from animal venoms and toxins
Crotalphine Molecular mechanisms involved in the analgesic action
Venom of Activation of CB2 cannabinoid receptors
release of endogenous Potent analgesic opioids long-lasting analgesic action (2-5 days) Activation of peripheral and opioid receptors
Crotalus intracellular signaling pathways activation Recent studies show involvement of PKC Activation of NO-cGMP-PKG which activates MAPKs and opening of potassium (Mechanism that may contribute to its long-lasting channels analgesic action)
Instituto Butantan | 29 Amblyomin-X ® An anti-cancer from Amblyomma cajannense
Experiments were performed in C57BL/6J mice
Amblyomin-X ® A Amblyomma cajennense
Scale-up Production
Tumor implants - Suspensions of B16F10 (2.5 x 104) cells subcutaneously injected
Kunitz type Inhibitor
. Partnership – União Química / BNDES / FAPESP
. Phase of Development – Pre-clinical and clinical studies Experimental model: Macroscopic aspect of nude dorsal Mia-PaCa-2 and SKMel-28 tumors ® = Patents (INPI, PCT)
Source: Chudzinski-Tavassi et al. Toxicon 2010 Instituto Butantan | 30 Lopap ® – Patents Application A prothombin activator from Lonomia obliqua
1- INPI (PI0200269-8, Brazil, 2002) Prothombin Activator 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). Antithrombotic Chudzinski-Tavassi AM, Reis CV, Ramos CR, Ho PL. Recombinant molecule Agent and its use as defibrinogenating/ antithrombotic agent and a kit for diagnosis of dysprothrombinemias PCT/BR2005/000171 Lopap Anti apoptotic 3- INPI (PI0504199-6, Brazil, 2005). activity Reis CV, Maria DA, Falsi M, Chudzinski-Tavassi AM. Anti-apoptotic activity of CNF 021.01 and pharmaceutical formulations
4- USPTO Peptide, compositions, and uses thereof. Chudzinski-Tavassi AM, USPTO Carrijo-Carvalho LC, Ventura JS, inventors. US patent application. 2009.
L. obliqua
Instituto Butantan | 31 Av. Vital Brasil, 1500 - Butantã São Paulo – SP Zip Code - 05503-900 (+ 55 11) 3726.7222 www.butantan.gov.br
Instituto Butantan | 32