biologicals biosimilars introduction - … set of methods for isolation, ... prokaryotic cell...
TRANSCRIPT
MPS2
Biologicals
BIOLOGICALS – BIOSIMILARS INTRODUCTION
DIRSCH 1
MPS2
Biologicals
Natural products
Biologicals
Phytopharmaceuticals
O
O O H
O
O O C O O H
S
N
C H 3
C H 3 C
O
N H C H 2
DIRSCH
PHARMAGOGNOSY
2
MPS2
Biologicals
• Biotechnology Production or modification of chemical compounds using organisms or parts of organisms in the context of industrial processes and use. (e.g. bread, Sauerkraut, cheese, beer, ....... vitamins, antibiotics, taxol etc. Pharmaceutical biotechnology)
• Genetic engineering / recombinant DNA technology
A set of methods for isolation, characterisation, amplification and recombination of DNA within and across species boundaries.
• Gene therapy
Transfer of genetic material to tissue cells in order to replace, switch off or complement defective genes.
RELEVANT DEFINITIONS
DIRSCH 3
MPS2
Biologicals
LITERATURE used (most in German) Dingermann – Winckler – Zündorf: Gentechnik – Biotechnik Dingermann – Zündorf - Hänsel: Pharmazeutische Biologie Müller-Esterl: Biochemie Löffler – Petrides: Biochemie & Pathobiochemie Alberts: Molekularbiologie der Zelle Web: http://www.deutscher-apotheker-verlag.de/DAZ/
http://www.rote-liste.de/online http://www.ema.europa.eu/ http://www.vfa.de/gentech http:/www.vfa-bio.de/biosimilars
DIRSCH 4
MPS2
Biologicals
RECOMBINANT PROTEINS – the principle of production: heterologous expression
Target organism
Cell containing gene of interest
information unit
genetically
modified
organism
"genetic engineering"
Control regions
Extraction and
purification
© Theo Dingermann DIRSCH Host-vector system 5
Biologicals
MPS2
Expression vector
Restriction digestion
DNA sequence encoding requested gene product
Recombinant Expression vector
Transformation and propagation of E. coli
Isolation of expressed protein DIRSCH 6
Cloning site DNA sequence encoding requested gene product
Biologicals
MPS2
Restriction enzyme or restriction endonuclease
DIRSCH 7
s
Recognition sequences
sticky ends
DNA fragments
blunt ends
Biologicals
MPS2
DIRSCH
DNA ligase
8
Sticky ends after EcoR I Restriction
Biologicals
MPS2
DIRSCH 9
intergenic sequences
protein-coding genes
non protein- coding genes
Transcripts with unknown function (ca. 21%)
Biologicals
MPS2
Eukaryotic cell Prokaryotic cell
DIRSCH 10
Nucleus
Addition of 5‘cap and 3‘Poly(A)tail
RNA splicing
primary DNA transcript
MPS2
Biologicals
Posttranslational modifications • Glycosylation • Disulfide bridges
• Cutting off signal peptides/sequences protein synthesis
Ribosome
Golgi apparatus
Endoplasmatic reticulum (ER)
DIRSCH 11
MPS2
Biologicals
Since eucaryotic mRNA has a poly-(A)-tail,
complementary oligo-d(T)n is an often used
primer. The enzyme reverse transcriptase (RT)
synthesizes from the 3‘- end of the mRNA a
cDNA copy: with ist endogenous Rnase activity
RT cuts mRNA into pieces. Starting with the 3‘-
end of the cDNA RT degrades mRNA with ist 5'-
3'-exonuclease activity and at the same time
replaces it by complementary DNA. After
ligation of linker DNA and restriction, cDNA can
be inserted into a suitabel DNA vector.
Synthesis of cDNA
DIRSCH 12
Biologicals
MPS2
Cloning of DNA
A) Cloning vector
B) PCR
DIRSCH 13
Multiplication/Colony
Plasmid isolation
Transformation in bacteria
restriciton site
known DNA sequences
DNA sequence of interest
Denaturation at 95°C
Primer hybridisation at 55°C
Polymerisation by Taq-Polymerase
Recombined plasmid vector
MPS2
Biologicals
DIRSCH
© http://newscenter.lbl.gov/news-
releases/2011/11/29/e-coli-make-three-fuels/ ©http://microbewiki.kenyon.edu/index.php/Sa
ccharomyces_cerevisiae_NEU2011
©http://en.m.wikipedia.org/wiki/File:Cho_cells
_adherend2.jpg ©https://www.dsmz.de/de/downloads/bilde
r/plant-cell-lines.html
Bacteria Mammals /
Mammalian cells Plants / plant cells Yeast
RECOMBINANT PROTEINS – host organisms / expression systems
14
Biologicals
MPS2
DIRSCH
Bacterial cell, e.g. E. coli
© WVG Stuttgart: Dingermann/Winckler/Zündorf: Gentechnik Biotechnik, 2011
transgenic bacterium
gene expression
recombinant protein in the periplasm
recombinant protein in inclusion bodies
isolation of recombinant protein
Cell lysis and isolation of inclusion bodies with the denatured protein
Osmotic destruction of the outer membrane and isolation of the native protein
Pros
• Non pathogenic • Entire genome has been sequenced • Extensive experience with bacterial genetics • Easily takes up DNA • Rapid growth • Easy handling • Inexpensive media • Often extremely high product yield
Cons
• No RNA processing • No posttranslational modifications • Proteins often in incorrect tertiary structure • Proteins often denatured („inclusion
bodies“) • Release of endotoxins (pyrogens)
15
Biologicals
MPS2
Pros
• Eucaryot • Rapid growth • Extensive experience with yeast genetics • Entire genome has been sequenced • Cells and their products are non pathogenic • No production of pyrogenic substances • Proteins are usually correctly folded • Often high product yield • Secretion into the culture medium • Some posttranslation modifications possible • Genetically recognized as safe (GRAS) • Easily takes up DNA • Easy handling • Inexpensive media
Cons
• Glycosylations differ largely from mammalian cells (Man6-9GlcNAc2; Hypermannosylation: ManxGlcNAc2)
• Few dominant markers, instead auxotrophy marker
DIRSCH
Baker‘s Yeasts, e.g. Saccharomyces cerevisiae
© WVG Stuttgart: Dingermann/Winckler/Zündorf: Gentechnik Biotechnik, 2011
Nucleus
Transcription
Rough ER
Golgi apparatus
Secretion into culture medium
Signal peptide
Glucan cell wall
16
Biologicals
MPS2
ER (human, yeast)
Golgi (human) Golgi (yeast)
adapted from © Michaela Bogner: Hefe-basierte Produktion von Antikörpern mit verbesserten Eigenschaften, 2010
Complex saccharide structure and terminal NANA
sugar residues of the „High Mannose-Type“
β-1,N-GlcNAc
β-1,4-GlcNAc
β-1,2-GlcNAc
β-1,4-Man
α-1,6-Man
α-1,2-Man
α-1,3-Man
β-1,4-Gal
α-1,3-NANA/α-2,6-NANA
DIRSCH 17
Biologicals
MPS2
PROTEIN GLYCOSYLATION affects …
• Charge
• Stability
• Solubility
• Folding
• Cell cell interaction
• Ligand recognition/binding
• Biological function
• Immunogenicity DIRSCH
18
Biologicals
MPS2
DIRSCH
Mammalian cells, e.g. CHO cells
© Springer: Dingermann, Hänsel, Zündorf: Pharmazeutische Biologie, 2002 © WVG Stuttgart: Dingermann/Winckler/Zündorf: Gentechnik Biotechnik, 2011
Fixed bed reactor
pump
beads with adherent cells
https://www.youtube.com/watch?v=aK9sNe8c48U
19
Pros Cons
• Close relation to „human system“ • Correct folding, even of complex proteins • Secretion into the culture medium • (Almost) authentic posttranslational
modifications
• High production costs (expensive media) • Slow growth • Relatively low protein yield • High investment costs for fermentation • Use of animal-derived medium components
(„Fetal calf serum“, FCS) • High storage costs for master cell lines • „Upscaling“ challenging • Possibly susceptable to human pathogens
(especially viruses)
Biologicals
MPS2
DIRSCH
Plants (Plant cells)
http://blogs.nature.com/news/2012/05/first-plant-made-drug-on-the-market.html
© WVG Stuttgart: Dingermann/Winckler/Zündorf: Gentechnik Biotechnik, 2011
Pros
• Extensive experience from traditional plant cultivation
• Self insemination • Regenerative ability • Inexpensive cultivation and storage • Variable „Upscaling“ by field cultivation • No human pathogens (especially viruses)
Cons
• Secondary cell wall • Polyploidy • Somaclonal variation • Low product yield • Incorrect glycosylation: simple N-glycans
without sialic acid, unusual links, e.g. 1-3 linked Fucose, Xylose(1-2)Mannose
• „Gene silencing“ • Recombinant protein is generally instable in the
cytosol • Specific regulatory requirements for „release“
of transgenic plants • No GLP/GMP conditions in field cultivation
20
MPS2
Biologicals
Proteins glowing under ultraviolet light on a type of plant used to produce the drug ZMapp. Credit Sean Gallup/Getty Images
DIRSCH 21
Biologicals
MPS2
DIRSCH
E. coli Yeast
Mammalian cells, such as CHO
complex, glycosylated proteins
„simple proteins“
(amino acid sequence) disulfide bridge
Formation of secondary, tertiary and quaternary structure
modification with sugar residues, lipid chains, sulfate or phosphate residues
© Springer: Dingermann, Hänsel, Zündorf: Pharmazeutische Biologie, 2002
22
Biologicals
MPS2
DIRSCH
The European Pharmacopoeia (Ph. Eur.) is Europe’s legal and scientific benchmark for pharmacopoeial standards which contribute to delivering high quality medicines in Europe and beyond. The Ph. Eur. is applicable in 37 European countries and used in over 100 countries worldwide. The European Pharmacopoeia is a single reference work for the quality control of medicines in the signatory states of the Convention on its elaboration. The official standards published within provide a legal and scientific basis for quality control during the development, production and marketing processes. They concern the qualitative and quantitative composition and the tests to be carried out on medicines, on the raw materials used in production of medicines and on the intermediates of synthesis. All producers of medicines and/or substances for pharmaceutical use must therefore apply these quality standards in order to market their products in the signatory states of the Convention.
23
MPS2
Biologicals
DIRSCH
EUROPEAN PHARMACOPOEIA 9th edition 2017
24
Products of Recombinant DNA Technology
DEFINITION Products of recombinant DNA technology are produced by genetic modification in which DNA coding for the required product is introduced, usually by means of a plasmid or a viral vector, into a suitable micro-organism or cell line, in which that DNA is expressed and translated into protein. The desired product is then recovered by extraction and purification. The cell or micro-organism before harbouring the vector is referred to as the host cell, and the stable association of the two used in the manufacturing process is referred to
as the host-vector system.
This monograph provides general requirements for the development and manufacture of products of recombinant DNA technology. These requirements are not necessarily comprehensive in a given case and requirements complementary or additional to those prescribed in this monograph may be imposed in an individual monograph or by the competent authority. The monograph is not applicable to modified live organisms that are intended to be used directly in man and animals, for example as live vaccines.
MPS2
Biologicals
DIRSCH
Products of recombinant DNA technology are produced by genetic modification ....
25
... in which DNA coding for the required product... ... is introduced, usually by means of a plasmid or a viral vector, ..... ... into a suitable micro-organism or cell line, in which that DNA is expressed and translated into protein. The desired product is then recovered by extraction and purification. .................... host-vector system
MPS2
Biologicals
DIRSCH 26
EUROPEAN PHARMACOPOEIA 9th edition 2017
PRODUCTION Production is based on a validated seed-lot system using a host-vector combination that has been shown to be suitable to the satisfaction of the competent authority. The seed-lot system uses a master cell bank and a working cell bank derived from the master seed lot of the host-vector combination. A detailed description of cultivation, extraction and purification steps and a definition of the production batch shall be established. Where products of recombinant DNA technology are manufactured using materials of human or animal origin, the requirements of chapter 5.1.7. Viral safety apply. The determination of the suitability of the host-vector combination and the validation of the seed-lot system include the following elements.
• Cloning and expression • Cell bank system • Validation of the cell banks • Control of the cells • Validation of the production
process
MPS2
Biologicals
DIRSCH 27
EUROPEAN PHARMACOPOEIA 9th edition 2017
CLONING AND EXPRESSION The suitability of the host-vector system, particularly as regards microbiological purity, is demonstrated by: Characterisation of the host cell, including source, phenotype and genotype, and of the cell-culture media; Documentation of the strategy for the cloning of the gene and characterisation of the recombinant vector, including: i. the origin and characterisation of the gene; ii. nucleotide-sequence analysis of the cloned gene and the flanking control regions of the
expression vector; the cloned sequences are kept to a minimum and all relevant expressed sequences are clearly identified and confirmed at the RNA level; the DNA sequence of the cloned gene is normally confirmed at the seed-lot stage, up to and beyond the normal level of population doubling for full-scale fermentation; in certain systems, for example, where multiple copies of the gene are inserted into the genome of a continuous cell line, it may be inappropriate to sequence the cloned gene at the production level; under these circumstances, Southern blot analysis of total cellular DNA or sequence analysis of the messenger RNA (mRNA) may be helpful, particular attention being paid to the characterisation of the expressed protein;
iii. the construction, genetics and structure of the complete expression vector. Characterisation of the host-vector system, including: i. mechanism of transfer of the vector into the host cells; ii. copy number, physical state and stability of the vector inside the host cell; iii. measures used to promote and control the expression.
MPS2
Biologicals
DIRSCH 28
EUROPEAN PHARMACOPOEIA 9th edition 2017
CELL-BANK SYSTEM
The master cell bank Is a homogeneous suspension of the original cells already transformed by the expression vector containing the desired gene, distributed in equal volumes into individual containers for storage (for example, in liquid nitrogen). In some cases it may be necessary to establish separate master cell banks for the expression vector and the host cells. The working cell bank Is a homogeneous suspension of the cell material derived from the master cell bank(s) at a finite passage level, distributed in equal volumes into individual containers for storage (for example, in liquid nitrogen). In both cell banks, all containers are treated identically during storage and, once removed from storage, the containers are not returned to the cell stock. The cell bank may be used for production at a finite passage level or for continuous-culture production.
Production at a finite passage level This cultivation method is defined by a limited number of passages or population doublings which must not be exceeded during production. The maximum number of cell doublings, or passage levels, during which the manufacturing process routinely meets the criteria described below must be stated.
Continuous-culture production By this cultivation method the number of passages or population doublings is not restricted from the beginning of production. Criteria for the harvesting as well as for the termination of production have to be defined by the manufacturer. Monitoring is necessary throughout the life of the culture; the required frequency and type of monitoring will depend on the nature of the production system and the product. Information is required on the molecular integrity of the gene being expressed and on the phenotypic and genotypic characteristics of the host cell after long-term cultivation. The acceptance of harvests for further processing must be clearly linked to the schedule of monitoring applied and a clear definition of a ‘batch’ of product for further processing is required.
MPS2
Biologicals
DIRSCH 29
EUROPEAN PHARMACOPOEIA 9th edition 2017
VALIDATION OF THE CELL BANKS Validation of the cell banks includes: i. stability by measuring viability and the retention of the vector; ii. identity of the cells by phenotypic features; iii. where appropriate, evidence that the cell banks are free from potentially
oncogenic or infective adventitious agents (viral, bacterial, fungal or mycoplasmal); special attention has to be given to viruses that can commonly contaminate the species from which the cell line has been derived; certain cell lines contain endogenous viruses, for example, retroviruses, which may not readily be eliminated; the expression of these organisms, under a variety of conditions known to cause their induction, shall be tested for;
iv. for mammalian cells, details of the tumorigenic potential of the cell bank shall be obtained.
CONTROL OF THE CELLS The origin, form, storage, use and stability at the anticipated rate of use must be documented in full for all cell banks under conditions of storage and recovery. New cell banks must be fully validated.
MPS2
Biologicals
DIRSCH
© Springer: Dingermann, Hänsel, Zündorf: Pharmazeutische Biologie, 2002
Culture liquid
Tangential Flow Filtration centrifugation (TFF)
Ultrafiltration (UF) Diafiltration (DF)
Anion exchange chromatography
Filtration
UF, DF
Cation exchange chromatography
A
UF, DF
Gel Permeation Chromatography
Bottling, labeling
Product
F
30
EUROPEAN PHARMACOPOEIA 9th edition 2017
VALIDATION OF THE PRODUCTION PROCESS Extraction and purification contaminating substances derived from the host cell or culture medium, including, in particular, virus particles, proteins, nucleic acids and excipients must be removed and/or inactivated Depletion factor, Inactivation factor Characterisation of the final bulk product identity, purity, potency, stability Production consistency
MPS2
Biologicals
PROTEIN ANALYTICS: gel filtration/size exclusion chromatography Separation of proteins by size
DIRSCH 31
MPS2
Biologicals
DIRSCH
PROTEIN ANALYTICS: gel filtration/ size exclusion chromatography Separation of proteins by size
32
sample buffer
small protein
large protein
MPS2
Biologicals
PROTEIN ANALYTICS: ion exchange chromatography Separation of proteins by charge
DIRSCH 33
stationary phase
stationary
phase Elution of protein with positive charge
Flo
w d
irec
tio
n
Protein with negative charge binds to stationary phase
MPS2
Biologicals
PROTEIN ANALYTICS: affinity chromatography Separation of proteins by specific binding properties
DIRSCH 34
matrix bound transcription faktor
free oligonucleotide
released transcription
factor
MPS2
Biologicals
DIRSCH 35
PROTEIN ANALYTICS: electrophoresis analyzes protein
mixtures qualitatively
SDS molecule
Polypeptide chain
Biologicals
MPS2
36
PROTEIN ANALYTICS: polyacrylamide gel Molecular sieve
DIRSCH
Polyacrylamide network
small proteins
sample well
sample
large proteins
Elec
tro
ph
ore
sis
Biologicals
MPS2
37
PROTEIN ANALYTICS: coomassie staining of a SDS gel
DIRSCH
MPS2
Biologicals
38
PROTEIN ANALYTICS: isoelectric focusing separates proteins by
differences in their isoelectric point
DIRSCH
negatively charged protein moves towards
the anode
neutral protein diffuses towards the kathode, becomes
negatively charged ... ... and moves back towards
the anode
isoelectric point of the protein
MPS2
Biologicals
39
PROTEIN ANALYTICS: Two-dimensional gel electroporesis
1st dimension
2nd dimension
basic
low molecular weight
high molecular weight
acidic
MPS2
Biologicals
40
SDS Gel with complex
protein mixture
Sandwich with Filter paper
soaked in buffer
Electrophoresis 1. Blocking 2. Primary antibody 3. Secondary antibody 4. Dyes
Dyed protein band
Membrane with transferred
proteins
Other proteins stay invisible
PROTEIN ANALYTICS: Western blot analysis
Biologicals
MPS2
41
colorless, soluble substrates
Secondary antibody conjugated with
alkaline phosphatase
membrane fixed protein
Primary antibody
MPS2
Biologicals
DIRSCH 42
PROTEIN ANALYTICS: Enzyme-linked immunosorbent assay (ELISA) quantifies proteins in complex mixtures
Well plate
Specific primary antibody
Other proteins
Analyte
calibration curve
sample absorption
sample concentration
Concentration
Colored product of the enzymatic reaction
Enzyme linked secondary antibody
MPS2
Biologicals
DIRSCH
BIOSIMILARS – Definition
What are biosimilars? Imitation products of already approved biopharmaceutical medicines which are no longer patent protected. In contrast to generics (imitation products of small molecule drugs), biosimilars are not identical but similar to the original product since the use of living cells / organisms for their production and their structural complexity gives the products a degree of variability which makes the production of identical copies impossible - especially when different production processes are used.
43
MPS2
Biologicals
DIRSCH
Generics Biosimilars
Character small molecule drugs „the molecule is the product“
complex molecules, proteins „the process is the product“
Pharmaceutical form predominantly oral Solutions for injection
Production mostly chemical synthesis • Obtained from living cells (complex matrix)
• Highly sensitive processes
Approval • simplified conditions • usually no safety and efficacy
studies required
complex test program, seperately determined for each substance class
Comparison to reference product
Identical active ingredient Proven comparability in quality, safety, efficacy
BIOSIMILARS versus GENERICS
adapted from © Sandoz Biopharmaceuticals 44
MPS2
Biologicals
© Dingermann/Zündorf PharmuZ 2012
EXAMPLES of BIOSIMILARS (incomplete list! Snap shot of 2013)
Omnitrope®
Valtropin®
Binocrit®
Epoetin alfa Hexal®
Abseamed®
Retacrit®
Silapo®
Biograstim®
Filgrastim Ratiopharm®
Ratiograstim®
Tevagrastim®
Zarzio®
Filgrastim Hexal®
Nivestim®
Somatropin
Somatropin
Epoetin zeta
Epoetin zeta
Epoetin alfa
Epoetin alfa
Epoetin alfa
Filgrastim
Filgrastim
Filgrastim
Filgrastim
Filgrastim
Filgrastim
Filgrastim
Sandoz
Bio Partners
Sandoz
Hexal
Medice
Hospira
STADA
CT Arzneimittel GmbH
Ratiopharm GmbH
Ratiopharm GmbH
Teva Generics GmbH
Sandoz
Hexal
Hospira
Genotropin®
Humatrope®
Eprex®
Eprex®
Eprex®
Eprex®
Eprex®
Neupogen®
Neupogen®
Neupogen®
Neupogen®
Neupogen®
Neupogen®
Neupogen®
12.04.2006
24.04.2006
28.08.2007
28.08.2007
28.08.2007
18.12.2007
18.12.2007
16.09.2008
16.09.2008
16.09.2008
16.09.2008
06.02.2009
06.02.2009
08.06.2010
Handelsname INN Vertreiber Referenzprodukt Zulassung
Human growth hormone
Erythropoetin
Granulocyte-colony stimulating factor (G-CSF)
Remicade®
Remicade®
Remsima®
Inflectra®
Infliximab
Infliximab
28.06.2013
28.06.2013
Celltrion
Hospira Monoclonal antibodies (against TNF) DIRSCH 45
MPS2
Biologicals
DIRSCH
BIOSIMILARS – reference products (originator products) also have biological variability:
Schiestl M et al, Nature Biotechnology 29, 310–312 (2011)
Molecular variations (within defined specification limits) also exist between batches. Changes in the production process can cause greater variations, which are reported to the regulatory authority for thorough examination and approval.
The analysis of various batches of rituximab showed different glycosylation patterns (left), depending on the expiry date, and thus different ADCC (antibody dependent cellular cytotoxicity) potencies (right). light blue dots: before dark blue dots: after a change in the manufacturing process
46
MPS2
Biologicals
DIRSCH
„Micro-heterogeneity“
Protein drugs, such as insulin or monoclonal antibodies, have molecular weights up to 1000 times larger than „small molecule“ drugs. As the molecular weight increases, the complexity of the molecule and the possibility of product heterogeneity increase.
BIOSIMILARS – complexity and product heterogeneity
© Isam Rais, PharmuZ 2012
complexity
47
synthetic drug biological drug
Monoclonal antibodies 150.000 Da
Acetyl salicylic acid 180 Da
Insulin 5700 Da
Complexity
IgG monoclonal antibody C6466H9982N1726O2024S40
MW = approx. 150.000 Da
N-terminal pyro-glutamate Methionine oxidation Deamidation Excision of C-terminal lysine Glycation Fragmentation Glycosylation
Examples of possible mikroheterogeneities:
MPS2
Biologicals
DIRSCH
BIOSIMILARS – Analytics
Overview of analytical methods for the characterization of proteins
Analytic principle Methods Application
Electrophoresis SDS-PAGE, Isoelectric Focusing (IEF), Capillary Gel Electrophoresis (CGE)
Purity (aggregates, fragments), charge heterogeneity
Chromatography RP-HPLC, Size Exclusion Chromatography (HP-SEC), Ion Exchange Chromatography (IEC), Hydrophobic Interaction Chromatography (HIC)
Purity, charge heterogeneity
Mass Spectrometry Peptide Mapping Identity, chemical modifications
Spectroscopy Circular Dichroism (CD), Differential Scanning Calorimetry (DSC)
Spatial structure
Bioassay, binding assay Cell based assay formats, Biacore, ELISA Identity, potency
adapted from © Isam Rais, PharmuZ 2012 48
MPS2
Biologicals
DIRSCH adapted from © Isam Rais, PharmuZ 2012
BIOSIMILARS – Development
The development of biosimilars is a is a step by step process. By analysis of originator batches, a Target Product Profile which serves as a target for process development can be created. On the basis of this TPP, a manufacturing process for a drug with similar properties is developed. Subsequently, the new drug is compared with the originator product in detailed studies.
development of a manufacturing process
Similarity/ comparability exercise
Characterization of the reference product batch
Definition of a target product profile (TPP)
Implementation of Quality by design (QbD)
Optimization of upstream and downstream
processes
Defined product profile of the target structure
49
MPS2
Biologicals
DIRSCH
BIOSIMILARS – Development – comparability exercise
Reference product (Biopharmaceutical comparator product)
Physicochemical and biological
characterization
Non-clinical PK/PD/safety
studies Clinical studies Quality
Structure of the biosimilar,
manufacturing process, ...
adapted from © Isam Rais, PharmuZ 2012
Each aspect of the product development must always be carried out in direct comparison to an approved reference product. Comparison with literature data alone is not acceptable.
50
MPS2
Biologicals
DIRSCH adapted from © Isam Rais, PharmuZ 2012
Physicochemical characterization
Biological characterization
Preclinical studies
PK/PD studies in humans
Clinical studies
Sim
ilari
ty
Co
mp
arab
ility
Val
idat
ion
BIOSIMILARS – Development – comparability exercise
A step by step approach is necessary in order to achieve the label „Similarity“. It starts with physicochemical „comparability“ and, as the amount of acquired data increases, the highest step of „Similarity“ is reached.
51
MPS2
Biologicals
DIRSCH
BIOSIMILARS – Regulatory approval
adapted from © Isam Rais, PharmuZ 2012
Extent and proportion of reporting and documentation requirements for a biosimilar in comparison to a new drug
52
MPS2
Biologicals
DIRSCH
BIOLOGICALS – BOSIMILARS – BIOIDENTICALS – BIOBETTERS
Dingermann & Zündorf: Biobetter – Kopien besser als das Original, PZ 06/2015: http://www.pharmazeutische-zeitung.de/index.php?id=56229
Biologicals – 1st and 2nd generation drugs Bioidenticals – Biologicals with different names that are actually identical because they were manufactured in the very same process Biosimilars – generic versions of a recombinant drug that are the closest possible replica of their originals. Ultimately, they are similar but not identical to the original. Biobetters (Biosuperiors) – copies or improved versions of a biological drug (for example with modified pharmacokinetic or -dynamic properties); see also „muteins“
2nd generation
drugs
identical to human structure
1st generation
drugs
technical limitations
53