Biotech Intro

BIOPROCESSING IN THE PHARMACEUTICAL

INDUSTRY (An Engineering and Technological Overview)

Erick Prado, P.E. 18-Sep-07

UNIVERSIDAD INTERAMERICANA, BAYAMON

THE INTRODUCTIONWhat is Biotechnology?

Traditional vs. Modern BiotechnologyPharmaceutical Industry at Glance

BIOPHARMACEUTICALSSmall Molecules vs. Macro-MoleculesBiotechnologies for API ManufacturingBiotechnology-based Pharmaceuticals ENGINEERING ASPECTS

cGMP RequirementsEquipment Design Considerations

Facility Design ConsiderationsCommissioning and Qualification

BIOPROCESSING TECHNOLOGIESTypical Biopharmaceutical ProcessUpstream Processing: Cultivation

Fermentation: Bio-reactionDownstream Processing: Recovery

Downstream Processing: PurificationFill and Finish Operations

WHAT’S NEXT?Future Directions

Knowledge-based Economy: The Bio-Island

SCIENCE BEHIND MODERN BIOTECHThe Chemistry of Life

The DNAGenetic Engineering

SEMINAR MAPPING

THE INTRODUCTION

What is Biotechnology?

Biotechnology

• Technology based on biology

• Biotechnology is the integration of natural sciences and engineering sciences in order to achieve the application of organisms, cells, parts thereof and molecular analogues for products and services

Disciplines of Biotechnology

Biotechnology Color Code

RED BIOTECHNOLOGY: Medicine / Pharmaceuticals• Gene therapy• Production of proteins, antibodies, and vaccines• Tissue engineering

WHITE BIOTECHNOLOGY: Chemical industry• Basic chemicals• Biopolymers• Biorefinery

GREEN BIOTECHNOLOGY: Agriculture• Crops improvement• Natural pesticides• Pest and disease resistance

• There are 20 different amino acids

• Within a gene, each specific sequence of three DNA bases (codons) directs the cell's protein-synthesizing machinery to add a specific amino acid

1930

1970

1990

1910191519201925

1935194019451950195519601965

197519801985

199520002005

19051900

CLASSICAL GENETIC

MICROBIAL GENETIC

GENE MANIPULATION

Genetic mapping

Transformation demonstrated

Molecular genetics

Development of technologies

Applications

TH

E H

ISTO

RY O

F G

EN

ETIC

S

Traditional vs. Modern Biotechnology

THE INTRODUCTION

Traditional vs. Modern Biotechnology

Traditional• Based on microorganisms• Cells are not manipulated• Production of antibiotics by fermentation

Modern• Based on microorganisms and cell culture• Cells are manipulated• Production of protein products based on recombinant

DNA and cell culture• Modern biotech refers to Molecular Biology, Molecular

Genetics and Genetic Engineering

Biotechnology

• Broad definition can include many traditional food preparation processes

– Beer and wine

– Leavened bread

• Usually refers to recombinant DNA or tissue culture based processes

– emphasizes modern technology

– generate genetically modified organisms

THE INTRODUCTION

Pharmaceutical Industry at Glance

Pharmaceutical Industry Main Customers

Medicines

Return

Documents

Patient

Shareholders

Regulatory Agencies

Pharmaceutical Industry

FDA OSHA EPA

From Molecule to Patient

DiscoveryResearch and Development

FDA Approval

API ManufactureDistribution

Therapeutic Target

Final Dosage ManufacturePatient

ACTIVEPHARMACEUTICA

LINGREDIENT

API

PROVIDES THE THERAPEUTIC EFFECT

ACTIVE PHARMACEUTICAL INGREDIENTS = DRUG SUBSTANCES

API Sources

Chemical Synthesis

Micro-organisms

Natural Sources

Fermentation Biotechnology

Extraction Purification

Chemical reactions

Synthetic APIs

Biological/Biotech APIs

Natural APIs

+API

DOSAGE FORM

EXCIPIENTS

INERTACTIVE

Pharmaceutical Commercial Chain

PRIMARY PHARMACEUTICAL

PRODUCTION

CHEMICAL SYNTHESIS

MICRO-ORGANISM

PLANT ANIMAL

TABLETS

CAPSULE

SYRUPS

SUSPENSIONS

EMULSIONS

CREAMS

OINTMENTS

SUPPOSITORIES

DROPS

SPRAYS

INJECTIONS

SECONDARY PHARMACEUTICAL

PRODUCTION

RAW MATERIALS

APIFINAL DOSAGE DISTRIBUTION

CHAIN

TABLETS

CAPSULE

SYRUPS

SUSPENSIONS

EMULSIONS

CREAMS

OINTMENTS

SUPPOSITORIES

DROPS

SPRAYS

INJECTIONS

SECONDARY PHARMACEUTICAL

PRODUCTION

RAW MATERIALS

API

PRIMARY PHARMACEUTICAL

PRODUCTION

CHEMICAL SYNTHESIS

MICRO-ORGANISM

PLANT ANIMAL

FINAL DOSAGE DISTRIBUTION

CHAIN

RATE-LIMITING STEP

R&D Costs for a Biopharmaceutical

Small Molecules vs.

Macro-Molecules

BIOPHARMACEUTICALS

Small vs. Macro Molecules

Small Molecule Products• Molecular weight in 10² order of magnitude• Produced by:

– Traditional chemical synthesis– Microorganisms (fermentation or biotransformation)– Extraction from plants and animals

• Most of the medicines today• N-heterocyclic compounds most important• Administration: oral, dermal, inhalation,

parenteral

Small vs. Macro Molecules (cont.)

Macro Molecule Products• Molecular weight in 10³ order of magnitude• Very fragile molecules• Obtained mainly from modern biotechnology• Most of the biopharmaceuticals today• Proteins and peptides most important• Administration: parenteral

Biotechnologyfor

APIs Manufacturing

BIOPHARMACEUTICALS

• Known as biotransformation and bioconversion• Production of small molecules by use of:

– Natural or modified isolated enzymes– Enzyme extracts– Whole-cell

• A starting material is converted by the enzyme in the desired product

Biocatalysis (enzymes)

• Known as microbial fermentation• Part of the traditional biotechnology• Used for the production of:

– Small molecules – Antibiotics– Peptides– Less complex big molecules

• Bioreactor called fermenter• Microbes are allowed to grow by:

– Supplying the appropriate nutrients – Maintaining the proper conditions (pH, temperature,

agitation, etc.)

Biosynthesis

• Old technology for product isolation:– Solvent extraction of the medium containing the

product

• Modern technologies for product isolation:– Membrane (reverse osmosis, ultra- and nanofiltration) – Chromatography

• Final purification by conventional crystallization

Biosynthesis (cont.)

• Use of animal or plant cells, removed from tissues

• Cells are allowed to grow by:– Supplying the appropriate nutrients – Maintaining the proper conditions (pH, temperature,

etc.)

• Used for the generation of high molecular weight proteins

Cell Cultures

• Contrary to microbes, cell culture can performs post-translational modifications required by large proteins production

• Typical post-translational modifications:– Protein folding– Disulfide linkages– Glycosylation– Carboxylation

Cell Cultures (cont.)

Biotechnology-based Pharmaceuticals

BIOPHARMACEUTICALS

Biopharmaceutical

• A therapeutic product created through the genetic manipulation of living things, including (but not limited to): – Proteins– Monoclonal Antibodies– Peptides

• Monoclonal Antibody (MAb)– Highly specific antibody that is derived from only one

clone of cells and recognize only one antigen– Antigen: agent that stimulates production of antibody

that will react specifically with it

Peptide Drugs

• Many hormones are actually small peptides (2-40 amino acids)

• Calcitonin (Calcimar, Miacalcin, 32 res.) – Thyroid hormone to enhance bone mass

• Oxytocin (Pitocin, 9 residues)– Pituitary hormone to stimulate labor

• Vasopressin (Pitressin, 9 residues)– Pituitary h. for antidiuretic/vasconstriction

Protein Pharmaceuticals

• Insulin (diabetes)

• Interferon (relapsing MS)

• Interferon (granulomatous)

• TPA (heart attack)

Protein Pharmaceuticals

• Actimmune (If )• Activase (TPA)• BeneFix (F IX)• Betaseron (If )• Humulin• Novolin• Pegademase (AD)

• Epogen• Regranex (PDGF)• Novoseven (F VIIa)• Intron-A• Neupogen• Pulmozyme• Infergen

Protein Pharmaceuticals

• Insulin Pigs or cattle (pancreas)

• Albumin Human blood (donated)

• HGH Human brains

• Factor VIII Human blood (donated)

• Calcitonin Salmon

• Anti-venom Horse of Goat blood

Protein Drug Original Source

Protein Pharmaceuticals

• Natural sources are often rare and expensive – Difficult to keep up with demand– Hard to isolate product– Lead to immune reactions (diff. species)– Viral & pathogen contamination

• Most protein pharmaceuticals today are produced recombinant– Cheaper, safer, abundant supply

Categories of Biopharmaceuticals

• Hematopoietic growth factors and coagulation factors – Stimulate production of certain blood cell types and

clotting agents– Epogen®,Neupoeng®, Benefix®, Neulasta®

• Interferons and cytokines for anti-infective and cancer therapy– Antiviral and anti-prolifertation activity– Avonex®, Betaseron®, Rebil®

Categories of Biopharmaceuticals (cont.)

• Hormones – Pre-recombinant from slaughtered animals– Humulin®,Glucagon®, Somatropin®, Synarel®

• Enzymes– Catalyze unique biochemical reactions with high

substrate specificity– Enzyme replacement, thrombolytich enzymes, DNAse

therapy, cancer therapy– Actovase®, Pulmosyme®, Oncaspar®

Categories of Biopharmaceuticals (cont.)

• Antibodies and Derivatives – Chimeric, Humanized, Fully human– Enbrel®, Remicade®, Rituxan®, Herceptin®

• Vaccines (for viral and bacterial infections)– Smallpox, polio, measles, mumps,rubella, chicken

pox, flu, rabies, hepatitis, etc.– TB, typhus, cholera, lyme, pertussis, tetanus,

diphtheria, pneumonia, etc.