qps biosimilar bioanalytical approaches
Post on 20-Aug-2015
2.803 Views
Preview:
TRANSCRIPT
Current Practice in Bioanalytical Methods for Biosimilars:
Case Study of Rituximab Assay Development for PK, ADA and Nab
Xun Wang, Ph.D.
Department of Translational MedicineQPS, LLC
Top 10 Prescription Drugs in 2016 (*)
(*) 8 out of the 10 top-selling drugs in the US are biologics. By 2016, some of which will face biosimilar entry.
Drug Name(Patent Expiry)
Innovator Company
Indication Mech. Of Action
WW Sales Billion ($)2009 →2016
Humira™ (2016) Abbott Rhematoid Arthritis TNFαMAb 5.6 → 10.1
Avastin™ (2019) Roche Colorectal Cancer VEGF MAb 5.7 → 8.9
Enbrel ™ (2012) Pfizer, Amgen & Takeda
Rhematoid Arthritis TNFα Inhibitor 6.5 → 7.3
Rituxan™ (2018) Roche Non-Hodgkins Lymphoma
Anti-CD20 MAb 5.6 → 6.8
Crestor ™ (2012) AstraZeneca/ Shionogi/Chiesi
Anti-Hyperlipidemics Small Molecule Chemistry
4.8 → 6.3
Herceptin™(2019) Roche Breast Cancer Her2/neu MAb 4.9 → 6.2
Remicade ™ (2014) JNJ Rhematoid Arthritis TNFα MAb 5.9 → 5.7
Lantus ™ (2018) Sanofi Aventis Diabetics Insulin 4.3 → 5.3
Advair ™ (2011) GSK Bronchodilators Small Molecule Chemistry
8.0 → 5.2
Prolia ™ (2013) Amgen/Daichii Sankyo/GSK
Osteoporosis Bone Calcium Reg. MAb
0.0 → 5.2
The biological product is highly similar to the reference product notwithstanding minor differences in clinically inactive components,” and that “there are no clinically meaningful differences between the biological product and the reference product in terms of the safety, purity, and potency of the product.”
Biosimilar
Section 7002(b)(3) of the Affordable Care Act, adding section 351(i)(2) of the PHS Act.
Biosimilar Industry Today
Market Intelligence$378M in July, 2010 – June, 2011Forecast to rise to $2.6B by 2015
Development Cost$2M – $4M (small molecule generic)$100M - $200M (large molecule biosimilar)
Key Players: Innovator companies, Emerging market
Major Alliances announced in recent yearsAmgen -Watson: Oncology antibody productsRoche - Emcure Pharmaceuticals (India CMO): Herceptin and Mabthera Merck - ParexelBiocon (India) - Mylan: Biosimilar monoclonal antibodies Pfizer - Biocon: Insulin and insulin analogs (The deal was prematurealy terminated in March)Momenta (Cambridge, MA) – Baxter International: Enoxparin plus 6 other unspecified biosimilarsBiogen Idec – Samsung (Korea)
FDA Guidance
Public Health Services Act (e.g., erythropoietin)
The Biologics Price Competition and Innovation Act of 2009 (BPCI).
Quality Considerations in Demonstrating Biosimilarity to a Reference Protein Product (February, 2012).
Scientific Considerations in Demonstrating Biosimilarity to a Reference Product (February, 2012).
Biosimilars: Questions and Answers Regarding Implementation of the Biologics Price Competition and Innovation Act of 2009 (February, 2012).
Bioanalytical Approaches
PK Assay:1 Assay vs 2 AssaysReference standardQCs
ADA/Nab Assays:1 Assays vs 2 AssaysPositive controlsComparable activity and sensitivity
QPS Validated Bioanalytical Methods for Biosimilar
Currently QPS has validated methods for the following biosimilars of market potential:
Anti-Factor IIa and Anti-Factor Xa assays for Enoxaparin (Activity Assay)Anti-Factor IIa and Anti-Factor Xa assays for Dalteparin (Activity Assay)Rituximab (MabtheraTM, RituxanTM): Human PK method (ELISA, Gyrolab)Rituximab Human ADA methods (MSD ECL)Rituximab Human Nab methods (Cell based functional)Bevacizumab (AvastinTM) Human PK method (ELISA)Darbepoietin Human PK method (ELISA)Trastuzumab (HerceptinTM) Human PK method (ELISA)Cetuximab (ErbituxTM) Human PK method (ELISA)FSH (PuregonTM, Gonal-FTM) (Immulite)
Case Study: Rituximab PK, ADA and Nab
Rituximab (Mabthera, Rituxan) is genetically modified monoclonal antibody that targets CD20 on B cell surface.
Treat solid tumor of lymphoid cells and rheumatoid arthritis with excessive or dysfunctional B cells.
The mechanisms of action is not fully understood.Direct signaling
Complement-dependent cytotoxicity
Antibody-directed cell-mediated cytotoxicity
Case Study: Rituximab PK
1 Assay
Reference standard: Innovator
QCs: Innovator and Biosimilar
PK Assay Development (ELISA)
Rituximab Concentration (ng/mL)
10 100 1000 100000
0.5
1
1.5
2
y = ( (A - D)/(1 + (x/C)^B ) ) + D: A B C D R^2
Standard Curve (Std: Conc. vs OD) 0.064 1.22 3542.079 3.361 0.999
Case Study: Rituximab PK
PK Assay Development (ELISA)*
Validation Items Results
Assay Range 100 - 5000 ng/mL (HUSE)
QC intra/Inter Precision 2.9% to 10.7%
QC intra/Inter Accuracy -8.8% to 9.5%
Matrix Selectivity (low and high QC) At least 85% within ±25% for solid tumor and RA serum lots
Dilution Linearity 500,000 ng/mL diluted up to 2,000–fold;
ADA Interference At least 1 µg/mL at low QC and 10 µg/mL at high QC
Stability in HUSE -200C/-700C/RT/FT
* Data shown here are from the Rituximab Innovator
Case Study: Rituximab PK
PK Assay Development (Gyrolab)
A high throughput assay platform with miniaturization and automation
Wide assay range: 90 – 60,000 ng/mL in HUSE
Validated for Innovator SA analysisStreptavidin-coated bead
Alexa labeled -anti human IgG
Ab
Biotinylated-Rat anti ID rituximab
Rituximab
Flow
X. Liu et. al. Journal of Immunological Method, 2012
Case Study: Rituximab ADA
2 Assays (Innovator and Biosimilar)
Assay format: Bridging with labeled Innovator or Biosimilar
Positive controls
Anti-Rituximab Id
Rabbit anti-Rituximab Polyclonal
Anti-Biosimilar Id
ADA Assay Development
Case Study: Rituximab ADA
ADA Assay Development (Innovator)*
Validation Items
Approaches Results
Screening cut point(SCC)
At least 50 individual ST or RA lots
1. Floating cut point2. Significantly different SCPs
between matrixes3. Similar between Innovator and BSI
Confirmatory cut point (CCP)
At least 28 individual ST or RA lots spiked with drug
Similar CCPs between innovator and BSI
PC Crossing Check Anti-Biosimilar/Innovator Abs in Innovator/BSI assay
PC showed similar activity in both innovator and BSI assays
Assay sensitivity Determined for all three positive control antibodies
Sensitivities are similar between innovator and BSI assays (< 20 ng/mL)
Drug Tolerance Determined at low and high PC (50, 400 ng/mL)
Low and high PC can tolerate at least 12 and 50 µg/mL Rituximab.
* Data shown here are from the Rituximab Innovator
Case Study: Rituximab Nab
2 Assays (Innovator and Biosimilar)
Assay format: Cell based
Binding?
Functional -- Go
Positive controls
Anti-Rituximab Id
Rabbit anti-Rituximab Polyclonal
Anti-Biosimilar Id
Nab Assay Development
Case Study: Rituximab Nab
Rituximab MOA:
Complement-dependent cytotoxicity (CDC)
Antibody-dependent cell-mediated cytotoxicity (ADCC)
ApoptosisA non-radioactive complement-dependent cytotoxicity assay for anti-CD20 monoclonal antibody (Gazzano-Santoro et. al., Journal
of Immunological Methods, 1997)
Nab Assay Development
http://minimednews.wordpress.com/
Case Study: Rituximab Nab
Method Development
Cells growth and density curve: WIL2-S
Rituximab dose killing curve: 70-80% killing
Rabbit complement dose curve
Incubation time
Matrix Evaluation (ST and RA): 1:100• Specificity• Selectivity
Compare the biosimilar to the innovator
PC cross check
Nab Assay Development
Case Study: Rituximab Nab
Nab Assay Development (Innovator)*
Validation Items
Approaches Results
Screening cut point(SCC)
At least 50 individual ST or RA lots
1. Floating cut point2. Significantly different SCPs
between matrixes3. Similar between Innovator and BSI
Immunodepletion At least 28 individual serum lots treated with Protein A/G
Similar between innovator and BSI
PC Crossing Check Anti-Biosimilar/innovator Abs in Innovator/BSI assay
PC showed similar activity in both innovator and BSI assays
Assay sensitivity Determined for all three positive control antibodies
Currently ongoing (~ 50 µg/mL)
Drug Tolerance Determined by High PC titering in the presence of various drug levels.
Can tolerate 10 µg/mL Rituximab at the estimated assay sensitivity
* Data shown here are from the Rituximab Innovator
Conclusion
PK Assay: One assay is sufficient to measure both innovator and biosimilar drugs.
Reference Standard: Innovator/BSI
QCs: Innovator/BSI
Immunogenicity (ADA and Nab) requires two assays, which are best to develop simultaneously and achieve:
Similar screening and confirmatory cut points
Similar sensitivity
Similar PC cross reactivity
Similar drug tolerance
Acknowledgement
Joy HeFrank LiuLaurelle CallisteChristina XiaYun Shen
Avery TolosaYinLing LiBreann BarkerRoni Weaver
Jiannian ZhouMargaret Ma
LingSing ChenHui ZhangHolly ShenChad Asher
top related