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REGULATORY STUDIES
SUPPORT
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Drug discovery services Regulatory Development
Selvita Services
• Target validation
• Hit identification
• Hit to lead
• Lead optimization
• Preclinical candidate nomination
• Broad target class and disease area expertise
Support for drug discovery in the areas of:
• Medicinal/Synthetic chemistry
• Assay development & screening
• In vitro pharmacology
• ADME/DMPK
• Structural biology
• CADD/ AI-driven drug discovery2
• Quality Control
• Release Analyses
• Impurity Studies
• Method Development
and Validation
Comparative studies of biosimilar products
• Pharmacodynamic studies
• Crystallography
• Proliferation studies in vitro
• Metabolic studies in vitro
• Process related impurities measurement
• Synthesis process optimization andparametrization
• Scale-up of processes & technology transfers
• Custom synthesis of NCEs, impurities/metabolites identification and synthesis
• Isotope labelling (2H, 13C, 15N)
• Computational chemistry
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Methoddevelopment
Methodvalidation
Methodtransfer
Routinetesting
Regulatory aspects support
Development and
optimization of the
method based on
Quality by Design
approach
Validation of the
method developed
in house or provided
by the customer,
based on ICH
guidelines
Transfer of fully validated „ready to use” method to the customer or receiving the validated method from customer’s side for routine testing at Selvita
QC testing of finished products, full battery of physical, chemical, microbiological and biological tests performed in accordance with the specification
Support of CMC process on customer side, filling quality module of CTD, batch certification (FP, APIs), providing CoAs and CoCs confirmed by QP
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Analytical and bioanalytical support of the drug developmentprocess
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CERTIFICATES
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EU-FDA mutual recognition agreement
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• From February 7, 2019 Polish regulatory authority is recognized by FDA as equal to the US GMP inspection.
• This means that the FDA can now rely on a Polish GMP inspection to replace their own inspections.
• FDA inspection will be not needed for the following products:
• human chemical pharmaceuticals;
• human biologicals including immunologicals and biotherapeutics;
• active pharmaceutical ingredients;
• intermediate products and bulk pharmaceuticals.
https://www.fda.gov/internationalprograms/agreements/ucm598735.htm
https://www.fda.gov/internationalprograms/agreements/ucm598735.htm
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GxP quality system – general overview
GMP GLP
FIRST CERTIFICATE 2009 2011
CERTIFICATE ISSUED BY The Chief Pharmaceutical Inspectorate The Bureau for Chemical Substances
CERTIFICATE SCOPE• physical and chemical testing• biological testing in quality control of medicines
• physical and chemical testing, mutagenicity studies
• chemical analyses and clinical chemistry testing• cytotoxicity tests of chemical substances• bioanalysis and pharmacokinetic studies
LAST INSPECTION/DEVIATIONS 4-5 Jun 2019 (critical 0, major 3, minor 8) 17-19 Sep 2019 (critical 0, major 2, minor 10)
NUMBER OF EXTERNAL/INTERNAL AUDITS Avg 10 external/4 internal audits per year
PERSONNEL IN GXP SYSTEM 85 in Quality Control, 5 in Quality Assurance (including 2 QPs)
DOCUMENTATION STRUCTURE Quality Manual, SMF, SOPs, Instructions and Analytical Methods, Laboratory Records, Raw Data
EQUIPMENT IN GXP SYSTEM 130 pieces of critical qualified equipment (OQ/PQ every year)
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ANALYTICAL AND
REGULATORY STUDIES
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Analytical Laboratory
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Development of new drugs - ADME and PK studies
Development of bioanalytical methods
Analytical method development and validation
Stability and release testing
Impurity synthesis and identification
Support in CMC projects
Scope of services
CERTIFIED
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Analytical services for the pharmaceutical industry
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• Assay/identity (HPLC, GC, potentiometric titration)
• Identity (MS, NMR, classical methods)
• Impurity profiling and Identification
• Stress studies and degradation profiles
• Stability testing - shelf-life determination, storage and handling requirements
• Cleaning validation
• Residual solvents determination
• Dissolution profiles – pharmaceutical equivalence
• Metal analysis using AAS, ICP techniques
• Microbial purity testing
• Excipient determination - reverse engineering
• CMC projects – analytical method establishment and validation, analytical support of synthesis and formulation stability testing, release analysis
• ADME (solubility, liofilicity, metabolic and plasma stability, metabolite profiling, permeability, plasma protein binding, cytochrome P450 inhibition)
• Study of physicochemical properties
(KF, osmolality, logD, pKa, DSC, melting/freezing point, IR, NMR, PSD, SEM-EDS, XRPD)
• Genotoxicity testing of impurities (performed by the Cell & Molecular Laboratory)
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Analytical Services for the Pharmaceutical Industry
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• Impurity identification
• In-process manufacturing/formulation support
• Analytical method validation/verification
• Characterization and qualification of reference standard
• Stability testing (for demo batches and GMP batches)
• GMP lot release testing or
• Analytical method transfer
ANALYTICAL SUPPORT FOR CHEMISTRY, MANUFACTURING AND CONTROLS (CMC) PURPOSE
Analytical method development and pre-validation:
assay, identity, purity, stress tests, residual solvents,
techniques: H(U)PLC-DAD/UV/CAD/ELSD/MS, GC(HS)-FID/MS
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Analytical Services for the Pharmaceutical Industry
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• Long-term stability testing
(including refrigerator, freezer and below -20⁰C)
• Intermediate stability testing
• Accelerated stability testing
• Forced degradation study
• Photo stability testing
• Other (freeze/thaw, in-use stability, open-pot stability)
STABILITY TESTING
STORAGE AND ANALYSES, FOR FORMULATIONS, DRUG SUBSTANCE AND DRUG PRODUCT
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Analytical Services for the Pharmaceutical Industry
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• Appearance
• Identification by HPLC-DAD, UV, HPLC-CAD,
GC-FID-HS, LC/MS , IR
• Uniformity of mass
• Loss on drying
• Hardness
• Disintegration time
• Dissolution
• Assay by HPLC-DAD, UV, HPLC-CAD, GC-FID-HS,
LC/MS
RELEASE AND SHELF-LIFE EVALUATION OF RAW MATERIALS, DRUG SUBSTANCES
AND DRUG PRODUCTS FOR SPECIFICATION COMPLIANCE
• Related Substances: identified and unidentified
impurities by HPLC-DAD, UV, HPLC-CAD,
GC-FID-HS, LC/MS
• Microbiology (microbial purity limits, bacteria, fungi,
E.coli)
• Analysis by ICP-OES, ICP-MS - catalyst residues
• Analysis by AAS - metal content
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Analytical Methods
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• For small and big (biosimilar) molecules
• According to the requirements of the respective
guidance
• ICH Topic Q2 (R1) Validation of Analytical Procedures:
Text and Methodology, CPMP/ICH/381/95
• U.S. Department of Health and Human Services,
Food and Drug Administration: Guidance for Industry,
Analytical Procedures and Methods Validation for Drugs
and Biologics, July 2015
• Committee for Medicinal Products for Human Use (CHMP):
Guideline on bioanalytical method validation, 21 July 2011,
EMEA/CHMP/EWP/192217/2009
• U.S. Department of Health and Human Services,
Food and Drug Administration: Guidance for Industry,
Bioanalytical Method Validation, May 2018
METHOD DEVELOPMENT, VERIFICATION, VALIDATION AND TRANSFER
• According to the general pharmacopoeial requirements (methods
and monographs); customer’s methods and specifications;
internal procedures
• Verification of analytical methods
• Optimization and development of analytical methods
• Pre-validation
• Validation
• Transfer
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Analytical Documentation
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• Report on analytical method development
• Report on pharmacopoeial analysis
• Report on analyses for specification compliance
(stability, release)
• Certificate of Analysis
• Parts of the Common Technical Document
• Analytical method – Standard Operating Procedure
• Validation protocol and report
• Transfer protocol and report
• Raw/integrated/calculated data
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Equipment (GLP/GMP)
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• GC(HS)/FID (Agilent)
• GC(HS)/FID (Shimadzu)
• GC/FID/MS (Agilent) - ESI/quadrupole
• GC(HS)/MS (Shimadzu) - ESI/quadrupole
• UHPLC/HPLC/MS (Bruker) - ESI/APCI/ion trap
• UHPLC/MS (Thermo Fisher Scientific) - ESI/APCI/TSQ
• UHPLC/MS (Thermo Fisher Scientific) - H-ESI/APCI Q/Orbitrap
• UHPLC/MS (Shimadzu) - ESI/APCI/QQQ
• UHPLC/MS (Agilent) - ESI/APCI/QQQ
• HPLC-VWD (Dionex/Thermo Scientific)
• UHPLC-DAD (Agilent)
• HPLC-DAD (Dionex/Thermo Scientific)
• HPLC/VWD/Corona (Dionex/Thermo Scientific) Detector (CAD)
• HPLC/PDA (Dionex/Thermo Scientific)
• HPLC/DAD/ELSD (Dionex/Thermo Scientific, Agilent)
• HPLC/VWD/RI (Dionex/Thermo Scientific)
• HPLC/UV/FLD (Dionex/Thermo Scientific)
• HPLC/DAD (Dionex/Thermo Scientific) - non-aqueous conditions
• IC (Dionex/Thermo Scientific)
• Capillary electrophoresis CESI 8000 Plus (Sciex)
• Isoelectric focusing electrophoresis (IEF), iCE3 (bio-techne)
• Spectrophotometer UV-VIS (Shimadzu, Agilent)
• Dissolution analysis SR8+ (Hanson Research)
• ICP-MS (Agilent), ICP-OES (Shimadzu), AAS (Thermo Fisher Scientific)
• FTIR (Thermo Fisher Scientific), NIR (Thermo Fisher Scientific)
• KF (Metrohm, Crison)
• Osmometer
• Particle analyser
• Disintegration DISI (Hanson Research), hardness (ERWEKA), friability (ERWEKA)
• Stability chambers (Memmert)
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Biopharmaceuticals and Biosimilars Services
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All studies are designed, optimized and performed based
on the EMA and ICH guidelines, as well as European
and US Pharmacopoeias.
Selvita has established an extensive set of in house tests
suggested by the ICH Q6B guideline to perform during
biopharmaceuticals’ characterization.
STRUCTURAL CHARACTERIZATION
PHYSICOCHEMICAL PROPERTIES
IN VITRO PHARMACODYNAMIC
STUDIES
BATCH-TO-BATCH CONSISTENCY
STUDIES
IN VITRO PROLIFERATION
STUDIES
PROCESS-AND-PRODUCT RELATED
IMPURITIES
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Biopharmaceuticals Analysis at Selvita
Types of biopharmaceuticals Assays perfomed at CMBD
Bioassay
Cell proliferation and cytotoxicity assays, Neutralization assays, Apotosis assay,cAMP assay
Binding assay
ELISA and SPR based
Identification assays
• SDS-PAGE with Silver Staining/Coomassie Brilliant Blue Staining underreducing and non-reducing conditions
• SDS-PAGE with Iodine Staining (detection of protein PEGylation)
• Western Blot analysis
• Iso Electrical Focusing with Comassie Brilliant Blue Staining
Purity assays
SDS-PAGE with Silver Staining/Coomassie Brilliant Blue Staining under reducing
and non-reducing conditions
Iso Electrical Focusing with Comassie Brilliant Blue Staining
Growth factors and hormones:• GM-CSF (Molgramostim)
(Human Granulocyte Macrophage Colony-StimulatingFactor)
• G-CSF (Filgrastim) and PEG-G-CSF (Pegfilgrastim)
(Human Granulocyte Colony-Stimulating Factor)
• PTH (Teriparatide)
(Human Parathyroid Hormone)
• Insulin and insulin analogs
Monoclonal antibodies:▪ Anti hVEGF (Anti Human Vascular Endothelial Growth Factor)
▪ Bevacizumab
▪ Ranibizumab
▪ Anti hTNF-α (Anti HumanTumor Necrosis Factor α)
▪ Etanercept (Fusion protein)
▪ Adalimumab
▪ Infliximab
▪ Anti hHER2 (Anti Human Epidermal Growth Factor Receptor 2)
▪ Trastuzumab
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Bioassay development and validation
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Selvita approach to in vitro bioassay development & validation (GMP)
In-vitro biossay development in accordance with USP ‘Design and development
of biological assays’.
Validation of non-compendial methods conducted according to the requirements of
ICH Topic Q2 (R1) Validation of Analytical Procedures: Text and Methodology,
CPMP/ICH/381/95, London, 1995.
Analysis of results is performed using PLA 3.0 Bioassay Software (Stegmann Systems
GmbH).
Exemplary scope of in vitro bioassay validation
• system suitability
• specificity
• repeatability (6 assays performed on the same day by single Scientist)
• accuracy (six potency levels of the sample: 175%, 150%, 130%, 100%,
70% and 50%, assay performed by 3 scientists on 3 different days)
• linearity and range
• intermediate precision (3 scientists)
• robustness
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• mAbs binding to target
antigen
(e.g. TNFα, VEGF)
including on-off kinetics
using SPR method
• Binding to representative
isoforms of the relevant
three Fc gamma receptors
(FcγRI, FcγRII and FcγRIII),
neonatal Fc receptor (FcRn)
and complement (C1q) using
SPR method
• Neutralization assays for
anti-TNFα mAbs:
− Inhibition of hTNFα-induced
cytotoxicity/apoptosis
− Inhibition of hTNFα-induced
expression of adhesion
molecules
• Neutralization assays for
anti-VEGF mAbs:
− Cell growth inhibition assay
• Antibody-dependent cell-
mediated cytotoxicity
(ADCC) assay
• Jurkat cells stably expressing
FcƴRIIIa and luciferase
reporter under NFAT control
• LDH-based method using
mTNF-CHO cells and CD16
(v/f)-NK-92 cells
• Complement-dependent
cytotoxicity (CDC) assay
In vitro Comparability Studies of Biosimilar mAbs
Binding to target antigen(s)
Binding to FcγRs and complement
Fab-associatedfunctions
Fc-associatedfunctions
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Comparability in vitro Studies of Biosimilar Insulin and Insulin Analogues
• IR-A and IR-B binding assay including on-off kinetics using SPRmethod
• IR binding assay using MST and FP methods
IR binding assays
• Radiometric glucose uptake assay in 3T3-L1 adipocytes
• Radiometric lipogenesis assay in 3T3-L1 cells
• Inhibition of stimulated lipolysis assay in 3T3-L1 adipocytes (FFA evaluation)
Metabolic assays
• Cell-based assay measuring phosphorylation/ dephosphorylation of IR-A, IR-B (stable cell lines) and IGF-1R(HepG2 and MCF7 cells) (AlphaLISA, ELISA)
• Cell-based assay measuring activation of IRS-1 protein, Akt1 and MAP kinase in HepG2 and MCF7 cells (ELISA)
Intrinsic activity
• Analysis of the mitogenic activity of insulin analogs in primary and established cell lines, e.g. SaOS-2, MCF7, MCF10A and HMEC
• Evaluation of cell viability using MTS, CellTiter-Glo® Luminescent Cell Viability Assay and BrdU assay
Mitogenic activity
• Identification of process-related impurities:
• Host Cell Proteins (HCPs)
• DNA Content
Impurities
Standard curve HCPcode: #41599
Log conc. Total HCP eq.[ng/ml]
OD
[40
5n
m]-
OD
[49
2n
m]
0.0 0.5 1.0 1.5 2.0
0.0
0.5
1.0
1.5
2.0
2.5
log(agonist) vs. response -- Variable slope (four parameters)
Best-fit values
Bottom
Top
LogEC50
HillSlope
EC50
Span
Std. Error
Top
LogEC50
HillSlope
95% Confidence Intervals
Top
LogEC50
HillSlope
EC50
Goodness of Fit
Degrees of Freedom
R square
Absolute Sum of Squares
Sy.x
Constraints
Bottom
Number of points
Analyzed
OD[405nm]-OD[492nm]
= 0.2640
4.766
2.080
0.9621
120.2
= 4.502
1.398
0.2427
0.09645
1.690 to 7.843
1.546 to 2.614
0.7498 to 1.174
35.15 to 411.4
11
0.9954
0.02160
0.04432
Bottom = 0.2640
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Equipment for Biosimilar Studies
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MicroscaleThermophoresis
(Monolith® NT.115, NanoTemperTechnologies)
GE Healthcare Biacore™ label-freeinteraction analysiswith Biacore T200
instrument
MicroBeta®2 (PerkinElmer) Microplate
ScintillationAnd Luminescence
Counter
Multi-mode Microplate Readers:• Synergy™2 (Biotek)• Synergy™ Neo2 (Biotek)• Spark (Tecan)
CBS Isothermal(Panasonic) Cell Lines Long Term
Storage System in Nitrogen Vapour
Phase
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/company/selvita/
SelvitaKrakow
/Biotechnology-Company/Selvita-SA
THANK YOU FOR ATTENTION!
/company/selvita/
SelvitaKrakow
/Biotechnology-Company/Selvita-SA
Get in touch with us: [email protected]