Engineering Conferences International ECI Digital Archives

Vaccine Technology IV Proceedings

Spring 5-21-2012 111 years producing immunobiologicals: New challenges Jorge Kalil Instituto Butantan

Follow this and additional works at: http://dc.engconfintl.org/vaccine_iv Part of the Biomedical Engineering and Bioengineering Commons

Recommended Citation Jorge Kalil, "111 years producing immunobiologicals: New challenges" in " Technology IV", B. Buckland, University College London, UK; J. Aunins, Janis Biologics, LLC; P. Alves , ITQB/IBET; K. Jansen, Wyeth Vaccine Research Eds, ECI Symposium Series, (2013). http://dc.engconfintl.org/vaccine_iv/6

This Conference Proceeding is brought to you for free and open access by the Proceedings at ECI Digital Archives. It has been accepted for inclusion in Vaccine Technology IV by an authorized administrator of ECI Digital Archives. For more information, please contact [email protected]. THE EFFORTS OF A PUBLIC INSTITUTE TO DEVELOP NEW PROF . J ORGE KALIL

PORTUGAL , 2012 AAGENDA

 OVERVIEW OF IMMUNIZATION PROGRAM IN AND INSTITUTO BUTANTAN

 BUTANTAN DEVELOPMENTS

Instituto Butantan | 1 AAGENDA

 OVERVIEW OF IMMUNIZATION PROGRAM IN BRAZIL AND INSTITUTO BUTANTAN

 BUTANTAN DEVELOPMENTS

Instituto Butantan | 2 THE IMPACTS OF VACCINATION IN BRAZIL Children vaccination coverage by type of vaccination

Vaccination coverage in Brazil % (2000 to 2010) 120

110

100 Over 70 million

90 children successfully vaccinated in two

80 decades Implementation of the MMR vaccine and DTP + Hib vaccine 70 (tetravalent) in 100% municipalities 60

50 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010

BCG MMR B Oral Polio Tetravalent (DTP+Hib) Rotavirus

Instituto Butantan | 3 THE IMPACTS OF VACCINATION IN BRAZIL Number of cases for – accidental and neonatal

Number of cases – Accidental and neonatal Tetanus

Accidental Neonatal (1990 – 2010) (1983 – 2010) Cases Cases 1,800 0.500 800 1,600 700 1,400 0.400 Elimination 600 Plan 1,200 0.300 500 Strengthening 1,000 actions in areas 400 800 of potential risk 0.200 Emergency Plan for 300 600 high-risk counties 200 400 0.100 200 100 0 0.000 0 90 91 92 93 94 95 96 97 98 99 00 01 02 03 04 05 06 07 08 09 10 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 00 01 02 03 04 05 06 07 08 09 10

Number of accidental tetanus per year Number of neonatal tetanus per year Incidence per 100K inhabitants

Source: Sinan/SVS/MS, data in 25/08/11Ministry of , Brazil , 2010. Instituto Butantan | 4 THE IMPACTS OF VACCINATION IN BRAZIL The number of severe cases and deaths due to influenza A H1N1 has been falling since March 2010

Number of severe cases and deaths due to influenza A H1N1 Brazil (2010)

80 79 12

70 65 65 10 59 Vaccination 60 campaign 8 50 47 47 42 43 40 6 34 Deaths Deaths

Severe Cases Cases Severe Severe 30 27 4 21 19 20 19 20 15 17 17 17 13 10 10 10 11 10 2 10 8

1 1 0 0 0 Weeks * * epidemiological week of onset of symptoms

Severe hospitalized cases of pandemic influenza (H1N1) Deaths from pandemic influenza (H1N1)

Source: Sinan/SVS, Ministry of Health, Brazil , 2010. Instituto Butantan | 5 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 | 6 NATIONAL IMMUNIZATION PROGRAM (PNI) IN BRAZIL

PNI – Founded in 1973

PNI – General Information of Brazil (2011)  Smallpox Eradication  Poliomyelitis  194 MM inhabitants  Measles (autocne transmission)  43 types of immunobiological

 26 Vaccines  13 sera from animal  Neonatal tetanus  4 sera from human Under  Accidental tetanus Control  77% produced in Brazil    ~ 300 MM doses of vaccines per year  Pertussis  30 K vaccination rooms  Hepatite B  Influenza  Expansion of national self-sufficiency  Pneumococcus

Source: SVS/Ministry of Health, National Immunizatation Program, Brazil, 2011. Instituto Butantan | 7 WHAT DO WE ARE ? BUTANTAN – A P UBLIC INSTITUTION OF THE STATE GOVERNMENT OF

 In 1901 Butantan was established to produce serum against the  , the first director, investigated against bites

 Currently, Butantan is the main public producer of vaccines, antivenoms, antitoxins in  Fully dedicated to develop scientific research and production of immunobiological products for public health

Instituto Butantan | 8 RESEARCH & DEVELOPMENT LABORATORIES

 ~21 scientific labs

 ~180 Researchers  85% are PhD

 1 Center  Multiple laboratories

 1 Hospital (10 hospital beds)

 1 Central Animal Facility

 Training programs (PAP)

 Graduate studies in Toxicology

 Masters and PhDs Instituto Butantan | 9 SCREENING OF BIOACTIVE COMPOUNDS OF ANIMAL

Pharmacological Activities

ANALGESIC ANTIINFLAMATORY ANTI -MICROBIOLOGICAL ANTI-COAGULANT ANTI-TUMORAL NERVOUS SYSTEM ACTION ANTI-VENOMS REACTIVITY ANTI-HYPERTENSION

Venom Composition by Transcriptomics and Proteomics

Instituto Butantan | Plasma fractioning by chromatography

Plasma fractioning by chromatography

Instituto Butantan | INDUSTRIAL COMPLEX

 7 Main Industrial Plants (Buildings)  Anaerobic vaccines ( tetanus and botulinic ) and Anatoxin Purification  Biological control  Aerobic Vaccine (Diphtheria and Pertussis )  Hepatitis  Influenza   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 | 12 WHAT DO WE DO ? NATIONAL SUPPLIERS OF VACCINES FOR THE MINISTRY OF HEALTH

‘Market Share’ per Suppliers (1) Products (Vaccines) (2010)

FUNED 3% FAP 4% DTP DT dT

Butantan Biomanguinhos 51% 42% Rabies

Note: Part of Butantan’s production was sent to other Influenza Institutes, such as Biomanguinhos. Not computed in the (Flu) analysis

1Source: Ministry of Health, 2010 Instituto Butantan | 13 WHAT DO WE DO ? NATIONAL SUPPLIERS OF ANTIVENOMS AND ANTITOXINS FOR THE MINISTRY OF HEALTH

‘Market Share’ per Suppliers (1) Products (Sera) (2010)

Snakes AV

IVB 25%

Butantan AV AV Caterpillar AV 56% FUNED 19%

Tetanus AT Diphitheria AT AntiRabies

Note: Part of Butantan’s production was sent to other Institutes, such as FUNED and CPPI. Not computed in the analysis AB & E

1Source: Ministry of Health, 2010 Instituto Butantan | 14 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 Visceral 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 – / Novartis - Siena / Recombinant BCG-Pertussis Albert Einstein College of Medicine

Instituto Butantan | 15 AAGENDA

 OVERVIEW OF IMMUNIZATION PROGRAM IN BRAZIL AND INSTITUTO BUTANTAN

 BUTANTAN DEVELOPMENTS

Instituto Butantan | 16 WHAT DO WE WANT TO DO ? Presentation and discussion of vaccines projects

Area Vaccines Projects 1

Vaccines - Research and  Pertussis low improvemen t  Adjuvant Bp MPLA  Recombinant onco BCG  Silica nanostructure mesoporous – vaccine antigens encapsulated Vaccines - Collaborative  Rotavirus (pentavalent) development  Dengue (tetravalent)  Streptococcus pneumoniae (cellular - SPWCV)  BCG-Pertussis Vaccines – Basics R&D  Leptospira

Techtransfer  Several vaccines under negociation

1 Not exhaustive

Instituto Butantan | 17 Pertussis low vaccine

Product - Pertussis low vaccine Butantan –  Composition  Challenges: – B.pertussis whole cell with lower content of – Scale-up LPS

 Production Technology  Objectives:

– Organic extraction of the cells to reduce LPS – To make available an alternative vaccine for content immunization of children, adolescent, pregnant women and adults  ~ 70% reduction of LPS

– “in line ” process without additional costs

 Phase of Development

– Pre-clinical studies performed in Butantan and in the Netherlands Institute Vaccine (NIV)

– Phase I (2012) – Brazil

Instituto Butantan | 18 Pertussis low vaccine - technical and scientific aspects New developments in Pertussis Vaccines with Appropriate technologies

® = Patents Whole Cell Pertussis Vaccine

Whole Cell Pertussis  ~ 70% reduction of LPS  Less reactogenic Low LPS content  Low cost

B. pertussis WCP WCP ® fermentation low Tangential Organic filtration extraction

Instituto Butantan | Adjuvant – Monophosphoryl lipid A ( Bp MPLA)

Product - Monophosphoryl lipid A ( Bp MPLA) Butantan –

 Composition  Challenges: – Bp MPLA 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 Bp MPLA – Clinical trial  Pandemic H1N1 + BpMPLA – Pre-clinical  Human rabies – Animal Study  Dog Leishimania – In development  Hepatites B + BpMPLA  Seazonal Influenza + BpMPLA

Instituto Butantan | 20 Recombinant BCG-Pertussis Neonate vaccine / Onco BCG for Bladder cancer

Product - Recombinant BCG – Pertussis Butantan –

 Composition  Challenges:

– Recombinant BCG strain expressing the S1 – To produce the vaccine by fermentation or subunit 1 of Pertussis static culture

– To perform the clinical trials  Production Technology

– The rBCG -Pertussis strain was produced without antibiotic resistance gene 1  Objectives:  Appropriate for use in humans – To immunize infants 0 – 2 months of age  Phase of Development – To make available a new vaccine for bladder – Production of GMP lots cancer

Nota: 1 Auxotrofic strain for lysine is complemented with a plasmid that expresses the deleted gene plus the heterologous gene – S1PT

Instituto Butantan | 21 Recombinant BCG-Pertussis - technical and scientific aspects Protection of neonate mice immunized with rBCG-S1PT against intracerebral challenge with B. pertussis

ONE DOSE AT DAY 5

100 Saline 100 Saline DPT BCG 80 80 BCG rBCG-S1PT rBCG-S1PT 60 60

40 40 % Survival Survival % % % Survival Survival % %

20 20

0 0 0 2 4 6 8 10 12 0 2 4 6 8 10 12 Days after Challenge Days after Challenge

Challenge dose at day 21: 10 6 CFU Challenge dose at day 21: 3x10 7 CFU

Source: Microbes Infect.10:198-202 (2008) Instituto Butantan | 22 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 | 23 Rotavirus Vaccine

Product – Pentavelent Rotavirus Vaccine Butantan –

 Composition  Challenges:

– Attenutated – 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 K doses) – Pentavalent low cost vaccine  Phase of Development – Phase I: 2010  Results: safe and immunogenic – Phase II: 2013

• Partnership – NIH / PATH / BNDES

Instituto Butantan | 24

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 III – Cell substrate: Vero cells  Equipment – Recombinant DNA technology  Plant – Chimeric  Maintenance of “The Global Solutions for – Nº lots produced: 06 (17 K doses) Infectious Disease” support – To define target population for immunization  Phase of Development – Production capacity x national and – Phase I and II: 2012/2013 international demand  Objective: • Partnership – NIH - DVI (Dr. Steve Whitehead) – Tetravalent low cost vaccine – BNDES / FAPESP

Instituto Butantan | 25 [email protected] Obrigado Thank you

Av. Vital Brasil, 1500 - Butantã São Paulo – SP Zip Code - 05503-900 (+ 55 11) 3726.7222 www.butantan.gov.br

Instituto Butantan | 26